#### Sample records for capture microdissection combined

1. Gene expression profiling of gastric cancer by microarray combined with laser capture microdissection

Ming-Shiang Wu; Yi-Shing Lin; Yu-Ting Chang; Chia-Tung Shun; Ming-Tsan Lin; Jaw-Town Lin

2005-01-01

AIM: To examine the gene expression profile of gastric cancer (GC) by combination of laser capture microdissection (LCM) and microarray and to correlate the profiling with histological subtypes. METHODS: Using LCM, pure cancer cells were procured from 45 cancerous tissues. After procurement of about 5 000 cells, total RNA was extracted and the quality of RNA was determined before further amplification and hybridization. One microgram of amplified RNA was converted to cDNA and hybridized to cDNA microarray. RESULTS: Among 45 cases, only 21 were qualified for their RNAs. A total of 62 arrays were performed. These included 42 arrays for cancer (21 cases with dyeswab duplication) and 20 arrays for non-tumorous cells (10 cases with dye-swab duplication) with universal reference. Analyzed data showed 504 genes were differentially expressed and could distinguish cancerous and non-cancerous groups with more than 99% accuracy. Of the 504 genes, trefoil factors 1, 2, and 3 were in the list and their expression patterns were consistent with previous reports. Immunohistochemical staining of trefoil factor 1 was also consistent with the array data. Analyses of the tumor group with these 504 genes showed that there were 3 subgroups of GC that did not correspond to any current classification system, including Lauren's classification. CONCLUSION: By using LCM, linear amplification of RNA, and cDNA microarray, we have identified a panel of genes that have the power to discriminate between GC and non-cancer groups. The new molecular classification and the identified novel genes in gastric carcinogenesis deserve further investigations to elucidate their dinicopathological significance.

2. Laser Capture Microdissection in Dentistry

Uraiwan Chokechanachaisakul; Tomoatsu Kaneko; Takashi Okiji; Reika Kaneko; Hideaki Suda; Jacques E. Nör

2010-01-01

Laser capture microdissection (LCM) allows for the microscopic procurement of specific cell types from tissue sections that can then be used for gene expression analysis. According to the recent development of the LCM technologies and methodologies, the LCM has been used in various kinds of tissue specimens in dental research. For example, the real-time polymerase-chain reaction (PCR) can be performed from the formaldehyde-fixed, paraffin-embedded, and immunostained sections. Thus, the advanc...

3. Laser Capture Microdissection in Dentistry

Uraiwan Chokechanachaisakul

2010-01-01

Full Text Available Laser capture microdissection (LCM allows for the microscopic procurement of specific cell types from tissue sections that can then be used for gene expression analysis. According to the recent development of the LCM technologies and methodologies, the LCM has been used in various kinds of tissue specimens in dental research. For example, the real-time polymerase-chain reaction (PCR can be performed from the formaldehyde-fixed, paraffin-embedded, and immunostained sections. Thus, the advance of immuno-LCM method allows us to improve the validity of molecular biological analysis and to get more accurate diagnosis in pathological field in contrast to conventional LCM. This paper is focused on the presentation and discussion of the existing literature that covers the fields of RNA analysis following LCM in dentistry.

4. Proteomic analysis of prolactinoma cells by immuno-laser capture microdissection combined with online two-dimensional nano-scale liquid chromatography/mass spectrometry

Chen Luping

2010-01-01

Full Text Available Abstract Background Pituitary adenomas, the third most common intracranial tumor, comprise nearly 16.7% of intracranial neoplasm and 25%-44% of pituitary adenomas are prolactinomas. Prolactinoma represents a complex heterogeneous mixture of cells including prolactin (PRL, endothelial cells, fibroblasts, and other stromal cells, making it difficult to dissect the molecular and cellular mechanisms of prolactin cells in pituitary tumorigenesis through high-throughout-omics analysis. Our newly developed immuno-laser capture microdissection (LCM method would permit rapid and reliable procurement of prolactin cells from this heterogeneous tissue. Thus, prolactin cell specific molecular events involved in pituitary tumorigenesis and cell signaling can be approached by proteomic analysis. Results Proteins from immuno-LCM captured prolactin cells were digested; resulting peptides were separated by two dimensional-nanoscale liquid chromatography (2D-nanoLC/MS and characterized by tandem mass spectrometry. All MS/MS spectrums were analyzed by SEQUEST against the human International Protein Index database and a specific prolactinoma proteome consisting of 2243 proteins was identified. This collection of identified proteins by far represents the largest and the most comprehensive database of proteome for prolactinoma. Category analysis of the proteome revealed a widely unbiased access to various proteins with diverse functional characteristics. Conclusions This manuscript described a more comprehensive proteomic profile of prolactinomas compared to other previous published reports. Thanks to the application of immuno-LCM combined with online two-dimensional nano-scale liquid chromatography here permitted identification of more proteins and, to our best knowledge, generated the largest prolactinoma proteome. This enlarged proteome would contribute significantly to further understanding of prolactinoma tumorigenesis which is crucial to the management of

5. Laser Capture Microdissection as a Tool to Study Tumor Stroma.

Bertos, Nicholas R; Park, Morag

2016-01-01

Laser capture microdissection (or LCM) allows for isolation of cells from specific tissue compartments, which can then be followed by DNA, RNA, and/or protein isolation and downstream characterization. Unlike other methods for cell isolation, LCM can be directed towards cells situated in specific anatomical contexts, and is therefore of significant value when investigating the tumor microenvironment, where localization is often key to function. Here, we present a summary of ways in which LCM can be utilized, as well as protocols for the isolation of tumor and tumor-associated stromal elements from frozen breast cancer samples, with a focus on preparation of samples for RNA characterization. PMID:27581011

6. Fusobacterium necrophorum determined as abortifacient in sheep by laser capture microdissection and fluorescence in situ hybridization

Boye, Mette; Aalbæk, Bent; Agerholm, Jørgen S.

2006-01-01

Fluorescent in situ hybridization (FISH) has been extensively used for identification of individual microbial cells within their natural environment. The present work describes the identification of Fusobacterium necrophorum in formalin-fixed tissue samples from three sets of ovine twins aborted at...... late pregnancy by a technique that combines laser capture microdissection (LCM) and fluorescent in situ hybridization (LCM-FISH). Cultural bacteriological examination had failed to identify an infectious agent but by histological examination, large colonies of bacteria associated with tissue...... inflammation were seen. In situ hybridization visualized the bacteria in the tissue samples and microcolonies closely associated with lesions were isolated by LCM. PCR-amplification and sequencing of 16S rRNA gene from the microdissected bacteria identified the organisms as Fusobacterium necrophorum. A r...

7. The cerebral microvasculature in schizophrenia: a laser capture microdissection study.

Laura W Harris

Full Text Available BACKGROUND: Previous studies of brain and peripheral tissues in schizophrenia patients have indicated impaired energy supply to the brain. A number of studies have also demonstrated dysfunction of the microvasculature in schizophrenia patients. Together these findings are consistent with a hypothesis of blood-brain barrier dysfunction in schizophrenia. In this study, we have investigated the cerebral vascular endothelium of schizophrenia patients at the level of transcriptomics. METHODOLOGY/PRINCIPAL FINDINGS: We used laser capture microdissection to isolate both microvascular endothelial cells and neurons from post mortem brain tissue from schizophrenia patients and healthy controls. RNA was isolated from these cell populations, amplified, and analysed using two independent microarray platforms, Affymetrix HG133plus2.0 GeneChips and CodeLink Whole Human Genome arrays. In the first instance, we used the dataset to compare the neuronal and endothelial data, in order to demonstrate that the predicted differences between cell types could be detected using this methodology. We then compared neuronal and endothelial data separately between schizophrenic subjects and controls. Analysis of the endothelial samples showed differences in gene expression between schizophrenics and controls which were reproducible in a second microarray platform. Functional profiling revealed that these changes were primarily found in genes relating to inflammatory processes. CONCLUSIONS/SIGNIFICANCE: This study provides preliminary evidence of molecular alterations of the cerebral microvasculature in schizophrenia patients, suggestive of a hypo-inflammatory state in this tissue type. Further investigation of the blood-brain barrier in schizophrenia is warranted.

8. Laser capture microdissection-based in vivo genomic profiling of wound keratinocytes identifies similarities and differences to squamous cell carcinoma

Pedersen, Tanja Xenia; Leethanakul, Chidchanop; Patel, Vyomesh;

2003-01-01

we present the first analysis of global changes in keratinocyte gene expression during skin wound healing in vivo, and compare these changes to changes in gene expression during malignant conversion of keratinized epithelium. Laser capture microdissection was used to isolate RNA from wound...... reepithelialization. Furthermore, the analyses revealed that the phenotypic resemblance of wound keratinocytes to squamous cell carcinoma is mimicked at the level of gene expression, but notable differences between the two tissue-remodeling processes were also observed. The combination of laser capture...

9. Laser capture microdissection enables cellular and molecular studies of tooth root development

Sun, Jian-Xun; Horst, Orapin V; Bumgarner, Roger; Lakely, Bryce; Somerman, Martha J; Zhang, Hai

2012-01-01

Epithelial–mesenchymal interactions (EMIs) are critical for tooth development. Molecular mechanisms mediating these interactions in root formation is not well understood. Laser capture microdissection (LCM) and subsequent microarray analyses enable large scale in situ molecular and cellular studies of root formation but to date have been hindered by technical challenges of gaining intact histological sections of non-decalcified mineralized teeth or jaws with well-preserved RNA. Here,we descri...

10. Laser capture microdissection and single-cell RT-PCR without RNA purification

Keays, Kathryne Melissa; Owens, Gregory P.; Ritchie, Alanna M.; Gilden, Donald H.; Burgoon, Mark P.

2005-01-01

Chronic infectious diseases of the central nervous system (CNS) are characterized by intrathecal synthesis of increased amounts of immunoglobulin G (IgG) directed against the agent that causes disease. In other inflammatory CNS diseases such as multiple sclerosis and CNS sarcoid, the targets of the humoral immune response are uncertain. To identify the IgGs expressed by individual CD38+ plasma cells seen in human brain sections, we merged the techniques of laser capture microdissection (LCM) ...

11. Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

Stephan Gehring; Edmond Sabo; Maryann E San Martin; Elizabeth M Dickson; Chao-Wen Cheng; Stephen H Gregory

2009-01-01

AIM: To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection (LCM). METHODS: Tissues are complex structures comprised of a heterogeneous population of interconnected cells. LCM offers a method of isolating a single cell type from specific regions of a tissue section. LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling is achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. RESULTS: mRNA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer cells derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant (2.5-fold, q value < 10) change in 493 genes. Based on this fold-change and a standardized PubMed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury.CONCLUSION: The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

12. Gene expression profiling of neurochemically defined regions of the human brain by in situ hybridization-guided laser capture microdissection

Bernard, R.; Kerman, I A; F. Meng; Evans, S J; Amrein, I.; Jones, E. G.; Bunney, W. E.; Akil, H; Watson, S. J.; Thompson, R C

2009-01-01

Laser capture microdissection (LCM) permits isolation of specific cell types and cell groups based upon morphology, anatomical landmarks and histochemical properties. This powerful technique can be used for region-specific dissection if the target structure is clearly delineated. However, it is difficult to visualize anatomical boundaries in an unstained specimen, while histological staining can complicate the microdissection process and compromise downstream processing and analysis. We now i...

13. Laser capture microdissection to identify septum-associated proteins in Aspergillus nidulans.

Zhang, Ying; Fischer, Reinhard; Teichert, Ines; Kück, Ulrich

2016-05-01

To spatially resolve genetic differences at the cellular level, the laser-capture microdissection technique was developed. With this method cells can be cut from tissues with a laser beam and analyzed for DNA, RNA or protein composition. Here we adapted the technique to isolate septal microtubule-organizing center (MTOC)-associated proteins in Aspergillus nidulans About 3000 septa were collected and subjected to peptide fingerprinting by mass-spectrometric analysis. We identified the microtubule polymerase AlpA and found it interacts with ApsB specifically at sMTOCs, suggesting that AlpA might be involved in the assembly or the functioning of this protein complex. PMID:26951366

14. Maintaining RNA integrity in a homogeneous population of mammary epithelial cells isolated by Laser Capture Microdissection

Helbling Jean-Christophe

2010-12-01

Full Text Available Abstract Background Laser-capture microdissection (LCM that enables the isolation of specific cell populations from complex tissues under morphological control is increasingly used for subsequent gene expression studies in cell biology by methods such as real-time quantitative PCR (qPCR, microarrays and most recently by RNA-sequencing. Challenges are i to select precisely and efficiently cells of interest and ii to maintain RNA integrity. The mammary gland which is a complex and heterogeneous tissue, consists of multiple cell types, changing in relative proportion during its development and thus hampering gene expression profiling comparison on whole tissue between physiological stages. During lactation, mammary epithelial cells (MEC are predominant. However several other cell types, including myoepithelial (MMC and immune cells are present, making it difficult to precisely determine the specificity of gene expression to the cell type of origin. In this work, an optimized reliable procedure for producing RNA from alveolar epithelial cells isolated from frozen histological sections of lactating goat, sheep and cow mammary glands using an infrared-laser based Arcturus Veritas LCM (Applied Biosystems® system has been developed. The following steps of the microdissection workflow: cryosectioning, staining, dehydration and harvesting of microdissected cells have been carefully considered and designed to ensure cell capture efficiency without compromising RNA integrity. Results The best results were obtained when staining 8 μm-thick sections with Cresyl violet® (Ambion, Applied Biosystems® and capturing microdissected cells during less than 2 hours before RNA extraction. In addition, particular attention was paid to animal preparation before biopsies or slaughtering (milking and freezing of tissue blocks which were embedded in a cryoprotective compound before being immersed in isopentane. The amount of RNA thus obtained from ca.150 to 250 acini

15. Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA

Li Ming-Chung

2006-04-01

Full Text Available Abstract Background Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E, Nissl Stain (NS, and for immunofluorescence (IF as well as with the plasma cell-revealing methyl green pyronin (MGP stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. Results The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. Conclusion RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

16. Pathway-focused PCR array profiling of enriched populations of laser capture microdissected hippocampal cells after traumatic brain injury.

Deborah R Boone

Full Text Available Cognitive deficits in survivors of traumatic brain injury (TBI are associated with irreversible neurodegeneration in brain regions such as the hippocampus. Comparative gene expression analysis of dying and surviving neurons could provide insight into potential therapeutic targets. We used two pathway-specific PCR arrays (RT2 Profiler Apoptosis and Neurotrophins & Receptors PCR arrays to identify and validate TBI-induced gene expression in dying (Fluoro-Jade-positive or surviving (Fluoro-Jade-negative pyramidal neurons obtained by laser capture microdissection (LCM. In the Apoptosis PCR array, dying neurons showed significant increases in expression of genes associated with cell death, inflammation, and endoplasmic reticulum (ER stress compared with adjacent, surviving neurons. Pro-survival genes with pleiotropic functions were also significantly increased in dying neurons compared to surviving neurons, suggesting that even irreversibly injured neurons are able to mount a protective response. In the Neurotrophins & Receptors PCR array, which consists of genes that are normally expected to be expressed in both groups of hippocampal neurons, only a few genes were expressed at significantly different levels between dying and surviving neurons. Immunohistochemical analysis of selected, differentially expressed proteins supported the gene expression data. This is the first demonstration of pathway-focused PCR array profiling of identified populations of dying and surviving neurons in the brain after TBI. Combining precise laser microdissection of identifiable cells with pathway-focused PCR array analysis is a practical, low-cost alternative to microarrays that provided insight into neuroprotective signals that could be therapeutically targeted to ameliorate TBI-induced neurodegeneration.

17. Identification of Site-Specific Stroke Biomarker Candidates by Laser Capture Microdissection and Labeled Reference Peptide

Tingting Lian

2015-06-01

Full Text Available The search to date for accurate protein biomarkers in acute ischemic stroke has taken into consideration the stage and/or the size of infarction, but has not accounted for the site of stroke. In the present study, multiple reaction monitoring using labeled reference peptide (LRP following laser capture microdissection (LCM is used to identify site-specific protein biomarker candidates. In middle cerebral artery occlusion (MCAO rat models, both intact and infarcted brain tissue was collected by LCM, followed by on-film digestion and semi-quantification using triple-quadrupole mass spectrometry. Thirty-four unique peptides were detected for the verification of 12 proteins in both tissue homogenates and LCM-captured samples. Six insoluble proteins, including neurofilament light polypeptide (NEFL, alpha-internexin (INA, microtubule-associated protein 2 (MAP2, myelin basic protein (MBP, myelin proteolipid protein (PLP and 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP, were found to be site-specific. Soluble proteins, such as neuron-specific enolase (NSE and ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1, and some insoluble proteins, including neurofilament heavy polypeptide (NEFH, glial fibrillary acidic protein (GFAP, microtubule-associated protein tau (MAPT and tubulin β-3 chain (TUBB3, were found to be evenly distributed in the brain. Therefore, we conclude that some insoluble protein biomarkers for stroke are site-specific, and would make excellent candidates for the design and analysis of relevant clinical studies in the future.

18. Recovery of high-quality RNA from laser capture microdissected human and rodent pancreas

Butler, Alexandra E.; Matveyenko, Aleksey V.; Kirakossian, David; Park, Johanna; Gurlo, Tatyana; Butler, Peter C.

2016-01-01

Laser capture microdissection (LCM) is a powerful method to isolate specific populations of cells for subsequent analysis such as gene expression profiling, for example, microarrays or ribonucleic (RNA)-Seq. This technique has been applied to frozen as well as formalin-fixed, paraffin-embedded (FFPE) specimens with variable outcomes regarding quality and quantity of extracted RNA. The goal of the study was to develop the methods to isolate high-quality RNA from islets of Langerhans and pancreatic duct glands (PDG) isolated by LCM. We report an optimized protocol for frozen sections to minimize RNA degradation and maximize recovery of expected transcripts from the samples using quantitative real-time polymerase chain reaction (RT-PCR) by adding RNase inhibitors at multiple steps during the experiment. This technique reproducibly delivered intact RNA (RIN values 6–7). Using quantitative RT-PCR, the expected profiles of insulin, glucagon, mucin6 (Muc6), and cytokeratin-19 (CK-19) mRNA in PDGs and pancreatic islets were detected. The described experimental protocol for frozen pancreas tissue might also be useful for other tissues with moderate to high levels of intrinsic ribonuclease (RNase) activity.

19. Optimization of laser capture microdissection and RNA amplification for gene expression profiling of prostate cancer

Vasmatzis George

2007-03-01

Full Text Available Abstract Background To discover prostate cancer biomarkers, we profiled gene expression in benign and malignant cells laser capture microdissected (LCM from prostate tissues and metastatic prostatic adenocarcinomas. Here we present methods developed, optimized, and validated to obtain high quality gene expression data. Results RNase inhibitor was included in solutions used to stain frozen tissue sections for LCM, which improved RNA quality significantly. Quantitative PCR assays, requiring minimal amounts of LCM RNA, were developed to determine RNA quality and concentration. SuperScript II™ reverse transcriptase was replaced with SuperScript III™, and SpeedVac concentration was eliminated to optimize linear amplification. The GeneChip® IVT labeling kit was used rather than the Enzo BioArray™ HighYield™ RNA transcript labeling kit since side-by-side comparisons indicated high-end signal saturation with the latter. We obtained 72 μg of labeled complementary RNA on average after linear amplification of about 2 ng of total RNA. Conclusion Unsupervised clustering placed 5/5 normal and 2/2 benign prostatic hyperplasia cases in one group, 5/7 Gleason pattern 3 cases in another group, and the remaining 2/7 pattern 3 cases in a third group with 8/8 Gleason pattern 5 cases and 3/3 metastatic prostatic adenocarcinomas. Differential expression of alpha-methylacyl coenzyme A racemase (AMACR and hepsin was confirmed using quantitative PCR.

20. Laser capture microdissection of bacterial cells targeted by fluorescence in situ hybridization

Schou, Kirstine Klitgaard; Mølbak, Lars; Jensen, Tim Kåre;

2005-01-01

Direct cultivation-independent sequence retrieval of unidentified bacteria from histological tissue sections has been limited by the difficulty of selectively isolating specific bacteria from a complex environment. Here, a new DNA isolation approach is presented for prokaryotic cells. By this met...... within the tissue with molecular analysis of 16S rRNA gene or other genes of interest. This method is widely applicable for the identification of noncultivable bacteria and their gene pool from formalin-fixed paraffin-embedded tissue samples.......Direct cultivation-independent sequence retrieval of unidentified bacteria from histological tissue sections has been limited by the difficulty of selectively isolating specific bacteria from a complex environment. Here, a new DNA isolation approach is presented for prokaryotic cells. By this...... method, a potentially pathogenic strain of the genus Brachyspira from formalin-fixed human colonic biopsies were visualized by fluorescence in situ hybridization (FISH) with a 16S rRNA-targeting oligonucleotide probe, followed by laser capture microdissection (LCM) of the targeted cells. Direct 16S r...

1. Laser capture microdissection of intestinal tissue from sea bass larvae using an optimized RNA integrity assay and validated reference genes.

Schaeck, M; De Spiegelaere, W; De Craene, J; Van den Broeck, W; De Spiegeleer, B; Burvenich, C; Haesebrouck, F; Decostere, A

2016-01-01

The increasing demand for a sustainable larviculture has promoted research regarding environmental parameters, diseases and nutrition, intersecting at the mucosal surface of the gastrointestinal tract of fish larvae. The combination of laser capture microdissection (LCM) and gene expression experiments allows cell specific expression profiling. This study aimed at optimizing an LCM protocol for intestinal tissue of sea bass larvae. Furthermore, a 3'/5' integrity assay was developed for LCM samples of fish tissue, comprising low RNA concentrations. Furthermore, reliable reference genes for performing qPCR in larval sea bass gene expression studies were identified, as data normalization is critical in gene expression experiments using RT-qPCR. We demonstrate that a careful optimization of the LCM procedure allows recovery of high quality mRNA from defined cell populations in complex intestinal tissues. According to the geNorm and Normfinder algorithms, ef1a, rpl13a, rps18 and faua were the most stable genes to be implemented as reference genes for an appropriate normalization of intestinal tissue from sea bass across a range of experimental settings. The methodology developed here, offers a rapid and valuable approach to characterize cells/tissues in the intestinal tissue of fish larvae and their changes following pathogen exposure, nutritional/environmental changes, probiotic supplementation or a combination thereof. PMID:26883391

2. DNA profiling of spermatozoa by laser capture microdissection and low volume-PCR.

Cai-xia Li

Full Text Available Genetic profiling of sperm from complex biological mixtures such as sexual assault casework samples requires isolation of a pure sperm population and the ability to analyze low abundant samples. Current standard procedure for sperm isolation includes preferential lysis of epithelial contaminants followed by collection of intact sperm by centrifugation. While effective for samples where sperm are abundant, this method is less effective when samples contain few spermatozoa. Laser capture microdissection (LCM is a proven method for the isolation of cells biological mixtures, even when found in low abundance. Here, we demonstrate the efficacy of LCM coupled with on-chip low volume PCR (LV-PCR for the isolation and genotyping of low abundance sperm samples. Our results indicate that this method can obtain complete profiles (13-16 loci from as few as 15 sperm cells with 80% reproducibility, whereas at least 40 sperm cells are required to profile 13-16 loci by standard 'in-tube' PCR. Further, LCM and LV-PCR of a sexual assault casework sample generated a DNA genotype that was consistent with that of the suspect. This method was unable, however, to analyze a casework sample from a gang rape case in which two or more sperm contributors were in a mixed population. The results indicate that LCM and LV-PCR is sensitive and effective for genotyping sperm from sperm/epithelial cell mixtures when epithelial lysis may be insufficient due to low abundance of sperm; LCM and LV-PCR, however, failed in a casework sample when spermatozoa from multiple donors was present, indicating that further study is necessitated.

3. DETECTION OF K-RAS AND P53 MUTATIONS IN SPUTUM SAMPLES OF LUNG CANCER PATIENTS USING LASER CAPTURE MICRODISSECTION MICROSCOPE AND MUTATION ANALYSIS

Detection of K-ras and p53 Mutations in Sputum Samples of Lung Cancer Patients Using Laser Capture Microdissection Microscope and Mutation AnalysisPhouthone Keohavong a,*, Wei-Min Gao a, Kui-Cheng Zheng a, Hussam Mady b, Qing Lan c, Mona Melhem b, and Judy Mumford d.<...

4. Transcriptome profiling of sheep granulosa cells and oocytes during early follicular development obtained by Laser Capture Microdissection

Bonnet Agnes

2011-08-01

Full Text Available Abstract Background Successful achievement of early folliculogenesis is crucial for female reproductive function. The process is finely regulated by cell-cell interactions and by the coordinated expression of genes in both the oocyte and in granulosa cells. Despite many studies, little is known about the cell-specific gene expression driving early folliculogenesis. The very small size of these follicles and the mixture of types of follicles within the developing ovary make the experimental study of isolated follicular components very difficult. The recently developed laser capture microdissection (LCM technique coupled with microarray experiments is a promising way to address the molecular profile of pure cell populations. However, one main challenge was to preserve the RNA quality during the isolation of single cells or groups of cells and also to obtain sufficient amounts of RNA. Using a new LCM method, we describe here the separate expression profiles of oocytes and follicular cells during the first stages of sheep folliculogenesis. Results We developed a new tissue fixation protocol ensuring efficient single cell capture and RNA integrity during the microdissection procedure. Enrichment in specific cell types was controlled by qRT-PCR analysis of known genes: six oocyte-specific genes (SOHLH2, MAEL, MATER, VASA, GDF9, BMP15 and three granulosa cell-specific genes (KL, GATA4, AMH. A global gene expression profile for each follicular compartment during early developmental stages was identified here for the first time, using a bovine Affymetrix chip. Most notably, the granulosa cell dataset is unique to date. The comparison of oocyte vs. follicular cell transcriptomes revealed 1050 transcripts specific to the granulosa cell and 759 specific to the oocyte. Functional analyses allowed the characterization of the three main cellular events involved in early folliculogenesis and confirmed the relevance and potential of LCM-derived RNA. Conclusions

5. Region-Specific In Situ Hybridization-Guided Laser-Capture Microdissection on Postmortem Human Brain Tissue Coupled with Gene Expression Quantification

Bernard, René; Burke, Sharon; Kerman, Ilan A.

2011-01-01

This chapter describes the procedure of in situ hybridization-guided laser-capture microdissection performed on postmortem human brain tissue. This procedure permits the precise collection of brain tissue within anatomically defined brain nuclei that is enriched with mRNA. The chapter emphasizes the specific handling of postmortem tissue and preservation of RNA integrity to ensure high-quality gene profiling. Downstream procedures including mRNA amplification, gene profiling using high-densit...

6. Identification of pro-angiogenic markers in blood vessels from stroked-affected brain tissue using laser-capture microdissection

Baldellou Maribel

2009-03-01

Full Text Available Abstract Background Angiogenesis correlates with patient survival following acute ischaemic stroke, and survival of neurons is greatest in tissue undergoing angiogenesis. Angiogenesis is critical for the development of new microvessels and leads to re-formation of collateral circulation, reperfusion, enhanced neuronal survival and improved recovery. Results Here, we have isolated active (CD105/Flt-1 positive and inactive (CD105/Flt-1 minus (n=5 micro-vessel rich-regions from stroke-affected and contralateral tissue of patients using laser-capture micro-dissection. Areas were compared for pro- and anti-angiogenic gene expression using targeted TaqMan microfluidity cards containing 46 genes and real-time PCR. Further analysis of key gene de-regulation was performed by immunohistochemistry to define localization and expression patterns of identified markers and de novo synthesis by human brain microvessel endothelial cells (HBMEC was examined following oxygen-glucose deprivation (OGD. Our data revealed that seven pro-angiogenic genes were notably up-regulated in CD105 positive microvessel rich regions. These were, beta-catenin, neural cell adhesion molecule (NRCAM, matrix metalloproteinase-2 (MMP-2, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1, hepatocyte growth factor-alpha (HGF-alpha, monocyte chemottractant protein-1 (MCP-1 and and Tie-2 as well as c-kit. Immunohistochemistry demonstrated strong staining of MMP-2, HGF-alpha, MCP-1 and Tie-2 in stroke-associated regions of active remodeling in association with CD105 positive staining. In vitro, OGD stimulated production of Tie-2, MCP-1 and MMP-2 in HBMEC, demonstrated a de novo response to hypoxia. Conclusion In this work we have identified concurrent activation of key angiogenic molecules associated with endothelial cell migration, differentiation and tube-formation, vessel stabilization and stem cell homing mechanisms in areas of revascularization. Therapeutic stimulation of these

7. Gene Expression Analysis of Neurons and Astrocytes Isolated by Laser Capture Microdissection from Frozen Human Brain Tissues.

Tagliafierro, Lidia; Bonawitz, Kirsten; Glenn, Omolara C; Chiba-Falek, Ornit

2016-01-01

Different cell types and multiple cellular connections characterize the human brain. Gene expression analysis using a specific population of cells is more accurate than conducting analysis of the whole tissue homogenate, particularly in the context of neurodegenerative diseases, where a specific subset of cells is affected by the different pathology. Due to the difficulty of obtaining homogenous cell populations, gene expression in specific cell-types (neurons, astrocytes, etc.) has been understudied. To leverage the use of archive resources of frozen human brains in studies of neurodegenerative diseases, we developed and calibrated a method to quantify cell-type specific-neuronal, astrocytes-expression profiles of genes implicated in neurodegenerative diseases, including Parkinson's and Alzheimer's diseases. Archive human frozen brain tissues were used to prepare slides for rapid immunostaining using cell-specific antibodies. The immunoreactive-cells were isolated by Laser Capture Microdissection (LCM). The enrichment for a particular cell-type of interest was validated in post-analysis stage by the expression of cell-specific markers. We optimized the technique to preserve the RNA integrity, so that the RNA was suitable for downstream expression analyses. Following RNA extraction, the expression levels were determined digitally using nCounter Single Cell Gene Expression assay (NanoString Technologies®). The results demonstrated that using our optimized technique we successfully isolated single neurons and astrocytes from human frozen brain tissues and obtained RNA of a good quality that was suitable for mRNA expression analysis. We present here new advancements compared to previous reported methods, which improve the method's feasibility and its applicability for a variety of downstream molecular analyses. Our new developed method can be implemented in genetic and functional genomic research of neurodegenerative diseases and has the potential to significantly

8. Laser capture microdissection of bacterial cells targeted by fluorescence in situ hybridization

Schou, Kirstine Klitgaard; Mølbak, Lars; Jensen, Tim Kåre; Lindboe, Christian Frederik; Boye, Mette

2005-01-01

Direct cultivation-independent sequence retrieval of unidentified bacteria from histological tissue sections has been limited by the difficulty of selectively isolating specific bacteria from a complex environment. Here, a new DNA isolation approach is presented for prokaryotic cells. By this......RNA gene PCR was performed from the dissected microcolonies, and the subsequent DNA sequence analysis identified the dissected bacterial cells as belonging to the Brachyspira aalborgi cluster 1. The advantage of this technique is the ability to combine the histological recognition of the specific bacteria...

