Expression of immunoglobulin kappa and lambda chains in mink.
The ratio of kappa and lambda chains of immunoglobulins varies significantly from one species to another. It has previously been thought that lambda was only type expressed in mink. We tested mink immunoglobulin light chains using two monoclonal antibodies G80 and G88. It has been shown that G80 and G88 specifically recognize two antigenically different subpopulations of the light chains. Immunochemical analysis of these subpopulations separated by affinity chromatography suggested that they represent lambda and kappa types of light chains, respectively. Screening of a mink cDNA library with monoclonal antibody G88 resulted in the isolation of clone pIGK-1 containing kappa chain-encoding sequence. The cDNA insert of pIGK-1 included most of the V segment, as well as the J, C and 3' untranslated sequences. Mink V kappa sequence shown the highest homology with the human V kappa II subgroup genes (76-79%). Mink C kappa sequence was 53-63% homologous to C kappa of other species. The striking feature of mink C kappa chain is the presence of glutamine in the C-terminal position. Southern blot analysis suggested that mink haploid genome has one C kappa gene and multiple V kappa genes. The kappa:lambda chain ratio in the 12 minks studied was, on the average, 46:54. The same ratio was observed for the kappa- and lambda-producing cells in the mesenteric lymph nodes. The five previously identified mink light chain allotypes were assigned to the lambda chains, thereby confirming that lambda chains in this species are additionally subdivided into several subtypes.


Advances in FDG PET probes in surgical oncology.
Whole-body positron emission tomography (WB-PET) has been developed as a sensitive, cost-effective method for imaging malignant disease. WB-PET provides complete body imaging with a single scanning approach for a variety of malignancies. With increasing clinical experience, the indications for its use have broadened. This article reviews current uses of the technology and discusses some potential applications, particularly the utility of a commercially available surgical gamma probe for detecting the gamma particles emitted in the decay process of FDG.


A computer evaluation of ventilation performance in a negative-pressure operating theater.
A negative-pressure operating theater is required to limit the spread of respiratory diseases in patients with severe acute respiratory syndrome, tuberculosis, avian influenza, or similar infectious diseases. In Hong Kong, we converted a conventional operating theater into a negative-pressure operating theater that has been in service for more than a year. In this article, we introduce its ventilation design and evaluate the airflow performance in relation to different combinations of medical lamp configurations and modes of launching infectious particles into the room air. We used a computational fluid dynamics technique for the numerical analysis. Our analyses showed that the airflow performance in the negative-pressure operating theater was satisfactory and comparable to the original positive-pressure design. The airflow pattern effectively controlled the dispersion of infectious particles. Our calculations demonstrated that the airflow contained the dispersion of infectious particles released from the patient sufficiently to protect the surgical team, and vice versa. Computational fluid dynamics can be used to assess airflow in a negative-pressure operating room and model the dispersion of infectious particles from the patient.


The Role of Neural and Genetic Processes in Learning to Read and Specific Reading Disabilities: Implications for Instruction.
To learn to read, the brain must repurpose neural systems for oral language and visual processing to mediate written language. We begin with a description of computational models for how alphabetic written language is processed. Next, we explain the roles of a dorsal sublexical system in the brain that relates print and speech, a ventral lexical system that develops the visual expertise for rapid orthographic processing at the word level, and the role of cognitive control networks that regulate attentional processes as children read. We then use studies of children, adult illiterates learning to read, and studies of poor readers involved in intervention, to demonstrate the plasticity of these neural networks in development and in relation to instruction. We provide a brief overview of the rapid increase in the field's understanding and technology for assessing genetic influence on reading. Family studies of twins have shown that reading skills are heritable, and molecular genetic studies have identified numerous regions of the genome that may harbor candidate genes for the heritability of reading. In selected families, reading impairment has been associated with major genetic effects, despite individual gene contributions across the broader population that appear to be small. Neural and genetic studies do not prescribe how children should be taught to read, but these studies have underscored the critical role of early intervention and ongoing support. These studies also have highlighted how structured instruction that facilitates access to the sublexical components of words is a critical part of training the brain to read.


Cardiovascular response to mental stress: dynamic and isometric exercise in hypertensive patients treated with prizidilol hydrochloride (SK&F 92657).
The cardiovascular effects of a new antihypertensive agent, prizidilol hydrochloride (SK&F 92657), have been evaluated in a group of 13 patients with mild to moderate essential hypertension, both at rest and during mental stress and isometric or dynamic exercise. After 6 weeks of therapy (200-300 mg of prizidilol b.i.d.) both systolic and diastolic blood pressure values at rest were significantly lower than those recorded before treatment and during placebo (with an average decrease of about 22/11 mm Hg in the supine position). Heart rate at rest was slightly reduced. Cardiovascular responses to dynamic exercise and mental stress were not prevented by prizidilol, but blood pressure and heart rate levels around which variation occurred were lowered to a significant extent. During isometric exercise prizidilol did not protect against a rise in diastolic blood pressure. Blood pressure and heart rate reduction both at rest and during stimulation were not related to plasma drug concentrations. Side-effects were very rare. It is concluded that prizidilol significantly reduces blood pressure both at rest and during mental stress and physical activity with only sight effects on heart rate, and with a very simple therapeutic scheme.


Biological mesh in hernia repair, abdominal wall defects, and reconstruction and treatment of pelvic organ prolapse: a review of the clinical evidence.
The use of biological meshes seems to be an innovation that influences all surgical fields. The existing evidence indicates that biological meshes cannot only be applied on the area of hernia repair, but also on cases of abdominal reconstruction and pelvic organ prolapse treatment, especially in the presence of contaminated or potentially contaminated surgical fields. Low risks of graft rejection, complications, and infection seem to be the main advantages of biological meshes in comparison with the synthetic nonabsorbable ones. Nevertheless, the elevated costs suggest that biological mesh implantation should be evaluated in correlation to the needs of each clinical case.


Giant Lipoma of the Anterior Neck Causing Dyspnea.
Lipomas are the most common benign mesenchymal tumors, composed of mature adipocytes, and may occur anywhere in the body where fatty tissue is present. Neck lipomas are rare and usually appear as solitary, small (<5 cm), and asymptomatic lesions located in the subcutis of posterior neck. Giant anterior neck lipomas are exceptionally rare and, other than cosmetic concerns, seldom present with respiratory symptoms. A 37-year-old female patient was referred for a long-standing anterior and right-sided neck swelling, which had markedly increased in size causing cosmetic disfigurement, neck discomfort, and transient episodes of dyspnea when lying on the side opposite the neck mass. Under general anesthesia, the lesion was completely dissected and removed through the elliptical cervical incision over the right supraclavicular fossa, while carefully preserving the surrounding neck structures. The postoperative recovery was uneventful and the patient was satisfied with the cosmetic outcome and relief of her respiratory symptoms.