9. Efficacy of laser capture microdissection plus RT-PCR technique in analyzing gene expression levels in human gastric cancer and colon cancer

Thymidylate synthase, dihydropyrimidine dehydrogenase, thymidine phosphorylase, and orotate phosphoribosyltransferase gene expressions are reported to be valid predictive markers for 5-fluorouracil sensitivity to gastrointestinal cancer. For more reliable predictability, their expressions in cancer cells and stromal cells in the cancerous tissue (cancerous stroma) have been separately investigated using laser capture microdissection. Thymidylate synthase, dihydropyrimidine dehydrogenase, thymidine phosphorylase, and orotate phosphoribosyltransferase mRNA in cancer cells and cancerous stroma from samples of 47 gastric and 43 colon cancers were separately quantified by reverse transcription polymerase chain reaction after laser capture microdissection. In both gastric and colon cancers, thymidylate synthase and orotate phosphoribosyltransferase mRNA expressions were higher (p < 0.0001, p <0.0001 respectively in gastric cancer and P = 0.0002, p < 0.0001 respectively in colon cancer) and dihydropyrimidine dehydrogenase mRNA expressions were lower in cancer cells than in cancerous stroma (P = 0.0136 in gastric cancer and p < 0.0001 in colon cancer). In contrast, thymidine phosphorylase mRNA was higher in cancer cells than in cancerous stroma in gastric cancer (p < 0.0001) and lower in cancer cells than in cancerous stroma in colon cancer (P = 0.0055). By using this method, we could estimate gene expressions separately in cancer cells and stromal cells from colon and gastric cancers, in spite of the amount of stromal tissue. Our method is thought to be useful for accurately evaluating intratumoral gene expressions

10. Rhodamine-RCA in vivo labeling guided laser capture microdissection of cancer functional angiogenic vessels in a murine squamous cell carcinoma mouse model

Bur Monica

2006-02-01

Full Text Available Abstract Background Cancer growth, invasion and metastasis are highly related to tumor-associated neovasculature. The presence and progression of endothelial cells in cancer is chaotic, unorganized, and angiogenic vessels are less functional. Therefore, not all markers appearing on the chaotic endothelial cells are accessible if a drug is given through the vascular route. Identifying endothelial cell markers from functional cancer angiogenic vessels will indicate the accessibility and potential efficacy of vascular targeted therapies. Results In order to quickly and effectively identify endothelial cell markers on the functional and accessible tumor vessels, we developed a novel technique by which tumor angiogenic vessels are labeled in vivo followed by Laser Capture Microdissection of microscopically isolated endothelial cells for genomic screening. Female C3H mice (N = 5 with established SCCVII tumors were treated with Rhodamine-RCA lectin by tail vein injection, and after fluorescence microscopy showed a successful vasculature staining, LCM was then performed on frozen section tissue using the PixCell II instrument with CapSure HS caps under the Rhodamine filter. By this approach, the fluorescent angiogenic endothelial cells were successfully picked up. As a result, the total RNA concentration increased from an average of 33.4 ng/ul +/- 24.3 (mean +/- S.D. to 1913.4 ng/ul +/- 164. Relatively pure RNA was retrieved from both endothelial and epithelial cells as indicated by the 260/280 ratios (range 2.22–2.47. RT-PCR and gene electrophoresis successfully detected CD31 and Beta-Actin molecules with minimal Keratin 19 expression, which served as the negative control. Conclusion Our present study demonstrates that in vivo Rhodamine RCA angiogenic vessel labeling provided a practical approach to effectively guide functional endothelial cell isolation by laser capture microdissection with fluorescent microscopy, resulting in high quality RNA and

11. Laser capture microdissection and cDNA array analysis of endometrium identify CCL16 and CCL21 as epithelial-derived inflammatory mediators associated with endometriosis

Jones Rebecca L

2007-05-01

Full Text Available Abstract Background Understanding the pathophysiology of chemokine secretion in endometriosis may offer a novel area of therapeutic intervention. This study aimed to identify chemokines differentially expressed in epithelial glands in eutopic endometrium from normal women and those with endometriosis, and to establish the expression profiles of key chemokines in endometriotic lesions. Methods Laser capture microdissection isolated epithelial glands from endometrial eutopic tissue from women with and without endometriosis in the mid-secretory phase of their menstrual cycles. Gene profiling of the excised glands used a human chemokine and receptor cDNA array. Selected chemokines were further examined using real-time PCR and immunohistochemistry. Results 22 chemokine/receptor genes were upregulated and two downregulated in pooled endometrial epithelium of women with endometriosis compared with controls. CCL16 and CCL21 mRNA was confirmed as elevated in some women with endometriosis compared to controls on individual samples. Immunoreactive CCL16 and CCL21 were predominantly confined to glands in eutopic and ectopic endometrium: leukocytes also stained. Immunoreactive CCL16 was overall higher in glands in ectopic vs. eutopic endometrium from the same woman (P Conclusion This study provides novel candidate molecules and suggests a potential local role for CCL16 and CCL21 as mediators contributing to the inflammatory events associated with endometriosis.

12. Senescent vs. non-senescent cells in the human annulus in vivo: Cell harvest with laser capture microdissection and gene expression studies with microarray analysis

Ingram Jane A

2010-01-01

Full Text Available Abstract Background Senescent cells are well-recognized in the aging/degenerating human disc. Senescent cells are viable, cannot divide, remain metabolically active and accumulate within the disc over time. Molecular analysis of senescent cells in tissue offers a special challenge since there are no cell surface markers for senescence which would let one use fluorescence-activated cell sorting as a method for separating out senescent cells. Methods We employed a novel laser capture microdissection (LCM design to selectively harvest senescent and non-senescent annulus cells in paraffin-embedded tissue, and compared their gene expression with microarray analysis. LCM was used to separately harvest senescent and non-senescent cells from 11 human annulus specimens. Results Microarray analysis revealed significant differences in expression levels in senescent cells vs non-senescent cells: 292 genes were upregulated, and 321 downregulated. Genes with established relationships to senescence were found to be significantly upregulated in senescent cells vs. non-senescent cells: p38 (MPAK14, RB-Associated KRAB zinc finger, Discoidin, CUB and LCCL domain, growth arrest and DNA-damage inducible beta, p28ING5, sphingosine-1-phosphate receptor 2 and somatostatin receptor 3; cyclin-dependent kinase 8 showed significant downregulation in senescent cells. Nitric oxidase synthase 1, and heat shock 70 kDa protein 6, both of which were significantly down-regulated in senescent cells, also showed significant changes. Additional genes related to cytokines, cell proliferation, and other processes were also identified. Conclusions Our LCM-microarray analyses identified a set of genes associated with senescence which were significantly upregulated in senescent vs non-senescent cells in the human annulus. These genes include p38 MAP kinase, discoidin, inhibitor of growth family member 5, and growth arrest and DNA-damage-inducible beta. Other genes, including genes

13. Laser capture microdissection followed by next-generation sequencing identifies disease-related microRNAs in psoriatic skin that reflect systemic microRNA changes in psoriasis

Løvendorf, Marianne B; Mitsui, Hiroshi; Zibert, John R;

2015-01-01

Psoriasis is a systemic disease with cutaneous manifestations. MicroRNAs (miRNAs) are small non-coding RNA molecules that are differentially expressed in psoriatic skin; however, only few cell- and region-specific miRNAs have been identified in psoriatic lesions. We used laser capture microdissec......Psoriasis is a systemic disease with cutaneous manifestations. MicroRNAs (miRNAs) are small non-coding RNA molecules that are differentially expressed in psoriatic skin; however, only few cell- and region-specific miRNAs have been identified in psoriatic lesions. We used laser capture...

14. Suspension fluorescence in situ hybridization (S-FISH) combined with automatic detection and laser microdissection for STR profiling of male cells in male/female mixtures

Vandewoestyne, Mado; Van Hoofstat, David; Van Nieuwerburgh, Filip; Deforce, Dieter

2009-01-01

Laser microdissection is a valuable tool for isolating specific cells from mixtures, such as male cells in a mixture with female cells, e.g., in cases of sexual assault. These cells can be stained with Y-chromosome-specific probes. We developed an automatic screening method to detect male cells after fluorescence in situ hybridization in suspension (S-FISH). To simulate forensic casework, the method was tested on female saliva after cataglottis (a kiss involving tongue-to-tongue contact) and ...

15. Laser capture microscopy

Curran, S.; McKay, J A; McLeod, H L; Murray, G I

2000-01-01

Human tissues are composed of complex admixtures of different cell types and their biologically meaningful analysis necessitates the procurement of pure samples of the cells of interest. Many approaches have been used in attempts to overcome this difficulty, including a variety of microdissection methods. This review concerns a recent advance in microdissection techniques, namely laser capture microdissection (LCM). The principle underlying this technique is outlined, and practical issues per...

16. CpG Methylation Analysis of HPV16 in Laser Capture Microdissected Archival Tissue and Whole Tissue Sections from High Grade Anal Squamous Intraepithelial Lesions: A Potential Disease Biomarker

Molano, Monica; Tabrizi, Sepehr N.; Garland, Suzanne M.; Roberts, Jennifer M.; Machalek, Dorothy A.; Phillips, Samuel; Chandler, David; Hillman, Richard J.; Grulich, Andrew E.; Jin, Fengyi; Poynten, I. Mary; Templeton, David J.; Cornall, Alyssa M.

2016-01-01

Incidence and mortality rates of anal cancer are increasing globally. More than 90% of anal squamous cell carcinomas (ASCC) are associated with human papillomavirus (HPV). Studies on HPV-related anogenital lesions have shown that patterns of methylation of viral and cellular DNA targets could potentially be developed as disease biomarkers. Lesion-specific DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissues from existing or prospective patient cohorts may constitute a valuable resource for methylation analysis. However, low concentrations of DNA make these samples technically challenging to analyse using existing methods. We therefore set out to develop a sensitive and reproducible nested PCR-pyrosequencing based method to accurately quantify methylation at 10 CpG sites within the E2BS1, E2BS2,3,4 and Sp1 binding sites in the viral upstream regulatory region of HPV16 genome. Methylation analyses using primary and nested PCR-pyrosequencing on 52 FFPE tissue [26 paired whole tissue sections (WTS) and laser capture microdissected (LCM) tissues] from patients with anal squamous intraepithelial lesions was performed. Using nested PCR, methylation results were obtained for the E2BS1, E2BS2,3,4 and Sp1 binding sites in 86.4% of the WTS and 81.8% of the LCM samples. Methylation patterns were strongly correlated within median values of matched pairs of WTS and LCM sections, but overall methylation was higher in LCM samples at different CpG sites. High grade lesions showed low methylation levels in the E2BS1 and E2BS2 regions, with increased methylation detected in the E2BS,3,4/Sp1 regions, showing the highest methylation at CpG site 37. The method developed is highly sensitive in samples with low amounts of DNA and demonstrated to be suitable for archival samples. Our data shows a possible role of specific methylation in the HPV16 URR for detection of HSIL. PMID:27529629

17. CpG Methylation Analysis of HPV16 in Laser Capture Microdissected Archival Tissue and Whole Tissue Sections from High Grade Anal Squamous Intraepithelial Lesions: A Potential Disease Biomarker.

Molano, Monica; Tabrizi, Sepehr N; Garland, Suzanne M; Roberts, Jennifer M; Machalek, Dorothy A; Phillips, Samuel; Chandler, David; Hillman, Richard J; Grulich, Andrew E; Jin, Fengyi; Poynten, I Mary; Templeton, David J; Cornall, Alyssa M

2016-01-01

Incidence and mortality rates of anal cancer are increasing globally. More than 90% of anal squamous cell carcinomas (ASCC) are associated with human papillomavirus (HPV). Studies on HPV-related anogenital lesions have shown that patterns of methylation of viral and cellular DNA targets could potentially be developed as disease biomarkers. Lesion-specific DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissues from existing or prospective patient cohorts may constitute a valuable resource for methylation analysis. However, low concentrations of DNA make these samples technically challenging to analyse using existing methods. We therefore set out to develop a sensitive and reproducible nested PCR-pyrosequencing based method to accurately quantify methylation at 10 CpG sites within the E2BS1, E2BS2,3,4 and Sp1 binding sites in the viral upstream regulatory region of HPV16 genome. Methylation analyses using primary and nested PCR-pyrosequencing on 52 FFPE tissue [26 paired whole tissue sections (WTS) and laser capture microdissected (LCM) tissues] from patients with anal squamous intraepithelial lesions was performed. Using nested PCR, methylation results were obtained for the E2BS1, E2BS2,3,4 and Sp1 binding sites in 86.4% of the WTS and 81.8% of the LCM samples. Methylation patterns were strongly correlated within median values of matched pairs of WTS and LCM sections, but overall methylation was higher in LCM samples at different CpG sites. High grade lesions showed low methylation levels in the E2BS1 and E2BS2 regions, with increased methylation detected in the E2BS,3,4/Sp1 regions, showing the highest methylation at CpG site 37. The method developed is highly sensitive in samples with low amounts of DNA and demonstrated to be suitable for archival samples. Our data shows a possible role of specific methylation in the HPV16 URR for detection of HSIL. PMID:27529629

18. Pre-Combustion Carbondioxide Capture in Integrated Gasification Combined Cycles

M. Zeki YILMAZOĞLU

2010-02-01

Full Text Available Thermal power plants have a significant place big proportion in the production of electric energy. Thermal power plants are the systems which converts heat energy to mechanical energy and also mechanical energy to electrical energy. Heat energy is obtained from combustion process and as a result of this, some harmful emissions, like CO2, which are the reason for global warming, are released to atmosphere. The contribution of carbondioxide to global warming has been exposed by the previous researchs. Due to this fact, clean energy technologies are growing rapidly all around the world. Coal is generally used in power plants and when compared to other fossil energy sources unit electricity production cost is less than others. When reserve rate is taken into account, coal may be converted to energy in a more efficient and cleaner way. The aim for using the clean coal technologies are to eradicate the harmful emissions of coal and to store the carbondioxide, orginated from combustion, in different forms. In line with this aim, carbondioxide may be captured by either pre-combustion, by O2/CO2 recycling combustion systems or by post combustion. The integrated gasification combined cycles (IGCC are available in pre-combustion capture systems, whereas in O2/CO2 recycling combustion systems there are ultrasuper critical boiler technologies and finally flue gas washing systems by amines exists in post combustion systems. In this study, a pre-combustion CO2 capture process via oxygen blown gasifiers is compared with a conventional power plant in terms of CO2 emissions. Captured carbondioxide quantity has been presented as a result of the calculations made throughout the study.

19. Linear combination reading program for capture gamma rays

Tanner, Allan B.

1971-01-01

This program computes a weighting function, Qj, which gives a scalar output value of unity when applied to the spectrum of a desired element and a minimum value (considering statistics) when applied to spectra of materials not containing the desired element. Intermediate values are obtained for materials containing the desired element, in proportion to the amount of the element they contain. The program is written in the BASIC language in a format specific to the Hewlett-Packard 2000A Time-Sharing System, and is an adaptation of an earlier program for linear combination reading for X-ray fluorescence analysis (Tanner and Brinkerhoff, 1971). Following the program is a sample run from a study of the application of the linear combination technique to capture-gamma-ray analysis for calcium (report in preparation).

20. 激光捕获显微切割技术诊断石蜡切片中早期胚胎成分与亲权关系的研究%Separation of chorionic villi from paraffin-embedded early abortion materials and confirmation of parent-child relationship by laser capture microdissection

李艮平; 李彩霞; 郝建文; 万立华; 胡兰; 韩俊平; 杨帆

2012-01-01

目的 通过激光捕获显微切割技术捕获石蜡切片中胚胎绒毛细胞,进行低体积扩增,实现胚胎个体分型,证实亲子关系.方法 通过激光捕获显微技术捕获石蜡切片上胚胎绒毛细胞,设置20、30、40个细胞数目组,分别捕获不同数目绒毛细胞,利用低体积扩增仪进行1.5μl体系PCR扩增,采用3130遗传分析仪得出胚胎个体分型图谱,并比较各细胞数目组胚胎个体识别的检出率、等位基因丢失率和非特异性扩增情况.最后进行实际案例应用.结果 捕获20个胚胎绒毛细胞即可获得胚胎个体分型,40个细胞数目组的胚胎分型检出率最高,等位基因丢失率和非特异性扩增率最低.20个细胞数目反之.结论 激光捕获显微切割技术结合低体积扩增技术可应用于石蜡切片,实现胚胎与母体成分的精确分离并确证亲权关系.%Objective It remains difficult to genotype the mixture of maternal and embryo from early abortion tissues to investigate forensic evidence and confirm parent-child relationship. Our study was aimed to confirm the parent-child relationship by using laser capture microdissection ( LCM) to capture the embryonic villi cells in paraffin-embedded section for genotyping embryonic individual via ultralow volume PCR amplification. Methods Embryonic villi cells in the paraffin sections was captured by LCM technology. With the setting of cell numbers at 20, 30 and 40 cells as a group, villus cells of different cell numbers were captured and subjected to PCR amplification at low volume of 1. 5 μl by using 3130 Genetic Analysis instrument to yield embryonic genotyping maps. The artificial alleles, detection rate and allelic dropout rate of each group were compared. The technology was also applied on an actual case. Results Embryonic villi cells in the paraffin sections were captured by LCM technology. With the setting of cell numbers at 20, 30 and 40 cells for group, villus cells of different cell

1. Laser capture.

Potter, S Steven; Brunskill, Eric W

2012-01-01

This chapter describes detailed methods used for laser capture microdissection (LCM) of discrete subpopulations of cells. Topics covered include preparing tissue blocks, cryostat sectioning, processing slides, performing the LCM, and purification of RNA from LCM samples. Notes describe the fine points of each operation, which can often mean the difference between success and failure. PMID:22639264

2. MICRODISSECTION OF MINIATURE PIG EAR

YI Haijin; GuoWeiwei; Chen Lei; Wu Na; Li Jiana; Ren Lili; Yang Shiming

2013-01-01

Objective To investigate the suitability of miniature pigs as an animal model for otological research. Methods Microdissection of the temporal bone was performed on 10 miniature pigs and recorded on photo-graphs. Results The morphology and measurement of the external, middle and inner ear and the lateral re-cess of the miniature pigs were obtained by microdissection. Conclusion Compared to traditional animal models, the miniature pig may be a better model for biomedical research because of its many similarities in physiological functions with humans. Similarities of the temporal bone structures, including the external, middle and inner ear and the lateral recess, between the miniature pig and human make the animal a poten-tially useful model for otological research.

3. Microdissection, mRNA amplification and microarray: a study of pleural mesothelial and malignant mesothelioma cells.

Mohr, Steve; Bottin, Marie-Claire; Lannes, Béatrice; Neuville, Agnès; Bellocq, Jean-Pierre; Keith, Gérard; Rihn, Bertrand Henri

2004-01-01

The studies of molecular alterations in tumor cells with microarrays are often hampered by inherent tissue heterogeneity. The emergence of Laser Capture Microdissection (LCM) allowed us to overcome this challenge since it gives selective access to cancer cells that are isolated from their native tissue environment. In this report, we microdissected mesothelial cells and malignant mesothelioma cells of ex vivo resected specimens using LCM. Amplified RNA from mesothelial and mesothelioma microdissected cells allowed us to measure global gene expression with 10 K-microarrays in four independent experiments. We screened 9850 annotated human genes, 1275 of which have satisfied our data analysis requirements. They included 302 overexpressed genes and 160 downregulated genes in mesothelioma microdissected cells as compared to mesothelial microdissected cells. Among them, the expression levels of eight genes, namely BF, FTL, IGFBP7, RARRES1, RARRES2, RBP1, SAT, and TXN according to HUGO nomenclature, were increased, whereas six: ALOX5AP, CLNS1A, EIF4A2, ELK3, REQ and SYPL, were found to be underexpressed in mesothelioma microdissected cells. The ferritin light polypeptide (FTL) gene overexpression was confirmed by real time quantitative PCR. Our approach allowed a comprehensive in situ examination of mesothelioma and provided an accurate way to find new marker genes that may be useful for diagnosis and treatment of malignant pleural mesothelioma. PMID:14987796

4. Counting with Combined Splitting and Capture-Recapture Methods

2011-01-01

We apply the splitting method to three well-known counting problems, namely 3-SAT, random graphs with prescribed degrees, and binary contingency tables. We present an enhanced version of the splitting method based on the capture-recapture technique, and show by experiments the superiority of this technique for SAT problems in terms of variance of the associated estimators, and speed of the algorithms.

5. Proteomic analysis of prolactinoma cells by immuno-laser capture microdissection combined with online two-dimensional nano-scale liquid chromatography/mass spectrometry

Chen Luping; Zhuang Dongxiao; Hou Ruiping; Yan Guoquan; Wu Jinsong; Liu Yingchao; Pang Qi; Zhu Jianhong

2010-01-01

Abstract Background Pituitary adenomas, the third most common intracranial tumor, comprise nearly 16.7% of intracranial neoplasm and 25%-44% of pituitary adenomas are prolactinomas. Prolactinoma represents a complex heterogeneous mixture of cells including prolactin (PRL), endothelial cells, fibroblasts, and other stromal cells, making it difficult to dissect the molecular and cellular mechanisms of prolactin cells in pituitary tumorigenesis through high-throughout-omics analysis. Our newly d...

6. Optimizing Frozen Sample Preparation for Laser Microdissection: Assessment of CryoJane Tape-Transfer System®.

Yelena G Golubeva

Full Text Available Laser microdissection is an invaluable tool in medical research that facilitates collecting specific cell populations for molecular analysis. Diversity of research targets (e.g., cancerous and precancerous lesions in clinical and animal research, cell pellets, rodent embryos, etc. and varied scientific objectives, however, present challenges toward establishing standard laser microdissection protocols. Sample preparation is crucial for quality RNA, DNA and protein retrieval, where it often determines the feasibility of a laser microdissection project. The majority of microdissection studies in clinical and animal model research are conducted on frozen tissues containing native nucleic acids, unmodified by fixation. However, the variable morphological quality of frozen sections from tissues containing fat, collagen or delicate cell structures can limit or prevent successful harvest of the desired cell population via laser dissection. The CryoJane Tape-Transfer System®, a commercial device that improves cryosectioning outcomes on glass slides has been reported superior for slide preparation and isolation of high quality osteocyte RNA (frozen bone during laser dissection. Considering the reported advantages of CryoJane for laser dissection on glass slides, we asked whether the system could also work with the plastic membrane slides used by UV laser based microdissection instruments, as these are better suited for collection of larger target areas. In an attempt to optimize laser microdissection slide preparation for tissues of different RNA stability and cryosectioning difficulty, we evaluated the CryoJane system for use with both glass (laser capture microdissection and membrane (laser cutting microdissection slides. We have established a sample preparation protocol for glass and membrane slides including manual coating of membrane slides with CryoJane solutions, cryosectioning, slide staining and dissection procedure, lysis and RNA extraction

7. USING TAILORED TRANQUILIZER COMBINATIONS TO REDUCE STRESS ASSOCIATED WITH LARGE UNGULATE CAPTURE AND TRANSLOCATION.

Wolfe, Lisa L; Miller, Michael W

2016-04-01

Capture and translocation are important tools for managing and studying large ungulates. Although widely used, many established field practices cause fear and stress in subject animals that can hamper overall effectiveness and safety. Over the last 10 years we have been exploring uses of tranquilizer combinations as adjuncts to wild ungulate capture and translocation work in Colorado, USA. Our approaches have been tailored to various field applications to reduce fear and stress, facilitate handling, and improve the overall success of capture and translocation for research or management purposes. For physical capture (drop net or helicopter-net gunning) with local release, combinations of midazolam and azaperone administered immediately upon capture provide transient tranquilization and muscle relaxation during manual restraint and handling to prevent hyperthermia and capture myopathy. For extended tranquilization (during transport and overnight holding), adding a sustained-release haloperidol formulation provides calming effects for at least 24-48 h. In our assessment, appropriate and adaptive use of these tranquilizer combinations benefits captured animals without impeding management or research goals. PMID:26845295

8. Landmine detection method combined with backscattering neutrons and capture {gamma}-rays from hydrogen

Takahashi, Yoshiyuki, E-mail: ytaka@rri.kyoto-u.ac.j [Graduate School of Energy Science, Kyoto University, Yoshida-honmachi, Sakyo-ku, Kyoto 606-8501 (Japan); Misawa, Tsuyoshi; Pyeon, Cheol Ho; Shiroya, Seiji [Research Reactor Institute, Kyoto University, Asashiro-nishi, Kumatori-cho, Sennan-gun, Osaka 590-0494 (Japan); Yoshikawa, Kiyoshi [Institute of Advanced Energy, Kyoto University, Gokashou, Uji-shi, Kyoto 611-0011 (Japan)

2011-07-15

The usefulness of the measurements of the backscattering neutron and 2.22 MeV capture {gamma}-ray from hydrogen in the landmine detection method is described in this paper. When the soil moisture content is increased, the reaction rates of both the neutron scattering reaction and capture reaction are increased. However, the backscattering neutrons are more influenced than the capture {gamma}-rays by the soil moisture before the reaction with the detector. The facts that the backscattering neutron method is useful in the dry soil case and that the capture {gamma}-ray method is effective in well-wet soil case are confirmed by the experiments and the calculations. The landmine detection efficiency is improved in various soil moisture conditions by combining the backscattering neutron method together with the capture {gamma}-ray method. The effectiveness of the pulse mode operation was confirmed numerically.

9. Captured streams and springs in combined sewers: a review of the evidence, consequences and opportunities.

2013-09-01

Captured streams and springs may be flowing in combined sewers, increasing clean baseflow in pipes and wastewater treatment works (WwTWs), reducing pipe capacity and increasing treatment costs. The UK water industry is aware of this in principle, but there has been no explicit discussion of this in the published literature, nor have there been any known attempts to manage it. Instead, the current focus is on the similar intrusion of groundwater infiltration through pipe cracks and joints. We have conducted a thorough review of literature and international case studies to investigate stream and spring capture, finding several examples with convincing evidence that this occurs. We identify three modes of entry: capture by conversion, capture by interception, and direct spring capture. Methods to identify and quantify capture are limited, but the experience in Zurich suggests that it contributed 7-16% of the baseflow reaching WwTWs. There are negative impacts for the water industry in capital and operational expenditure, as well as environmental and social impacts of loss of urban streams. For a typical WwTW (Esholt, Bradford) with 16% of baseflow from captured streams and springs, we conservatively estimate annual costs of £ 2 million to £ 7 million. A detailed case study from Zurich is considered that has successfully separated captured baseflow into daylighted streams through the urban area, with multiple economic, environmental and social benefits. We conclude that there is a strong case for the UK water industry to consider captured streams and springs, quantify them, and assess the merits of managing them. PMID:23810838

10. Extracellular protease mRNAs are predominantly expressed in the stromal areas of microdissected mouse breast carcinomas

Pedersen, Tanja Xenia; Pennington, Caroline J; Almholt, Kasper;

2005-01-01

cells. We have now used laser capture microdissection and real-time PCR to quantify the mRNA expression of components of matrix-degrading proteolytic systems in cancer and stromal areas of mouse mammary tumors genetically induced by the polyoma virus middle T (PyMT) antigen. We examined the mRNA levels...

11. Microdissection testicular sperm extraction: an update

Dabaja, Ali A; Schlegel, Peter N.

2012-01-01

Patients with non-obstructive azoospermia (NOA) were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate. In the last two decades, the advent of intracytoplasmic sperm injection (ICSI), and the application of various testicular sperm retrieval techniques, including fine needle aspiration (FNA), conventional testicular sperm extraction (TESE) and microdissection testicular sperm extraction (micro-TESE) have revolutionized treatment in this gr...

12. Processing of Microdissected Tissue for Molecular Analysis

sprotocols

2014-01-01

Author: National Cancer Institute These methods were successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor the following protocol for their own research objectives and tissue under study. **More than 10,000 Cells** If the amount of microdissected material is substantial (>10,000 cells) then any of the standard procedures for isolating DNA are acceptable. **Less than 10,000 Cells** If the n...

13. Extended Field Laser Confocal Microscopy (EFLCM): Combining automated Gigapixel image capture with in silico virtual microscopy

Confocal laser scanning microscopy has revolutionized cell biology. However, the technique has major limitations in speed and sensitivity due to the fact that a single laser beam scans the sample, allowing only a few microseconds signal collection for each pixel. This limitation has been overcome by the introduction of parallel beam illumination techniques in combination with cold CCD camera based image capture. Using the combination of microlens enhanced Nipkow spinning disc confocal illumination together with fully automated image capture and large scale in silico image processing we have developed a system allowing the acquisition, presentation and analysis of maximum resolution confocal panorama images of several Gigapixel size. We call the method Extended Field Laser Confocal Microscopy (EFLCM). We show using the EFLCM technique that it is possible to create a continuous confocal multi-colour mosaic from thousands of individually captured images. EFLCM can digitize and analyze histological slides, sections of entire rodent organ and full size embryos. It can also record hundreds of thousands cultured cells at multiple wavelength in single event or time-lapse fashion on fixed slides, in live cell imaging chambers or microtiter plates. The observer independent image capture of EFLCM allows quantitative measurements of fluorescence intensities and morphological parameters on a large number of cells. EFLCM therefore bridges the gap between the mainly illustrative fluorescence microscopy and purely quantitative flow cytometry. EFLCM can also be used as high content analysis (HCA) instrument for automated screening processes

14. Laser capture microdissection of gonads from juvenile zebrafish

Jørgensen, Anne; Nielsen, John; Morthorst, Jane Ebsen;

2009-01-01

Background: Investigating gonadal gene expression is important in attempting to elucidate the molecular mechanism of sex determination and differentiation in the model species zebrafish. However, the small size of juvenile zebrafish and correspondingly their gonads complicates this type of...... investigation. Furthermore, the lack of a genetic sex marker in juvenile zebrafish prevents pooling gonads from several individuals. The aim of this study was to establish a method to isolate the gonads from individual juvenile zebrafish allowing future investigations of gonadal gene expression during sex...... was adjusted and optimised to isolate juvenile zebrafish gonads. Results: The juvenile zebrafish gonad is not morphologically distinguishable when using dehydrated cryosections on membrane slides and a specific staining method is necessary to identify the gonads. The protocol setup in this study...

15. Combination of the prompt neutron capture method with other neutron methods for substance elemental content analysis

Neutron analysis method of determining element composition have found wide range of applications in industry thanks to different types of interaction of neutron with substances /1/. With the aim of widening the range of problems to be solved, on the basis of the device /2/ for determining the element content of substance, possibilities of combining the method based on the use of neutron capture gamma-ray spectrometry with other neutron methods, in particular neutron activation analysis and neutron absorption analysis were studied. In this radionuclide source (252Cf) with the yield of 1,5 x 107 neutron/sec is used. By means of using neutron capture gamma radiation spectrometry the possibilities of determining some elements (H, B, N, S etc. ), which are not determined by very widely used method, activation analysis. These elements can be determined by both the semiconductor and scintillation detectors with parameters fitting the manufacturing requirements. And for a number of elements ( B, Cl, Cd, Sm, Gd) very high limits of determination ( up to 10- 5 %) are possible using semiconductor Ge (Li) -detectors with high resolution. Possibility of determination of some 'well' activated elements ( K, Al, Fe, Mn, Ti, Sc etc.) in samples of ore and products of their processing using the neutron-activation analysis. For 1 hour of irradiation on the experimental device quite accurate analytical peak, of these elements are obtained, allowing to determine them qualitatively. However, with decreasing neutron yield of radionuclide source it becomes more difficult to achieve the necessary parameters both in neutron capture and activation analysis. Experimental works on determination of some elements with large cross-sections of capture ( B, Cd, Sm ) by absorption of neutrons in the investigated substance, i.e. using the neutron absorption analysis method with absence of other large capture cross section elements in the samples being studied. (author)

16. Microfluidics: The future of microdissection TESE?

Samuel, Raheel; Badamjav, Odgerel; Murphy, Kristin E; Patel, Darshan P; Son, Jiyoung; Gale, Bruce K; Carrell, Douglas T; Hotaling, James M

2016-06-01

Non-obstructive azoospermia (NOA) is a severe form of infertility accounting for 10% of infertile men. Microdissection testicular sperm extraction (microTESE) includes a set of clinical protocols from which viable sperm are collected from patients (suffering from NOA), for intracytoplasmic sperm injection (ICSI). Clinical protocols associated with the processing of a microTESE sample are inefficient and significantly reduce the success of obtaining a viable sperm population. In this review we highlight the sources of these inefficiencies and how these sources can possibly be removed by microfluidic technology and single-cell Raman spectroscopy. PMID:27104311

17. Transient studies of an Integrated Gasification Combined Cycle (IGCC) plant with CO2 capture

Bhattacharyya, D.; Turton, R.; Zitney, S.

2010-01-01

Next-generation coal-fired power plants need to consider the option for CO2 capture as stringent governmental mandates are expected to be issued in near future. Integrated gasification combined cycle (IGCC) plants are more efficient than the conventional coal combustion processes when the option for CO2 capture is considered. However, no IGCC plant with CO2 capture currently exists in the world. Therefore, it is important to consider the operability and controllability issues of such a plant before it is commercially built. To facilitate this objective, a detailed plant-wide dynamic simulation of an IGCC plant with 90% CO2 capture has been developed in Aspen Plus Dynamics{reg_sign}. The plant considers a General Electric Energy (GEE)-type downflow radiant-only gasifier followed by a quench section. A two-stage water gas shift (WGS) reaction is considered for conversion of CO to CO2. A two-stage acid gas removal (AGR) process based on a physical solvent is simulated for selective capture of H2S and CO2. Compression of the captured CO2 for sequestration, an oxy-Claus process for removal of H2S and NH3, black water treatment, and the sour water treatment are also modeled. The tail gas from the Claus unit is recycled to the SELEXOL unit. The clean syngas from the AGR process is sent to a gas turbine followed by a heat recovery steam generator. This turbine is modeled as per published data in the literature. Diluent N2 is used from the elevated-pressure ASU for reducing the NOx formation. The heat recovery steam generator (HRSG) is modeled by considering generation of high-pressure, intermediate-pressure, and low-pressure steam. All of the vessels, reactors, heat exchangers, and the columns have been sized. The basic IGCC process control structure has been synthesized by standard guidelines and existing practices. The steady state results are validated with data from a commercial gasifier. In the future grid-connected system, the plant should satisfy the environmental

18. Microdissection of shoot meristem functional domains.

Lionel Brooks

2009-05-01

Full Text Available The shoot apical meristem (SAM maintains a pool of indeterminate cells within the SAM proper, while lateral organs are initiated from the SAM periphery. Laser microdissection-microarray technology was used to compare transcriptional profiles within these SAM domains to identify novel maize genes that function during leaf development. Nine hundred and sixty-two differentially expressed maize genes were detected; control genes known to be upregulated in the initiating leaf (P0/P1 or in the SAM proper verified the precision of the microdissections. Genes involved in cell division/growth, cell wall biosynthesis, chromatin remodeling, RNA binding, and translation are especially upregulated in initiating leaves, whereas genes functioning during protein fate and DNA repair are more abundant in the SAM proper. In situ hybridization analyses confirmed the expression patterns of six previously uncharacterized maize genes upregulated in the P0/P1. P0/P1-upregulated genes that were also shown to be downregulated in leaf-arrested shoots treated with an auxin transport inhibitor are especially implicated to function during early events in maize leaf initiation. Reverse genetic analyses of asceapen1 (asc1, a maize D4-cyclin gene upregulated in the P0/P1, revealed novel leaf phenotypes, less genetic redundancy, and expanded D4-CYCLIN function during maize shoot development as compared to Arabidopsis. These analyses generated a unique SAM domain-specific database that provides new insight into SAM function and a useful platform for reverse genetic analyses of shoot development in maize.

19. Microdissecção e captura a laser na investigação do gene TP53 em tecidos incluídos em parafina Laser-capture microdissection for TP53 gene analysis in paraffin-embedded tissues

2007-02-01

20. Sample preparation for laser-microdissection of soybean shoot apical meristem

Wong, Chui E; Mohan B. Singh; Bhalla, Prem L

2012-01-01

The shoot apical meristem houses stem cells responsible for the continuous formation of aerial plant organs including leaves and stems throughout the life of plants. Laser-microdissection in combination with high-throughput technology such as next generation sequencing permits an in-depth analysis of molecular events associated with specific cell type of interest. Sample preparation is the most critical step in ensuring good quality RNA to be extracted from samples following laser-microdissec...

1. Capture and immobilisation of aardvark (Orycteropus afer using different drug combinations : research communication

P.J. Nel

2000-07-01

Full Text Available Nine aardvarks (Orycteropus afer were captured in the southern Free State, South Africa, for the placement of abdominal radio transmitters. Five combinations of ketamine hydrochloride with xylazine hydrochloride, midazolam or medetomidine hydrochloride were used to induce anaesthesia. In some cases the level of anaesthesia was maintained with 1.5 % halothane. A mixture of ketamine hydrochloride and medetomidine hydrochloride was found to be most effective. Atipamizole reversed the affects of medetomidine hydrochloride, resulting in a smooth and full recovery within 8 minutes. The immobilisation and subsequent anaesthesia of these animals on cold winter nights resulted in hypothermia, and keeping the animals warm was essential to the success of the procedures undertaken. Reversal of the sedative medetomidine hydrochloride proved to be important, because animals that were released before they were fully conscious took refuge in their burrows so that care was impossible.

2. Sample preparation for laser-microdissection of soybean shoot apical meristem

Chui E. Wong

2012-10-01

Full Text Available The shoot apical meristem houses stem cells responsible for the continuous formation of aerial plant organs including leaves and stems throughout the life of plants. Laser-microdissection in combination with high-throughput technology such as next generation sequencing permits an in-depth analysis of molecular events associated with specific cell type of interest. Sample preparation is the most critical step in ensuring good quality RNA to be extracted from samples following laser-microdissection. Here, we optimized the sample preparation for a major legume crop, soybean. We used Farmer’s solution as a fixative and paraffin as the embedding medium for soybean shoot apical meristem tissue without the use of any specialized equipment. Shorter time for tissue fixation (two days was found to be critical for the preservation of RNA in soybean shoot apical meristem. We further demonstrated the utility of this method for different tissues derived from soybean and rice. The method outlined here shall facilitate studies on crop plants involving laser-microdissection.

3. A laser microdissection-based workflow for FFPE tissue microproteomics: Important considerations for small sample processing.

Longuespée, Rémi; Alberts, Deborah; Pottier, Charles; Smargiasso, Nicolas; Mazzucchelli, Gabriel; Baiwir, Dominique; Kriegsmann, Mark; Herfs, Michael; Kriegsmann, Jörg; Delvenne, Philippe; De Pauw, Edwin

2016-07-15

Proteomic methods are today widely applied to formalin-fixed paraffin-embedded (FFPE) tissue samples for several applications in research, especially in molecular pathology. To date, there is an unmet need for the analysis of small tissue samples, such as for early cancerous lesions. Indeed, no method has yet been proposed for the reproducible processing of small FFPE tissue samples to allow biomarker discovery. In this work, we tested several procedures to process laser microdissected tissue pieces bearing less than 3000 cells. Combined with appropriate settings for liquid chromatography mass spectrometry-mass spectrometry (LC-MS/MS) analysis, a citric acid antigen retrieval (CAAR)-based procedure was established, allowing to identify more than 1400 proteins from a single microdissected breast cancer tissue biopsy. This work demonstrates important considerations concerning the handling and processing of laser microdissected tissue samples of extremely limited size, in the process opening new perspectives in molecular pathology. A proof of the proposed method for biomarker discovery, with respect to these specific handling considerations, is illustrated using the differential proteomic analysis of invasive breast carcinoma of no special type and invasive lobular triple-negative breast cancer tissues. This work will be of utmost importance for early biomarker discovery or in support of matrix-assisted laser desorption/ionization (MALDI) imaging for microproteomics from small regions of interest. PMID:26690073

4. The combined effect of electroporation and borocaptate in boron neutron capture therapy for murine solid tumors

10B-Enriched borocaptate (BSH) was administered intraperitoneally to SCCVII tumor-bearing C3H/He mice. Electroporation (EP) was conducted by using a tweezers-type electrode. The 10B contents in tumors were measured by prompt γ-ray spectrometry. The colony formation assay was applied to investigate the antitumor effects of boron neutron capture therapy (BNCT) and thereby to estimate the intratumor localization of BSH. The 10B concentrations in tumors decreased with time following BSH administration, falling to 5.4(±0.1) ppm at 3 h, whereas EP treatment (3 repetitions) 15 min after BSH injection delayed the clearance of BSH from tumors, and the 10B level remained at 19.4(±0.9) ppm at 3 h. The effect of BNCT increased with the 10B concentration in tumors, and the combination with EP showed a remarkably large cell killing effect even at 3 h after BSH injection. The effect of BNCT, i.e., slope coefficient of the cell survival curve of tumors, without EP was proportional to tumor 10B level (r=0.982), and that of BSH-BNCT combined with EP lay close to the same correlation line. However, tumors subjected to EP after BSH injection did not show high radiosensitivity when irradiated after conversion to a single cell suspension by enzymatic digestion. This indicates that the increase of the BNCT effect by EP was a consequence of enclosure of BSH in the interstitial space of tumor tissue and not within tumor cells. This is different from a previous in vitro study. The combination of EP and BNCT may be clinically useful, if a procedure to limit EP to the tumor region becomes available or if an alternative similar method is employed. (author)

5. Laser microdissection and microarray analysis of Tuber melanosporum ectomycorrhizas reveal functional heterogeneity between mantle and Hartig net compartments.

Hacquard, Stéphane; Tisserant, Emilie; Brun, Annick; Legué, Valérie; Martin, Francis; Kohler, Annegret

2013-06-01

The ectomycorrhizal (ECM) symbiosis, a mutualistic plant-fungus association, plays a fundamental role in forest ecosystems by enhancing plant growth and by providing host protection from root diseases. The cellular complexity of the symbiotic organ, characterized by the differentiation of structurally specialized tissues (i.e. the fungal mantle and the Hartig net), is the major limitation to study fungal gene expression in such specific compartments. We investigated the transcriptional landscape of the ECM fungus Tuber melanosporum during the major stages of its life cycle and we particularly focused on the complex symbiotic stage by combining the use of laser capture microdissection and microarray gene expression analysis. We isolated the fungal/soil (i.e. the mantle) and the fungal/plant (i.e. the Hartig net) interfaces from transverse sections of T. melanosporum/Corylus avellana ectomycorrhizas and identified the distinct genetic programmes associated with each compartment. Particularly, nitrogen and water acquisition from soil, synthesis of secondary metabolites and detoxification mechanisms appear to be important processes in the fungal mantle. In contrast, transport activity is enhanced in the Hartig net and we identified carbohydrate and nitrogen-derived transporters that might play a key role in the reciprocal resources' transfer between the host and the symbiont. PMID:23379715

6. Combining hybrid cars and synthetic fuels with electricity generation and carbon capture and storage

We examined the co-evolution of the transportation, and electricity and heat generation sectors in the Netherlands until 2040 using a MARKAL bottom-up cost optimisation model. All scenario variants investigated indicate a switch away from crude oil-based diesel and petrol for transportation. Lowest overall CO2 abatement cost is achieved by accommodating transportation first and using relatively expensive options for emissions reduction in electricity generation if needed. Biomass and carbon capture and storage (CCS) are used to full potential. Transportation CO2 emissions are reduced by switching to ethanol or bio-based synthetic fuels combined with CCS, and series hybrid cars if needed. Depending on the availability of biomass and carbon storage capacity, electricity is produced from biomass, coal with CCS, or wind complemented with natural gas. Indirect greenhouse gas emissions rise to 34-54% of national emissions in 2040. The difference in annual investment required between the scenario variants with and without CO2 emissions reductions of 68% by 2040 is 4-7 billion euro/year, or 0.5-1.2% of projected GDP. Investment costs are mostly determined by the cost of cars and electricity generation capacity. We observe competition for limited biomass supply and CO2 storage capacity between the transportation and power sectors.

7. Enhancing Protein Capture Using a Combination of Nanoyeast Single-Chain Fragment Affinity Reagents and Alternating Current Electrohydrodynamic Forces.

Vaidyanathan, Ramanathan; Rauf, Sakandar; Grewal, Yadveer S; Spadafora, Lauren J; Shiddiky, Muhammad J A; Cangelosi, Gerard A; Trau, Matt

2015-12-01

New high-performance detection technologies and more robust protein capture agents can be combined to both rapidly and specifically capture and detect protein biomarkers associated with disease in complex biological samples. Here we demonstrate the use of recently developed recombinant affinity reagents, namely nanoyeast-scFv, in combination with alternating current electrohydrodynamic (ac-EHD)-induced shear forces, to enhance capture performance during protein biomarker analysis. The use of ac-EHD significantly improves fluid transport across the capture domain, resulting in enhanced sensor-target interaction and simultaneous displacement of nonspecific molecules from the electrode surface. We demonstrate this simple proof-of-concept approach for the capture and detection of Entamoeba histolytica antigens from disinfected stool, within a span of 5 min using an ac-EHD microfluidic device. Under an ac-EHD field, antigens were captured on a nanoyeast-scFv immobilized device and subsequently detected using a quantum dot conjugated antibody. This immunosensor specifically detected antigen in disinfected stool with low background noise at concentrations down to 58.8 fM with an interassay reproducibility (%RSD of n = 3) gold electrodes in disinfected stool. We predict this rapid and sensitive approach using these stable affinity reagents may offer a new methodology to detect protein disease biomarkers from biological matrices. PMID:26551436

8. Microdissection testicular sperm extraction: an update

Ali A Dabaja; Peter N Schlegel

2013-01-01

Patients with non-obstructive azoospermia (NOA) were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate.In the last two decades,the advent of intracytoplasmic sperm injection (ICSI),and the application of various testicular sperm retrieval techniques,including fine needle aspiration (FNA),conventional testicular sperm extraction (TESE) and microdissection testicular sperm extraction (micro-TESE) have revolutionized treatment in this group of men.Because most men with NOA will have isolated regions of spermatogenesis within the testis,studies have illustrated that sperm can be retrieved in most men with NOA,including Klinefelter's syndrome (KS),prior history of chemotherapy and cryptorchidism.Micro-TESE,when compared with conventional TESE has a higher sperm retrieval rate (SRR) with fewer postoperative complications and negative effects on testicular function.In this article,we will compare the efficacy of the different procedures of sperm extraction,discuss the medical treatment and the role of testosterone optimization in men with NOA and describe the micro-TESE surgical technique.Furthermore,we will update our overall experience to allow counseling on the prognosis of sperm retrieval for the specific subsets of NOA.

9. Gestational diabetic transcriptomic profiling of microdissected human trophoblast.

Bari, Muhammad Furqan; Ngo, Sherry; Bastie, Claire C; Sheppard, Allan M; Vatish, Manu

2016-04-01

Gestational diabetes mellitus (GDM), the most common metabolic complication of pregnancy, is influenced by the placenta, and its prevalence directly increases with obesity. Therefore, to define the aetiology of GDM requires that the confounding influence of obesity and the heterogeneous nature of the placenta impairing accurate quantitative studies be accounted for. Using laser capture microdissection (LCM), we optimized RNA extraction from human placental trophoblast, the metabolic cellular interface between mother and foetus. This allowed specific transcriptomic profiling of trophoblast isolated from GDM, and obese and normal human placentae. Genome-wide gene expression analysis was performed on the RNA extracted from the trophoblast of GDM and obese and normal placentae. Forty-five differentially expressed genes (DEGs) specifically discriminated GDM from matched obese subjects. Two genes previously linked with GDM, pregnancy specific beta-1 glycoprotein 6 (PSG6) and placental system A sodium-dependent transporter system (SLC38A1), were significantly increased in GDM. A number of these DEGs (8 ubiquitin-conjugating enzymes (UBE) splice variants (UBE2D3 variants 1, 3, 4, 5, 6, 7, and 9) and UBE2V1 variant 4)) were involved in RNA processing and splicing, and a significant number of the DEGs, including the UBE variants, were associated with increased maternal fasting plasma glucose.It is concluded that DEGs discriminating GDM from obese subjects were pinpointed. Our data indicate a biological link between genes involved in RNA processing and splicing, ubiquitination, and fasting plasma glucose in GDM taking into account obesity as the confounder. PMID:26869332

10. Microdissection of gonadal tissues for gene expression analyses

Jørgensen, Anne; Dalgaard, Marlene Danner; Sonne, Si Brask

2011-01-01

, but in most cases a more specific staining is required to identify which cells to microdissect. We have established two staining protocols for frozen sections (1) Oil red O, which stains lipid droplet in fat cells and steroid-producing cells and (2) NBT BCIP, which stains cells expressing an alkaline...... protocols could also be used in other species and for other cell types containing lipid droplets or expressing alkaline phosphatase. Both protocols ensure a morphology that enables microdissection of single cells with RNA quality sufficient for subsequent gene expression analysis. However, RNA yields after......Laser microdissection permits isolation of specific cell types from tissue sections or cell cultures. This may be beneficial when investigating the role of specific cells in a complex tissue or organ. In tissues with easily distinguishable morphology, a simple hematoxylin staining is sufficient...

11. 激光捕获显微分离技术分离多人混合精斑%DNA profiling of spermatozoa mixture by low volume PCR combined with laser capture microdissection

杨帆; 曾宪海; 丰蕾; 徐程; 赵禾苗; 李彩霞; 胡兰

2014-01-01

目的 利用激光捕获显微分离技术(LCM)分离多供体混合精斑.方法 将3名自愿者供给的精液混合后,制备成混合精斑,并涂于覆膜玻片上.用PALM激光捕获显微分离系统切割并捕获样本中的单个精子,利用低体积扩增技术(LV-PCR)扩增并进行检测,并对分型结果进行统计.结果 联用LCM和低体积扩增技术捕获样本中单个精子细胞进行扩增,可获得每位自愿者的Y-STR分型,分型成功率为73.15％.结论 激光捕获显微分离联用低体积扩增技术可完成多供体混合精斑的分离检验.

12. An energetic analysis of CO2 capture on a gas turbine combining flue gas recirculation and membrane separation

Post-combustion Carbon Capture and Storage (CCS) is currently intensively investigated as a key issue for the mitigation of greenhouse gases emissions. A very large number of studies is dedicated to coal power plants. In this paper, the possibility to achieve carbon capture on a gas turbine, based on a combination of flue gas recycle and membrane separation is reported. Membrane processes are effectively known to offer attractive performances in terms of energy efficiency, as soon as concentrated and/or pressure mixtures have to be treated. Two different flow schemes have been simulated and compared: flue gas recycle with air combustion and flue gas recycle with an oxygen enriched feed mixture. The energy requirement of the different processes, expressed in GJ (thermal basis) per ton of recovered CO2, and the size of the membrane capture process (expressed in m2 of membrane area) have been systematically estimated for different membrane separation performances. It is shown that an overall energy requirement down to 2.6 GJ per ton can possibly be achieved when optimal operating conditions, based on oxygen enriched air (OEA) combustion together with a highly selective membrane (CO2/N2 selectivity of 200) are combined. Additional possibilities in order to minimise the energy penalty of the process are discussed. -- Highlights: ► A carbon capture process for gas turbine has been investigated for the first time, with membrane separation unit. ► Air combustion systematically induces CO2 capture specific energy requirement far above alternative capture processes. ► Remarkably, a very low energy requirement can be achieved (down to 2.6 GJ/ton) with Oxygen Enriched Air combustion. ► Target membrane selectivities and optimal oxygen content for combustion have been identified.

13. Combining different mRNA capture methods to analyze the transcriptome: analysis of the Xenopus laevis transcriptome.

Michael D Blower

Full Text Available mRNA sequencing (mRNA-seq is a commonly used technique to survey gene expression from organisms with fully sequenced genomes. Successful mRNA-seq requires purification of mRNA away from the much more abundant ribosomal RNA, which is typically accomplished by oligo-dT selection. However, mRNAs with short poly-A tails are captured poorly by oligo-dT based methods. We demonstrate that combining mRNA capture via oligo-dT with mRNA capture by the 5' 7-methyl guanosine cap provides a more complete view of the transcriptome and can be used to assay changes in mRNA poly-A tail length on a genome-wide scale. We also show that using mRNA-seq reads from both capture methods as input for de novo assemblers provides a more complete reconstruction of the transcriptome than either method used alone. We apply these methods of mRNA capture and de novo assembly to the transcriptome of Xenopus laevis, a well-studied frog that currently lacks a finished sequenced genome, to discover transcript sequences for thousands of mRNAs that are currently absent from public databases. The methods we describe here will be broadly applicable to many organisms and will provide insight into the transcriptomes of organisms with sequenced and unsequenced genomes.

14. Solvents for CO2 capture. Structure-activity relationships combined with vapour-liquid-equilibrium measurements

Mergler, Y.L.; Rumley-Van Gurp, R.; Brasser, P.; Koning, M.C. de; Goetheer, E.L.V.

2011-01-01

In this study a systematic approach was chosen to test and characterize amine systems for CO2 capture. Vapour-liquid-equilibrium measurements were performed on a homologue series of amines, with ethylene amine as base structure. Various functional groups were used that ranged in chemical and physica

15. Advanced modeling and simulation of integrated gasification combined cycle power plants with CO2-capture

The objective of this thesis is to provide an extensive description of the correlations in some of the most crucial sub-processes for hard coal fired IGCC with carbon capture (CC-IGCC). For this purpose, process simulation models are developed for four industrial gasification processes, the CO-shift cycle, the acid gas removal unit, the sulfur recovery process, the gas turbine, the water-/steam cycle and the air separation unit (ASU). Process simulations clarify the influence of certain boundary conditions on plant operation, performance and economics. Based on that, a comparative benchmark of CC-IGCC concepts is conducted. Furthermore, the influence of integration between the gas turbine and the ASU is analyzed in detail. The generated findings are used to develop an advanced plant configuration with improved economics. Nevertheless, IGCC power plants with carbon capture are not found to be an economically efficient power generation technology at present day boundary conditions.

16. Pre-combustion CO2 capture: Analysis of integrated reforming combined cycle

Nord, Lars Olof

2010-01-01

This thesis presents processes for reducing CO2 emissions from natural gas (NG) power plants, which could help attenuate the rise in atmospheric temperature. Objectives for the thesis work were process design and integration of NG pre-combustion CO2 capture plants, and evalution, through process simulation, of the concepts. An important aspect of the evaluation was investigation of plant flexibility, specifically off-design analysis. Contributions comprised detailed heat recovery steam genera...

17. Pre-combustion CO2 capture: Analysis of integrated reforming combined cycle

Nord, Lars Olof

2010-01-01

This thesis presents processes for reducing CO2 emissions from natural gas (NG) power plants, which could help attenuate the rise in atmospheric temperature. Objectives for the thesis work were process design and integration of NG pre-combustion CO2 capture plants, and evalution, through process simulation, of the concepts. An important aspect of the evaluation was investigation of plant flexibility, specifically off-design analysis. Contributions comprised detailed heat recovery steam genera...

18. Microdissection of black widow spider silk-producing glands.

Jeffery, Felicia; La Mattina, Coby; Tuton-Blasingame, Tiffany; Hsia, Yang; Gnesa, Eric; Zhao, Liang; Franz, Andreas; Vierra, Craig

2011-01-01

Modern spiders spin high-performance silk fibers with a broad range of biological functions, including locomotion, prey capture and protection of developing offspring. Spiders accomplish these tasks by spinning several distinct fiber types that have diverse mechanical properties. Such specialization of fiber types has occurred through the evolution of different silk-producing glands, which function as small biofactories. These biofactories manufacture and store large quantities of silk proteins for fiber production. Through a complex series of biochemical events, these silk proteins are converted from a liquid into a solid material upon extrusion. Mechanical studies have demonstrated that spider silks are stronger than high-tensile steel. Analyses to understand the relationship between the structure and function of spider silk threads have revealed that spider silk consists largely of proteins, or fibroins, that have block repeats within their protein sequences. Common molecular signatures that contribute to the incredible tensile strength and extensibility of spider silks are being unraveled through the analyses of translated silk cDNAs. Given the extraordinary material properties of spider silks, research labs across the globe are racing to understand and mimic the spinning process to produce synthetic silk fibers for commercial, military and industrial applications. One of the main challenges to spinning artificial spider silk in the research lab involves a complete understanding of the biochemical processes that occur during extrusion of the fibers from the silk-producing glands. Here we present a method for the isolation of the seven different silk-producing glands from the cobweaving black widow spider, which includes the major and minor ampullate glands [manufactures dragline and scaffolding silk], tubuliform [synthesizes egg case silk], flagelliform [unknown function in cob-weavers], aggregate [makes glue silk], aciniform [synthesizes prey wrapping and egg

19. Evaluation of the dose enhancement of combined 10B + 157Gd Neutron Capture Therapy (NCT)

An innovative molecule, GdBLDL, for boron neutron capture therapy (BNCT) has been developed and its effectiveness as a BNCT carrier is currently under evaluation using in vivo experiments on small animal tumour models. The molecule contains both 10B (the most commonly used NCT agent) and 157Gd nuclei. 157Gd is the second most studied element to perform NCT, mainly thanks to its high cross section for the capture of low-energy neutrons. The main drawback of 157Gd neutron capture reaction is the very short range and low-energy secondary charged particles (Auger electrons), which requires 157Gd to be very close to the cellular DNA to have an appreciable biological effect. Treatment doses were calculated by Monte Carlo simulations to ensure the optimised tumour irradiation and the sparing of the healthy organs of the irradiated animals. The enhancement of the absorbed dose due to the simultaneous presence of 10B and 157Gd in the experimental set-up was calculated and the advantage introduced by the presence of 157Gd was discussed. (authors)

20. Antibodies and antiestrogens combined with boron for use in the neutron capture therapy

The ZR-75-1 cell line developed from a mammary carcinoma was chosen to characterise the binding of antiestrogen U23.469-M to the cell, which was subsequently compared to that of a derivative combined with boron. It was found that the original U23.469-M showed antiestrogenic activity, while this effect was largely abolished after the substance had been modified using b-decachloro-o-carborane. In this study, boron-conjugated antibodies were produced in order to find out whether those modified immunoglobulins would be suitable to bind sufficient quantities of boron to the tumour cells. It was calculated by experts on radiation biology that a minimum of 1000 boron atoms is required for a tumour-specific antibody to be therapeutically effective. When oxidated dextran of a molecular weight of 33 kD was used as a linking molecule, a reproducible method could be developed that permitted more than 1000 boron atoms to be bound per antibody. In one of the monoclonal antibodies tested here a combination with boron could, however, only be achieved at the expense of complete inactivation. A model was developed allowing to significantly increase the number modified antibodies attached to any one tumour cell. The cell binding experiments and radioimmunoassays then carried out were able to show that the number of antibodies bound to tumour cells can be increased to different degrees, depending on the monoclonal antibody used in each case. (orig./MG)

1. Marker chromosome 21 identified by microdissection and FISH

Sun, Y.; Palmer, C.G. [Indiana Univ. School of Medicine, Indianapolis, IN (United States); Rubinstein, J. [Univ. Affiliated Cincinnati Center for Developmental Disorders, OH (United States)] [and others

1995-03-27

A child without Downs syndrome but with developmental delay, short stature, and autistic behavior was found to be mosaic 46,XX/47,XX,+mar(21) de novo. The marker was a small ring or dot-like chromosome. Microdissection of the marker was performed. The dissected fragments were biotinylated with sequence-independent PCR as a probe pool for fluorescence in situ hybridization (FISH). FISH results suggested an acrocentric origin of the marker. Subsequent FISH with {alpha}-satellite DNA probes for acrocentric chromosomes and chromosome-specific 21 and 22 painting probes confirmed its origin from chromosome 21. 14 refs., 3 figs.

2. Simulation and exergetic evaluation of CO2 capture in a solid-oxide fuel-cell combined-cycle power plant

Highlights: • An exergetic analysis is used to identify the thermodynamic irreversibilities of a power plant. • The plant includes a solid-oxide fuel-cell unit and CO2 capture. • Additional power generated in the fuel-cell unit enhances the power output of the plant. • The power plant results in a high efficiency compared both to conventional and other CO2 capture plants. • High irreversibilities are found for the solid-oxide fuel cell. - Abstract: The incorporation of fuel cells into power plants can enhance the operational efficiency and facilitate the separation and capture of emissions. In this paper a fuel-cell unit, consisting of solid-oxide fuel-cell stacks, a pre-reformer, and an afterburner is incorporated into a combined-cycle power plant with CO2 capture. The thermodynamic performance of the plant is examined using an exergetic analysis and it is compared with a conventional combined-cycle power plant (reference plant) without CO2 capture, as well as with other plants with CO2 capture. The inefficiencies of the chemical reactions taking place in the fuel-cell unit are found to be the main source of exergy destruction among the plant components. However, the additional power generated in the fuel-cell stacks and the afterburner enhances the overall efficiency and compensates for the energy needed for the capture and compression of the carbon dioxide. When compared with the reference plant and with alternative capture technologies, the solid-oxide fuel-cell plant with CO2 capture operates more efficiently and appears to be a thermodynamically promising approach for carbon capture

3. Low-Energy Solvents For Carbon Dioxide Capture Enabled By A Combination Of Enzymes And Vacuum Regeneration

Salmon, Sonja [Novozymes North America, Inc., Franklinton, NC (United States); House, Alan [Novozymes North America, Inc., Franklinton, NC (United States); Liu, Kun [Univ. of Kentucky, Lexington, KY (United States); Frimpong, Reynolds [Univ. of Kentucky, Lexington, KY (United States); Liu, Kunlei [Univ. of Kentucky, Lexington, KY (United States); Freeman, Charles [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Whyatt, Greg [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Slater, Jonathan [Doosan Babcock, Renfew (United Kingdom); Fitzgerald, David [Doosan Babcock, Renfew (United Kingdom)

2015-08-31

An integrated bench-scale system combining the attributes of the bio-renewable enzyme carbonic anhydrase (CA) with low-enthalpy CO2 absorption solvents and vacuum regeneration was designed, built and operated for 500 hours using simulated flue gas. The objective was to develop a CO2 capture process with improved efficiency and sustainability when compared to NETL Case 10 monoethanolamine (MEA) scrubbing technology. The use of CA accelerates inter-conversion between dissolved CO2 and bicarbonate ion to enhance CO2 absorption, and the use of low enthalpy CO2 absorption solvents makes it possible to regenerate the solvent at lower temperatures relative to the reference MEA-based solvent. The vacuum regeneration-based integrated bench-scale system operated successfully for an accumulated 500 hours using aqueous 23.5 wt% K2CO3-based solvent containing 2.5 g/L enzyme to deliver an average 84% CO2 capture when operated with a 20% enzyme replenishment rate per ~7 hour steady-state run period. The total inlet gas flow was 30 standard liters per minute with 15% CO2 and 85% N2. The absorber temperature was 40°C and the stripper operated under 35 kPa pressure with an approximate 77°C stripper bottom temperature. Tests with a 30°C absorber temperature delivered >90% capture. On- and off-line operational measurements provided a full process data set, with recirculating enzyme, that allowed for enzyme replenishment and absorption/desorption kinetic parameter calculations. Dissolved enzyme replenishment and conventional process controls were demonstrated as straightforward approaches to maintain system performance. Preliminary evaluation of a novel flow-through ultrasonically enhanced regeneration system was also conducted, yet resulted in CO2 release within the range of temperature-dependent release, and further work would be needed to validate the benefits of ultrasonic enhanced stripping. A full technology assessment was completed in which four techno-economic cases for

4. FAST-FISH with laser beam microdissected DOP-PCR probe distinguishes the sex chromosomes of Silene latifolia.

Hobza, Roman; Lengerova, Martina; Cernohorska, Halina; Rubes, Jiri; Vyskot, Boris

2004-01-01

We present an improved FISH strategy for differentiating the sex chromosomes of the dioecious model plant, Silene latifolia. Fixed mitotic protoplasts were dropped on a polyethylene naphthalate membrane, the X or Y chromosomes were isolated using nitrogen laser beam microdissection, catapulted by laser pressure, and amplified by DOP-PCR. A modified FAST-FISH protocol based on a short hybridization time combined with a low concentration of probe was used. The success of this approach is demonstrated by the differential labeling of the X and Y chromosomes and it could represent a quick method for comparing organization of plant genomes. PMID:15125638

5. Competitiveness of CO2 capture from an industrial solid oxide fuel cell combined heat and power system in the early stage of market introduction

Kuramochi, T.; Turkenburg, W.C.; Faaij, A.P.C.