Targeted mutagenesis in cotton (Gossypium hirsutum L.) using the CRISPR/Cas9 system.
The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 system has been widely used for genome editing in various plants because of its simplicity, high efficiency and design flexibility. However, to our knowledge, there is no report on the application of CRISPR/Cas9-mediated targeted mutagenesis in cotton. Here, we report the genome editing and targeted mutagenesis in upland cotton (Gossypium hirsutum L., hereafter cotton) using the CRISPR/Cas9 system. We designed two guide RNAs to target distinct sites of the cotton Cloroplastos alterados 1 (GhCLA1) and vacuolar H+-pyrophosphatase (GhVP) genes. Mutations in these two genes were detected in cotton protoplasts. Most of the mutations were nucleotide substitutions, with one nucleotide insertion and one substitution found in GhCLA1 and one deletion found in GhVP in cotton protoplasts. Subsequently, the two vectors were transformed into cotton shoot apexes through Agrobacterium-mediated transformation, resulting in efficient target gene editing. Most of the mutations were nucleotide deletions, and the mutation efficiencies were 47.6-81.8% in transgenic cotton plants. Evaluation using restriction-enzyme-PCR assay and sequence analysis detected no off-target mutations. Our results indicated that the CRISPR/Cas9 system was an efficient and specific tool for targeted mutagenesis of the cotton genome.


Intraventricular Meningioma Resection with Postoperative Ischemia of the Lateral Geniculate Nucleus.
Intraventricular meningiomas comprise 0.5%-3% of intracranial meningiomas. They often cause obstructive hydrocephalus and commonly are treated with surgical resection or stereotactic radiosurgery. To describe the surgical approaches and resection techniques needed to approach intraventricular tumors while highlighting the eloquent anatomy and blood supply surrounding the ventricular system to avoid neurological injury. Two cases of left atrial intraventricular meningiomas that were complicated by postoperative lateral geniculate nucleus ischemia and resultant temporary contralateral quadrantanopia are described. The safe surgical approaches to the atrium of lateral ventricles as well as the anatomy and blood supply to the lateral geniculate nucleus and optic radiation are discussed and illustrated. In both cases, the patients had complete resection of their tumors. They both ultimately made a good recovery after transient visual deficits. These cases provide useful demonstrations of eloquent anatomy of the ventricular walls and visual pathway anatomy. Care must be taken to avoid visual pathways along the lateral ventricle wall and the nearby arterial supply of the lateral geniculate nucleus from the choroidal arteries when resecting intraventricular tumors.


Prooxidative activities of 10 phenazine derivatives relative to that of clofazimine.
The objective of this study was to investigate the relationship between the antimycobacterial properties of the antileprosy drug clofazimine and its stimulatory effect on the release of reactive oxidants by polymorphonuclear leukocytes by using a variety of phenazine derivatives. The effects of these compounds on myeloperoxidase-mediated iodination, luminol-enhanced chemiluminescence, and the release of superoxide anion by polymorphonuclear leukocytes were investigated. Dissociation of the antimycobacterial and prooxidative effects of clofazimine was possible by manipulation of the chemical group in position 2 of the phenazine molecule. When nitrogen-containing substituents in this position were replaced by oxygen, the mode of the prooxidative action of the compounds was altered.


Fibroblast-Cytophilic and HeLa-Cytotoxic Dual Function Carbon Nanoribbon Network Platform.
Carbon nanomaterials have emerged as a promising material in cancer diagnosis and therapy. Carbon nanomaterials/nanostructures (C-C molecular structure) act as a carrier/skeleton and require further surface modification through functionalization with chemicals or biomolecules to attain cell response. We report the synthesis of a novel carbon nanoribbon network (CNRN) platform that possesses a combination of C-C and C-O bond architecture. The bioactive CNRN showed enhanced ability for cell adhesion. Most importantly, it induced opposite cell responses from healthy cells and cancerous cells, cytophilic to fibroblasts but cytotoxic to HeLa cells. Ultrafast laser ionization under ambient conditions transforms nonbioresponsive C-C bond of graphite to C-C and C-O bonds, forming a self-assembled CNRN platform. The morphology, nanochemistry, and functionality on modulating fibroblast and HeLa adhesion and proliferation of the fabricated CNRN platforms were investigated. The results of in vitro studies suggested that the CNRN platforms not only attracted but also actively accelerated the adhesion and proliferation of both fibroblasts and HeLa cells. The proliferation rate of fibroblasts and HeLa cells is 91 and 98 times greater compared with that of a native graphite substrate, respectively. The morphology of the cells over a period of 24 to 48 h revealed that the CNRN platform induced an apoptosis-like cytotoxic function on HeLa cells, whereas fibroblasts experienced a cytophilic effect and formed a tissuelike structure. The degree of cytotoxic or cytophilic effect can be further enhanced by adjusting parameters such as the ratio of C-C bonds to C-O bonds, the nanoribbon width, and the nanovoid porosity of the CNRN platforms, which could be tuned by careful control of laser ionization. In a nutshell, for the first time, pristine carbon nanostructures free from biochemical functionalization demonstrate dual function, cytophilic to fibroblast cells and cytotoxic to HeLa cells.


Cooperative pathogenicity in cystic fibrosis: Stenotrophomonas maltophilia modulates Pseudomonas aeruginosa virulence in mixed biofilm.
The present study was undertaken in order to understand more about the interaction occurring between S. maltophilia and P. aeruginosa, which are frequently co-isolated from CF airways. For this purpose, S. maltophilia RR7 and P. aeruginosa RR8 strains, co-isolated from the lung of a chronically infected CF patient during a pulmonary exacerbation episode, were evaluated for reciprocal effect during planktonic growth, adhesion and biofilm formation onto both polystyrene and CF bronchial cell monolayer, motility, as well as for gene expression in mixed biofilms. P. aeruginosa significantly affected S. maltophilia growth in both planktonic and biofilm cultures, due to an inhibitory activity probably requiring direct contact. Conversely, no effect was observed on P. aeruginosa by S. maltophilia. Compared with monocultures, the adhesiveness of P. aeruginosa on CFBE41o- cells was significantly reduced by S. maltophilia, which probably acts by reducing P. aeruginosa's swimming motility. An opposite trend was observed for biofilm formation, confirming the findings obtained using polystyrene. When grown in mixed biofilm with S. maltophilia, P. aeruginosa significantly over-expressed aprA, and algD-codifying for protease and alginate, respectively-while the quorum sensing related rhlR and lasI genes were down-regulated. The induced alginate expression by P. aeruginosa might be responsible for the protection of S. maltophilia against tobramycin activity we observed in mixed biofilms. Taken together, our results suggest that the existence of reciprocal interference of S. maltophilia and P. aeruginosa in CF lung is plausible. In particular, S. maltophilia might confer some selective "fitness advantage" to P. aeruginosa under the specific conditions of chronic infection or, alternatively, increase the virulence of P. aeruginosa thus leading to pulmonary exacerbation.


N,N'-Dioxide/nickel(ii)-catalyzed asymmetric Diels-Alder reaction of cyclopentadiene with 2,3-dioxopyrrolidines and 2-alkenoyl pyridines.
A chiral N,N'-dioxide/Ni(OTf)2 complex-catalyzed asymmetric Diels-Alder reaction of cyclopentadiene with 2,3-dioxopyrrolidines and 2-alkenoyl pyridines has been achieved. The corresponding chiral bridged compounds were obtained in high yields with excellent dr and ee values (up to 97% yield, 95 : 5 dr and 97% ee).