2011-01-01

In this article, it was investigated whether potentially low-cost CO2 capture from SOFC systems could enhance the penetration of SOFC in the energy market in a highly carbon-constrained society in the mid-term future (up to year 2025). The application of 5 MWe SOFC systems for industrial combined he

6. Combining gas phase electron capture and IRMPD action spectroscopy to probe the electronic structure of a metastable reduced organometallic complex containing a non-innocent ligand.

Katari, Madanakrishna; Payen de la Garanderie, Eleonore; Nicol, Edith; Steinmetz, Vincent; van der Rest, Guillaume; Carmichael, Duncan; Frison, Gilles

2015-10-21

Combining electron capture dissociation mass spectrometry and infrared multiple photon dissociation action spectroscopy allows the formation, selection and characterisation of reduced metal complexes containing non-innocent ligands. Zinc complexes containing diazafluorenone ligands have been studied and the localisation of the single electron on the metal atom in the mono-ligated complex has been demonstrated. PMID:26051020

7. Microdissection and polymerase chain reaction amplification of genomic DNA from histological tissue sections.

Moskaluk, C A; Kern, S.E.

1997-01-01

Polymerase chain reaction (PCR)-based assays are being used increasingly to study the molecular genetic changes that occur in minute cellular lesions that are identified in histological sections. It is often desirable to microdissect the cells of interest in a lesion, isolating them from surrounding normal tissue to obtain the purest representation of genomic DNA possible. We present here an optimized microdissection and DNA extraction protocol that reliably produces PCR-amplifiable DNA from ...

8. Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

DeCarlo, Correne A.; Escott, Nicholas G.; Werner, Julieta; Robinson, Kerry; Lambert, Paul F.; Law, R. David; Zehbe, Ingeborg

2008-01-01

Optimal sample handling techniques for tissue preparation and storage, RNA extraction and quantification, and target gene detection are crucial for reliable gene expression analysis. Methods for measuring low-expressing genes, such as interferons, in human cervical samples are not described in the scientific literature. To detect interferon mRNA in human cervical samples we obtained normal and dysplastic frozen and formalin-fixed cervical biopsies from colposcopy. Histopathological diagnosis ...

9. Identification of novel immune and barrier genes in atopic dermatitis by laser capture micro-dissection

Esaki, H.; Ewald, David Adrian; Ungar, B.;

2014-01-01

BACKGROUND: The molecular signature of atopic dermatitis (AD) lesions is associated with TH2 and TH22 activation and epidermal alterations. However, the epidermal and dermal AD transcriptomes and their respective contributions to abnormalities in respective immune and barrier phenotypes are unknown...... normal skin from healthy volunteers, followed by gene expression (microarrays and real-time PCR) and immunostaining studies. RESULTS: Our study identified novel immune and barrier genes, including the IL-34 cytokine and claudins 4 and 8, and showed increased detection of key AD genes usually undetectable...... immune molecules and enabling detection of gene products usually not detected on arrays....

10. Identification of novel immune and barrier genes in atopic dermatitis by means of laser capture microdissection

Esaki, Hitokazu; Ewald, David Adrian; Ungar, Benjamin;

2015-01-01

Background : The molecular signature of atopic dermatitis (AD) lesions is associated with T(H)2 and T(H)22 activation and epidermal alterations. However, the epidermal and dermal AD transcriptomes and their respective contributions to abnormalities in respective immune and barrier phenotypes are...... with AD and normal skin from healthy volunteers, followed by gene expression (microarrays and real-time PCR) and immunostaining studies. Results : Our study identified novel immune and barrier genes, including the IL-34 cytokine and claudins 4 and 8, and showed increased detection of key AD genes...... key barrier or immune molecules and enabling detection of gene products usually not detected on arrays....

11. Capturing Thoughts, Capturing Minds?

Nielsen, Janni

2004-01-01

Think Aloud is cost effective, promises access to the user's mind and is the applied usability technique. But 'keep talking' is difficult, besides, the multimodal interface is visual not verbal. Eye-tracking seems to get around the verbalisation problem. It captures the visual focus of attention....... However, it is expensive, obtrusive and produces huge amount of data. Besides, eye-tracking do not give access to user's mind. Capturing interface/cursor tracking may be cost effective. It is easy to install, data collection is automatic and unobtrusive and replaying the captured recording to the user and...

12. High-Throughput Microdissection for Next-Generation Sequencing

Rosenberg, Avi Z.; Armani, Michael D.; Fetsch, Patricia A.; Xi, Liqiang; Pham, Tina Thu; Raffeld, Mark; Chen, Yun; O’Flaherty, Neil; Stussman, Rebecca; Blackler, Adele R.; Du, Qiang; Hanson, Jeffrey C.; Roth, Mark J.; Filie, Armando C.; Roh, Michael H.; Emmert-Buck, Michael R.; Hipp, Jason D.; Tangrea, Michael A.

2016-01-01

Precision medicine promises to enhance patient treatment through the use of emerging molecular technologies, including genomics, transcriptomics, and proteomics. However, current tools in surgical pathology lack the capability to efficiently isolate specific cell populations in complex tissues/tumors, which can confound molecular results. Expression microdissection (xMD) is an immuno-based cell/subcellular isolation tool that procures targets of interest from a cytological or histological specimen. In this study, we demonstrate the accuracy and precision of xMD by rapidly isolating immunostained targets, including cytokeratin AE1/AE3, p53, and estrogen receptor (ER) positive cells and nuclei from tissue sections. Other targets procured included green fluorescent protein (GFP) expressing fibroblasts, in situ hybridization positive Epstein-Barr virus nuclei, and silver stained fungi. In order to assess the effect on molecular data, xMD was utilized to isolate specific targets from a mixed population of cells where the targets constituted only 5% of the sample. Target enrichment from this admixed cell population prior to next-generation sequencing (NGS) produced a minimum 13-fold increase in mutation allele frequency detection. These data suggest a role for xMD in a wide range of molecular pathology studies, as well as in the clinical workflow for samples where tumor cell enrichment is needed, or for those with a relative paucity of target cells. PMID:26999048

13. ERα and ERK1/2 MAP kinase expression in microdissected stromal and epithelial endometrial cells

Our previous published data detected higher expression of total and active mitogen activated protein kinase (MAPK) in the epithelial vs. stromal cells of the endometrium. In the present work we compared the expression of ERK1/2 MAPK and estrogen receptor α (ERα) in epithelial versus stromal cells in benign human endometrial tissues. Laser capture microdissection was used to separate glandular epithelium and stromal cells from six frozen, proliferative phase endometrial specimens. Total and phosphorylated levels for ERK1/2 and ERα were measured by quantitation of signals from Western blots using specific antibodies against the active and total forms of ERK1/2 and against ERα. When the level of the proteins was quantitated and normalized to p actin from micro dissected stroma and epithelium, no significant difference was detected in the levels of these proteins between the two tissue compartments. There was a trend toward higher expression in the stroma vs. epithelium, respectively (active ERK1/2 0.45 ± 0.17 vs. 0.2 ± 0.65; total ERK1/2 0.54 ± 0.35 vs. 0.28 ± 0.23; ERα 0.82 ± 0.28 vs. 0.54 ± 0.18; n = 6). These data demonstrate that there are comparable levels of ERα (P = 0.41), total ERK1/2 (P = 0.18) and active ERK1/2 (P = 0.13) in the stroma and epithelium of proliferative phase endometrium with a trend toward higher expression of these proteins in the stromal compartment.

14. Using genic sequence capture in combination with a syntenic pseudo genome to map a deletion mutant in a wheat species.

Gardiner, Laura-Jayne; Gawroński, Piotr; Olohan, Lisa; Schnurbusch, Thorsten; Hall, Neil; Hall, Anthony

2014-12-01

Mapping-by-sequencing analyses have largely required a complete reference sequence and employed whole genome re-sequencing. In species such as wheat, no finished genome reference sequence is available. Additionally, because of its large genome size (17 Gb), re-sequencing at sufficient depth of coverage is not practical. Here, we extend the utility of mapping by sequencing, developing a bespoke pipeline and algorithm to map an early-flowering locus in einkorn wheat (Triticum monococcum L.) that is closely related to the bread wheat genome A progenitor. We have developed a genomic enrichment approach using the gene-rich regions of hexaploid bread wheat to design a 110-Mbp NimbleGen SeqCap EZ in solution capture probe set, representing the majority of genes in wheat. Here, we use the capture probe set to enrich and sequence an F2 mapping population of the mutant. The mutant locus was identified in T. monococcum, which lacks a complete genome reference sequence, by mapping the enriched data set onto pseudo-chromosomes derived from the capture probe target sequence, with a long-range order of genes based on synteny of wheat with Brachypodium distachyon. Using this approach we are able to map the region and identify a set of deleted genes within the interval. PMID:25205592

15. Improvement of Capture Compound Mass Spectrometry Technology (CCMS) for the Profiling of Human Kinases by Combination with 2D LC-MS/MS

Fischer, Jenny J.; Graebner, Olivia; Dreger, Mathias; Glinski, Mirko; Baumgart, Sabine; Koester, Hubert

2011-01-01

An increasingly popular and promising field in functional proteomics is the isolation of proteome subsets based on small molecule-protein interactions. One platform approach in this field are Capture Compounds that contain a small molecule of interest to bind target proteins, a photo-activatable reactivity function to covalently trap bound proteins, and a sorting function to isolate captured protein conjugates from complex biological samples for direct protein identification by liquid chromatography/mass spectrometry (nLC-MS/MS). In this study we used staurosporine as a selectivity group for analysis in HepG2 cells derived from human liver. In the present study, we combined the functional isolation of kinases with different separation workflows of automated split-free nanoflow liquid chromatography prior to mass spectrometric analysis. Two different CCMS setups, CCMS technology combined with 1D LC-MS and 2D LC-MS, were compared regarding the total number of kinase identifications. By extending the chromatographic separation of the tryptic digested captured proteins from 1D LC linear gradients to 2D LC we were able to identify 97 kinases. This result is similar to the 1D LC setup we previously reported but this time 4 times less input material was needed. This makes CCMS of kinases an even more powerful tool for the proteomic profiling of this important protein family. PMID:21941435

16. Hydrate-based CO2 (carbon dioxide) capture from IGCC (integrated gasification combined cycle) synthesis gas using bubble method with a set of visual equipment

The hydrate-based carbon dioxide (CO2) capture from the integrated gasification combined cycle (IGCC) synthesis gas using the bubble method is investigated with a set of visual equipment in this work. The gas bubble is created with a bubble plate on the bottom of the equipment. By the visual equipment, the hydrate formation and the hydrate shape are visually captured. With the move of the gas bubble from the bottom to the top of the reactor, gas hydrate forms firstly from the gas–liquid boundary around the bubble, then the hydrate gradually grows up and piles up in the bottom side of the bubble to form a hydrate particle. The gas hydrate shape is affected by the gas flow rate. The hydrate is acicular crystal at the low gas flow rate while the hydrate is fine sand-like crystal at the high gas flow rate. The bubble size and the gas flow rate have an obvious impact on the hydrate-based CO2 separation process. The experimental results show the gas bubble of 50 μm and the gas flow rate of 6.75 mL/min/L are ideal for CO2 capture from IGCC synthesis gas under the condition of 3.0 MPa and 274.15 K. -- Highlights: ► A set of visual equipment and bubble method are used for hydrate-based CO2 capture. ► CO2-TBAB gas hydrate formation process and shape are captured clearly. ► Gas bubble of 50 μm and gas flow rate of 6.75 mL/min/L are the optimum process.

17. Exergy analysis of a hydrogen fired combined cycle with natural gas reforming and membrane assisted shift reactors for CO2 capture

Highlights: ► Exergy analysis of NGCC with CCS. ► WGS-MR: exergetically efficient technology for CCS, less than 2% total exergy losses. ► 10% of total exergy dissipation in the ATR. ► Optimization of ATR operation and CO2 stream treatment. - Abstract: Hydrogen production from fossil fuels together with carbon capture has been suggested as a means of providing a carbon free power. The paper presents a comparative exergetic analysis performed on the hydrogen production from natural gas with several combinations of reactor systems: (a) oxy or air fired autothermal reforming with subsequent water gas shift reactor and (b) membrane reactor assisted with shift catalysts. The influence of reactor temperature and pressure as well as operating parameter steam-to-carbon ratio, is also studied exergetically. The results indicate optimal power plant configurations with CO2 capture, or hydrogen delivery for industrial applications.

18. Multiphase, multicomponent simulations and experiments of reactive flow, relevant for combining geologic CO2 sequestration with geothermal energy capture

Saar, Martin O.

2011-11-01

Understanding the fluid dynamics of supercritical carbon dioxide (CO2) in brine- filled porous media is important for predictions of CO2 flow and brine displacement during geologic CO2 sequestration and during geothermal energy capture using sequestered CO2 as the subsurface heat extraction fluid. We investigate multiphase fluid flow in porous media employing particle image velocimetry experiments and lattice-Boltzmann fluid flow simulations at the pore scale. In particular, we are interested in the motion of a drop (representing a CO2 bubble) through an orifice in a plate, representing a simplified porous medium. In addition, we study single-phase/multicomponent reactive transport experimentally by injecting water with dissolved CO2 into rocks/sediments typically considered for CO2 sequestration to investigate how resultant fluid-mineral reactions modify permeability fields. Finally, we investigate numerically subsurface CO2 and heat transport at the geologic formation scale.

19. Understanding the effect of side groups in ionic liquids on carbon-capture properties: a combined experimental and theoretical effort

Yan, Fangyong [Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engingeering; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Materials Sciences Division; Lartey, Michael [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); Damodaran, Krishnan [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); Univ. of Pittsburgh, PA (United States). Dept. of Chemistry; Albenze, Erik [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); URS Corporation, South Park, PA (United States); Thompson, Robert L. [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); URS Corporation, South Park, PA (United States); Kim, Jihan [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Materials Science Div.; Harancyzk, Maciel [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Computational Research Div.; Nulwala, Hunaid B. [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); Carnegie Mellon Univ., Pittsburgh, PA (United States); Luebke, David R. [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); Smit, Berend [Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engingeering; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Materials Sciences Division

2013-01-01

Ionic liquids are an emerging class of materials with applications in a variety of fields. Steady progress has been made in the creation of ionic liquids tailored to specific applications. However, the understanding of the underlying structure–property relationships has been slower to develop. As a step in the effort to alleviate this deficiency, the influence of side groups on ionic liquid properties has been studied through an integrated approach utilizing synthesis, experimental determination of properties, and simulation techniques. To achieve this goal, a classical force field in the framework of OPLS/Amber force fields has been developed to predict ionic liquid properties accurately. Cu(I)-catalyzed click chemistry was employed to synthesize triazolium-based ionic liquids with diverse side groups. Values of densities were predicted within 3% of experimental values, whereas self-diffusion coefficients were underestimated by about an order of magnitude though the trends were in excellent agreement, the activation energy calculated in simulation correlates well with experimental values. The predicted Henry coefficient for CO{sub 2} solubility reproduced the experimentally observed trends. This study highlights the importance of integrating experimental and computational approaches in property prediction and materials development, which is not only useful in the development of ionic liquids for CO{sub 2} capture but has application in many technological fields.

20. Linking microbial community structure to biogeochemical function in coastal marine sediments: Stable isotope probing combined with magnetic bead capture

T. Miyatake

2011-01-01

Microbial community structure and its biogeochemical functions in marine sediments can be successfully linked by using the improved Mag-SIP method in combination with other approaches. In this thesis, we were able to provide detailed information on the microorganisms responsible for the utilization

1. Combined infrared multiphoton dissociation and electron-capture dissociation using co-linear and overlapping beams in Fourier transform ion cyclotron resonance mass spectrometry.

Mihalca, Romulus; van der Burgt, Yuri E M; McDonnell, Liam A; Duursma, Marc; Cerjak, Iliya; Heck, Albert J R; Heeren, Ron M A

2006-01-01

A novel set-up for Fourier transform ion cyclotron resonance mass spectrometry (FTICR) is reported for simultaneous infrared multiphoton dissociation (IRMPD) and electron-capture dissociation (ECD). An unmodified electron gun ensures complete, on-axis overlap between the electron and the photon beams. The instrumentation, design and implementation of this novel approach are described. In this configuration the IR beam is directed into the ICR cell using a pneumatically actuated mirror inserted into the ion-optical path. Concept validation was made using different combinations of IRMPD and ECD irradiation events on two standard peptides. The ability to perform efficient IRMPD, ECD and especially simultaneous IRMPD and ECD using lower irradiation times is demonstrated. The increase in primary sequence coverage, with the combined IRMPD and ECD set-up, also increases the confidence in peptide and protein assignments. PMID:16705647

2. Boron determination in liver tissue by combining quantitative neutron capture radiography (QNCR) and histological analysis for BNCT treatment planning at the TRIGA Mainz.

Schütz, C; Brochhausen, C; Altieri, S; Bartholomew, K; Bortolussi, S; Enzmann, F; Gabel, D; Hampel, G; Kirkpatrick, C J; Kratz, J V; Minouchehr, S; Schmidberger, H; Otto, G

2011-09-01

The typical primary malignancies of the liver are hepatocellular carcinoma and cholangiocarcinoma, whereas colorectal liver metastases are the most frequently occurring secondary tumors. In many cases, only palliative treatment is possible. Boron neutron capture therapy (BNCT) represents a technique that potentially destroys tumor tissue selectively by use of externally induced, locally confined secondary particle irradiation. In 2001 and 2003, BNCT was applied to two patients with colorectal liver metastases in Pavia, Italy. To scrutinize the rationale of BNCT, a clinical pilot study on patients with colorectal liver metastases was carried out at the University of Mainz. The distribution of the (10)B carrier (p-borono-phenylalanine) in the liver and its uptake in cancerous and tumor-free tissue were determined, focusing on a potential correlation between the uptake of p-borono-phenylalanine and the biological characteristics of cancerous tissue. Samples were analyzed using quantitative neutron capture radiography of cryosections combined with histological analysis. Methodological aspects of the combination of these techniques and results from four patients enrolled in the study are presented that indicate that the uptake of p-borono-phenylalanine strongly depends on the metabolic activity of cells. PMID:21692653

3. Linking microbial community structure to biogeochemical function in coastal marine sediments: Stable isotope probing combined with magnetic bead capture

Stal, L.J.; Boschker, H.T.S.; Miyatake, T.

2011-01-01

Microbial community structure and its biogeochemical functions in marine sediments can be successfully linked by using the improved Mag-SIP method in combination with other approaches. In this thesis, we were able to provide detailed information on the microorganisms responsible for the utilization of major substrates in marine sediments. It was shown that a number of important groups in the microbial community played a broader role than expected. Some surprising results are that oxygenic pho...

4. Laser-assisted Microdissection (LAM) as a Tool for Transcriptional Profiling of Individual Cell Types.

Florez Rueda, Ana Marcela; Grossniklaus, Ueli; Schmidt, Anja

2016-01-01

The understanding of developmental processes at the molecular level requires insights into transcriptional regulation, and thus the transcriptome, at the level of individual cell types. While the methods described here are generally applicable to a wide range of species and cell types, our research focuses on plant reproduction. Plant cultivation and seed production is of crucial importance for human and animal nutrition. A detailed understanding of the regulatory networks that govern the formation of the reproductive lineage (germline) and ultimately of seeds is a precondition for the targeted manipulation of plant reproduction. In particular, the engineering of apomixis (asexual reproduction through seeds) into crop plants promises great improvements, as it leads to the formation of clonal seeds that are genetically identical to the mother plant. Consequently, the cell types of the female germline are of major importance for the understanding and engineering of apomixis. However, as the corresponding cells are deeply embedded within the floral tissues, they are very difficult to access for experimental analyses, including cell-type specific transcriptomics. To overcome this limitation, sections of individual cells can be isolated by laser-assisted microdissection (LAM). While LAM in combination with transcriptional profiling allows the identification of genes and pathways active in any cell type with high specificity, establishing a suitable protocol can be challenging. Specifically, the quality of RNA obtained after LAM can be compromised, especially when small, single cells are targeted. To circumvent this problem, we have established a workflow for LAM that reproducibly results in high RNA quality that is well suitable for transcriptomics, as exemplified here by the isolation of cells of the female germline in apomictic Boechera. In this protocol, procedures are described for tissue preparation and LAM, also with regard to RNA extraction and quality control

5. Gene recovery microdissection (GRM) a process for producing chromosome region-specific libraries of expressed genes

Christian, A T; Coleman, M A; Tucker, J D

2001-02-08

Gene Recovery Microdissection (GRM) is a unique and cost-effective process for producing chromosome region-specific libraries of expressed genes. It accelerates the pace, reduces the cost, and extends the capabilities of functional genomic research, the means by which scientists will put to life-saving, life-enhancing use their knowledge of any plant or animal genome.

6. Real-time quantitative PCR of microdissected paraffin-embedded breast carcinoma

Gjerdrum, Lise Mette; Sorensen, Boe Sandahl; Kjeldsen, Eigil;

2004-01-01

We studied the feasibility of using real-time quantitative PCR to determine HER-2 DNA amplification and mRNA expression in microdissected formalin-fixed, paraffin-embedded breast tumors and compared this with standard immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) methods...

7. The status of Tsukuba BNCT trial: BPA-based boron neutron capture therapy combined with X-ray irradiation

Yamamoto, T., E-mail: tetsu_tsukuba@yahoo.co.jp [Department of Neurosurgery, Graduate School of Comprehensive Human Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba (Japan)] [Department of Radiation Oncology, Graduate School of Comprehensive Human Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba (Japan); Nakai, K. [Department of Neurosurgery, Graduate School of Comprehensive Human Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba (Japan); Nariai, T. [Department of Neurosurgery, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo (Japan); Kumada, H.; Okumura, T.; Mizumoto, M.; Tsuboi, K. [Department of Radiation Oncology, Graduate School of Comprehensive Human Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba (Japan); Zaboronok, A.; Ishikawa, E.; Aiyama, H.; Endo, K.; Takada, T.; Yoshida, F.; Shibata, Y.; Matsumura, A. [Department of Neurosurgery, Graduate School of Comprehensive Human Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba (Japan)

2011-12-15

The phase II trial has been prepared to assess the effectiveness of BPA (250 mg/kg)-based NCT combined with X-ray irradiation and temozolomide (75 mg/m{sup 2}) for the treatment of newly diagnosed GBM. BPA uptake is determined by {sup 18}F-BPA-PET and/or {sup 11}C-MET-PET, and a tumor with the lesion to normal ratio of 2 or more is indicated for BNCT. The maximum normal brain point dose prescribed was limited to 13.0 Gy or less. Primary end point is overall survival.

8. Rate-based modelling of combined SO2 removal and NH3 recycling integrated with an aqueous NH3-based CO2 capture process

Highlights: • A rigorous, rate-based model for an NH3–CO2–SO2–H2O system was developed. • Model predictions are in good agreement with pilot plant results. • >99.9% of SO2 was captured and >99.9% of slipped ammonia was reused. • The process is highly adaptable to the variations of SO2/NH3 level, temperatures. - Abstract: To reduce the costs of controlling emissions from coal-fired power stations, we propose an advanced and effective process of combined SO2 removal and NH3 recycling, which can be integrated with the aqueous NH3-based CO2 capture process to simultaneously achieve SO2 and CO2 removal, NH3 recycling and flue gas cooling in one process. A rigorous, rate-based model for an NH3–CO2–SO2–H2O system was developed and used to simulate the proposed process. The model was thermodynamically and kinetically validated by experimental results from the open literature and pilot-plant trials, respectively. Under typical flue gas conditions, the proposed process has SO2 removal and NH3 reuse efficiencies of >99.9%. The process is strongly adaptable to different scenarios such as high SO2 levels in flue gas, high NH3 levels from the CO2 absorber and high flue gas temperatures, and has a low energy requirement. Because the process simplifies flue gas desulphurisation and resolves the problems of NH3 loss and SO2 removal, it could significantly reduce the cost of CO2 and SO2 capture by aqueous NH3

9. An integrated power generation system combining solid oxide fuel cell and oxy-fuel combustion for high performance and CO2 capture

An integrated power generation system combining solid oxide fuel cell (SOFC) and oxy-fuel combustion technology is proposed. The system is revised from a pressurized SOFC-gas turbine hybrid system to capture CO2 almost completely while maintaining high efficiency. The system consists of SOFC, gas turbine, oxy-combustion bottoming cycle, and CO2 capture and compression process. An ion transport membrane (ITM) is used to separate oxygen from the cathode exit air. The fuel cell operates at an elevated pressure to facilitate the use of the ITM, which requires high pressure and temperature. The remaining fuel at the SOFC anode exit is completely burned with oxygen at the oxy-combustor. Almost all of the CO2 generated during the reforming process of the SOFC and at the oxy-fuel combustor is extracted from the condenser of the oxy-combustion cycle. The oxygen-depleted high pressure air from the SOFC cathode expands at the gas turbine. Therefore, the expander of the oxy-combustion cycle and the gas turbine provides additional power output. The two major design variables (steam expander inlet temperature and condenser pressure) of the oxy-fuel combustion system are determined through parametric analysis. There exists an optimal condenser pressure (below atmospheric pressure) in terms of global energy efficiency considering both the system power output and CO2 compression power consumption. It was shown that the integrated system can be designed to have almost equivalent system efficiency as the simple SOFC-gas turbine hybrid system. With the voltage of 0.752 V at the SOFC operating at 900 oC and 8 bar, system efficiency over 69.2% is predicted. Efficiency penalty due to the CO2 capture and compression up to 150 bar is around 6.1%.

10. 激光显微捕获口腔上皮细胞的DNA分型%DNA Genotyping of Oral Epithelial Cells by Laser Capure Microdissection

顾丽华; 张晨; 陈连康; 郑会芬; 程莉; 周怀谷

2006-01-01

目的探索激光显微技术(laser capture microdissection system,LCM)捕获口腔上皮细胞,并进行STR-DNA分型检测的方法.方法用VERITAS显微切割仪红外低能激光显微捕获一定数量口腔上皮细胞,进行Profiler Plus试剂盒STR复合扩增,检测DNA基因型.结果7～8个口腔上皮细胞能成功获得STR-DNA分型.3～4个口腔上皮细胞不能成功获得STR-DNA分型.结论激光显微捕获作为一种分离单个细胞的新技术,对于微量口腔上皮细胞的STR-DNA分型是可行的.

11. A novel procedure for the quantitative analysis of metabolites, storage products and transcripts of laser microdissected seed tissues of Brassica napus

2011-06-01

Full Text Available Abstract Background The biology of the seed is complicated by the extensive non-homogeneity (spatial gradients in gene expression, metabolic conversions and storage product accumulation. The detailed understanding of the mechanisms underlying seed growth and storage therefore requires the development of means to obtain tissue-specific analyses. This approach also represents an important priority in the context of seed biotechnology. Results We provide a guideline and detailed procedures towards the quantitative analysis of laser micro-dissected (LM tissues in oilseed rape (Brassica napus. This includes protocols for laser microdissection of the seed, and the subsequent extraction and quantitative analysis of lipids, starch and metabolites (sugars, sugar phosphates, nucleotides, amino acids, intermediates of glycolysis and citric acid cycle. We have also developed a protocol allowing the parallel analysis of the transcriptome using Brassica-specific microarrays. Some data are presented regarding the compartmentation of metabolites within the oilseed rape embryo. Conclusion The described methodology allows for the rapid, combined analysis of metabolic intermediates, major storage products and transcripts in a tissue-specific manner. The protocols are robust for oilseed rape, and should be readily adjustable for other crop species. The suite of methods applied to LM tissues represents an important step in the context of both the systems biology and the biotechnology of oilseeds.

12. Study of the CO2 capture with physical absorption in an electricity production system based on coal gasification, with an integrated combined cycle

The reduction of greenhouse gas emissions call for new strategies toward the use of fossil fuels. The study is aimed to assess the possibility of reducing CO2 emission from an Integrated Gasification Combined Cycle plant. A comparative study of the CO2 capture processes retained in literature (MEA-MDEA, AMP, N-Methyl-Pyrrolidone and methanol) led to the choice of the physical absorption by methanol. The simulation of the IGCC with CO2 capture was carried out with the Aspen Plus software. Integrations of the chosen process, and of the carbon monoxide conversion process in the IGCC, were carried out upstream of the combustion turbine. The results showed that the performance in term of the power output and electric efficiency decrease appreciably. A parametric study was led in order to specify the influence of the vapor rate introduced for the CO conversion; absorption rate; regeneration operating pressure and to determine the optimum conditions for separation. With the aim to improve the calculation of the hydrogen solubility in methanol and to study its influence, measurements of hydrogen solubility in methanol for five isotherms 248; 263; 278; 291 and 308 K were obtained. The experimental technique consisted in a static-analytic method with liquid phase sampling and analysis by a gas chromatography. The parameters of the binary interaction coefficient of the thermodynamic model were adjusted on these results. It appears that this adjustment involves a very little variation on the electric efficiency. The fall of the performances remains however very important. A final efficiency loss of 8,8 % was found in comparison with a standard IGCC. (author)

13. Mini-incision microdissection testicular sperm extraction: a useful technique for men with cryptozoospermia.

Alrabeeah, K; Witmer, J; Ruiz, S; AlMalki, A; Phillips, S; Zini, A

2016-03-01

Microdissection testicular sperm extraction (micro-TESE) was developed to minimize the testicular injury associated with multiple open TESEs. We sought to evaluate a mini-incision micro-TESE in men with cryptozoospermia and non-obstructive azoospermia (NOA). We conducted a retrospective study of 26 consecutive men with NOA and cryptozoospermia who underwent a primary (first) micro-TESE between March 2015 and August 2015. Final assessment of sperm recovery (reported on the day of intra-cytoplasmic sperm injection (ICSI)) was recorded as (i) successful (available spermatozoa for ICSI) or (ii) unsuccessful (no spermatozoa for ICSI). The decision to perform a mini-incision micro-TESE (with limited unilateral micro-dissection) or standard/extensive (with unilateral or bilateral micro-dissection) was guided by the intra-operative identification of sperm recovery (≥5 spermatozoa) from the first testicle. Overall, sperm recovery was successful in 77% (20/26) of the men. In 37% of the men (8/26), the mini-incision micro-TESE was successful (positive sperm recovery). The remaining 18 men required a standard (extensive) microdissection: 61% (11/18) underwent a unilateral and 39% (7/18) a bilateral micro-TESE. We found that 90% (9/10) of the men with cryptozoospermia and 63% (10/16) of the men with NOA underwent a unilateral (mini or standard micro-TESE). The mini-incision micro-TESE allowed for successful sperm recovery in 60% (6/10) of the men with cryptozoospermia and 13% (2/16) of the men with NOA. The data demonstrate that a mini-incision micro-TESE together with rapid intra-operative assessment and identification of spermatozoa recovery can be useful in men undergoing microTESE, particularly, men with cryptozoospermia. PMID:26743017

14. Beyond laser microdissection technology: follow the yellow brick road for cancer research

Legres, Luc G; Janin, Anne; Masselon, Christophe; Bertheau, Philippe

2014-01-01

Normal biological tissues harbour different populations of cells with intricate spacial distribution patterns resulting in heterogeneity of their overall cellular composition. Laser microdissection involving direct viewing and expertise by a pathologist, enables access to defined cell populations or specific region on any type of tissue sample, thus selecting near-pure populations of targeted cells. It opens the way for molecular methods directed towards well-defined populations, and provides...

15. The Development of Chromosome Microdissection and Microcloning Technique and its Applications in Genomic Research

Zhou, Ruo-Nan; Hu, Zan-Min

2007-01-01

The technique of chromosome microdissection and microcloning has been developed for more than 20 years. As a bridge between cytogenetics and molecular genetics, it leads to a number of applications: chromosome painting probe isolation, genetic linkage map and physical map construction, and expressed sequence tags generation. During those 20 years, this technique has not only been benefited from other technological advances but also cross-fertilized with other techniques. Today, it becomes a p...