Identification and quantification of betalains from the fruits of 10 mexican prickly pear cultivars by high-performance liquid chromatography and electrospray ionization mass spectrometry.
Qualitative and quantitative analyses of betalain pigments in 10 cultivars/lines of prickly pear (Opuntia spp.) fruit grown in Mexico were conducted with reverse phase high-performance liquid chromatography-diode array detection (HPLC-DAD) coupled with electrospray mass spectrometry (ESI-MS). Betacyanins and betaxanthins were identified by comparison with the UV/vis and mass spectrometric characteristics as well as the retention times of semisynthesized reference betaxanthins. Data revealed that the ratio and concentration of betalain pigments are responsible for the color in the different cultivars, showing the highest betalains content in the fruit of purple colored Camuesa (O. robusta Wendl.) (8.1 mg/g dry fruit), which is comparable to that found in red beet Beta vulgaris L. ssp. Var. Pablo) (8.6 mg/g dry tissue). Yellow betalains were absent in Reyna (O. alba-carpa) prickly pear cultivar. A total of 24 known/unknown betalains were present in the prickly pear fruit samples studied, including 18 betaxanthins and 6 betacyanins. Our results indicate that prickly pear cultivars can be considered as a potential source of yellow and red natural colorants.


Modulation of the intestinal environment, innate immune response, and barrier function by dietary threonine and purified fiber during a coccidiosis challenge in broiler chicks.
Coccidiosis is a major contributor to economic losses in the poultry industry due to its detrimental effects on growth performance and nutrient utilization. We hypothesized that the combined effects of supplemental dietary Thr and purified fiber may modulate the intestinal environment and positively affect intestinal immune responses and barrier function in broiler chicks infected with Eimeria maxima. A Thr-deficient basal diet (3.1 g of Thr/kg of diet) was supplemented with 70 g/kg of silica sand (control) or high-methoxy pectin and 1 of 2 concentrations of Thr (1.8 or 5.3 g/kg of diet; 4 diets total), and fed to chicks from hatch to d 16 posthatch. On d 10 posthatch, chicks received 0.5 mL of distilled water or an acute dose of Eimeria maxima (1.5 × 10(3) sporulated oocytes) with 6 replicate pens of 6 chicks per each of 8 treatment combinations (4 diets and 2 inoculation states). Body weight gain, feed intake, and G:F increased (P < 0.01) with addition of 5.3 g of Thr/kg of diet. Eimeria maxima schizonts were present only in intestinal tissue sampled from infected birds (P < 0.01). Weights of cecal digesta were highest (P < 0.01) in pectin-fed birds, and ceca with the heaviest weights also had the highest concentrations of total short-chain fatty acids. Expression of interleukin-12 in ileal mucosa was highest (P < 0.01) in infected birds receiving the control diet with 5.3 g of supplemental Thr/kg. In cecal tonsils, interferon-γ expression was highest in infected birds receiving the control diet (fiber × infection, P < 0.05); interferon-γ expression was lowest in infected birds fed the high Thr diet (Thr × infection, P < 0.05). There were no differences due to infection or Thr supplementation for cytokine expression in birds fed pectin-containing treatments. Overall, we conclude that although pectin has some protective function against coccidiosis, Thr supplementation had the greatest effect on intestinal immune response and maintenance of near normal growth in young broiler chicks infected with E. maxima.


Prediction of Recurrent Ischemic Stroke Using Registry Data and Machine Learning Methods: The Erlangen Stroke Registry.
There have been multiple efforts toward individual prediction of recurrent strokes based on structured clinical and imaging data using machine learning algorithms. Some of these efforts resulted in relatively accurate prediction models. However, acquiring clinical and imaging data is typically possible at provider sites only and is associated with additional costs. Therefore, we developed recurrent stroke prediction models based solely on data easily obtained from the patient at home. Data from 384 patients with ischemic stroke were obtained from the Erlangen Stroke Registry. Patients were followed at 3 and 12 months after first stroke and then annually, for about 2 years on average. Multiple machine learning algorithms were applied to train predictive models for estimating individual risk of recurrent stroke within 1 year. Double nested cross-validation was utilized for conservative performance estimation and models' learning capabilities were assessed by learning curves. Predicted probabilities were calibrated, and relative variable importance was assessed using explainable artificial intelligence techniques. The best model achieved the area under the curve of 0.70 (95% CI, 0.64-0.76) and relatively good probability calibration. The most predictive factors included patient's family and housing circumstances, rehabilitative measures, age, high calorie diet, systolic and diastolic blood pressures, percutaneous endoscopic gastrotomy, number of family doctor's home visits, and patient's mental state. Developing fairly accurate models for individual risk prediction of recurrent ischemic stroke within 1 year solely based on registry data is feasible. Such models could be applied in a home setting to provide an initial risk assessment and identify high-risk patients early.


Antibody Modified Nanoparticle-Mediated Delivery of miR-124 Regulates Apoptosis via Repression the Stat3 Signal in Mycobacterial-Infected Microglia.
The mechanism of Mycobacterium tuberculosis (M.tb) evasion of host cell remains elusive. Several microRNAs that are involved in this complex process were identified. miRNA interference-based therapeutics represents an attractive challenge and shows huge potential for disorder treatment. In this study, we found that miR-124-3p expression is significantly decreased in microglia after Mycobacterium marinum (M.m) infection. To achieve better target transfection effect, a CD11b antibody and PEI modified nanoparticles-based Nano platform had been developed. This system was equipped by conjugation of miRNA-124-3p onto the surface of nanoparticles with a PEI/CD11b antibody coating. Transfection with miR-124-3p promoted microglia apoptosis through upregulation of Caspase3 or downregulation of Bcl-2 and Bcl-xl. More importantly, transfection with miR-124-3p inhibitor increases the mycobacterium proliferation in microglia. Based on the above, we further found miRNA-124-3p to bind to 3'untranslated region of Stat3, resulting in the downregulation of its protein to trigger cells apoptosis through Stat3-related pathway. As such, our research might provide new insights towards target delivering miRNA through the bold-brain barrier (BBB) and exploiting highly effective anti-tuberculous meningitis drugs. Taken together, our findings suggest how Mycobacterium can manipulate host miRNA expression to regulate cell survival for its own proliferation, and highlight the importance to develop novel therapeutic strategies against tuberculous meningitis.


Dissolution kinetics and mechanisms at dolomite-water interfaces: effects of electrolyte specific ionic strength.
Elucidating dissolution kinetics and mechanisms at carbonate mineral-water interfaces is essential to many environmental and geochemical processes, including geologic CO(2) sequestration in deep aquifers. In the present work, effects of background electrolytes on dolomite (CaMg(CO(3))(2)) reactivity were investigated by measuring step dissolution rates using in situ hydrothermal atomic force microscopy (HAFM) at 90 °C. Cleaved surfaces of dolomite were exposed to sodium chloride and tetramethylammonium chloride (TMACl) aqueous solutions with ionic strengths (I) ranging from 0 to 0.77 m at pH 4 and pH 9. HAFM results demonstrated that dolomite step retreat rates increased with increasing solution ionic strength and decreasing pH. Comparison of [481] and [441] steps revealed that the anisotropy of [481] and [441] step speeds became significant as solution ionic strength increased, with NaCl exerting more pronounced effects than TMACl for the same I. To interpret the different trends observed for NaCl and TMACl, a dissolution mechanism involving orientation-dependent ion adsorption and consequent edge free energy changes is proposed.


Tim-3 expression on peripheral monocytes and CD3+CD16/CD56+natural killer-like T cells in patients with chronic hepatitis B.
Hepatitis B virus (HBV) infection is one of the major causes of chronic liver inflammation. Tim-3 acts as a negative regulatory molecule and plays a critical role in immune tolerance. In the current study, we investigated Tim-3 expression on peripheral monocytes and CD3+CD16/CD56+ natural killer like T (NKT-like) cells in chronic hepatitis B (CHB) patients. Peripheral blood mononuclear cells (PBMCs) were isolated from 52 CHB patients and 60 healthy controls. Tim-3+CD14+ cells and Tim-3+CD3+CD16/CD56+ cells were analyzed by flow cytometry. Results showed that expression of Tim-3 was significantly increased on both the monocytes and NKT-like cells in CHB patients than in controls (P = 0.002 and P < 0.001, respectively). Tim-3 levels on monocytes and NKT-like cells were further upregulated in patients with acute-on-chronic liver failure (ACLF). In addition, we assessed the correlation of Tim-3 expression with levels of alanine aminotransferase (ALT) and tumor necrosis factor alpha (TNF-α). Data revealed that Tim-3 expression on both monocytes and NKT-like cells was positively correlated with level of ALT (r = 0.59, P < 0.001, and r = 0.60, P < 0.001, respectively), whereas Tim-3 expression on NKT-like cells was negatively correlated with serum level of TNF-α (r = -0.54, P < 0.001) in CHB patients. Our results suggest that Tim-3 may play important roles in the pathogenesis of CHB.


The association of HLA-C and ERAP1 polymorphisms in early and late onset psoriasis and psoriatic arthritis patients of Hungary.
Single nucleotide polymorphisms (SNPs) of the HLA-C and ERAP1 genes were recently determined to contribute to psoriasis susceptibility. However, data regarding the association of these genes with specific subgroups of psoriasis are scarce. To examine the possible association of the HLA-C and ERAP-1 polymorphisms with early and late onset psoriasis and psoriatic arthritis. Five ERAP1 SNPs and two HLA-C SNPs were genotyped in 105 psoriatic arthritis patients, 214 cutaneous psoriasis patients and 200 healthy individuals. Haplotypes were constructed for three ERAP1 SNPs (rs17482078, rs10050860, rs30187), and interaction between HLA-Cw*0602 and ERAP1 was also analysed. The HLA-Cw*0602 rs10484554 SNP was found to be a strong susceptibility factor for early onset cutaneous psoriasis and early onset psoriatic arthritis. ERAP1 SNPs (rs10050860, rs17482078, rs27525) appear to have a protective function for early onset psoriatic arthritis. The haplotype B was identified as a susceptibility factor for late onset psoriatic arthritis. In HLA-C positive individuals the rs27524 ERAP1 SNP was associated with a significantly increased risk of psoriatic arthritis development, whereas the rs27525 ERAP1 SNP had the opposite effect. These results suggest that the HLA-C and ERAP1 genes contribute to the pathogenesis of psoriasis and psoriatic arthritis in an age-dependent manner.


Development and evaluation of a novel user interface for reviewing clinical microbiology results.
Microbiology laboratory results are complex and cumbersome to review. We sought to develop a new review tool to improve the ease and accuracy of microbiology results review. We observed and informally interviewed clinicians to determine areas in which existing microbiology review tools were lacking. We developed a new tool that reorganizes microbiology results by time and organism. We conducted a scenario-based usability evaluation to compare the new tool to existing legacy tools, using a balanced block design. The average time-on-task decreased from 45.3 min for the legacy tools to 27.1 min for the new tool (P < .0001). Total errors decreased from 41 with the legacy tools to 19 with the new tool (P = .0068). The average Single Ease Question score was 5.65 (out of 7) for the new tool, compared to 3.78 for the legacy tools (P < .0001). The new tool scored 88 ("Excellent") on the System Usability Scale. The new tool substantially improved efficiency, accuracy, and usability. It was subsequently integrated into the electronic health record and rolled out system-wide. This project provides an example of how clinical and informatics teams can innovative alongside a commercial Electronic Health Record (EHR).


Echinacoside regulates PI3K/AKT/HIF-1α/VEGF cross signaling axis in proliferation and apoptosis of breast cancer.
Echinacoside (ECH) is a natural anti-cancer compound and is of great value in cancer treatment. However, the mechanism underlying this effect on breast cancer (BC) was unclear. To explore the mechanism of ECH treating BC by network pharmacology and experimental validation. Several databases were searched to screen potential targets of ECH and obtain information on targets related to BC. STRING was applied to construct a Protein-protein interaction (PPI) network. DAVID was applied for Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Gene Expression Profiling Interactive Analysis (GEPIA) was searched for the relationship between the expression profile and overall survival of major targets in normal breast and BC tissues. Finally, the results of network pharmacology analysis were validated by experiments. Seventeen targets of ECH overlapped with targets in BC. Ten hub targets were determined through PPI. By GO and KEGG analysis 15 entries and 25 pathways were obtained, in which phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) played greater roles. Validation of key targets in the GEPIA database showed that PIK3R1 and PIK3CD remained consistent with the results of the study. Experiments in vitro showed ECH inhibited proliferation, induced apoptosis and reduced mRNA levels and protein expression of PI3K, AKT, hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor A (VEGFA) in MCF-7 cells. Furthermore, experiments in vivo revealed that ECH significantly reduced tumor growth, promoted apoptosis and decreased the related mRNA levels and protein expression, suggesting ECH works on BC by regulating PI3K/AKT/HIF-1α/VEGF signaling pathway. In summary, ECH played an important role in anti-BC by regulating PI3K/AKT/HIF-1α/VEGF signaling pathway. Furthermore, ECH had multi-target and multi-pathway effects, which may be a promising natural compound for treating BC.


Voltage-Driven Conformational Switching with Distinct Raman Signature in a Single-Molecule Junction.
Precisely controlling well-defined, stable single-molecule junctions represents a pillar of single-molecule electronics. Early attempts to establish computing with molecular switching arrays were partly challenged by limitations in the direct chemical characterization of metal-molecule-metal junctions. While cryogenic scanning probe studies have advanced the mechanistic understanding of current- and voltage-induced conformational switching, metal-molecule-metal conformations are still largely inferred from indirect evidence. Hence, the development of robust, chemically sensitive techniques is instrumental for advancement in the field. Here we probe the conformation of a two-state molecular switch with vibrational spectroscopy, while simultaneously operating it by means of the applied voltage. Our study emphasizes measurements of single-molecule Raman spectra in a room-temperature stable single-molecule switch presenting a signal modulation of nearly 2 orders of magnitude.