16. CO{sub 2} Capture Properties of Alkaline-earth Metal Oxides and Hydroxides: A Combined Density Functional Theory and Lattice Phonon Dynamics Study

Duan, Yuhua; Sorescu, Dan

2010-01-01

By combining density functional theory and lattice phonon dynamics, the thermodynamic properties of CO{sub 2} absorption/desorption reactions with alkaline earth metal oxides MO and hydroxides M(OH){sub 2} (where M = Be,Mg,Ca,Sr,Ba) are analyzed. The heats of reaction and the chemical potential changes of these solids upon CO{sub 2} capture reactions have been calculated and used to evaluate the energy costs. Relative to CaO, a widely used system in practical applications, MgO and Mg(OH){sub 2} systems were found to be better candidates for CO{sub 2} sorbent applications due to their lower operating temperatures (600–700 K). In the presence of H{sub 2}O, MgCO{sub 3} can be regenerated into Mg(OH){sub 2} at low temperatures or into MgO at high temperatures. This transition temperature depends not only on the CO{sub 2} pressure but also on the H{sub 2}O pressure. Based on our calculated results and by comparing with available experimental data, we propose a general computational search methodology which can be used as a general scheme for screening a large number of solids for use as CO{sub 2} sorbents.

17. CO2 capture properties of alkaline earth metal oxides and hydroxides: A combined density functional theory and lattice phonon dynamics study

Duan, Yuhua [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States); Sorescu, Dan C. [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States)

2010-01-01

By combining density functional theory and lattice phonon dynamics, the thermodynamic properties of CO2 absorption/desorption reactions with alkaline earth metal oxides MO and hydroxides M(OH)2 (where M = Be, Mg, Ca, Sr, Ba) are analyzed. The heats of reaction and the chemical potential changes of these solids upon CO2 capture reactions have been calculated and used to evaluate the energy costs. Relative to CaO, a widely used system in practical applications, MgO and Mg(OH)2 systems were found to be better candidates for CO2 sorbent applications due to their lower operating temperatures (600-700 K). In the presence of H2O, MgCO3 can be regenerated into Mg(OH)2 at low temperatures or into MgO at high temperatures. This transition temperature depends not only on the CO2 pressure but also on the H2O pressure. Based on our calculated results and by comparing with available experimental data, we propose a general computational search methodology which can be used as a general scheme for screening a large number of solids for use as CO2 sorbents.

18. CO(2) capture properties of alkaline earth metal oxides and hydroxides: A combined density functional theory and lattice phonon dynamics study.

Duan, Yuhua; Sorescu, Dan C

2010-08-21

By combining density functional theory and lattice phonon dynamics, the thermodynamic properties of CO(2) absorption/desorption reactions with alkaline earth metal oxides MO and hydroxides M(OH)(2) (where M=Be,Mg,Ca,Sr,Ba) are analyzed. The heats of reaction and the chemical potential changes of these solids upon CO(2) capture reactions have been calculated and used to evaluate the energy costs. Relative to CaO, a widely used system in practical applications, MgO and Mg(OH)(2) systems were found to be better candidates for CO(2) sorbent applications due to their lower operating temperatures (600-700 K). In the presence of H(2)O, MgCO(3) can be regenerated into Mg(OH)(2) at low temperatures or into MgO at high temperatures. This transition temperature depends not only on the CO(2) pressure but also on the H(2)O pressure. Based on our calculated results and by comparing with available experimental data, we propose a general computational search methodology which can be used as a general scheme for screening a large number of solids for use as CO(2) sorbents. PMID:20726653

19. In vivo gene expression profiling of human intestinal epithelial cells: analysis by laser microdissection of formalin fixed tissues

2008-05-01

Full Text Available Abstract Background The small intestinal epithelium mediates vital functions of nutrient absorption and host defense. The spatial organization of the epithelial cells along the crypt-villus axis segregates them into regions of specialized function. However, the differences in transcriptional programming and the molecular machinery that governs the migration, adhesion, and differentiation of intestinal epithelial cell lineages in humans remain under-explored. To increase our understanding of these mechanisms, we have evaluated gene expression patterns of ileal epithelial cells isolated by laser capture microdissection from either the villus epithelial or crypt cell regions of healthy human small intestinal mucosa. Expression profiles in villus and crypt epithelium were determined by DNA microarray, quantitative real-time PCR, and immunohistochemistry based methods. The expression levels of selected epithelial biomarkers were also compared between gastrointestinal tissues. Results Previously established biomarkers as well as a novel and distinct set of genes believed to be linked to epithelial cell motility, adhesion, and differentiation were found to be enriched in each of the two corresponding cell populations (GEO accession: GSE10629. Additionally, high baseline expression levels of innate antimicrobials, alpha defensin 5 (HD5 and regenerating islet-derived 3 alpha (Reg3A, were detected exclusively within the small bowel crypt, most notably in the ileum in comparison to other sites along the gastrointestinal tract. Conclusion The elucidation of differential gene expression patterns between crypt and villus epithelial cell lineages in human ileal tissue provides novel insights into the molecular machinery that mediates their functions and spatial organization. Moreover, our findings establish an important framework of knowledge for future investigations of human gastrointestinal diseases.

20. Local and Global Impacts of Carbon Capture and Storage Combined with Enhanced Oil Recovery in Four Depleted Oil Fields, Kern County, California

Gillespie, J.; Jordan, P. D.; Goodell, J. A.; Harrington, K.; Jameson, S.

2015-12-01

Depleted oil reservoirs are attractive targets for geologic carbon storage (GCS) because they possess proven trapping mechanisms and large amounts of data pertaining to production and reservoir geometry. In addition, CO2 enhanced oil recovery (EOR) can improve recovery of the remaining oil at recovery factors of 6 to 20% of original oil in place in appropriate reservoirs. CO2 EOR increases the attractiveness of depleted oil and gas reservoirs as a starting point for CCS because the CO2 becomes a commodity that can be purchased by field operators for EOR purposes thereby offsetting the costs of CO2 capture at the power plant. In California, Kern County contains the largest oil reservoirs and produces 76% of California's oil. Most of the production at depths suitable for CCS combined with CO2 EOR comes from three reservoirs: the Vedder and Temblor formations and the Stevens Sandstone of the Monterey Formation. These formations were evaluated for GCS and CO2 EOR potential at the North and South Coles Levee (Stevens Sandstone), Greeley (Vedder) and McKittrick (Temblor) fields. CO2 EOR could be expected to produce an additional 150 million bbls of oil. The total storage space created by pre- and post-EOR fluid production for all three reservoirs is approximately 104 million metric tons (MMT). Large fixed sources in California produce 156 MMT/yr of CO2, and sources in Kern County produce 26 MMT/yr (WESTCARB, 2012). Therefore, the fields could store about four years of local large fixed source emissions and about two thirds of statewide emissions. However, from a global perspective, burning the additional oil produced by CO2 EOR would generate an additional 65 MMT of CO2 if not captured. This would result in a net reduction of greenhouse gas of only 39 MMT rather than the full 104 MMT. If the water produced along with the oil recovered during CO2 EOR operations is not reinjected into the reservoir, the storage space could be much higher.

1. Combining coal gasification, natural gas reforming, and external carbonless heat for efficient production of gasoline and diesel with CO2 capture and sequestration

Highlights: • Several systems are presented which convert NG, coal, and carbonless heat to fuel. • Using nuclear heat can reduce the direct fossil fuel consumption by up to 22%. • The use of CCS depended on the carbon tax: above $20-30/t is sufficient to use CCS. • CTL is only the most economical when the price of NG is more than$5 /MMBtu. • Compared to a traditional CTL plant, total CO2 emission can be reduced up to 79%. - Abstract: In this paper, several novel polygeneration systems are presented which convert natural gas, coal, and a carbonless heat source such as high-temperature helium to gasoline and diesel. The carbonless heat source drives a natural gas reforming reaction to produce hydrogen rich syngas, which is mixed with coal-derived syngas to produce a syngas blend ideal for the Fischer–Tropsch reaction. Simulations and techno-economic analyses performed for 16 different process configurations under a variety of market conditions indicate significant economic and environmental benefits. Using a combination of coal, gas, and carbonless heat, it is possible to reduce CO2 emissions (both direct and indirect) by 79% compared to a traditional coal-to-liquids process, and even achieve nearly zero CO2 emissions when carbon capture and sequestration technology is employed. Using a carbonless heat source, the direct fossil fuel consumption can be reduced up to 22% and achieve a carbon efficiency up to 72%. Market considerations for this analysis include prices of coal, gas, high-temperature helium, gasoline, and CO2 emission tax rates. The results indicate that coal-only systems are never the most economical choice, unless natural gas is more than 5 /MMBtu 2. Combined infrared multiphoton dissociation and electron capture dissociation with a hollow electron beam in Fourier transform ion cyclotron resonance mass spectrometry. Tsybin, Youri O; Witt, Matthias; Baykut, Gökhan; Kjeldsen, Frank; Håkansson, Per 2003-01-01 An electron injection system based on an indirectly heated ring-shaped dispenser cathode has been developed and installed in a 7 Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. This new hardware design allows high-rate electron capture dissociation (ECD) to be carried out by a hollow electron beam coaxial with the ion cyclotron resonance (ICR) trap. Infrared multiphoton dissociation (IRMPD) can also be performed with an on-axis IR-laser beam passing through a hole at the centre of the dispenser cathode. Electron and photon irradiation times of the order of 100 ms are required for efficient ECD and IRMPD, respectively. As ECD and IRMPD generate fragments of different types (mostly c, z and b, y, respectively), complementary structural information that improves the characterization of peptides and proteins by FTICR mass spectrometry can be obtained. The developed technique enables the consecutive or simultaneous use of the ECD and IRMPD methods within a single FTICR experimental sequence and on the same ensemble of trapped ions in multistage tandem (MS/MS/MS or MS(n)) mass spectrometry. Flexible changing between ECD and IRMPD should present advantages for the analysis of protein digests separated by liquid chromatography prior to FTICRMS. Furthermore, ion activation by either electron or laser irradiation prior to, as well as after, dissociation by IRMPD or ECD increases the efficiency of ion fragmentation, including the w-type fragment ion formation, and improves sequencing of peptides with multiple disulfide bridges. The developed instrumental configuration is essential for combined ECD and IRMPD on FTICR mass spectrometers with limited access into the ICR trap. PMID:12872281 3. Laser Microdissection of Grapevine Leaves Reveals Site-Specific Regulation of Transcriptional Response to Plasmopara viticola. Lenzi, Luisa; Caruso, Carla; Bianchedi, Pier Luigi; Pertot, Ilaria; Perazzolli, Michele 2016-01-01 Grapevine is one of the most important fruit crops in the world, and it is highly susceptible to downy mildew caused by the biotrophic oomycete Plasmopara viticola. Gene expression profiling has been used extensively to investigate the regulation processes of grapevine-P. viticola interaction, but all studies to date have involved the use of whole leaves. However, only a small fraction of host cells is in contact with the pathogen, so highly localized transcriptional changes of infected cells may be masked by the large portion of non-infected cells when analyzing the whole leaf. In order to understand the transcriptional regulation of the plant reaction at the sites of pathogen infection, we optimized a laser microdissection protocol and analyzed the transcriptional changes in stomata cells and surrounding areas of grapevine leaves at early stages of P. viticola infection. The results indicate that the expression levels of seven P. viticola-responsive genes were greater in microdissected cells than in whole leaves, highlighting the site-specific transcriptional regulation of the host response. The gene modulation was restricted to the stomata cells and to the surrounding areas of infected tissues, indicating that the host response is mainly located at the infection sites and that short-distance signals are implicated. In addition, due to the high sensitivity of the laser microdissection technique, significant modulations of three genes that were completely masked in the whole tissue analysis were detected. The protocol validated in this study could greatly increase the sensitivity of further transcriptomic studies of the grapevine-P. viticola interaction. PMID:26546320 4. Alteration of gene expression during nasopharyngeal carcinogenesis revealed by oligonucleotide microarray after microdissection of tumor tissue and normal epithelia from nasopharynx LIU Zhong-qi; TIAN Yong-quan; HU Yong-fang; LI Xiao-ling; MA Fu-rong; LI Gui-yuan 2009-01-01 Background Microarray and microdissection techniques were being used for many applications to study the carcinogenesis of some human tumors. But seldom studies had hitherto combined these two techniques to study carcinogenesis mechanism of nasopharyngeal carcinoma (NPC). To identify a set of genes involved in the carcinogenesis and development of NPC, we used the microdissected homogeneous NPC tissue cells and the pure normal epithelium pillar cells to construct the whole human genome expression profiles.Methods We preserved the tissue samples from nasopharynx of 18 patients (including 13 samples of NPC and 5 samples of normal or inflammatory mucous tissue samples from nasopharynx) in RNAlater Stabilization Reagent. The tissue samples were microdissected to harvest the homogeneous tissue cells, then total RNA was isolated from them. The sufficient antisense RNA (aRNA) was amplified from these total RNA. HG-U133.Plus.2.0 GeneChip was used to construct the human whole genome expression profiling of each sample. Differential patterns of expression of genes correlated with the carcinogenesis, classification and progression of NPC were identified with comparing the expression profiling data respectively in leave one out cross-validation analysis. Correlation between aRNA expression measured by the microarrays and semi-quantitative reverse transcription polymerase chain reaction (sqRT-PCR) were also ascertained, and found that hybridization results were validated in all of the 18 patients.Results Differential patterns of expression of 127 genes correlated with the carcinogenesis (A P value less than 0.001 with the 2-fold differentiated expression between case group and control group) of the NPG were filtered. The top most up-regulated and down-regulated 8 genes by the way of permutation test were also selected and listed in the paper. Expression of genes E2F6 and TSPAN-1 was identified using aRNA by sqRT-PCR and showed that there was significant difference between the 5. A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling Tauris, Birgitte; Borg, Søren; Gregersen, Per L; 2009-01-01 Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression p... 6. APPLICATION OF LASER CAPTURE MICRODISSECTION AND PROTEIN MICROARRAY TECHNOLOGIES IN THE MOLECULAR ANALYSIS OF AIRWAY INJURY FOLLOWING POLLUTION PARTICLE EXPOSURE Understanding the mechanisms by which various types of air pollution particles (PM) mediate adverse health effects would provide biological plausibility to epidemiological associations of increased rates of morbidity and mortality. The majority of information regarding the means ... 7. Transcriptome differentiation along the dorso-ventral axis in laser-captured microdissected rat hippocampal granular cell layer Christensen, T.; Bisgaard, C.F.; Nielsen, Henrik Bjørn; 2010-01-01 Several findings suggest a functional and anatomical differentiation along the dorso-ventral axis of the hippocampus. Lesion studies in rats have indicated that the dorsal hippocampus preferentially plays a role in spatial learning and memory, while the ventral hippocampus is involved in anxiety...... ventral granular cell layer with a false discovery rate below 5% and with a relative change in gene expression level of 20% or more. From this pool of genes 45 genes were more than two-fold regulated, 13 genes being dorsally enriched and 32 genes being ventrally enriched. Moreover, cluster analysis based... 8. Transcriptome profiling of sheep granulosa cells and oocytes during early follicular development obtained by Laser Capture Microdissection Bonnet Agnes; Bevilacqua Claudia; Benne Francis; Bodin Loys; Cotinot Corinne; Liaubet Laurence; Sancristobal Magali; Sarry Julien; Terenina Elena; Martin Patrice; Tosser-Klopp Gwenola; Mandon-Pepin Beatrice 2011-01-01 Abstract Background Successful achievement of early folliculogenesis is crucial for female reproductive function. The process is finely regulated by cell-cell interactions and by the coordinated expression of genes in both the oocyte and in granulosa cells. Despite many studies, little is known about the cell-specific gene expression driving early folliculogenesis. The very small size of these follicles and the mixture of types of follicles within the developing ovary make the experimental st... 9. Chromosomal rearrangements in cattle and pigs revealed by chromosome microdissection and chromosome painting Yerle Martine 2003-11-01 Full Text Available Abstract A pericentric inversion of chromosome 4 in a boar, as well as a case of (2q-;5p+ translocation mosaicism in a bull were analysed by chromosome painting using probes generated by conventional microdissection. For the porcine inversion, probes specific for p arms and q arms were produced and hybridised simultaneously on metaphases of a heterozygote carrier. In the case of the bovine translocation, two whole chromosome probes (chromosome 5, and derived chromosome 5 were elaborated and hybridised independently on chromosomal preparations of the bull who was a carrier of the mosaic translocation. The impossibility of differentiating chromosomes 2 and der(2 from other chromosomes of the metaphases did not allow the production of painting probes for these chromosomes. For all experiments, the quality of painting was comparable to that usually observed with probes obtained from flow-sorted chromosomes. The results obtained allowed confirmation of the interpretations proposed with G-banding karyotype analyses. In the bovine case, however, the reciprocity of the translocation could not be proven. The results presented in this paper show the usefulness of the microdissection technique for characterising chromosomal rearrangements in species for which commercial probes are not available. They also confirmed that the main limiting factor of the technique is the quality of the chromosomal preparations, which does not allow the identification of target chromosomes or chromosome fragments in all cases. 10. Analysis of the TP53 gene in laser-microdissected glioblastoma vasculature. Kulla, Andres; Burkhardt, Karim; Meyer-Puttlitz, Birgit; Teesalu, Tambet; Asser, Toomas; Wiestler, Otmar D; Becker, Albert J 2003-04-01 Malignant transformation of human gliomas is accompanied by extensive proliferation of stromal blood vessels. Recent data suggest mesenchymal transdifferentiation of neoplastic cells in various human cancers, including colon and breast cancer as well as gliosarcoma. In this study, we have analyzed proliferating stromal blood vessels in glioblastoma multiforme for the presence of mutations in the tumor suppressor gene TP53. Using tissue arrays derived from glioblastoma specimens, cases with significant immunohistochemical p53 accumulation were selected for molecular genetic detection of TP53 mutations in exons 5 to 8. None of the tumors included in this series displayed properties of gliosarcoma. Proliferating glomeruloid stromal vessels were isolated by laser microdissection from paraffin sections. In six cases, single-strand conformation polymorphism analysis for mutations of the TP53 gene in stromal blood vessels compared with adjacent tumor cells and subsequent DNA sequencing of the resulting DNA fragments were carried out. Glioblastoma cells of these cases exhibited TP53 mutations in exons 5, 7 and 8. None of these tumors showed TP53 mutations in microdissected samples from glomeruloid vessels. The absence of TP53 mutations in vascular stromal components of glioblastoma multiforme supports the hypothesis that microvascular proliferations originate from the tumor stroma and are not derived from transdifferentiated glioblastoma cells. PMID:12624785 11. In situ phosphorylation of proteins in MCTs microdissected from rat kidney: Effect of AVP Homma, S.; Gapstur, S.M.; Yusufi, N.K.; Dousa, T.P. (Mayo Clinic and Foundation, Rochester, MN (USA)) 1988-04-01 Adenosine 3{prime},5{prime}-cyclic monophosphate (cAMP)-dependent protein phosphorylation is considered a key step in the cellular action of vasopressin (AVP) to regulate water permeability in collecting tubules. However, the proteins serving as a substrate(s) for phosphorylation in undisrupted cells have not yet been identified. In the present study, the authors developed a method for investigation of in situ phosphorylation of microdissected segments of medullary collecting tubules (MCT) from rat kidney. Incubation of microdissected MCT segments with low concentrations of saponin, semipermeabilization, increased permeability of the membrane for ATP but did not allow leakage of macromolecules such as lactate dehydrogenase. This treatment also did not cause major disruption of cell structure, or impairment of AVP-sensitive adenylate cyclase. Incubation of semipermeabilized MCT with {gamma}-({sup 32}P)ATP resulted in corporation of {sup 32}P{sub i} into two major protein bands detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis and subsequent autoradiography. Similar incubation of tubules disrupted by hyposmotic solutions and a stronger detergent Triton X-100 resulted in {sup 32}P{sub i} incorporation into multiple protein bands. These findings demonstrate a novel method for identification of endogenous protein substrate(s) for cAMP-dependent protein kinase and other protein kinases and phosphatases that are probably involved in post-cAMP steps in the cellular action of AVP in the intact cells of collecting tubules. 12. In situ phosphorylation of proteins in MCTs microdissected from rat kidney: Effect of AVP Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein phosphorylation is considered a key step in the cellular action of vasopressin (AVP) to regulate water permeability in collecting tubules. However, the proteins serving as a substrate(s) for phosphorylation in undisrupted cells have not yet been identified. In the present study, the authors developed a method for investigation of in situ phosphorylation of microdissected segments of medullary collecting tubules (MCT) from rat kidney. Incubation of microdissected MCT segments with low concentrations of saponin, semipermeabilization, increased permeability of the membrane for ATP but did not allow leakage of macromolecules such as lactate dehydrogenase. This treatment also did not cause major disruption of cell structure, or impairment of AVP-sensitive adenylate cyclase. Incubation of semipermeabilized MCT with γ-[32P]ATP resulted in corporation of 32Pi into two major protein bands detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis and subsequent autoradiography. Similar incubation of tubules disrupted by hyposmotic solutions and a stronger detergent Triton X-100 resulted in 32Pi incorporation into multiple protein bands. These findings demonstrate a novel method for identification of endogenous protein substrate(s) for cAMP-dependent protein kinase and other protein kinases and phosphatases that are probably involved in post-cAMP steps in the cellular action of AVP in the intact cells of collecting tubules 13. Construction of a DNA library from chromosome 4 of rice （Oryza sativa） by microdissection MAOYINGWEI; SIYUANLIANG; 等 1998-01-01 A simple method to create a chromosome-specific DNA librqary of rice,including microdissection,amplification,charterization and cloning,is described.Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR).The PCR products were labeled as probes with DIG-11-dUTP using the random priming method.Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4.A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed.Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42% contained repetitive sequences.The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice. 14. Anatomy of the temporomandibular joint in the cat: a study by microdissection, cryosection and vascular injection. Arredondo, Jorge; Agut, Amalia; Rodríguez, María Jesús; Sarriá, Ricardo; Latorre, Rafael 2013-02-01 The minute anatomy of the temporomandibular joint (TMJ) is of great clinical relevance in cats owing to a high number of lesions involving this articulation. However, the precise anatomy is poorly documented in textbooks and scientific articles. The aim of this study was to describe, in detail, the TMJ anatomy and its relationship with other adjacent anatomical structures in the cat. Different anatomical preparations, including vascular and articular injection, microdissection, cryosection and plastination, were performed in 12 cadaveric cats. All TMJ anatomical structures were identified and described in detail. A thorough understanding of the TMJ anatomy is essential to understand the clinical signs associated with TMJ disorders, to locate lesions precisely and to accurately interpret the results in all diagnostic imaging techniques. PMID:23015066 15. Outcome of microdissection TESE compared with conventional TESE in non-obstructive azoospermia: a systematic review. Deruyver, Y; Vanderschueren, D; Van der Aa, F 2014-01-01 Retrieval of spermatozoa is unfortunately still only successful in a subset of patients suffering from non-obstructive azoospermia (NOA) by conventional testicular sperm extraction (TESE). Microdissection TESE may have some theoretical benefits over conventional TESE, but uncertainty exists about its superiority. The objective of this systematic review was therefore to compare the efficacy and safety of microTESE with conventional TESE in men with NOA. The systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis statement. Literature was searched for studies comparing outcome of conventional TESE with microdissection TESE. Primary outcome was sperm retrieval rate (SRR). Secondary outcomes were clinical predictors of sperm retrieval as well as complication rate. Of 62 articles, a total of seven studies were included in the final analysis. Overall SRR was significantly higher in the microTESE group in comparison with conventional TESE in five of these studies. Overall sperm retrieval ranged from 16.7 to 45% in the conventional TESE vs. 42.9 to 63% in the microTESE group. A sub-analysis of the SRR according to testicular histology was available in four of the selected articles. MicroTESE in men with Sertoli cell only syndrome and hypospermatogenesis carried a small but significant more favourable outcome according to, respectively, two and one of the studies. Correlation of serum follicle stimulating hormone and testicular volume with positive outcome was variable. Fewer complications were observed on ultrasound examination after microTESE procedure. Clinical randomized studies comparing microTESE with conventional TESE in NOA are still lacking to date. Pseudo-randomized prospective data, however, show more favourable sperm retrieval in NOA for microTESE, especially in histological patterns of patchy spermatogenesis such as Sertoli cell only syndrome. However, in patients with uniform histological patterns 16. Combined electron capture and infrared multiphoton dissociation for multistage MS/MS in a Fourier transform ion cyclotron resonance mass spectrometer. Håkansson, Kristina; Chalmers, Michael J; Quinn, John P; McFarland, Melinda A; Hendrickson, Christopher L; Marshall, Alan G 2003-07-01 We have mounted a permanent on-axis dispenser cathode electron source inside the magnet bore of a 9.4-T Fourier transform ion cyclotron resonance mass spectrometer. This configuration allows electron capture dissociation (ECD) to be performed reliably on a millisecond time scale. We have also implemented an off-axis laser geometry that enables simultaneous access to ECD and infrared multiphoton dissociation (IRMPD). Optimum performance of both fragmentation techniques is maintained. The analytical utility of performing either ECD or IRMPD on a given precursor ion population is demonstrated by structural characterization of several posttranslationally modified peptides: IRMPD of phosphorylated peptides results in few backbone (b- and y-type) cleavages, and product ion spectra are dominated by neutral loss of H3PO4. In contrast, ECD provides significantly more backbone (c- and z*-type) cleavages without loss of H3PO4. For N-glycosylated tryptic peptides, IRMPD causes extensive cleavage of the glycosidic bonds, providing structural information about the glycans. ECD cleaves all backbone bonds (except the N-terminal side of proline) in a 3-kDa glycopeptide with no saccharide loss. However, only a charge-reduced radical species and some side chain losses are observed following ECD of a 5-kDa glycopeptide from the same protein. An MS3 experiment involving IR laser irradiation of the charge-reduced species formed by electron capture results in extensive dissociation into c- and z-type fragment ions. Mass-selective external ion accumulation is essential for the structural characterization of these low-abundance (modified) peptides. PMID:12964777 17. Investigation of Partamona helleri (Apidae, Meliponini) B chromosome origin. An approach by microdissection and whole chromosome painting Martins, Cinthia; Diniz, Debora; Sobrinho-Scudeler, Patricia; Foresti, Fausto; Campos, Lucio; Costa, Marco 2012-01-01 The stingless bee Partamona helleri in southeast Brazil shows the regular chromosome number 2n = 34 and a variable number of up to four minute B1 or B2 chromosomes. Previous cytogenetic analyses have indicated morphological similarities between the B1 chromosome and chromosome segments in the regular karyotype. In this study, microdissection and chromosome painting were employed along with C banding, NOR banding, and base-specific fluorochrome staining to investigate the origin of the B1 chro... 18. Biogenic Amines in Microdissected Brain Regions of Drosophila melanogaster Measured with Micellar Electrokinetic Capillary Chromatography – Electrochemical Detection Kuklinski, Nicholas J.; Berglund, E. Carina; Engelbrektsson, Johan; Ewing, Andrew G. 2010-01-01 Micellar electrokinetic chromatography with electrochemical detection has been used to quantify biogenic amines in microdissected Drosophila melanogaster brains and brain regions. The effects of pigment from the relatively large fly eyes on the separation have been examined to find that the red pigment from the compound eye masks much of the signal from biogenic amines. The brains of white mutant flies, which have characteristically low pigment in the eyes, have a significantly simplified sep... 19. Synthesis of PBAD-lipiodol nanoparticles for combination treatment with boric acid in boron neutron capture therapy for hepatoma in-vitro This study attempted to increase BNCT efficiency for hepatoma by a combined treatment of phenylboric acid derivative entrapped lipiodol nanoparticles (PBAD-L nanoparticles) with boric acid. The size of PBAD-L nanoparticles were 400-750 nm at the boron concentrations of 0.3-2.7 mg/ml. After 24 hours the boron concentration in PBAD-L nanoparticles treated human hepatoma HepG2 cells was 112 ppm, while that in rat liver Clone 9 cells was 52 ppm. With the use of 25 μg B/ml boric acid, after 6 hours the boron concentration in HepG2 and Clone 9 cells were 75 ppm and 40 ppm, respectively. In a combined treatment, boron concentration in HepG2 cells which were treated with PBAD-L nanoparticles for 18 hours and then combined with boric acid for 6 hours was 158 ppm. After neutron irradiation, the surviving fraction of HepG2 cells treated with PBAD-L nanoparticles was 12.6%, while that in the ones with a combined treatment was 1.3%. In conclusion, the combined treatment provided a higher boron concentration in HepG2 cells than treatments with either PBAD-L nanoparticles or boric acid, resulting in a higher therapeutic efficacy of BNCT in hepatoma cells. (author) 20. Laser microdissection-based analysis of the Y sex chromosome of the Antarctic fish Chionodraco hamatus (Notothenioidei, Channichthyidae Ennio Cocca 2015-02-01 Full Text Available Microdissection, DOP-PCR amplification and microcloning were used to study the large Y chromosome of Chionodraco hamatus, an Antarctic fish belonging to the Notothenioidei, the dominant component of the Southern Ocean fauna. The species has evolved a multiple sex chromosome system with digametic males showing an X1YX2 karyotype and females an X1X1X2X2 karyotype. Fluorescence in situ hybridization, performed with a painting probe made from microdissected Y chromosomes, allowed a deeper insight on the chromosomal rearrangement, which underpinned the fusion event that generated the Y. Then, we used a DNA library established by microdissection and microcloning of the whole Y chromosome of Ch. hamatus for searching sex-linked sequences. One clone provided preliminary information on the presence on the Y chromosome of the CHD1 gene homologue, which is sex-linked in birds but in no other vertebrates. Several clones from the Y-chromosome mini-library contained microsatellites and transposable elements, one of which mapped to the q arm putative fusion region of the Y chromosome. The findings confirm that interspersed repetitive sequences might have fostered chromosome rearrangements and the emergence of the Y chromosome in Ch. hamatus. Detection of the CHD1 gene in the Y sex-determining region could be a classical example of convergent evolution in action. 1. PRELIMINARY TECHNICAL AND ECONOMIC FEASIBILITY STUDY ON THE INTEGRATION OF A PROCESS UTILIZING LOW-ENERGY SOLVENTS FOR CARBON DIOXIDE CAPTURE ENABLED BY A COMBINATION OF ENZYMES AND ULTRASONICS WITH A SUBCRITICAL PC POWER PLANT Swaminathan, Saravanan; Kuczynska, Agnieszka; Hume, Scott; Mulgundmath, Vinay; Freeman, Charles; Bearden, Mark; Remias, Joe; Ambedkar, Balraj; Salmon, Sonja; House, Alan 2012-11-01 The results of the preliminary techno-economic assessment for integrating a process utilizing low-energy solvents for carbon dioxide (CO2) capture enabled by a combination of enzymes and ultrasonics with a subcritical pulverized coal (PC) power plant are presented. Four cases utilizing the enzyme-activated solvent are compared using different methodologies of regeneration against the DOE/NETL reference MEA case. The results are shown comparing the energy demand for post-combustion CO2 capture and the net higher heating value (HHV) efficiency of the power plant integrated with the post-combustion capture (PCC) plant. A levelized cost of electricity (LCOE) assessment was performed showing the costs of the options presented in the study. The key factors contributing to the reduction of LCOE were identified as enzyme make-up rate and the capability of the ultrasonic regeneration process. The net efficiency of the integrated PC power plant with CO2 capture changes from 24.9% with the reference Case 10 plant to between 24.34% and 29.97% for the vacuum regeneration options considered, and to between 26.63% and 31.41% for the ultrasonic regeneration options. The evaluation also shows the effect of the critical parameters on the LCOE, with the main variable being the initial estimation of enzyme dosing rate. The LCOE (/MWh) values range from 112.92 to 125.23 for the vacuum regeneration options and from 108.9 to 117.50 for the ultrasonic regeneration cases considered in comparison to 119.6 for the reference Case 10. A sensitivity analysis of the effect of critical parameters on the LCOE was also performed. The results from the preliminary techno-economic assessment show that the proposed technology can be investigated further with a view to being a viable alternative to conventional CO2 scrubbing technologies.