Transcriptional regulation of the TGF-beta 2 gene in choriocarcinoma cells and breast carcinoma cells: differential utilization of Cis-regulatory elements.
Previous studies have shown that the transcription of the TGF-beta 2 gene is controlled by at least one negative and two positive regulatory regions in differentiated cells derived from both embryonal carcinoma cells and embryonic stem cells. The use of TGF-beta 2 promoter/reporter gene constructs has also identified a CRE/ATF motif near the TATA box that appears to heavily influence the transcription of the TGF-beta 2 gene. In this study, two choriocarcinoma cell lines, JAR and JEG-3, and the breast cancer cell line, MCF-7, were used to determine whether differences exist in the transcriptional regulation of the TGF-beta 2 gene. We demonstrated that both similarities and differences exist in the transcriptional regulation of this gene. Common to all cells examined to date, the positive regulatory region just upstream of the TATA box contains an essential CRE/ATF motif that binds at least one transcription factor, ATF-1, in gel mobility shift assays. However, we did not detect ATF-2 binding to this site with any of the nuclear extracts used. We also determined that the effect of the region between -187 and -78 (relative to the transcription start site) is cell type dependent. Previous studies have shown that this region acts to reduce the activity of the TGF-beta 2 promoter in differentiated cells derived from embryonal carcinoma cells and embryonic stem cells. In direct contrast, this region acts as a strong positive regulatory region in JAR, JEC-3, and MCF-7 cells. The mechanisms responsible for these differing effects remain to be established. Interestingly, this region does not appear to contain sequence motifs that bind known transcription factors. Thus, this region is likely to bind one or more novel transcription factors or contain novel recognition sites for known transcription factors.


Interaction of surfactant protein A with cellular myosin.
The goal of the current investigation was to characterize, purify, and identify the proteins that bind surfactant protein A (SP-A). Several polypeptides were purified by SP-A affinity chromatography, and the 200 kD major polypeptide that reacted with SP-A on ligand blots was purified further by preparative SDS-PAGE. Protein sequencing of proteolytically derived subfragments of this polypeptide gave sequences that corresponded completely with nonmuscle (cellular) myosin heavy chain. The 200 kD polypeptide was then found to be immunoreactive with antibodies against cellular myosin. A smaller polypeptide of 135 kD also binds SP-A and appears to be a proteolytic fragment of the 200 kD peptide. The ability of SP-A to bind myosin was confirmed in a microtiter well assay and was found to be concentration dependent. We speculated that the physiologic relevance of the interaction of SP-A with myosin might be to facilitate clearance of myosin from the alveolar subphase following its release during lung injury. In support of this hypothesis, we found that there were detectable levels of myosin in lavage fluid and that SP-A could indeed enhance uptake and degradation of myosin by alveolar macrophages.


A dual-response fluorescent probe for N2H4 and viscosity in living cells and zebrafish to evaluate liver injury.
Hydrazide drugs can cause severe drug-induced liver injury owing to the enzymatic release of N2H4 in the liver. Also, changes in cellular viscosity are associated with liver damage. Thus, simultaneous monitoring of changes in N2H4 levels and viscosity can be used to evaluate the side effects of hydrazide drugs. Herein, we firstly reported a near-infrared fluorescent probe (FNN), which contains 1,8-naphthalimide as the fluorophore and a chalcone moiety as the responsive receptor, for sensitively detecting intracellular viscosity and N2H4. FNN showed a fast 'turn-on' fluorescence response to N2H4 with excellent selectivity. Additionally, FNN could selectively track viscosity without interference from polarity, pH, and other active species. Furthermore, imaging experiments suggested that FNN could be successfully applied in living cells and zebrafish larvae and embryos, which is of great importance for effectively assessing the degree of liver injury.


Recombinant human interferon-alpha A treatment of an experimental cutaneous herpes simplex virus type 1 infection of guinea pigs.
Recombinant human interferon-alpha A (rIFN-alpha A) was evaluated for therapeutic efficacy against an experimental dorsal cutaneous herpes simplex virus type 1 (HSV-1) infection of guinea pigs. Human IFN has activity in this species. Animals were treated either systemically (i.m.) or topically with different formulations of rIFN-alpha A. Therapy was initiated 24 h before (-24 h), 30 min after (+0.5 h), or 24 h following (+24 h) virus inoculation, and treatment was continued for 3-5 days. Efficacy was evaluated on day 4 following infection. Treatment with rIFN-alpha A given systemically beginning at -24 h was effective, reducing lesion number, total lesion area, and lesion virus titer by 64%, 85%, and 98% respectively (p less than 0.001). Efficacy diminished with delay in initiation of i.m. therapy. Topical rIFN-alpha A formulations were generally ineffective, showing only a marginal effect with therapy initiated -24 h or +0.5 h and no effect when treatment was delayed to +24 h. In summary, rIFN-alpha A is effective against cutaneous HSV-1 infection in this model if therapy is initiated prophylactically and if drug delivery is assured by systemic injection. Topical application of rIFN-alpha A showed little therapeutic effect. The large molecular weight of IFN likely retards its percutaneous delivery.


Gender difference in airway hyperresponsiveness in smokers with mild COPD. The Lung Health Study.
Methacholine bronchoprovocation challenge testing was successfully completed in 5,662 participants (3,556 men and 2,106 women) at the time they were randomized into the Lung Health Study, a multicenter trial designed to evaluate early intervention in chronic obstructive pulmonary disease (COPD). All participants were smokers between the ages of 35 and 60 yr who had mild COPD. The male:female prevalence of a positive challenge (PC20FEV1) was 25%:48% and 63%:87% at a PC20FEV1 of < or = 5 mg/ml (AHR5) and < or = 25 mg/ml (AHR25), respectively. This analysis explores these marked gender differences in airway hyperresponsiveness (AHR). Relative risks (RR) for predictors of AHR and the 95% confidence intervals (95% CI) were estimated using semiparametric Cox proportional-hazards models. The initial model controlled for age, gender, smoking history, height, and weight. The RR (95% CI) for female gender was 1.75 (1.60, 1.92). When the measured baseline FEV1 was added to the model as a surrogate for airway caliber, the RR for female gender decreased to 1.06 (0.96, 1.18). Thus, in this population of middle-aged smokers with mild COPD, the high prevalence of AHR appears to be associated with a decrease in airway caliber. The higher prevalence of AHR noted in women is due to their having a smaller airway caliber than their male counterparts.