2. Homogeneous liquid-liquid extraction combined with gas chromatography-electron capture detector for the determination of three pesticide residues in soils

Wang Xuedong [School of Environmental Science and Public Health, Wenzhou Medical College, Wenzhou 325035 (China)], E-mail: zjuwxd@yahoo.com.cn; Zhao Xinna; Liu Xiujuan; Li Yanyan; Fu Lingyan; Hu Jia [Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, Central China Normal University, Wuhan 430079 (China); Huang Changjiang [School of Environmental Science and Public Health, Wenzhou Medical College, Wenzhou 325035 (China)

2008-07-14

In this study, a new method was developed for analyzing malathion, cypermethrin and lambda-cyhalothrin from soil samples by using homogeneous liquid-liquid extraction (HLLE) and gas chromatography with electron capture detector (GC-ECD). Acetone was used as extraction solvent for the extraction of target pesticides from soil samples. When the extraction process was finished, the target analytes in the extraction solvent were rapidly transferred from the acetone extract to carbon tetrachloride, using HLLE. Under the optimum conditions, linearity was obtained in the range of 0.05-40 {mu}g kg{sup -1} for malathion, 0.04-10 {mu}g kg{sup -1} for lambda-cyhalothrin and 0.05-50 {mu}g kg{sup -1} for cypermethrin, respectively. Coefficients of correlation (r{sup 2}) ranged from 0.9993 to 0.9998. The repeatability was carried out by spiking soil samples at concentration levels of 2.5 {mu}g kg{sup -1} for lambda-cyhalothrin, and 10 {mu}g kg{sup -1} for malathion and cypermethrin, respectively. The relative standard deviations (RSDs) varied between 2.3 and 9.6% (n = 3). The limits of detection (LODs), based on signal-to-noise ratio (S/N) of 3, varied between 0.01 and 0.04 {mu}g kg{sup -1}. The relative recoveries of three pesticides from soil A1, A2 and A3 at spiking levels of 2.5, 5 and 10 {mu}g kg{sup -1} were in the range of 82.20-91.60%, 88.90-110.5% and 77.10-98.50%, respectively. In conclusion, the proposed method can be successfully applied for the determination of target pesticide residues in real soil samples.

3. Molecular Analysis of the B Microchromosome in Steindachnerina insculpta (Characiformes: Curimatidae) by Microdissection.

Sampaio, Tatiane R; Gouveia, Juceli G; da Silva, Carlos R M; Dias, Ana L; da Rosa, Renata

2015-01-01

B chromosomes are additional elements to standard karyotypes observed in different species of fishes, especially in Curimatidae. However, despite studies demonstrating the occurrence of Bs, little is known about their origin and evolution. To better understand the genomic composition and evolutionary processes involving B chromosomes, microdissection of B microchromosomes in Steindachnerina insculpta was conducted. Chromosome painting revealed the totally hybridized B and markings on A chromosomes both in S. in sculpta and in Cyphocharax spilotus, demonstrating a strong homology between these different species. In specimens of C. modestus, which do not have Bs, the signals were observed on A chromosomes. Cloning and sequencing of some B fragments revealed that the B microchromosome in S. insculpta is composed of repetitive elements, homologous to the DIRS-4 LTR retrotransposon of Xenopus (Silurana) tropicalis. FISH with clone pSi48 with the DIRS-4 retroelement revealed signals on all A chromosomes in the 2 species and also on the B, suggesting the insertion of repetitive elements in these species. PMID:25999244

4. Optimization of spermatozoa detection using immunofluorescent staining and laser micro-dissection.

Ping, Yueh Shyang; Chan, Xavier Liang Shun; Goh, Sze Kae; Syn, Christopher Kiu Choong

2015-10-01

The present study evaluated the use of an immunofluorescence-based assay for the microscopic detection of human spermatozoa, following which the fluorescence-labelled spermatozoa could be excised with a laser micro-dissection system. The Sperm Hy-Liter™ PI kit was able to detect spermatozoa from as little as 20nL of semen. No interference or non-specificity were observed when the kit was used on semen mixed with various body fluids such as blood and urine, as well as when semen was spiked onto different types of fabric. Good results could also be obtained with rectal samples which contain auto-fluorescent fecal materials through the use of dual FITC/PI filters. We also developed a method for concurrent testing of two protein biomarkers of semen (semenogelin and prostate-specific antigen) and detection of spermatozoa. This approach would maximize the evidential value from a single piece of sexual assault exhibit. The results also showed that staining by Sperm Hy-Liter™ PI does not interfere with DNA recovery, facilitating the generation of clear male DNA profiles from dissected spermatozoa, thereby making profile interpretation less complex. In summary, Sperm Hy-Liter™ PI staining was demonstrated to be sensitive, robust and specific. PMID:26338669

5. ERα and ERK1/2 MAP kinase expression in microdissected stromal and epithelial endometrial cells

Said Abu Alkhair Mohamed

2014-03-01

Total and phosphorylated levels for ERK1/2 and ERα were measured by quantitation of signals from Western blots using specific antibodies against the active and total forms of ERK1/2 and against ERα. When the level of the proteins was quantitated and normalized to β actin from microdissected stroma and epithelium, no significant difference was detected in the levels of these proteins between the two tissue compartments. There was a trend toward higher expression in the stroma vs. epithelium, respectively (active ERK1/2 0.45 ± 0.17 vs. 0.2 ± 0.65; total ERK1/2 0.54 ± 0.35 vs. 0.28 ± 0.23; ERα 0.82 ± 0.28 vs. 0.54 ± 0.18; n = 6. These data demonstrate that there are comparable levels of ERα (P = 0.41, total ERK1/2 (P = 0.18 and active ERK1/2 (P = 0.13 in the stroma and epithelium of proliferative phase endometrium with a trend toward higher expression of these proteins in the stromal compartment.

6. Design, modeling and control of a piezoelectric ultrasonic microdissection technique for the molecular analysis of tissue

Molecular techniques are transforming our understanding of cellular function and disease. However, accurate molecular analysis methods will be limited if the input DNA, RNA or protein is not derived from a pure population of cells or is contaminated by the wrong cells. The modeling and control of the piezoelectric actuator, with an objective application towards ultrasonic vibration cutting (UVC), is addressed in this paper. The piezoelectric actuator is used in realizing the fast and precise movements of the developed UVC so as to procure a pure population of targeted cells from tissue sections for subsequent pathology analysis with precision and without causing a large deformation. To address the nonlinearities and uncertainties of the piezoelectric actuator, an adaptive controller based on a hysteresis model is proposed to yield robust control performance. A multilayer piezoelectric actuator is used to actuate a sharp needle vibrating at high frequency and low amplitude to cut the tissue. Experimental results showed that the embedded tissue can be quickly and precisely cut with this ultrasonic vibration microdissection method

7. HER-2/neu amplification testing in breast cancer by Multiplex Ligation-dependent Probe Amplification: influence of manual- and laser microdissection

Accurate assessment of HER-2/neu status is crucial for proper prognostic information and to offer direct appropriate treatment for breast cancer patients. Next to immunohistochemistry (IHC) to evaluate HER2 protein overexpression, a second line gene amplification test is generally deemed necessary for cases with equivocal protein expression. Recently, a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), was introduced as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer. MLPA was previously shown to correlate well with IHC and in situ hybridization (ISH), but a low tumor percentage in the tissue tested could negatively affect the accuracy of MLPA results. To examine this, MLPA was repeated in 42 patients after serial H&E section guided manual dissection with a scalpel and after laser microdissection of the tumor. Both dissection techniques led to higher HER2 gene copy number ratios and thereby made MLPA more quantitative. Concordance between MLPA and ISH improved from 61% to 84% after manual microdissection and to 90% after laser microdissection. Manual and laser microdissection similarly increase the dynamic range of MLPA copy number ratios which is a technical advantage. As clinically a dichotomization between normal and amplified suffices and MLPA is relatively unsensitive to tumor content, microdissection before MLPA may not be routinely necessary but may be advisable in case of very low tumor content (≤30%), when MLPA results are equivocal, or when extensive ductal carcinoma in situ is present. Since differences between manual and laser microdissection were small, less time consuming manual microdissection appears to be sufficient

8. Solid-Fueled Pressurized Chemical Looping with Flue-Gas Turbine Combined Cycle for Improved Plant Efficiency and CO{sub 2} Capture

Liu, Kunlei; Chen, Liangyong; Zhang, Yi; Richburg, Lisa; Simpson, James; White, Jay; Rossi, Gianalfredo

2013-12-31

The purpose of this document is to report the final result of techno-economic analysis for the proposed 550MWe integrated pressurized chemical looping combustion combined cycle process. An Aspen Plus based model is delivered in this report along with the results from three sensitivity scenarios including the operating pressure, excess air ratio and oxygen carrier performance. A process flow diagram and detailed stream table for the base case are also provided with the overall plant energy balance, carbon balance, sulfur balance and water balance. The approach to the process and key component simulation are explained. The economic analysis (OPEX and CAPX) on four study cases via DOE NETL Reference Case 12 are presented and explained.

9. Proteomic analysis of hippocampal dentate granule cells in frontotemporal lobar degeneration: Application of laser capture technology.

YairM.Gozal

2011-04-01

Full Text Available Frontotemporal lobar degeneration (FTLD is the most common cause of dementia with pre-senile onset, accounting for as many as 20% of cases. A common subset of FTLD cases is characterized by the presence of ubiquitinated inclusions in vulnerable neurons (FTLD-U. While the pathophysiological mechanisms underlying neurodegeneration in FTLD-U have not yet been elucidated, the presence of inclusions in this disease indicates enhanced aggregation of one or several proteins. Moreover, these inclusions suggest altered expression, processing, or degradation of proteins during FTLD-U pathogenesis. Thus, one approach to understanding disease mechanisms is to delineate the molecular changes in protein composition in FTLD-U brain. Using a combined approach consisting of laser capture microdissection (LCM and high resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS, we identified 1252 proteins in hippocampal dentate granule cells excised from three post-mortem FTLD-U and three unaffected control cases processed in parallel. Additionally, we employed a labeling-free quantification technique to compare the abundance of the identified proteins between FTLD-U and control cases. Quantification revealed 54 proteins with selective enrichment in FTLD-U, including TAR DNA binding protein 43 (TDP-43, a recently identified component of ubiquitinated inclusions. Moreover, 19 proteins were selectively decreased in FTLD-U. Subsequent immunohistochemical analysis of TDP-43 and three additional protein candidates suggests that our proteomic profiling of FTLD-U dentate granule cells reveals both inclusion-associated proteins and non-aggregated disease-specific proteins. Application of LCM is a valuable tool in the molecular analysis of complex tissues, and its application in the proteomic characterization of neurodegenerative disorders such as FTLD-U may be used to identify proteins altered in disease.

10. Distribution of androgen receptor in microdissected brain areas of the female baboon (Papio cynocephalus).

Handa, R J; Roselli, C E; Resko, J A

1988-03-29

We measured androgen receptors in the brain and pituitary of 4 female baboons (Papio cynocephalus) by the in vitro binding of methyltrienolone (R1881) to cytosols from 17 brain subregions as well as anterior and posterior pituitaries. High levels of AR were detected in anterior (22.1 +/- 7.1 (S.E.M.) fmol/mg protein) and posterior pituitary (12.6 +/- 3.3 fmol/mg protein). In brain tissue, the highest androgen receptor levels were found in the infundibular nucleus/median eminence (9.4 +/- 2.3 fmol/mg protein), ventromedial nucleus (6.3 +/- 1.7 fmol/mg protein) and periventricular area (4.9 +/- 1.3 fmol/mg protein). Saturation analysis of anterior pituitary and brain tissue (pool of hypothalamic, preoptic area, amygdala and septum remaining after microdissection of brain nuclei) showed that [3H]R1881 binds to the androgen receptor with high specificity and affinity (Kd = 1.25 x 10(-10) M, 0.45 x 10(-10) M, in anterior pituitary and HPA cytosol, respectively). Serum testosterone levels were low in all animals (0.59 +/- 0.26 ng/ml). With these data we described the quantitative distribution of androgen receptor in the pituitary and in specific brain nuclei in a species of nonhuman primate. The distribution is similar in many respects to that described in the male rat and the data suggest a conservation of androgen receptor distribution across species. PMID:3259151

11. Delineation of a complex karyotypic rearrangement by microdissection and CGH in a family affected with split foot

Weimer, J.; Kiechle, M.; Wiedemann, U.; Tonnies, H; Neitzel, H; Ruhenstroth, E.; Ovens-Raeder, A.; Arnold, N.

2000-01-01

We report on a male patient and members of his family with additional material in chromosome 3. This derivative chromosome 3 was transmitted from his mother who had a complex rearrangement between chromosomes 2, 3, and 7. It was possible to delineate her chromosomal rearrangement by microdissection and reverse painting and to exclude these aberrations from being responsible for neonatal deaths and several abortions in this family. Two members of this family suffer from ectrodactyly or split h...

12. The use of laser microdissection in the identification of suitable reference genes for normalization of quantitative real-time PCR in human FFPE epithelial ovarian tissue samples.

Jing Cai

Full Text Available Quantitative real-time PCR (qPCR is a powerful and reproducible method of gene expression analysis in which expression levels are quantified by normalization against reference genes. Therefore, to investigate the potential biomarkers and therapeutic targets for epithelial ovarian cancer by qPCR, it is critical to identify stable reference genes. In this study, twelve housekeeping genes (ACTB, GAPDH, 18S rRNA, GUSB, PPIA, PBGD, PUM1, TBP, HRPT1, RPLP0, RPL13A, and B2M were analyzed in 50 ovarian samples from normal, benign, borderline, and malignant tissues. For reliable results, laser microdissection (LMD, an effective technique used to prepare homogeneous starting material, was utilized to precisely excise target tissues or cells. One-way analysis of variance (ANOVA and nonparametric (Kruskal-Wallis tests were used to compare the expression differences. NormFinder and geNorm software were employed to further validate the suitability and stability of the candidate genes. Results showed that epithelial cells occupied a small percentage of the normal ovary indeed. The expression of ACTB, PPIA, RPL13A, RPLP0, and TBP were stable independent of the disease progression. In addition, NormFinder and geNorm identified the most stable combination (ACTB, PPIA, RPLP0, and TBP and the relatively unstable reference gene GAPDH from the twelve commonly used housekeeping genes. Our results highlight the use of homogeneous ovarian tissues and multiple-reference normalization strategy, e.g. the combination of ACTB, PPIA, RPLP0, and TBP, for qPCR in epithelial ovarian tissues, whereas GAPDH, the most commonly used reference gene, is not recommended, especially as a single reference gene.

13. High quality RNA isolation from Aedes aegypti midguts using laser microdissection microscopy

Gobert Geoffrey N

2011-05-01

Full Text Available Abstract Background Laser microdissection microscopy (LMM has potential as a research tool because it allows precise excision of target tissues or cells from a complex biological specimen, and facilitates tissue-specific sample preparation. However, this method has not been used in mosquito vectors to date. To this end, we have developed an LMM method to isolate midgut RNA using Aedes aegypti. Results Total RNA was isolated from Ae. aegypti midguts that were either fresh-frozen or fixed with histological fixatives. Generally, fresh-frozen tissue sections are a common source of quality LMM-derived RNA; however, our aim was to develop an LMM protocol that could inactivate pathogenic viruses by fixation, while simultaneously preserving RNA from arbovirus-infected mosquitoes. Three groups (10 - 15 mosquitoes per group of female Ae. aegypti at 24 or 48-hours post-blood meal were intrathoracically injected with one of seven common fixatives (Bouin's, Carnoy's, Formoy's, Cal-Rite, 4% formalin, 10% neutral buffered formalin, or zinc formalin to evaluate their effect on RNA quality. Total RNA was isolated from the fixed abdomens using a Trizol® method. The results indicated that RNA from Carnoy's and Bouin's fixative samples was comparable to that of fresh frozen midguts (control in duplicate experiments. When Carnoy's and Bouin's were used to fix the midguts for the LMM procedure, however, Carnoy's-fixed RNA clearly showed much less degradation than Bouin's-fixed RNA. In addition, a sample of 5 randomly chosen transcripts were amplified more efficiently using the Carnoy's treated LMM RNA than Bouin's-fixed RNA in quantitative real-time PCR (qRT-PCR assays, suggesting there were more intact target mRNAs in the Carnoy's fixed RNA. The yields of total RNA ranged from 0.3 to 19.0 ng per ~3.0 × 106 μm2 in the LMM procedure. Conclusions Carnoy's fixative was found to be highly compatible with LMM, producing high quality RNA from Ae. aegypti midguts while

14. Synovectomy by Neutron capture

The Synovectomy by Neutron capture has as purpose the treatment of the rheumatoid arthritis, illness which at present does not have a definitive curing. This therapy requires a neutron source for irradiating the articulation affected. The energy spectra and the intensity of these neutrons are fundamental since these neutrons induce nuclear reactions of capture with Boron-10 inside the articulation and the freely energy of these reactions is transferred at the productive tissue of synovial liquid, annihilating it. In this work it is presented the neutron spectra results obtained with moderator packings of spherical geometry which contains in its center a Pu239 Be source. The calculations were realized through Monte Carlo method. The moderators assayed were light water, heavy water base and the both combination of them. The spectra obtained, the average energy, the neutron total number by neutron emitted by source, the thermal neutron percentage and the dose equivalent allow us to suggest that the moderator packing more adequate is what has a light water thickness 0.5 cm (radius 2 cm) and 24.5 cm heavy water (radius 26.5 cm). (Author)

15. Semi-automatic laser beam microdissection of the Y chromosome and analysis of Y chromosome DNA in a dioecious plant, Silene latifolia

Silene latifolia has heteromorphic sex chromosomes, the X and Y chromosomes. The Y chromosome, which is thought to carry the male determining gene, was isolated by UV laser microdissection and amplified by degenerate oligonucleotide-primed PCR. In situ chromosome suppression of the amplified Y chromosome DNA in the presence of female genomic DNA as a competitor showed that the microdissected Y chromosome DNA did not specifically hybridize to the Y chromosome, but-hybridized to all chromosomes. This result suggests that the Y chromosome does not contain Y chromosome-enriched repetitive sequences. A repetitive sequence in the microdissected Y chromosome, RMY1, was isolated while screening repetitive sequences in the amplified Y chromosome. Part of the nucleotide sequence shared a similarity to that of X-43.1, which was isolated from microdissected X chromosomes. Since fluorescence in situ hybridization analysis with RMY1 demonstrated that RMY1 was localized at the ends of the chromosome, RMY1 may be a subtelomeric repetitive sequence. Regarding the sex chromosomes, RMY1 was detected at both ends of the X chromosome and at one end near the pseudoautosomal region of the Y chromosome. The different localization of RMY1 on the sex chromosomes provides a clue to the problem of how the sex chromosomes arose from autosomes

16. Video Screen Capture Basics

Dunbar, Laura

2014-01-01

This article is an introduction to video screen capture. Basic information of two software programs, QuickTime for Mac and BlueBerry Flashback Express for PC, are also discussed. Practical applications for video screen capture are given.

17. Comparative transcriptional survey between laser-microdissected cells from laminar abscission zone and petiolar cortical tissue during ethylene-promoted abscission in citrus leaves

2009-10-01

Full Text Available Abstract Background Abscission is the cell separation process by which plants are able to shed organs. It has a great impact on the yield of most crop plants. At the same time, the process itself also constitutes an excellent model to study cell separation processes, since it occurs in concrete areas known as abscission zones (AZs which are composed of a specific cell type. However, molecular approaches are generally hampered by the limited area and cell number constituting the AZ. Therefore, detailed studies at the resolution of cell type are of great relevance in order to accurately describe the process and to identify potential candidate genes for biotechnological applications. Results Efficient protocols for the isolation of specific citrus cell types, namely laminar abscission zone (LAZ and petiolar cortical (Pet cells based on laser capture microdissection (LCM and for RNA microextraction and amplification have been developed. A comparative transcriptome analysis between LAZ and Pet from citrus leaf explants subjected to an in-vitro 24 h ethylene treatment was performed utilising microarray hybridization and analysis. Our analyses of gene functional classes differentially represented in ethylene-treated LAZ revealed an activation program dominated by the expression of genes associated with protein synthesis, protein fate, cell type differentiation, development and transcription. The extensive repertoire of genes associated with cell wall biosynthesis and metabolism strongly suggests that LAZ layers activate both catabolic and anabolic wall modification pathways during the abscission program. In addition, over-representation of particular members of different transcription factor families suggests important roles for these genes in the differentiation of the effective cell separation layer within the many layers contained in the citrus LAZ. Preferential expression of stress-related and defensive genes in Pet reveals that this tissue is

Some data are presented for radiative electron capture by fast moving ions. The radiative electron capture spectrum is shown for O8+ in Ag, along with the energy dependence of the capture cross-section. A discrepancy between earlier data, theoretical prediction, and the present data is pointed out. (3 figs) (U.S.)

19. Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ

Sonne, Si Brask; Dalgaard, Marlene D; Nielsen, John Erik;

2009-01-01

Microarray and RT-PCR based methods are important tools for analysis of gene expression; however, in tissues containing many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser microdis......Microarray and RT-PCR based methods are important tools for analysis of gene expression; however, in tissues containing many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser......-containing cells, which is useful for isolation of the androgen-producing Leydig cells. Both protocols retain a morphology that is compatible with laser microdissection and yield RNA of a quality suitable for PCR and microarray analysis....

20. Microdissection of additional chromosome in common wheat-Th. intermedium TAI-27 and screening of its special probe

田靫; 卢一凡; 邓继先; 李滨; 张学勇; 刘广田

2000-01-01

It was identified that there were 4 St chromosomes derived from Th, intermedium in common wheat-Th. intermedium alien additional line TAI-27 by in situ hybridization. Two St chromosomes added to wheat chromosome in TAI-27 as well as two of them replaced two of 42 in wheat chromosomes. This indicates that TAI-27 is not only an alien additional line, but also a replacing line. The additional chromosomes in TAI-27 were microdissected and a microcloning library was constructed. A special probe of Th. intermedium was obtained from a microcloning library. The sequence analysis indicated that there were no homology with Genebank data. This makes it possible to screen genes with the disease-resistance, adversity-tolerance and fine character from Th.intermedium.

1. Capture Their Attention: Capturing Lessons Using Screen Capture Software

Drumheller, Kristina; Lawler, Gregg

2011-01-01

When students miss classes for university activities such as athletic and academic events, they inevitably miss important class material. Students can get notes from their peers or visit professors to find out what they missed, but when students miss new and challenging material these steps are sometimes not enough. Screen capture and recording…

2. Laser capture microdissection coupled with RNA-seq reveals that short cuboidal trophoblast cells of the porcine placenta possess a transcriptome consistent with a migratory cell type

The porcine placenta is classified as epitheliochorial with a diffuse distribution of chorionic villi that do not invade maternal tissues. Thus, the maternal and fetal bloodstreams are separated by six distinct tissue layers. The porcine fetal-maternal interface (FMI) is substantially folded to incr...

3. Laser capture microdissection as a tool to evaluate human papillomavirus genotyping and methylation as biomarkers of persistence and progression of anal lesions

Cornall, Alyssa M; Roberts, Jennifer M; Molano, Monica; Machalek, Dorothy A; Phillips, Samuel; Hillman, Richard J; Grulich, Andrew E; Jin, Fengyi; Poynten, I Mary; Templeton, David J; Garland, Suzanne M; Tabrizi, Sepehr N.

2015-01-01

Introduction Anal squamous cell carcinoma is preceded by persistent infection with high-risk human papillomavirus (HPV) and the cancer precursor, high-grade squamous intraepithelial lesion (HSIL). Detection of specific HPV genotypes and HPV-related biomarkers may be an option for primary anal screening. However, more data on the natural history of HPV-related anal lesions are required. The outcomes from this study will enhance our understanding of the clinical and biological behaviour of HPV-...

4. Phylogeny and distribution of an unknown Treponema sp. associated with porcine colitis by using in situ hybridization and laser capture microdissection (LCM)

Mølbak, Lars; Schou, Kirstine Klitgaard; Jensen, Tim Kåre;

Helical-shaped bacteria resembling Spirochaetes commonly are present in the gastrointestinal tract of animals and humans. Culturing of Spirochaetes is in general fastidious and not always successful. Here, a new DNA isolation approach for prokaryotic cells in formalin-fixed paraffin-embedded tissue...... samples is applied for the identification of an unknown Spirochaete. The tissue samples from colon were obtained from a previous study (1). Sectioning, in situ hybridization, LCM, DNA extraction, PCR reaction and sequencing were done according to Klitgaard et al. (2). The unknown invasive Spirochaetes...

5. Enhanced image capture through fusion

Burt, Peter J.; Hanna, Keith; Kolczynski, Raymond J.

1993-01-01

Image fusion may be used to combine images from different sensors, such as IR and visible cameras, to obtain a single composite with extended information content. Fusion may also be used to combine multiple images from a given sensor to form a composite image in which information of interest is enhanced. We present a general method for performing image fusion and show that this method is effective for diverse fusion applications. We suggest that fusion may provide a powerful tool for enhanced image capture with broad utility in image processing and computer vision.

6. Overriding auditory attentional capture.

Dalton, Polly; Lavie, Nilli

2007-02-01

Attentional capture by color singletons during shape search can be eliminated when the target is not a feature singleton (Bacon & Egeth, 1994). This suggests that a "singleton detection" search strategy must be adopted for attentional capture to occur. Here we find similar effects on auditory attentional capture. Irrelevant high-intensity singletons interfered with an auditory search task when the target itself was also a feature singleton. However, singleton interference was eliminated when the target was not a singleton (i.e., when nontargets were made heterogeneous, or when more than one target sound was presented). These results suggest that auditory attentional capture depends on the observer's attentional set, as does visual attentional capture. The suggestion that hearing might act as an early warning system that would always be tuned to unexpected unique stimuli must therefore be modified to accommodate these strategy-dependent capture effects. PMID:17557587

7. Neutron Capture Nucleosynthesis

Kiss, Miklos

2016-01-01

Heavy elements (beyond iron) are formed in neutron capture nucleosynthesis processes. We have proposed a simple unified model to investigate the neutron capture nucleosynthesis in arbitrary neutron density environment. We have also investigated what neutron density is required to reproduce the measured abundance of nuclei assuming equilibrium processes. We found both of these that the medium neutron density has a particularly important role at neutron capture nucleosynthesis. About these resu...

8. Overriding auditory attentional capture

Dalton, Polly; Lavie, Nilli

2007-01-01

Attentional capture by color singletons during shape search can be eliminated when the target is not a feature singleton (Bacon & Egeth, 1994). This suggests that a "singleton detection" search strategy must be adopted for attentional capture to occur. Here we find similar effects on auditory attentional capture. Irrelevant high-intensity singletons interfered with an auditory search task when the target itself was also a feature singleton. However, singleton interference was eliminated when ...

9. Neutron Capture Nucleosynthesis

Kiss, Miklos

2016-01-01

Heavy elements (beyond iron) are formed in neutron capture nucleosynthesis processes. We have proposed a simple unified model to investigate the neutron capture nucleosynthesis in arbitrary neutron density environment. We have also investigated what neutron density is required to reproduce the measured abundance of nuclei assuming equilibrium processes. We found both of these that the medium neutron density has a particularly important role at neutron capture nucleosynthesis. About these results most of the nuclei can formed at medium neutron capture density environment e.g. in some kind of AGB stars. Besides these observations our model is capable to use educational purpose.

10. Detection of clonal B cells in microdissected reactive lymphoproliferations: possible diagnostic pitfalls in PCR analysis of immunoglobulin heavy chain gene rearrangement

Zhou, X.G.; Sandvej, K.; Gregersen, Niels;

1999-01-01

Aims-To evaluate the specificity of standard and fluorescence based (GENESCAN) polymerase chain reaction (PCR) immunoglobulin heavy chain (IgH) gene rearrangement analysis in complete and microdissected paraffin wax embedded sections from lymphoid proliferations. Methods-PCR IgH gene rearrangement...... analysis of whole sections and microdissected fragments (n = 62) from paraffin wax embedded reactive lymph nodes (n = 6) and tonsils (n = 3). Amplificant analysis used both standard methods and automated high resolution fluorescence based quantification and size determination using GENESCAN software...... rearrangement analysis is a sensitive and specific method for demonstrating B cell clonality in whole paraffin wax embedded sections. However, oligoclonal and monoclonal rearrangement patterns are regularly encountered in small tissue fragments from otherwise unremarkable reactive lymphoproliferations, possibly...

11. Improved method for extraction and detection of Helicobacter pylori DNA in formalin-fixed paraffin embedded gastric biopsies using laser micro-dissection

María Fernanda Loayza; Fernando Xavier Villavicencio; Stephanie Carolina Santander; Manuel Baldeón; Lourdes Karina Ponce; Iván Salvador; Nicolás Vivar Díaz

2014-01-01

To assess the molecular events exerted by Helicobacter pylori interacting directly with gastric epithelial cells, an improved procedure for microbial DNA isolation from stained hematoxilin-eosin gastric biopsies was developed based on laser micro-dissection (LM) [1]. Few articles have described the use of LM to select and detect H. pylori genome from formalin-fixed paraffin embedded gastric tissue [2]. To improve the yield and quality of DNA isolated from H. pylori contacting intestinal epith...

12. Chromosomal rearrangements in cattle and pigs revealed by chromosome microdissection and chromosome painting

Yerle Martine; Ducos Alain; Pinton Alain

2003-01-01

Abstract A pericentric inversion of chromosome 4 in a boar, as well as a case of (2q-;5p+) translocation mosaicism in a bull were analysed by chromosome painting using probes generated by conventional microdissection. For the porcine inversion, probes specific for p arms and q arms were produced and hybridised simultaneously on metaphases of a heterozygote carrier. In the case of the bovine translocation, two whole chromosome probes (chromosome 5, and derived chromosome 5) were elaborated and...

13. A Technical Assessment of the Utility of Reverse Phase Protein Arrays for the Study of the Functional Proteome in Non-microdissected Human Breast Cancers.

Hennessy, Bryan T

2010-12-01

INTRODUCTION: The lack of large panels of validated antibodies, tissue handling variability, and intratumoral heterogeneity potentially hamper comprehensive study of the functional proteome in non-microdissected solid tumors. The purpose of this study was to address these concerns and to demonstrate clinical utility for the functional analysis of proteins in non-microdissected breast tumors using reverse phase protein arrays (RPPA). METHODS: Herein, 82 antibodies that recognize kinase and steroid signaling proteins and effectors were validated for RPPA. Intraslide and interslide coefficients of variability were <15%. Multiple sites in non-microdissected breast tumors were analyzed using RPPA after intervals of up to 24 h on the benchtop at room temperature following surgical resection. RESULTS: Twenty-one of 82 total and phosphoproteins demonstrated time-dependent instability at room temperature with most variability occurring at later time points between 6 and 24 h. However, the 82-protein functional proteomic "fingerprint" was robust in most tumors even when maintained at room temperature for 24 h before freezing. In repeat samples from each tumor, intratumoral protein levels were markedly less variable than intertumoral levels. Indeed, an independent analysis of prognostic biomarkers in tissue from multiple tumor sites accurately and reproducibly predicted patient outcomes. Significant correlations were observed between RPPA and immunohistochemistry. However, RPPA demonstrated a superior dynamic range. Classification of 128 breast cancers using RPPA identified six subgroups with markedly different patient outcomes that demonstrated a significant correlation with breast cancer subtypes identified by transcriptional profiling. CONCLUSION: Thus, the robustness of RPPA and stability of the functional proteomic "fingerprint" facilitate the study of the functional proteome in non-microdissected breast tumors.

14. Comparison of Whole Genome Amplification Methods for Analysis of DNA Extracted from Microdissected Early Breast Lesions in Formalin-Fixed Paraffin-Embedded Tissue

Nona Arneson; Juan Moreno; Vladimir Iakovlev; Arezou Ghazani; Keisha Warren; David McCready; Igor Jurisica; Done, Susan J.