Nucleation and Crystal Growth in a Vitrification Solution Tested for Organ Cryopreservation by Vitrification.
Nucleation and crystal growth are investigated for vitrification solution VS41A (dimethyl sulfoxide, formamide, and 1,2-propanediol) in an aqueous carrier solution giving, when added to these three cryoprotectants, a concentration of other solutes in the whole solution the same as that in Euro-Collins, with a 55% (w/v) cryoprotectant concentration. This concentration is assumed to achieve physical properties under 1 atmosphere similar to those of solution VS4 used under 1000 atmospheres. The thermal range and the kinetics of nucleation and crystal growth are investigated by DSC through different thermal treatments. It is found that the nucleation thermal range is below -90 degrees C and that of crystal growth is above -85 degrees C for a relatively long experimental time. The nucleation density is also studied through direct observations by cryomicroscopy and is related to the amount of crystallization calorimetrically recorded. The effect of storage below the glass transition shows the possibility of a slow increase in nucleation below the glass transition, as already observed by other authors for different aqueous solutions. Isothermal crystallization is analyzed within the Johnson-Mehl-Avrami model for temperatures above -75 degrees C. The corresponding samples have been cooled and warmed at the same rate of 40 degrees C/min and calculations give, at constant nuclei numbers, an activation energy of 9.3 +/- 0.3 kcal/mol and the Avrami exponent n = 2.2 +/- 0.05. This shows a two-dimensional crystal growth as observed by cryomicroscopy. The estimated critical warming rate relevant to the preservation of rabbit kidneys by vitrification is 270 degrees C/min with or without an increase in the nucleus density during storage. The present results support the possibility of using VS4 solution for vitrification of rabbit kidneys if pressure is not a limiting factor. Copyright 1993, 1999 Academic Press.


Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize.
An amaranth ( Amaranthus hypochondriacus) 11S globulin cDNA, encoding one of the most important storage proteins (amarantin) of the seed, with a high content of essential amino acids, was used in the transformation of CIMMYT tropical maize genotype. Constructs contained the amarantin cDNA under the control of a tissue-specific promoter from rice glutelin-1 ( osGT1) or a constitutive ( CaMV 35S) promoter with and without the first maize alcohol dehydrogenase intron ( AdH). Southern-blot analysis confirmed the integration of the amarantin cDNA, and copy number ranged from one to more than ten copies per maize genome. Western-blot and ultracentrifugation analyses of transgenic maize indicate that the expressed recombinant amarantin precursors were processed into the mature form, and accumulated stably in maize endosperm. Total protein and some essential amino acids of the best expressing maize augmented 32% and 8-44%, respectively, compared to non-transformed samples. The soluble expressed proteins were susceptible to digestion by simulated gastric and intestinal fluids, and it is suggested that they show no allergenic activity. These findings demonstrate the feasibility of using genetic engineering to improve the amino acid composition of grain crops.


Mucociliary activities in fetal rabbits.
There have been many morphological investigations on the generation of respiratory epithelium. However, the mucociliary activity of fetal respiratory epithelium has never yet been discussed. In the present work, ciliary activity and mucociliary function in the nose and the trachea of fetal rabbits were studied and, moreover, their respiratory epithelial cells were observed by scanning and transmission electron microscopy. Ciliary activity as noted on the 26th day of fetal life (day 26) for the first time in both of the nasal and the tracheal region and this activity was already equivalent to that in adult rabbits. Mucociliary transport function in either region was first noted on day 27. A quantitative as well as a qualitative immaturity of the respiratory epithelium was recognized on the last day of observation (day 29). The quantitative immaturity is characterized by 1) the ratio of ciliated to non-ciliated cells being lower than in the adult epithelium, 2) each ciliated cell possessing about three-fifths as many cilia as those of a full-grown cell, and 3) some cilia being smaller than full-grown ones and the qualitative immaturity by the directional disorder of the basal foot. No differences were observed between a cilium on day 25 or earlier, another on day 26 or later, and a full-grown cilium. It is suggested that cilia of the respiratory epithelium are morphologically prepared for motion and are activated on day 26 by changes in their surroundings, and that the poorer mucociliary transport in fetuses despite an almost normal ciliary beating is due in fairly large part to their qualitative immaturity (directional disorder of the basal foot).


Distensibility of single capillaries and venules in the rat and frog mesentery.
The distensibility characteristics of single microvessels in the mesentery of frogs and rats has been determined by a new technique capable of resolving a change in radius of about 0.01 micron in vivo. Two water insoluble immiscible oils were micro-injected into occluded segments of the vessels. Since the volume of the oil remained constant, the position of the interface at different applied pressures could be used to determine changes in vessel diameter. The stress/strain relationship of capillaries and venules in the frog mesentery and venules in the rat mesentery were determined and found to be similar to each other. Vessel diameters increased by up to 15% over the whole pressure range used but in a markedly non-linear manner, stiffening at higher strains. It is suggested that the basement membrane has the appropriate properties to explain this behaviour.


Malignant mixed müllerian tumor of primary mesenteric origin associated with a synchronous ovarian cancer: case report and literature review.
Malignant mixed müllerian tumor (MMMT) is a rare tumor in females and extragenital MMMT is even more so. We report a patient with MMMT primarily in the mesentery with synchronous ovarian cancer. In the English literature, 42 cases of extragenital MMMT have been reported other than the presented case, and this is only the second MMMT arising from the mesentery. Furthermore, among the cases reviewed, MMMTs tend to be associated with synchronous or metachronous colonic cancer or gynecologic tumors originating from the müllerian duct, including ovarian tumors, fallopian tube cancer, endometrial cancer, cervical cancer, and serous carcinoma of the peritoneum (14 out of 43 patients; 32.6%). The risk factors for MMMT include obesity, nulliparity, exogenous estrogen, and long-term tamoxifen use. The prognosis of MMMT is catastrophic and the treatment is based on the experience of those of uterine sarcomas, which is composed of operation, radiotherapy and chemotherapy.


Temperature and water activity minima for growth of spoilage moulds from meat.
Five species of fungi were isolated from mould spoilage on meat other than black spot. 'White spot' colonies yielded Chrysosporium pannorum or an Acremonium sp.; 'whiskers' colonies yielded Thamnidium elegans or Mucor racemosus, and blue-green colonies yielded Penicillium corylophilum . Chrysosporium pannorum was moderately xerotolerant with a minimum growth temperature of -5 degrees C. The Acremonium sp. and P. corylophilum showed a similar level of xerotolerance but had a minimum growth temperature of -2 degrees C. Mucor racemosus was no more xerotolerant than many spoilage bacteria and did not grow below -1 degree C, but grew rapidly at 3 degrees C and above. Thamnidium elegans grew at -7 degrees C on supercooled medium and an intrinsic minimum growth temperature of -10 degrees C was indicated. However, the low xerotolerance of this species precluded growth on frozen media below -5 degrees C. It seems therefore that -5 degrees C is the practical limiting temperature for mould growth on meat, and mould spoilage usually indicates that surfaces of freezer stored meats have approached and possibly exceeded 0 degrees C.