2012-01-01

To understand cancer progression, it is desirable to study the earliest stages of its development, which are often microscopic lesions. Array comparative genomic hybridization (aCGH) is a valuable high-throughput molecular approach for discovering DNA copy number changes; however, it requires a relatively large amount of DNA, which is difficult to obtain from microdissected lesions. Whole genome amplification (WGA) methods were developed to increase DNA quantity; however their reproducibility...

15. Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ

Si Brask Sonne; Dalgaard, Marlene D; John Erik Nielsen; Hoei-Hansen, Christina E.; Ewa Rajpert-De Meyts; Lise Mette Gjerdrum; Henrik Leffers

2009-01-01

Udgivelsesdato: May 2009 Microarray and RT-PCR based methods are important tools for analysis of gene expression; however, in tissues containing many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser microdissection technology allows for enrichment of specific cell types. However, when the cells are not morphologically distinguishable, it is necessary to use a specific staining...

16. Carbon Capture and Storage

Benson, S.M.; Bennaceur, K.; Cook, P.; Davison, J.; Coninck, H. de; Farhat, K.; Ramirez, C.A.; Simbeck, D.; Surles, T.; Verma, P.; Wright, I.

2012-01-01

Emissions of carbon dioxide, the most important long-lived anthropogenic greenhouse gas, can be reduced by Carbon Capture and Storage (CCS). CCS involves the integration of four elements: CO 2 capture, compression of the CO2 from a gas to a liquid or a denser gas, transportation of pressurized CO 2

17. CAPTURED India Country Evaluation

O'Donoghue, R.; Brouwers, J.H.A.M.

2012-01-01

This report provides the findings of the India Country Evaluation and is produced as part of the overall CAPTURED End Evaluation. After five years of support by the CAPTURED project the End Evaluation has assessed that results are commendable. I-AIM was able to design an approach in which health fol

18. Direct thermal neutron capture

We discuss the direct-capture theory pertaining to primary electric dipole (E1) transitions following slow-neutron capture. For light nuclides that we have studied (including 9Be, 12C, 13C, 24Mg, 25Mg, 26Mg, 32S, 33S, 34S, 40Ca, and 44Ca), estimates of direct-capture cross sections using optical-model potentials with physically realistic parameters, are in reasonable agreement with the data. Minor disagreements that exist are consistent with extrapolations to light nuclides of generally accepted formulations of compound-nucleus capture. We also discuss the channel-capture approximation which is, in general, a good representation of these cross sections in heavier nuclei particularly if the scattering lengths are not different from the corresponding potential radii. We also draw attention to cases where the use of this formula leads to inaccurate predictions. 9 refs., 1 fig., 2 tab

19. Muon capture in hydrogen

In this lecture I will mainly speak about the recent results obtained by the Saclay-CERN-Bologna (SCB) collaboration on the muon capture rate at rest in liquid hydrogen.In the first talk I shall first briefly remind the theoretical description of the capture process, then describe the experimental difficulties and procedures and finally present the results obtained. In the second talk I shall compare them to other capture experiments on the proton and present the informations that are provided by muon capture in hydrogen, especially for the PCAC hypothesis, the second class currents and the μ-e universality. Finally I shall present the possible muon capture experiments on the proton that could still be undertaken in order to improve our present knowledge of the subject

20. Virtual microdissection identifies distinct tumor- and stroma-specific subtypes of pancreatic ductal adenocarcinoma

Moffitt, Richard A.; Marayati, Raoud; Flate, Elizabeth L.; Volmar, Keith E.; Loeza, S. Gabriela Herrera; Hoadley, Katherine A.; Rashid, Naim U.; Williams, Lindsay A.; Eaton, Samuel C.; Chung, Alexander H.; Smyla, Jadwiga K.; Anderson, Judy M.; Kim, Hong Jin; Bentrem, David J.; Talamonti, Mark S.; Iacobuzio-Donahue, Christine A.; Hollingsworth, Michael A.; Yeh, Jen Jen

2016-01-01

Pancreatic ductal adenocarcinoma (PDAC) remains a lethal disease with a 5-year survival of 4%. A key hallmark of PDAC is extensive stromal involvement, which makes capturing precise tumor-specific molecular information difficult. Here, we have overcome this problem by applying blind source separation to a diverse collection of PDAC gene expression microarray data, which includes primary, metastatic, and normal samples. By digitally separating tumor, stroma, and normal gene expression, we have identified and validated two tumor-specific subtypes including a “basal-like” subtype which has worse outcome, and is molecularly similar to basal tumors in bladder and breast cancer. Furthermore, we define “normal” and “activated” stromal subtypes which are independently prognostic. Our results provide new insight into the molecular composition of PDAC which may be used to tailor therapies or provide decision support in a clinical setting where the choice and timing of therapies is critical. PMID:26343385

1. Environment assisted electron capture

Electrons scattering off isolated atoms or positive atomic ions may be captured in the non-resonant photorecombination or resonant dielectronic recombination processes with the excess energy carried off by a photon. We demonstrate that in a medium a new interatomic process becomes important, where in the electron capture by an atom or atomic ion the energy is transferred to a neigbouring species ionising it. We discuss different manifestations of this interatomic Coulombic electron capture and estimate its efficiency for a number of realistic systems.

2. Tissue-specific laser microdissection of the Brassica napus funiculus improves gene discovery and spatial identification of biological processes

Chan, Ainsley C.; Khan, Deirdre; Girard, Ian J.; Becker, Michael G.; Millar, Jenna L.; Sytnik, David; Belmonte, Mark F.

2016-01-01

The three primary tissue systems of the funiculus each undergo unique developmental programs to support the growth and development of the filial seed. To understand the underlying transcriptional mechanisms that orchestrate development of the funiculus at the globular embryonic stage of seed development, we used laser microdissection coupled with RNA-sequencing to produce a high-resolution dataset of the mRNAs present in the epidermis, cortex, and vasculature of the Brassica napus (canola) funiculus. We identified 7761 additional genes in these tissues compared with the whole funiculus organ alone using this technology. Differential expression and enrichment analyses were used to identify several biological processes associated with each tissue system. Our data show that cell wall modification and lipid metabolism are prominent in the epidermis, cell growth and modification occur in the cortex, and vascular tissue proliferation and differentiation occur in the central vascular strand. We provide further evidence that each of the three tissue systems of the globular stage funiculus are involved in specific biological processes that all co-ordinate to support seed development. The identification of genes and gene regulators responsible for tissue-specific developmental processes of the canola funiculus now serves as a valuable resource for seed improvement research. PMID:27194740

3. Towards the finer mapping of facioscapulohumeral muscular dystrophy at 4q35: Construction of a laser microdissection library

Upadhyaya, M.; Osborn, M.; Maynard, J. [Institute of Medical Genetics, Wales (United Kingdom)] [and others

1995-06-19

Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal-dominant disorder which has been mapped to the 4q35 region. In order to saturate this distal 4q region with DNA markers, a laser-based chromosomal microdissection and microcloning procedure was used to construct a genomic library from the distal 20% of chromosome 4, derived from a single human metaphase spread. Of the 100 microclones analyzed from this library, 94 clones contained inserts sized from 80-800 bp, with an average size of 340 bp. Less than 20% of these clones hybridized to human repeat sequences. Seventy-two single-copy clones were further characterized by Southern blot hybridization against a DNA panel of somatic cell hybrids, containing various regions of chromosome 4. Forty-two clones mapped to chromosome 4, of which 8 clones mapped into the relevant 4q35 region. Twenty of these chromosome 4-specific clones were screened against {open_quotes}zoo-blots{close_quotes}; 11 clones, of which 3 mapped to 4q35, identified conserved sequences. This is the first report to describe the isolation of potential expressed sequences derived from the FSHD region. These chromosome region-specific microclones will be useful in the construction of the physical map of the region, the positional cloning of potential disease-associated genes, and the identification of additional polymorphic markers from within the distal 4q region. 47 refs., 6 figs., 1 tab.

4. Isolation of expressed sequences encoded by the human Xq terminal portion using microclone probes generated by laser microdissection

Yokoi, H.; Kang, X. (Univ. of Ottawa Faculty of Medicine, Ontario (Canada)); Hadano, Shinji; Kogi, Mieko; Wakasa, Kyo (Tokai Univ. School of Medicine, Kanagawa (Japan)); Ikeda, J.E. (Univ. of Ottawa Faculty of Medicine, Ontario (Canada) Tokai Univ. School of Medicine, Kanagawa (Japan) National Institute of Agrobiological Resources, Ibaraki (Japan))

1994-04-01

The genes that cause a variety of neurologic and neuromuscular disorders have been mapped to the distal region of Xq. In an effort to isolate genes from this area, a regional genomic library of the distal 30% of Xq was constructed from a single metaphase spread by means of laser microdissection and single unique primer-polymerase chain reaction. Using pooled probes of 1000 clones from the genomic library, human brain cDNA libraries were screened for expressed sequences encoded by this region. From the 250,000 cDNA clones screened so far, 10 nonoverlapping sequences that mapped back to the target portion were isolated. The complete nucleotide sequences of these cDNA clones have been determined. Analysis of the sequences indicates that none has significant similarity to previously characterized primate genes. One sequence mapping to Xq27.3-qter contained an open reading frame of 281 amino acids and was expressed in every tissue tested. This gene, as well as others isolated in this manner, may prove to be a candidate gene for heritable disorders mapping to this region. 36 refs., 6 figs., 2 tabs.

5. Neutrinoless double electron capture

Kotila, J; Iachello, F

2015-01-01

Direct determination of the neutrino mass is at the present time one of the most important aims of experimental and theoretical research in nuclear and particle physics. A possible way of detection is through neutrinoless double electron capture, $0\ 6. Activists versus Captured Regulators Daubanes, Julien; Rochet, Jean-Charles 2013-01-01 We analyze the consequences of activism in a regulated industry where the regulator has been captured by the industry. Unlike ordinary economic agents, activists are insensitive to monetary incentives. Moreover, they are less well informed than regulators and their actions generate dead-weight costs. Yet we find that activism may increase social welfare because it disciplines captured regulators and reduces the social cost of imperfect regulatory systems. 7. Muon capture by tritium The muon capture rate is computed with realistic wave function for the initial tritium nuclei (Faddeev equations on configuration space with realistic potentials), and plane wave approximation for the final three neutrons, with the effective Hamiltonian of Fujii and Primakoff for muon capture and via a non energy weighted sum rule. Such a forbidden transition is hoped to be a probe for exchange current contributions 8. Muon capture at PSI Winter, Peter 2010-01-01 Measuring the rate of muon capture in hydrogen provides one of the most direct ways to study the axial current of the nucleon. The MuCap experiment uses a negative muon beam stopped in a time projection chamber operated with ultra-pure hydrogen gas. Surrounded by a decay electron detector, the lifetime of muons in hydrogen can be measured to determine the singlet capture rate Lambda_s to a final precision of 1%. The capture rate determines the nucleon's pseudoscalar form factor g_p. A first result, g_p = 7.3 +- 1.1, has been published and the final analysis of the full statistics will reduce the error by a factor of up to 3. Muon capture on the deuteron probes the weak axial current in the two-nucleon system. Within the framework of effective field theories the calculation of such two-nucleon processes involving the axial current requires the knowledge of one additional low energy constant which can be extracted from the doublet capture rate Lambda_d. The same constant then allows to model-independently calcu... 9. Laser microdissection and genetic manipulation technologies to probe lignin heterogeneity and configuration in plant cell walls. Corea, Oliver R A; Ki, Chanyoung; Cardenas, Claudia L; Davin, Laurence B; Lewis, Norman G 2012-01-01 Single and multiple T-DNA knockouts of genes encoding arogenate dehydratases (ADTs) in Arabidopsis were obtained in homozygous form. These were analyzed for potential differences in lignin contents and compositions, as well as for distinct phenotypes over growth and development. Of these different lines, distinct reductions in lignin contents were obtained, with those having different G:S ratios depending upon the combination of ADT genes being knocked out. Results from pyrolysis GC/MS analyses indicated that differential carbon flux occurred into the vascular bundles (vb) and interfascicular fibers (if). These results provide additional new insight into factors controlling lignin heterogeneity and configuration. PMID:22843403 10. Muon Capture in Deuterium Ricci, P; Mosconi, B; Smejkal, J 2009-01-01 Model dependence of the capture rates of the negative muon capture in deuterium is studied starting from potential models and the weak two-body meson exchange currents constructed in the tree approximation and also from an effective field theory. The tree one-boson exchange currents are derived from the hard pion chiral Lagrangians of the$N \\Delta \\pi \\rho \\omega a_1$system. If constructed in conjunction with the one-boson exchange potentials, the capture rates can be calculated consistently. On the other hand, the effective field theory currents, constructed within the heavy baryon chiral perturbation theory, contain a low energy constant$\\hat d ^R$that cannot be extracted from data at the one-particle level nor determined from the first principles. Comparative analysis of the results for the doublet transition rate allows us to extract the constant$\\hat d ^R$. 11. The application of laser microdissection to in planta gene expression profiling of the maize anthracnose stalk rot fungus Colletotrichum graminicola. Tang, Weihua; Coughlan, Sean; Crane, Edmund; Beatty, Mary; Duvick, Jon 2006-11-01 Laser microdissection (LM) offers a potential means for deep sampling of a fungal plant-pathogen transcriptome during the infection process using whole-genome DNA microarrays. The use of a fluorescent protein-expressing fungus can greatly facilitate the identification of fungal structures for LM sampling. However, fixation methods that preserve both tissue histology and protein fluorescence, and that also yield RNA of suitable quality for microarray applications, have not been reported. We developed a microwave-accelerated acetone fixation, paraffin-embedding method that fulfills these requirements and used it to prepare mature maize stalk tissues infected with an Anemonia majano cyan fluorescent protein-expressing isolate of the anthracnose stalk rot fungus Colletotrichum graminicola. We successfully used LM to isolate individual maize cells associated with C. graminicola hyphae at an early stage of infection. The LM-derived RNA, after two-round linear amplification, was of sufficient quality and quantity for global expression profiling using a fungal microarray. Comparing replicated LM samples representing an early stage of stalk cell infection with samples from in vitro-germinated conidia, we identified 437 and 370 C. graminicola genes showing significant up- or downregulation, respectively. We confirmed the differential expression of several representative transcripts by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) and documented extensive overlap of this dataset with a PCR-subtraction library enriched for C. graminicola transcripts in planta. Our results demonstrate that LM is feasible for in planta pathogen expression profiling and can reveal clues about fungal genes involved in pathogenesis. The method in this report may be advantageous for visualizing a variety of cellular features that depend on a high degree of histochemical preservation and RNA integrity prior to LM. PMID:17073306 12. Outcomes of Micro-Dissection TESE in Patients with Non-Mosaic Klinefelter’s Syndrome without Hormonal Treatment Hakan Ozveri 2015-02-01 Full Text Available Background: Klinefelter syndrome (KS is the most common sex chromosomal disorder in males and historically patients have been labeled as sterile. After the introduction of microdissection testicular sperm extraction (micro-TESE, successful sperm retrievals for intracytoplasmic sperm injection (ICSI have been reported. Materials and Methods: A retrospective study was undertaken on ten patients with non-mosaic KS undergoing micro-TESE for ICSI. The testicular volume and FSH and LH levels of each patient were measured. Karyotypes were confirmed by analyzing peripheral lymphocyte metaphases. Physical examination of the external genitalia was performed in all patients to rule out any co-existing anomaly. Micro-TESE was performed in order to investigate the presence of seminiferous tubules which may contain spermatozoa. When testicular spermatozoa were found in micro-TESE, ICSI was performed. Embryos were evaluated for further development. Fertilization was considered to have occurred after the visualization of the two pro-nuclei stage of the oocyte 24 hours after the intracytoplasmic injection of the motile spermatozoa. Pregnancy was confirmed by visualization of an intrauterine gestational sac under ultrasonographic examination. Results: Testicular biopsy revealed motile spermatozoa in 6 of 9 patients (66.6 %. Fertilization rate per embryo-transfer was 40%. One patient was able to conceive and fathered a healthy boy weights 3410 g at the 39th week of gestation. Conclusion: Retrieval of testicular spermatozoa by micro-TESE is possible for azoospermic men with KS when assisted reproductive techniques are applied. For patients with KS who want to conceive, assisted reproductive techniques (ART should be recommended. 13. US Spacesuit Knowledge Capture Chullen, Cinda; Thomas, Ken; McMann, Joe; Dolan, Kristi; Bitterly, Rose; Lewis, Cathleen 2011-01-01 The ability to learn from both the mistakes and successes of the past is vital to assuring success in the future. Due to the close physical interaction between spacesuit systems and human beings as users, spacesuit technology and usage lends itself rather uniquely to the benefits realized from the skillful organization of historical information; its dissemination; the collection and identification of artifacts; and the education of those in the field. The National Aeronautics and Space Administration (NASA), other organizations and individuals have been performing United States (U.S.) Spacesuit Knowledge Capture since the beginning of space exploration. Avenues used to capture the knowledge have included publication of reports; conference presentations; specialized seminars; and classes usually given by veterans in the field. More recently the effort has been more concentrated and formalized whereby a new avenue of spacesuit knowledge capture has been added to the archives in which videotaping occurs engaging both current and retired specialists in the field presenting technical scope specifically for education and preservation of knowledge. With video archiving, all these avenues of learning can now be brought to life with the real experts presenting their wealth of knowledge on screen for future learners to enjoy. Scope and topics of U.S. spacesuit knowledge capture have included lessons learned in spacesuit technology, experience from the Gemini, Apollo, Skylab and Shuttle programs, hardware certification, design, development and other program components, spacesuit evolution and experience, failure analysis and resolution, and aspects of program management. Concurrently, U.S. spacesuit knowledge capture activities have progressed to a level where NASA, the National Air and Space Museum (NASM), Hamilton Sundstrand (HS) and the spacesuit community are now working together to provide a comprehensive closed-looped spacesuit knowledge capture system which includes 14. Marker-Free Human Motion Capture Grest, Daniel Human Motion Capture is a widely used technique to obtain motion data for animation of virtual characters. Commercial optical motion capture systems are marker-based. This book is about marker-free motion capture and its possibilities to acquire motion from a single viewing direction. The focus of...... this book is on the optimization framework, which can be applied to every pose estimation problem of articulated objects. The motion function is formed with a combination of kinematic chains. This formulation leads to a Nonlinear Optimization problem and is solved with gradient-based methods, which are...... compared with respect to their efficiency. A new contribution is the inclusion of second order motion derivatives within the pose estimation. The pose estimation step requires correspondences between known model of the person and observed data. Computer Vision techniques are used to combine multiple types... 15. Using laser micro-dissection and qRT-PCR to analyze cell type-specific gene expression in Norway spruce phloem Nina E. Nagy 2014-04-01 Full Text Available The tangentially oriented polyphenolic parenchyma (PP and radially organized ray parenchyma in the phloem are central in the defense of conifer stems against insects and pathogens. Laser micro-dissection enables examination of cell-specific defense responses. To examine induced defense responses in Norway spruce stems inoculated with the necrotrophic blue-stain fungus Ceratocystis polonica, RNA extracted from laser micro-dissected phloem parenchyma and vascular cambium was analyzed using real-time RT-PCR (qRT-PCR to profile transcript levels of selected resistance marker genes. The monitored transcripts included three pathogenesis-related proteins (class IV chitinase (CHI4, defensin (SPI1, peroxidase (PX3, two terpene synthesis related proteins (DXPS and LAS, one ethylene biosynthesis related protein (ACS, and a phenylalanine ammonia-lyase (PAL. Three days following inoculation, four genes (CHI4, PAL, PX3, SPI1 were differentially induced in individual cell and tissue types, both close to the inoculation site (5 mm above and, to a lesser degree, further away (10 mm above. These resistance marker genes were all highly induced in ray parenchyma, supporting the important role of the rays in spruce defense propagation. CHI4 and PAL were also induced in PP cells and in conducting secondary phloem tissues. Our data suggests that different cell types in the secondary phloem of Norway spruce have overlapping but not fully redundant roles in active host defense. Furthermore, the study demonstrates the usefulness of laser micro-dissection coupled with qRT-PCR to characterize gene expression in different cell types of conifer bark. 16. U-238 neutron-capture gamma-cascade generation and transport simulation for capture-tank response (final report). Doctoral thesis Rosener, T.J. 1992-05-07 A computer analysis has been performed to evaluate the energy dependent response of a capture tank to the gamma rays emitted in the cascades of the excited U-239 nucleus. This model determines the energies of the gamma-ray cascades, the order of emission of the gamma rays in a cascade, and the gamma-ray multiplicity of the cascades using Monte Carlo techniques. A capture tank responds to the combined effect of the various gamma rays emitted in the cascade. Examined is the energy deposition in a capture tank by the cascades generated in resonant (surface) capture and off-resonant (volumetric) capture, with and without internal conversion. Off-resonant capture deposits, on the average, less energy than resonant capture, due to self-shielding of the gamma-rays in the capture sample. Internal conversion has negligible effect on the average cascade energy deposited in the capture tank. Gamma-ray cascade, Capture tank, U-238 neutron capture. 17. CAPTURED End Evaluation Synthesis Report Brouwers, J.H.A.M. 2012-01-01 This report provides the findings of the Synthesis Study of the CAPTURED Evaluation and is produced as part of the overall CAPTURED End Evaluation. After five years of support by the CAPTURED project the three CAPTURED partners have achieved commendable results. Ten lessons learned are formulated th 18. Theory of electron capture The historical development of a theoretical model for the electron-capture detector, sometimes called the 'classical' model, is presented and the model as it stands today is described. This is followed by a comparison of the model with experimental data obtained with other techniques since the early development of the theory and finally a section on correlations and predictions is presented. (Auth.) 19. Membrane-Cryogenic Post-Combustion Carbon Capture of Flue Gases from NGCC Scholes, Colin A.; Minh T. Ho; Dianne E. Wiley 2016-01-01 Membrane gas separation for carbon capture has traditionally been focused on high pressure applications, such as pre-combustion capture and natural gas sweetening. Recently a membrane-cryogenic combined process has been shown to be cost competitive for post-combustion capture from coal fired power stations. Here, the membrane-cryogenic combined process is investigated for application to post-combustion carbon capture from the flue gas of a Natural Gas Combined Cycle (NGCC) process. This proce... 20. Thermal neutron capture cross sections of tellurium isotopes New values for thermal neutron capture cross sections of the tellurium isotopes 122Te, 124Te, 125Te, 126Te, 128Te, and 130Te are reported. These values are based on a combination of newly determined partial g-ray cross sections obtained from experiments on targets contained natural Te and gamma intensities per capture of individual Te isotopes. Isomeric ratios for the thermal neutron capture on the even tellurium isotopes are also given 1. Thermal neutron capture cross sections of tellurium isotopes New values for thermal neutron capture cross sections of the tellurium isotopes 122Te,124Te,125Te,126Te,128Te, and 130Te are reported. These values are based on a combination of newly determined partial γ-ray cross sections obtained from experiments on targets contained natural Te and γ intensities per capture of individual Te isotopes. Isomeric ratios for the thermal neutron capture on the even tellurium isotopes are also given 2. Thermal neutron capture cross sections of tellurium isotopes Tomandl, I.; Honzatko, J.; von Egidy, T.; Wirth, H.-F.; Belgya, T.; Lakatos, M.; Szentmiklosi, L.; Revay, Zs.; Molnar, G.L.; Firestone, R.B.; Bondarenko, V. 2004-03-01 New values for thermal neutron capture cross sections of the tellurium isotopes 122Te, 124Te, 125Te, 126Te, 128Te, and 130Te are reported. These values are based on a combination of newly determined partial g-ray cross sections obtained from experiments on targets contained natural Te and gamma intensities per capture of individual Te isotopes. Isomeric ratios for the thermal neutron capture on the even tellurium isotopes are also given. 3. Gadolinium neutron capture therapy Gadolinium neutron capture therapy makes use of photons and electrons produced by nuclear reactions between gadolinium and lower-energy neutrons which occur within the tumor. The results of our studies have shown that its radiation effect is mostly of low LET and that the electrons are the significant component in the over-all dose. The dose from gadolinium neutron capture reactions does not seem to increase in proportion to the gadolinium concentration, and the Gd-157 concentration of about 100 μg/ml appears most optimal for therapy. Close contact between gadolinium and the cell is not necessarily required for cell inactivation, however, the effect of electrons released from intracellular gadolinium may be significant. Experimental studies on tumor-bearing mice and rabbits have shown that this is a very promising modality though further improvements in gadolinium delivery to tumors are needed. (author) 4. Supernova electron capture rates Martínez-Pinedo, G 1999-01-01 We have calculated the Gamow-Teller strength distributions for the ground states and low lying states of several nuclei that play an important role in the precollapse evolution of supernova. The calculations reproduce the experimental GT distributions nicely. The GT distribution are used to calculate electron capture rates for typical presupernova conditions. The computed rates are noticeably smaller than the presently adopted rates. The possible implications for the supernova evolution are discussed. 5. Inland capture fisheries Welcomme, Robin L.; Cowx, Ian G.; Coates, David; Béné, Christophe; Funge-Smith, Simon; Halls, Ashley; Lorenzen, Kai 2010-01-01 The reported annual yield from inland capture fisheries in 2008 was over 10 million tonnes, although real catches are probably considerably higher than this. Inland fisheries are extremely complex, and in many cases poorly understood. The numerous water bodies and small rivers are inhabited by a wide range of species and several types of fisher community with diversified livelihood strategies for whom inland fisheries are extremely important. Many drivers affect the fisheries, including inter... 6. Novel markers for differentiation of lobular and ductal invasive breast carcinomas by laser microdissection and microarray analysis Invasive ductal and lobular carcinomas (IDC and ILC) are the most common histological types of breast cancer. Clinical follow-up data and metastatic patterns suggest that the development and progression of these tumors are different. The aim of our study was to identify gene expression profiles of IDC and ILC in relation to normal breast epithelial cells. We examined 30 samples (normal ductal and lobular cells from 10 patients, IDC cells from 5 patients, ILC cells from 5 patients) microdissected from cryosections of ten mastectomy specimens from postmenopausal patients. Fifty nanograms of total RNA were amplified and labeled by PCR and in vitro transcription. Samples were analysed upon Affymetrix U133 Plus 2.0 Arrays. The expression of seven differentially expressed genes (CDH1, EMP1, DDR1, DVL1, KRT5, KRT6, KRT17) was verified by immunohistochemistry on tissue microarrays. Expression of ASPN mRNA was validated by in situ hybridization on frozen sections, and CTHRC1, ASPN and COL3A1 were tested by PCR. Using GCOS pairwise comparison algorithm and rank products we have identified 84 named genes common to ILC versus normal cell types, 74 named genes common to IDC versus normal cell types, 78 named genes differentially expressed between normal ductal and lobular cells, and 28 named genes between IDC and ILC. Genes distinguishing between IDC and ILC are involved in epithelial-mesenchymal transition, TGF-beta and Wnt signaling. These changes were present in both tumor types but appeared to be more prominent in ILC. Immunohistochemistry for several novel markers (EMP1, DVL1, DDR1) distinguished large sets of IDC from ILC. IDC and ILC can be differentiated both at the gene and protein levels. In this study we report two candidate genes, asporin (ASPN) and collagen triple helix repeat containing 1 (CTHRC1) which might be significant in breast carcinogenesis. Besides E-cadherin, the proteins validated on tissue microarrays (EMP1, DVL1, DDR1) may represent novel 7. Proliferating cell nuclear antigen in gonad and associated storage tissue of the Pacific oyster Crassostrea gigas: seasonal immunodetection and expression in laser microdissected tissues. Franco, Alban; Jouaux, Aude; Mathieu, Michel; Sourdaine, Pascal; Lelong, Christophe; Kellner, Kristell; Heude Berthelin, Clothilde 2010-04-01 To understand the processes involved in tissue remodeling associated with the seasonal reproductive cycle of the oyster Crassostrea gigas, we used immunodetection and expression measurements of proliferating cell nuclear antigen (PCNA). The expression of the PCNA gene was measured by real-time polymerase chain reaction in the whole gonadal area compared with laser microdissected gonad and storage tissue. Results underlined the advantage of the laser microdissection approach to detect expression, mainly for early stages of spermatogenesis. In the storage tissue, PCNA expression was reduced in the gonadal tubules, but immunolabeled hemocytes and vesicular cells were detected when the storage tissue was being restored. In the gonadal tubules, the PCNA gene was more highly expressed in males than in females. As soon as spermatogenesis was initiated, PCNA expression showed a high and constant level. In females, the expression level increased gradually until the ripe stage. The immunological approach established the involvement of peritubular cells in gonadal tubule expansion during early gametogenesis. In both sexes, gonial mitosis was immunodetected throughout the reproductive cycle. In males, the occurrence of two types of spermatogonia was ascertained by differential immunolabeling, and intragonadal somatic cell proliferation was noted. As expected, immunolabeling was never observed from stage II spermatocytes to spermatozoa. In females, positively stained cells were detected from oogonia to growing oocytes with various labeled intracellular locations. PMID:20151153 8. Redistribution of ionotropic glutamate receptors detected by laser microdissection of the rat dentate gyrus 48 h following LTP induction in vivo. Jeremy T T Kennard Full Text Available The persistence and input specificity of long-term potentiation (LTP make it attractive as a mechanism of information storage. In its initial phase, both in vivo and in vitro studies have shown that LTP is associated with increased membrane localization of AMPA receptor subunits, but the molecular basis of LTP maintenance over the long-term is still unclear. We have previously shown that expression of AMPA and NMDA receptor subunits is elevated in whole homogenates prepared from dentate gyrus 48 h after LTP induction in vivo. In the present study, we utilized laser microdissection (LMD techniques to determine whether AMPA and NMDA receptor upregulation occurs specifically in the stimulated regions of the dentate gyrus dendritic arbor. Receptor proteins GluN1, GluA1 and GluA2, as well as postsynaptic density protein of 95 kDa and tubulin were detected by Western blot analysis in microdissected samples. Gradients of expression were observed for GluN1 and GluA2, decreasing from the inner to the outer zones of the molecular layer, and were independent of LTP. When induced at medial perforant path synapses, LTP was associated with an apparent specific redistribution of GluA1 and GluN1 to the middle molecular layer that contains these synapses. These data indicate that glutamate receptor proteins are delivered specifically to dendritic regions possessing LTP-expressing synapses, and that these changes are preserved for at least 48 h. 9. PRELIMINARY ENVIRONMENTAL, HEALTH AND SAFETY RISK ASSESSMENT ON THE INTEGRATION OF A PROCESS UTILIZING LOW-ENERGY SOLVENTS FOR CARBON DIOXIDE CAPTURE ENABLED BY A COMBINATION OF ENZYMES AND VACUUM REGENERATION WITH A SUBCRITICAL PC POWER PLANT Fitzgerald, David; Vidal, Rafael; Russell, Tania; Babcock, Doosan; Freeman, Charles; Bearden, Mark; Whyatt, Greg; Liu, Kun; Frimpong, Reynolds; Lu, Kunlei; Salmon, Sonja; House, Alan; Yarborough, Erin 2014-12-31 The results of the preliminary environmental, health and safety (EH&S) risk assessment for an enzyme-activated potassium carbonate (K2CO3) solution post-combustion CO2 capture (PCC) plant, integrated with a subcritical pulverized coal (PC) power plant, are presented. The expected emissions during normal steady-state operation have been estimated utilizing models of the PCC plant developed in AspenTech’s AspenPlus® software, bench scale test results from the University of Kentucky, and industrial experience of emission results from a slipstream PCC plant utilizing amine based solvents. A review of all potential emission species and their sources was undertaken that identified two credible emission sources, the absorber off-gas that is vented to atmosphere via a stack and the waste removed from the PCC plant in the centrifuge used to reclaim enzyme and solvent. The conditions and compositions of the emissions were calculated and the potential EH&S effects were considered as well as legislative compliance requirements. Potential mitigation methods for emissions during normal operation have been proposed and solutions to mitigate uncontrolled releases of species have been considered. The potential emissions were found to pose no significant EH&S concerns and were compliant with the Federal legislation reviewed. The limitations in predicting full scale plant performance from bench scale tests have been noted and further work on a larger scale test unit is recommended to reduce the level of uncertainty. 10. Rapid and sensitive detection of fipronil and its metabolites in edible oils by solid-phase extraction based on humic acid bonded silica combined with gas chromatography with electron capture detection. Peng, Xi-Tian; Li, Yu-Nan; Xia, Hong; Peng, Li-Jun; Feng, Yu-Qi 2016-06-01 Solid-phase extraction based on humic acid bonded silica followed by gas chromatography with electron capture detection was developed to determine fipronil and its metabolites in edible oil. To achieve the best extraction performance, we systematically investigated a series of solid-phase extraction parameters. Under the optimized conditions, the method was validated according to linearity, recovery, and precision. Good linearities were obtained with R(2) more than 0.9996 for all analytes. The limits of detection were between 0.3 and 0.5 ng/g, and the recoveries ranged from 83.1 to 104.0% at three spiked concentrations with intra- and interday relative standard deviation values less than 8.7%. Finally, the proposed method was applied to determine fipronil and its metabolites in 11 edible oil samples taken from Wuhan markets. Fipronil was detectable in four samples with concentrations ranging from 3.0 to 5.2 ng/g. In China, the maximum residue limits of fipronil in some vegetables and maize are 20 and 100 ng/g (GB/T 2763-2014), respectively. The residues of fipronil and its metabolites in commercial edible oils might exhibit some potential threat to human health as a result of high consumption of edible oil as part of daily intake. PMID:27280701 11. Lunar Sulfur Capture System Project National Aeronautics and Space Administration — The Lunar Sulfur Capture System (LSCS) is an innovative method to capture greater than 90 percent of sulfur gases evolved during thermal treatment of lunar soils.... 12. Capturing the Daylight Dividend Peter Boyce; Claudia Hunter; Owen Howlett 2006-04-30 Capturing the Daylight Dividend conducted activities to build market demand for daylight as a means of improving indoor environmental quality, overcoming technological barriers to effective daylighting, and informing and assisting state and regional market transformation and resource acquisition program implementation efforts. The program clarified the benefits of daylight by examining whole building systems energy interactions between windows, lighting, heating, and air conditioning in daylit buildings, and daylighting's effect on the human circadian system and productivity. The project undertook work to advance photosensors, dimming systems, and ballasts, and provided technical training in specifying and operating daylighting controls in buildings. Future daylighting work is recommended in metric development, technology development, testing, training, education, and outreach. 13. Robust automated knowledge capture. Stevens-Adams, Susan Marie; Abbott, Robert G.; Forsythe, James Chris; Trumbo, Michael Christopher Stefan; Haass, Michael Joseph; Hendrickson, Stacey M. Langfitt 2011-10-01 This report summarizes research conducted through the Sandia National Laboratories Robust Automated Knowledge Capture Laboratory Directed Research and Development project. The objective of this project was to advance scientific understanding of the influence of individual cognitive attributes on decision making. The project has developed a quantitative model known as RumRunner that has proven effective in predicting the propensity of an individual to shift strategies on the basis of task and experience related parameters. Three separate studies are described which have validated the basic RumRunner model. This work provides a basis for better understanding human decision making in high consequent national security applications, and in particular, the individual characteristics that underlie adaptive thinking. 14. Particle capture device Jayne, John T.; Worsnop, Douglas R. 2016-02-23 In example embodiments, particle collection efficiency in aerosol analyzers and other particle measuring instruments is improved by a particle capture device that employs multiple collisions to decrease momentum of particles until the particles are collected (e.g., vaporized or come to rest). The particle collection device includes an aperture through which a focused particle beam enters. A collection enclosure is coupled to the aperture and has one or more internal surfaces against which particles of the focused beam collide. One or more features are employed in the collection enclosure to promote particles to collide multiple times within the enclosure, and thereby be vaporized or come to rest, rather than escape through the aperture. 15. Enhancing stock returns using hedged dividend capture Terry L. Zivney; John H. Ledbetter; James P. Hoban Jr 2006-01-01 Purpose – This paper aims to explore the potential use of a dividend capture strategy by individual investors. This strategy arises from the 2003 tax law changes which lowered tax rates on dividends received, while leaving the short-term tax rates on capital losses unchanged. In addition, leverage can be used in combination with an aggressive call-writing strategy to receive a multiple of the tax-advantaged dividend yield without a corresponding increase in risk. Design/methodology/approach -... 16. STR profiling of epithelial cells identified by X/Y-FISH labelling and laser microdissection using standard and elevated PCR conditions. Lynch, Laura; Gamblin, Amelia; Vintiner, Sue; Simons, Joanne L 2015-05-01 During the investigation of allegations of sexual assault, samples are frequently encountered that contain DNA from a female and a male donor. These may represent contributions of DNA from the complainant and potentially, the offender. Many semen stained samples successfully undergo DNA analysis and interpretation using a differential extraction method that separates sperm from the epithelial cells present in the stain. However, for those mixed cell samples that contain only epithelial cells, separation of any male cells from female cells is problematic. This paper describes the application of fluorescent in situ hybridisation (FISH) for the gender identification of epithelial cells and subsequent recovery of target cells using laser microdissection (LMD). The profiling results obtained from samples of known cell numbers using the Identifiler™ multiplex at standard 28-cycle PCR conditions and, when cell numbers are low, the SGM Plus™ multiplex at elevated 34-cycle PCR conditions (also known as Low Copy Number DNA analysis (LCN)) are described. PMID:25555139 17. Use of laser microdissection for the construction of Humulus japonicus Siebold et Zuccarini, 1846 (Cannabaceae sex chromosome-specific DNA library and cytogenetics analysis Nickolay Yakovin 2014-12-01 Full Text Available Dioecy is relatively rare among plant species, and distinguishable sex chromosomes have been reported in few dioecious species. The multiple sex chromosome system (XX/XY1Y2 of Humulus japonicus Siebold et Zuccarini, 1846 differs from that of other members of the family Cannabaceae, in which the XX/XY chromosome system is present. Sex chromosomes of H. japonicus were isolated from meiotic chromosome spreads of males by laser microdissection with the P.A.L.M. MicroLaser system. The chromosomal DNA was directly amplified by degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR. Fast fluorescence in situ hybridization (FAST-FISH using a labeled, chromosome-specific DOP-PCR product as a probe showed preferential hybridization to sex chromosomes. In addition, the DOP-PCR product was used to construct a short-insert, H. japonicus sex chromosomes-specific DNA library. The randomly sequenced clones showed that about 12% of them have significant homology to H. lupulus and 88% to Cannabis sativa Linnaeus, 1753 sequences from GenBank database. Forty-four percent of the sequences show homology to plant retroelements. It was concluded that laser microdissection is a useful tool for isolating the DNA of sex chromosomes of H. japonicus and for the construction of chromosome-specific DNA libraries for the study of the structure and evolution of sex chromosomes. The results provide the potential for identifying unique or sex chromosome-specific sequence elements in H. japonicus and could aid in the identification of sex chromosome-specific repeat and coding regions through chromosome isolation and genome complexity reduction. 18. Lunar Sulfur Capture System Berggren, Mark; Zubrin, Robert; Bostwick-White, Emily 2013-01-01 The Lunar Sulfur Capture System (LSCS) protects in situ resource utilization (ISRU) hardware from corrosion, and reduces contaminant levels in water condensed for electrolysis. The LSCS uses a lunar soil sorbent to trap over 98 percent of sulfur gases and about two-thirds of halide gases evolved during hydrogen reduction of lunar soils. LSCS soil sorbent is based on lunar minerals containing iron and calcium compounds that trap sulfur and halide gas contaminants in a fixed-bed reactor held at temperatures between 250 and 400 C, allowing moisture produced during reduction to pass through in vapor phase. Small amounts of Earth-based polishing sorbents consisting of zinc oxide and sodium aluminate are used to reduce contaminant concentrations to one ppm or less. The preferred LSCS configuration employs lunar soil beneficiation to boost concentrations of reactive sorbent minerals. Lunar soils contain sulfur in concentrations of about 0.1 percent, and halogen compounds including chlorine and fluorine in concentrations of about 0.01 percent. These contaminants are released as gases such as H2S, COS, CS2,HCl, and HF during thermal ISRU processing with hydrogen or other reducing gases. Removal of contaminant gases is required during ISRU processing to prevent hardware corrosion, electrolyzer damage, and catalyst poisoning. The use of Earth-supplied, single-use consumables to entirely remove contaminants at the levels existing in lunar soils would make many ISRU processes unattractive due to the large mass of consumables relative to the mass of oxygen produced. The LSCS concept of using a primary sorbent prepared from lunar soil was identified as a method by which the majority of contaminants could be removed from process gas streams, thereby substantially reducing the required mass of Earth-supplied consumables. The LSCS takes advantage of minerals containing iron and calcium compounds that are present in lunar soil to trap sulfur and halide gases in a fixedbed reactor 19. Neutron Capture and Neutron Halos A.Mengoni; Otsuka, T; Nakamura, T.(International Center for Elementary Particle Physics and Department of Physics, The University of Tokyo, Tokyo, Japan); Ishihara, M. 1996-01-01 The connection between the neutron halo observed in light neutron rich nuclei and the neutron radiative capture process is outlined. We show how nuclear structure information such as spectroscopic factors and external components of the radial wave function of loosely bound states can be derived from the neutron capture cross section. The link between the direct radiative capture and the Coulomb dissociation process is elucidated. 20. Resource capture by single leaves Long, S.P. 1992-05-01 Leaves show a variety of strategies for maximizing CO{sub 2} and light capture. These are more meaningfully explained if they are considered in the context of maximizing capture relative to the utilization of water, nutrients and carbohydrates reserves. There is considerable variation between crops in their efficiency of CO{sub 2} and light capture at the leaf level. Understanding of these mechanisms indicate some ways in which efficiency of resource capture could be level cannot be meaningfully considered without simultaneous understanding of implications at the canopy level. 36 refs., 5 figs., 1 tab. 1. Neutron capture therapy The overall state of the art related with neutron capture therapy(NCT) is surveyed. Since the field related with NCT is very wide, it is not intended to survey all related subjects in depth. The primary objective of this report is to help those working for the installation of a NCT facility and a PGNAA(prompt gamma ray neutron activation analysis) system for the boron analysis understand overall NCT at Hanaro. Therefore, while the parts of reactor neutron source and PGNAA are dealt in detail, other parts are limited to the level necessary to understand related fields. For example, the subject of chemical compound which requires intensive knowledge on chemistry, is not dealt as a separated item. However, the requirement of a compound for NCT, currently available compounds, their characteristics, etc. could be understood through this report. Although the subject of cancer treated by NCT is out of the capability of the author, it is dealt focussing its characteristics related with the success of NCT. Each detailed subject is expected to be dealt more detail by specialists in future. This report would be helpful for the researchers working for the NCT to understand related fields. (author). 128 refs., 3 tabs., 12 figs 2. Gadolinium as a Neutron Capture Therapy Agent Shih, Jing-Luen Allen The clinical results of treating brain tumors with boron neutron capture therapy are very encouraging and researchers around the world are once again making efforts to develop this therapeutic modality. Boron-10 is the agent receiving the most attention for neutron capture therapy but ^{157}Gd is a nuclide that also holds interesting properties of being a neutron capture therapy agent. The objective of this study is to evaluate ^{157}Gd as a neutron capture therapy agent. In this study it is determined that tumor concentrations of about 300 mug ^{157}Gd/g tumor can be achieved in brain tumors with some FDA approved MRI contrast agents such as Gd-DTPA and Gd-DOTA, and up to 628 mug ^{157 }Gd/g tumor can be established in bone tumors with Gd-EDTMP. Monte Carlo calculations show that with only 250 ppm of ^{157}Gd in tumor, neutron capture therapy can deliver 2,000 cGy to a tumor of 2 cm diameter or larger with 5 times 10^{12} n/cm ^2 fluence at the tumor. Dose measurements which were made with films and TLD's in phantoms verified these calculations. More extended Monte Carlo calculations demonstrate that neutron capture therapy with Gd possesses comparable dose distribution to B neutron capture therapy. With 5 times 10^{12 } n/cm^2 thermal neutrons at the tumor, Auger electrons from the Gd produced an optical density enhancement on the films that is similar to the effect caused by about 300 cGy of Gd prompt gamma dose which will further enhance the therapeutic effects. A technique that combines brachytherapy with Gd neutron capture therapy has been evaluated. Monte Carlo calculations show that 5,000 cGy of prompt gamma dose can be delivered to a treatment volume of 40 cm^3 with a 3-plane implant of a total of 9 Gd needles. The tumor to normal tissue advantage of this method is as good as ^{60} Co brachytherapy. Measurements of prompt gamma dose with films and TLD-700's in a lucite phantom verify the Monte Carlo evaluation. A technique which displays the Gd 3. Electric glass capturing markets Wikman, K.; Wikstroem, T. 1996-11-01 Electric glass has found its place on the construction market. In public buildings, electrically heatable windows are becoming the leading option for large glass walls. Studies on detached houses, both new and renovated, show that floor heating combined with electrically heatable windowpanes is the best choice with respect to residents comfort. (orig.) 4. Capturing the Coma 2005-01-01 This image shows comet Tempel 1, as seen by the Deep Impact spacecraft on June 21, 2005. It was taken using the clear filter of the spacecraft's medium resolution imager camera. The spacecraft was 11,564,081.7 kilometers (7,185,920 miles) away from the comet. Twelve images were combined together, and a logarithmic stretch was applied to enhance the coma of the comet. 5. Thomas-forbidden particle capture Carter, John H.; Lieber, Michael 2010-01-01 At high energies, in particle-capture processes between ions and atoms, classical kinematic requirements show that generally double collision Thomas processes dominate. However, for certain mass-ratios these processes are kinematically forbidden. This paper explores the possibility of capture for such processes by triple or higher order collision processes. 6. Neutron capture reactions at DANCE The Detector for Advanced Neutron Capture Experiments (DANCE) is a 4π BaF2 array consisting of 160 active detector elements. The primary purpose of the array is to perform neutron capture cross section measurements on small (> or approx.100 μg) and/or radioactive (241,243Am, neutron capture and neutron-induced fission cross sections and capture-to-fission ratio (α = σγ/σf) for 235U using a new fission-tagging detector as well as neutron capture cross sections for several astrophysics branch-point nuclei. Results from several of these measurements will be presented along with a discussion of additional physics information that can be extracted from the DANCE data 7. Materials For Gas Capture, Methods Of Making Materials For Gas Capture, And Methods Of Capturing Gas Polshettiwar, Vivek 2013-06-20 In accordance with the purpose(s) of the present disclosure, as embodied and broadly described herein, embodiments of the present disclosure, in one aspect, relate to materials that can be used for gas (e.g., CO.sub.2) capture, methods of making materials, methods of capturing gas (e.g., CO.sub.2), and the like, and the like. 8. Power Plant with CO2 Capture based on PSA Cycle Stene, Henrik Sørskår; Moen, Ole Marius 2014-01-01 Two coal-fired power plants with CO2 capture by Pressure Swing Adsorption (PSA) havebeen modeled and simulated. The two power plants considered were IntegratedGasification Combined Cycle (IGCC) and conventional Pulverized Coal Combustion (PCC). Amathematical model of the PSA process for each of the power plants was developed and thegoal was to evaluate the feasibility of PSA as a technology for decarbonisation. Theperformance with CO2 capture by PSA was compared to a reference plant without C... 9. Effective Fingerprint Quality Estimation for Diverse Capture Sensors Dong Sun Park; Sook Yoon; Shan Juan Xie; Jinwook Shin 2010-01-01 Recognizing the quality of fingerprints in advance can be beneficial for improving the performance of fingerprint recognition systems. The representative features to assess the quality of fingerprint images from different types of capture sensors are known to vary. In this paper, an effective quality estimation system that can be adapted for different types of capture sensors is designed by modifying and combining a set of features including orientation certainty, local orientation quality an... 10. Electron capture and stellar collapse In order, to investigate the function of electron capture in the phenomenon of pre-supernovae gravitacional collapse, an hydrodynamic caculation was carried out, coupling capture, decay and nuclear reaction equation system. A star simplified model (homogeneous model) was adopted using fermi ideal gas approximation for tthe sea of free electrons and neutrons. The non simplified treatment from quasi-static evolution to collapse is presented. The capture and beta decay rates, as wellas neutron delayed emission, were calculated by beta decay crude theory, while the other reaction rates were determined by usual theories. The preliminary results are presented. (M.C.K.) 11. Pre-Combustion Carbondioxide Capture in Integrated Gasification Combined Cycles M. Zeki YILMAZOĞLU 2010-01-01 Thermal power plants have a significant place big proportion in the production of electric energy. Thermal power plants are the systems which converts heat energy to mechanical energy and also mechanical energy to electrical energy. Heat energy is obtained from combustion process and as a result of this, some harmful emissions, like CO2, which are the reason for global warming, are released to atmosphere. The contribution of carbondioxide to global warming has been exposed by the previous res... 12. Teaching Knowledge Management by Combining Wikis and Screen Capture Videos Makkonen, Pekka; Siakas, Kerstin; Vaidya, Shakespeare 2011-01-01 Purpose: This paper aims to report on the design and creation of a knowledge management course aimed at facilitating student creation and use of social interactive learning tools for enhanced learning. Design/methodology/approach: The era of social media and web 2.0 has enabled a bottom-up collaborative approach and new ways to publish work on the… 13. Lunar Sulfur Capture System Project National Aeronautics and Space Administration — The Lunar Sulfur Capture System (LSCS) is an innovative method to recover sulfur compounds from lunar soil using sorbents derived primarily from in-situ resources.... 14. Value Capture for Transportation Finance Zhirong (Jerry) Zhao; Michael Iacono; David Levinson 2009-01-01 As vehicles become more fuel-efficient and overall levels of travel stagnate in response to increases in fuel prices, conventional sources of revenue for transportation finance such as taxes on motor fuels have been put under increasing pressure. One potential replacement as a source of revenue is a set of policies collectively referred to as value capture policies. In contrast to fuel taxes and other instruments that impose charges on users of transportation networks, value capture policies ... 15. Toward transformational carbon capture systems Miller, David C. [National Energy Technology Laboratory, U.S. Dept. of Energy, Pittsburgh PA (United States); Litynski, John T. [Office of Fossil Energy, U.S. Dept. of Energy, Washington DC (United States); Brickett, Lynn A. [National Energy Technology Laboratory, U.S. Dept. of Energy, Pittsburgh PA (United States); Morreale, Bryan D. [National Energy Technology Laboratory, U.S. Dept. of Energy, Pittsburgh PA (United States) 2015-10-28 This paper will briefly review the history and current state of Carbon Capture and Storage (CCS) research and development and describe the technical barriers to carbon capture. it will argue forcefully for a new approach to R&D, which leverages both simulation and physical systems at the laboratory and pilot scales to more rapidly move the best technoogies forward, prune less advantageous approaches, and simultaneously develop materials and processes. 16. Direct Capture at Low Energies Balogh, W.; Bieber, R.; Oberhummer, H.; Rauscher, T.; Kratz, K.-L.; Mohr, P; Staudt, G.; Sharma, M. M. 1994-01-01 The importance of direct capture for (n,$\\gamma\$)--reactions on intermediate-- and heavy--mass target nuclei occuring in the s-- and r--process is investigated. It is shown that the direct mechanism is non--negligible for magic and neutron rich target nuclei. For some double magic and neutron rich nuclei in the r--process direct capture is even the dominant reaction mechanism.

17. Context Capture in Software Development

Antunes, Bruno; Correia, Francisco; Gomes, Paulo

2011-01-01

The context of a software developer is something hard to define and capture, as it represents a complex network of elements across different dimensions that are not limited to the work developed on an IDE. We propose the definition of a software developer context model that takes into account all the dimensions that characterize the work environment of the developer. We are especially focused on what the software developer context encompasses at the project level and how it can be captured. T...

18. Modified micro-dissection method to isolate and cultivate human pubic dermal papilla cells%改良显微解剖法分离阴部毛乳头细胞

庄晓晟; 范卫新

2013-01-01

Objective: To establish a proper method for isolating pubic dermal papilla cells. Methods: The dermal papilla cells in scalp and pubic area were isolated by enzyme digestion, micro-dissection and modified micro-dissection method, respectively. The pubic dermal papilla cells were identified by alkaline phosphatase staining and SMA-α immunofluorescence staining. Results: Based on microscopic observation and adherent rales, enzyme digestion method yielded more cells and higher adherent rates for isolating scalp cells. The adherent rate of pubic dermal papilla cells isolated by modified micro-dissection method was significantly higher than that isolated by micro-dissection only (P< 0.005). The pubic dermal papilla cells showed positive for alkaline phosphatase and SMA-α immunofluorescence staining. Conclusion: Modified micro-dissection methods is a suitable method for isolating dermal papilla cells from special site with thicker subcutaneous fat and a lower density of hair follicles.%目的:探讨适合阴部毛乳头细胞的分离培养法.方法:分别用二步酶法,显微解剖法,改良显微解剖法分离纯化人头皮毛乳头细胞及阴部毛乳头细胞,并鉴定培养的阴部毛乳头细胞.结果:通过镜下观及对贴壁率的统计,二步酶法分离人头皮毛乳头产量高,细胞易贴壁.而改良显微解剖法分离阴部毛乳头贴壁率显著高于单纯显微解剖法(P < 0.005),细胞迁出快.阴部毛乳头细胞碱性磷酸酶染色及平滑肌肌动蛋白(SMA)-α免疫荧光染色阳性.结论:改良显微解剖法适合用于皮下脂肪厚、毛囊密度低的特殊部位毛乳头细胞的分离培养.

19. CO2 capture RD & D proceedings in China Huaneng Group

Jinyi Wang; Shisen Xu

2014-01-01

CO2 capture is an important carbon management route to mitigate the greenhouse gas emission in power sector. In recent years, China Huaneng Group (CHNG) has paid more attention on CO2 capture technology development and launched a series of R&D and demonstration projects. In the area of pre-combustion CO2 capture technology, GreenGen project initiated by CHNG is the first integrated gasification combined cycle (IGCC) power plant in China. Located in Tianjin, GreenGen aims at the development, demonstration and promotion of a near-zero emissions power plant. An IGCC plant of 250 MW has successfully passed full-scale trial operation. In the next phase, a pre-combustion CO2 capture unit will be integrated into the system. Pre-combustion process based on coal chemical process has been developed with lower costs successfully. Regarding to post-combustion CO2 capture (PCC), in 2008, CHNG built a 3,000 tpa CO2 capture plant, which was the first CO2 capture demonstration plant in China. In 2009, CHNG launched a PCC project in Shanghai with a capture capacity of 120,000 tpa CO2. Recently, Huaneng Clean Energy Research Institute (CERI) and Powerspan formed a joint venture, Huaneng-CERI-Powerspan (HCP). HCP has completed the technology qualification program to supply carbon capture technology for the CO2 capture Mongstad project. Besides these activities mentioned above, feasibility studies and system design for large scale PCC system, have been undertaken by CERI and its partners from Australia, US and Europe.

20. Possible interspecific origin of the B chromosome of Hypsiboas albopunctatus (Spix, 1824) (Anura, Hylidae), revealed by microdissection, chromosome painting, and reverse hybridisation

Gruber, Simone Lilian; Diniz, Débora; Sobrinho-Scudeler, Patrícia Elda; Fausto Foresti; Haddad, Célio Fernando Baptista; Kasahara, Sanae

2014-01-01

Abstract The B chromosome in the hylid Hypsiboas albopunctatus (2n = 22 + B) is small, almost entirely composed of C-positive heterochromatin, and does not pair with any chromosome of the A complement. B probe, obtained by microdissection and DOP-PCR amplification, was used to search for homology between the B and regular chromosomes of H. albopunctatus and of the related species H. raniceps (Cope, 1862). Reverse hybridisation was also carried out in the investigation. The B probe exclusively painted the supernumerary, not hybridising any other chromosomes in H. albopunctatus, but all H. raniceps chromosomes showed small labelling signals. This result might be an indication that differences exist between the repetitive sequences of A and B chromosomes of H. albopunctatus, and that the chromosomes of H. raniceps and the heterochromatin of the B chromosome of H. albopunctatus are enriched with the same type of repetitive DNA. In meiotic preparations, the B labelled about 30% of scored spermatids, revealing a non-mendelian inheritance, and the painted B in micronucleus suggests that the supernumerary is eliminated from germ line cells. Although our results could suggest an interespecific origin of the B at first sight, further analysis on its repetitive sequences is still necessary. Nevertheless, the accumulation of repetitive sequences, detected in another species, even though closely related, remains an intriguing question. PMID:25349670

1. Possible interspecific origin of the B chromosome of Hypsiboas albopunctatus (Spix, 1824 (Anura, Hylidae, revealed by microdissection, chromosome painting, and reverse hybridisation

Simone Gruber

2014-08-01

Full Text Available The B chromosome in the hylid Hypsiboas albopunctatus (2n = 22 + B is small, almost entirely composed of C-positive heterochromatin, and does not pair with any chromosome of the A complement. B probe, obtained by microdissection and DOP-PCR amplification, was used to search for homology between the B and regular chromosomes of H. albopunctatus and of the related species H. raniceps (Cope, 1862. Reverse hybridisation was also carried out in the investigation. The B probe exclusively painted the supernumerary, not hybridising any other chromosomes in H. albopunctatus, but all H. raniceps chromosomes showed small labelling signals. This result might be an indication that differences exist between the repetitive sequences of A and B chromosomes of H. albopunctatus, and that the chromosomes of H. raniceps and the heterochromatin of the B chromosome of H. albopunctatus are enriched with the same type of repetitive DNA. In meiotic preparations, the B labelled about 30% of scored spermatids, revealing a non-mendelian inheritance, and the painted B in micronucleus suggests that the supernumerary is eliminated from germ line cells. Although our results could suggest an interespecific origin of the B at first sight, further analysis on its repetitive sequences is still necessary. Nevertheless, the accumulation of repetitive sequences, detected in another species, even though closely related, remains an intriguing question.

2. Methodological study of quantitative detection of Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA%免疫磁珠捕获联合PCR-ELISA定量检测结核分枝杆菌的方法学研究

王震; 龚玉华; 钱彩娣; 孙春红; 周丽萍; 傅行礼; 邵启祥

2014-01-01

Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads® ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .%目的：采用免疫磁珠捕获（IMC）联合双内标PCR-ELISA（IMC-PCR-ELISA）技术，建立定量检测结核分枝杆菌的方法。方法制备能够特异性捕获结核分枝杆菌的免疫磁珠（Dynabeads®）。并根据结核分枝杆菌M tp40基因序列以及结核分枝杆菌复合体群IS6110序列，设计2对特异性引物（上游引物的5′端用生物素修饰），以及2条与PCR扩增片段等长的内参照片段（其与扩增模板在引物区的序列相同）和3套捕获探针（3′端用地高辛标记）。先通过免疫磁珠特异性捕获结核分枝杆菌，再联合双内标PCR

3. Capture reactions at low energies

Some few body capture reaction studies at low energies are reviewed. The emphasis is on the use of polarized beams. New calculations of n+d and p+d capture had just recently been completed by the Los Alamos group (Friar, Gibson, and Payne). Their results are in good agreement with existing data, and emphasize the importance of meson exchange current effects in these processes at thermal energies. The effects of M1 radiation can be seen in the vector analyzing power of the p+d radiative capture reaction from its interference with E1 radiation. The vector analyzing powers of the rvec p+T → 4He + γ reaction can be used to search for M1 radiation in 4He. Low energy data show a substantial b1 coefficient which can only be accounted for with a substantial M1 contribution. The author is also working on studying the rvec d+T and rvec d+3He capture reactions in the region of the fusion resonance. His (published) rvec d+T capture data indicate that the reaction goes via ∼95% S = 3/2, E1 capture with small S = 1/2, E1 and small M1 admixtures. In the course of obtaining the d+T and d+3He data, he realized that he could use his spectra to obtain the ground state widths of 5He and 5Li. Finally, he will discuss the d+d reaction at very low energies. The simplest picture of this reaction dominated by S wave capture to the D-state below 1 MeV

4. CHAOTIC CAPTURE OF NEPTUNE TROJANS

Neptune Trojans (NTs) are swarms of outer solar system objects that lead/trail planet Neptune during its revolutions around the Sun. Observations indicate that NTs form a thick cloud of objects with a population perhaps ∼10 times more numerous than that of Jupiter Trojans and orbital inclinations reaching ∼25 deg. The high inclinations of NTs are indicative of capture instead of in situ formation. Here we study a model in which NTs were captured by Neptune during planetary migration when secondary resonances associated with the mean-motion commensurabilities between Uranus and Neptune swept over Neptune's Lagrangian points. This process, known as chaotic capture, is similar to that previously proposed to explain the origin of Jupiter's Trojans. We show that chaotic capture of planetesimals from an ∼35 Earth-mass planetesimal disk can produce a population of NTs that is at least comparable in number to that inferred from current observations. The large orbital inclinations of NTs are a natural outcome of chaotic capture. To obtain the ∼4:1 ratio between high- and low-inclination populations suggested by observations, planetary migration into a dynamically excited planetesimal disk may be required. The required stirring could have been induced by Pluto-sized and larger objects that have formed in the disk.

5. Body and Face Animation Based on Motion Capture

Xiaoting Wang

2011-03-01

Full Text Available This paper introduces the motion capture technology and its use in computer animation. Motion capture is a powerful aid in computer animation and a supplement to the traditional key-frame animation. We use professional cameras to record the body motion and facial expression of the actor and then manipulate the data in software to eliminate some occlusion and confusion errors. As to data that is still not satisfying, we use data filter to smooth the motion by cutting some awry frames. Then we import the captured data into Motionbuilder to adjust the motion and preview the real-time animation. At last in Maya we combine the motion data and character model, let the captured data drive the character and add the scene model and music to export the whole animation. In the course of computer animation, we use this method to design the animation of military boxing, basketball playing, folk dancing and facial expression.

6. Proton capture by magnetic monopoles

In the Kazama-Yang approximation, the lowest monopole-proton bound states have binding energies of 938 MeV, 263 keV, 105 eV, and 0.04 eV. The cross section for radiative capture to these states is for velocities β = 10-5 - 10-3 found to be of the order of 10-28 - 10-26 cm2. For the state that has a binding energy of 263 keV, the capture length in water is 171 x (β/10-4)sup(0.48) m. Observation of photons from the capture process would indicate the presence of monopoles. (orig.)

7. The Economics of Carbon Capture

Islegen Ozge; Reichelstein Stefan J

2009-01-01

As carbon capture technologies are likely to allow power plants to reduce emissions by up to 90 percent, Ozge Islegen and Stefan Reichelstein of Stanford Business School feel that plants will choose to implement such technologies to avoid paying even modest emissions permit prices.

8. Prey capture by harbor porpoises

Miller, Lee; Verfuss, Ursula

2009-01-01

source levels of clicks decrease by about half (6dB) for each halving of distance to the prey. The sequence ends in a terminal, high repetition rate buzz (~500 clicks/s) just before capturing the prey (a video will be shown). The temporal sequence differs from that of beaked whales, but is like that of...

9. Prey capture by harbor porpoises

Miller, Lee

2008-01-01

) that progressively decrease when closing on an object.  After detecting the prey, the click interval stabilizes and then becomes progressively shorter while approaching the prey.  The sequence ends in a terminal, high repetition rate buzz (~500 clicks/s) just before capturing the prey (a video will be...

10. Resource Complementarity and Value Capture in Firm Acquisitions

Grimpe, Christoph; Hussinger, Katrin

2014-01-01

exclusion and freedom to operate in R&D. We argue that such property rights hold preemptive power allowing firms to capture the value from combining complementary technologies and to realize gains from trade in strategic factor markets. Our results for a sample of 1,428 acquisitions indicate that...
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