Lack of 'acid reversal' of myofibrillar adenosine triphosphatase in masticatory muscle fibres of rhesus monkeys.
Myofibrillar adenosine triphosphatase (ATPase) activity was demonstrated in sections of masseter and temporalis muscles and of selected limb muscles of adult rhesus monkeys. Incubations were performed either with no pre-treatment or after prior incubation in alkaline media (pH 10.2-10.4) or acidic media (pH 3.8-4.6). Without pre-treatment, fibres having high or low ATPase activity were observed in limb and masticatory muscles. Following alkaline pre-incubation the difference between high and low ATPase of limb muscle fibres is accentuated, whereas pre-incubation in acidic media (pH 4.3) results in inhibition of high and potentiation of low ATPase activities ('acid reversal'). While pre-incubation of masticatory muscle sections at pH 10.2 accentuates differences in ATPase activity, pre-incubation at pH 10.4 abolishes ATPase activity. In contrast, masticatory muscle fibres showed no reversal of ATPase activity following acidic pre-incubation (pH 4.3). Pre-incubation at pH 3.8 abolished the ATPase activity of both limb and masticatory muscle fibres. The biochemical basis for the differences in ATPase histochemistry between masticatory and limb muscles is not known.


Thioredoxins in cardiovascular disease.
Key thioredoxin (Trx) system components are nicotinamide adenine dinucleotide phosphate (NADPH), Trx reductase (TrxR), and Trx. TrxR catalyzes disulfide reduction in Trx with NADPH as cofactor. Because Trx is an antioxidant, oxidative stress results in an increase in Trx, which has a reduced disulfide component. If Trx is suppressed, oxidative stress in higher. In contrast a decrease in oxidative stress is associated with low Trx levels. Trx is involved in inflammation, apoptosis, embryogenesis, and cardiovascular disease (CVD). This review focuses on the Trx system in CVD. Abnormal Trx binding occurs in mouse familial combined hyperlipidemia; however, this has not been confirmed in humans. Congestive heart failure is a manifestation of many CVDs, which may be improved by attenuating oxidative stress through the suppression of Trx and decreased reactive oxygen species. Angiotensin II is associated with hypertension and other CVDs, and its receptor blockade results in decreased oxidative stress with reduced Trx levels. Inflammation is a major causative factor of CVDs, and myocarditis as an example, is associated with increased Trx levels. Vascular endothelial dysfunction has an association with CVD. This dysfunction is alleviated by hormone replacement therapy, which involves decreased oxidative stress and Trx levels. Diabetes mellitus has a major association with CVDs; increase in Trx levels may reflect insulin resistance. Identification of Trx system abnormalities may lead to innovative approaches to treat multiple CVDs and other pathologies.


Cultural influences and patient behaviour: some experiences in the paediatric ward of a Nigerian hospital.
In this paper an attempt is made to analyse the incompatibilities between the professional and the layman's ideology in terms of models of illness and their treatment. From an interview with paediatricians at the Ile-Ife Teaching Hospitals Complex in Nigeria, we found out that the conflicting encounters experienced could be divided into 2 groups: opposition as a result of religious ideology, held especially from the Jehovah's Witnesses Sect; and opposition as a result of some child-rearing practices that were being strictly upheld by the mothers of the patients. From the cases cited, we noted that the adherence to some religious ideology was so strong that some patients were willing to withdraw from hospital treatment when it conflicted with a tenet of their religion. Of particular relevance here, was the fact that a religious leader of this sect agreed to a transfusion after he had been assured of secrecy. In this particular case, it was obvious that he feared 'losing face' among his followers. We felt that a more flexible relationship between the doctor and patient would reduce the tension; hence a case was made for a transcultural perspective based on an adequate knowledge of behavioural sciences.


[Effects of urologic surgery on lymphocyte subpopulations].
We studied the postoperative development of lymphocytes, T cells and lymphocyte subpopulations, in peripheral blood in 45 patients. Twenty-five of them were subjected to a TUR of a bladder tumor, 10 to a transvesical prostatectomy, 6 to a pyelolithotomy and 4 to a nephrectomy for renal atrophy secondary to lithiasis. We confirm a significant postoperative drop in the lymphocyte count and in the percentages of OKT 3+ (total T cells), OKT 4+ (helper T cells) and OKT 8+ cells (cytotoxic/suppressor T cells) 2-4 hours after the operation. Its duration varies according to the type of operation carried out. Despite the slight differences observed in the postoperative development of the lymphocyte subpopulations, at no time in the study did we report significant postoperative variations in the OKT 4+/OKT 8+ quotient.


Pulmonary embolism: diagnosis with electron-beam CT.
Contrast material-enhanced electron-beam computed tomography (CT) (100-msec scan time) was performed to image the pulmonary vasculature in 86 patients, each suspected of having a pulmonary embolism (PE). Thromboembolic material was demonstrated in 39 patients; no emboli were demonstrated in 47. In 25 patients, angiographic or pathologic proof was available. There were 19 proved positive CT scans, four proved negative scans, one false-negative scan, and one false-positive scan. In the 21 patients with CT and angiographic correlation, evidence of thromboembolic disease was seen in 88 vascular zones. Fifty-three zones were positive for PE at both CT and angiography. Eighteen zones were positive for PE at angiography alone, and 17 zones were positive at CT alone. Electron-beam CT is a potentially effective noninvasive means of diagnosing PE.


Multicopy STS1 restores both protein transport and ribosomal RNA stability in a new yeast sec23 mutant allele.
A thermosensitive mutant was selected on the basis of its resistance after a heat shock to the lethal effects of lomofungin, a drug that inhibits RNA synthesis. We demonstrate that the single mutation conferring thermosensitivity and lomofungin resistance is heteroallelic to sec23-1. This new allele of sec23 is designated sec23-11. The yeast SEC23 gene has previously been reported to function in protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. After a 1-min shift to elevated temperature, a sec23-1 mutant becomes defective in vesicle formation from the ER. We show here that upon a shift to high temperature, both sec23 alleles confer defects in ER-to-Golgi transport, and both mutants stop accumulating nascent RNA after 20 to 30 min. Transcription and early maturation of ribosomal RNA, the major RNA species, are normal in a sec23-11 mutant, but the matured ribosomal RNA is then degraded. This ribosomal RNA instability may reflect improper assembly of ribosomal subunits, due to failure to localize essential factors such as ribosomal proteins to the nucleus. In addition to the sec23 mutants, other ER-to-Golgi secretion mutants exhibit a strong RNA biosynthetic defect, and we conclude that continued post-ER protein transport, not just a functional SEC23 gene product (Sec23p), is required for ribosomal RNA stability. We also identified an extragenic multicopy suppressor of the sec23-11 mutant, the essential STS1 gene ("Sec Twenty-three Suppressor 1"). Besides restoring partial temperature resistance to the sec23-11 mutant, overexpression of STS1 also overcomes both the RNA synthesis and the protein transport blocks in this mutant. In contrast, multicopy STS1 exerts only small effects on the sec23-1 mutant. The predicted STS1 gene product (Sts1p) is a low-abundance protein of 36,500 daltons, with no signal peptide and no hydrophobic stretch of sufficient length to span a membrane. Like Sec23p, Sts1p is found in a cytosolic compartment. These features suggest that Sts1p may play a catalytic or regulatory role in conjunction with the more abundant Sec23p.


DDB1 and CUL4 associated factor 11 (DCAF11) mediates degradation of Stem-loop binding protein at the end of S phase.
In eukaryotes, bulk histone expression occurs in the S phase of the cell cycle. This highly conserved system is crucial for genomic stability and proper gene expression. In metazoans, Stem-loop binding protein (SLBP), which binds to 3' ends of canonical histone mRNAs, is a key factor in histone biosynthesis. SLBP is mainly expressed in S phase and this is a major mechanism to limit bulk histone production to the S phase. At the end of S phase, SLBP is rapidly degraded by proteasome, depending on two phosphorylations on Thr 60 and Thr 61. Previously, we showed that SLBP fragment (aa 51-108) fused to GST, is sufficient to mimic the late S phase (S/G2) degradation of SLBP. Here, using this fusion protein as bait, we performed pull-down experiments and found that DCAF11, which is a substrate receptor of CRL4 complexes, binds to the phosphorylated SLBP fragment. We further confirmed the interaction of full-length SLBP with DCAF11 and Cul4A by co-immunoprecipitation experiments. We also showed that DCAF11 cannot bind to the Thr61/Ala mutant SLBP, which is not degraded at the end of S phase. Using ectopic expression and siRNA experiments, we demonstrated that SLBP expression is inversely correlated with DCAF11 levels, consistent with the model that DCAF11 mediates SLBP degradation. Finally, we found that ectopic expression of the S/G2 stable mutant SLBP (Thr61/Ala) is significantly more toxic to the cells, in comparison to wild type SLBP. Overall, we concluded that CRL4-DCAF11 mediates the degradation of SLBP at the end of S phase and this degradation is essential for the viability of cells.


A guideline for the outpatient management of glycaemic control in people with cancer.
Individuals with cancer are at increased risk of developing new-onset diabetes mellitus and hyperglycaemia, and an estimated 20% of people with cancer already have an underlying diagnosis of diabetes mellitus. People with both cancer and diabetes may have an increased risk of toxicities, hospital admissions and morbidity, with hyperglycaemia potentially attenuating the efficacy of chemotherapy often secondary to dose reductions and early cessation. Numerous studies have demonstrated that hyperglycaemia is prognostic of worse overall survival and risk of cancer recurrence. These guidelines aim to provide the oncology/haemato-oncology and diabetes multidisciplinary teams with the tools to manage people with diabetes commencing anti-cancer/glucocorticoid therapy, as well as identifying individuals without a known diagnosis of diabetes who are at risk of developing hyperglycaemia and new-onset diabetes.


Cellular Localization and Trafficking of the Human ABCG1 Transporter.
We have developed a suitable heterologous cell expression system to study the localization, trafficking, and site(s) of function of the human ABCG1 transporter. Increased plasma membrane (PM) and late endosomal (LE) cholesterol generated by ABCG1 was removed by lipoproteins and liposomes, but not apoA-I. Delivery of ABCG1 to the PM and LE was required for ABCG1-mediated cellular cholesterol efflux. ABCG1 LEs frequently contacted the PM, providing a collisional mechanism for transfer of ABCG1-mobilized cholesterol, similar to ABCG1-mediated PM cholesterol efflux to lipoproteins. ABCG1-mobilized LE cholesterol also trafficked to the PM by a non-vesicular pathway. Transfer of ABCG1-mobilized cholesterol from the cytoplasmic face of LEs to the PM and concomitant removal of cholesterol from the outer leaflet of the PM bilayer by extracellular acceptors suggests that ABCG1 mobilizes cholesterol on both sides of the lipid bilayer for removal by acceptors. ABCG1 increased uptake of HDL into LEs, consistent with a potential ABCG1-mediated cholesterol efflux pathway involving HDL resecretion. Thus, ABCG1 at the PM mobilizes PM cholesterol and ABCG1 in LE/LYS generates mobile pools of cholesterol that can traffic by both vesicular and non-vesicular pathways to the PM where it can also be transferred to extracellular acceptors with a lipid surface.


Historical perspectives in the aetiology of ankylosing spondylitis.
A brief historical review of the aetiology of ankylosing spondylitis is presented. A recurrent theme has been diathesis and infection. Diathesis has now been pinpointed to the 6th chromosome in the form of HLA-B27, but, as yet, we know little of any infective aetiology for ankylosing spondylitis itself.


An unusual heparin-binding peptide from the carboxy-terminal hep-2 region of fibronectin.
A synthetic 22 residue peptide, N22W, with sequence NVSPPRRARVTDATETTITISW, derived from the amino terminus of type III module 13 in the carboxy-terminal hep-2 domain of fibronectin, was found to exhibit unusual heparin binding properties. Titration of fluoresceinamine-labeled heparin (FA-heparin) with N22W at 25 degrees C and pH 7.4 in 0.02 M Tris buffer containing 0.15 M NaCl (TBS) produced a cooperative sigmoidal increase in fluorescence polarization anisotropy with half-saturation near 2.5 microM. The increase in anisotropy was even greater than that produced by the much larger 30-kDa hep-2 fragment of fibronectin and saturation was achieved at lower concentration. Simply deleting the C-terminal Trp from the peptide abolished its heparin-binding activity as did deletion of residues TETTITIS or mutation of the RR doublet to SS. Further analysis suggested that peptide-peptide interactions mediated by the carboxy-terminal region of N22W play an important role in its binding to heparin. A branched tetrameric peptide containing four copies of N21S caused a nearly hyperbolic increase in anisotropy of FA-heparin with an apparent Kd of 0.3 microM in TBS, 10-fold lower than that of the monomer or of the parent domain from which the peptide was derived. The results illustrate that peptide-peptide interactions can lead to stronger binding by allowing multiple points of contact with the negatively charged polysaccharide.


KRIBB3, a novel microtubule inhibitor, induces mitotic arrest and apoptosis in human cancer cells.
KRIBB3 (5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl) isoxazole) inhibited cancer cell growth in vitro and in vivo. Flow cytometry studies showed that KRIBB3 caused cell cycle arrest at the G(2)/M phase and subsequent apoptosis. This was confirmed as accumulation of Cyclin B1 and cleavage of poly(ADP-ribose) polymerase (PARP) were detected. While transient inhibition by KRIBB3 led to reversible mitotic arrest, prolonged exposure to KRIBB3-induced apoptosis. Co-immunoprecipitation experiments showed that KRIBB3 initially induced association of inhibitory Mad2 with p55CDC (mammalian homologue of CDC20), an activator of APC/C (anaphase-promoting complex/cyclosome), suggesting that the mitotic spindle checkpoint was activated by KRIBB3. However, the level of this inhibitory complex of Mad2 with p55CDC was gradually decreased 24 h after KRIBB3 treatment, and was hardly detectable after 48 h, indicating some slipping of the mitotic checkpoint. Consistent with these observations, KRIBB3 activated the mitotic spindle checkpoint by disrupting the microtubule cytoskeleton. KRIBB3 was proven to be a tubulin inhibitor using in vitro polymerization assays and in vivo indirect immunofluorescence staining. The temporal pattern of Bax activation by KRIBB3 was similar to PARP cleavage, suggesting that Bax is a mediator of KRIBB3-dependent apoptosis. Furthermore, when KRIBB3 was administered intraperitoneally into nude mice at 50 mg/kg or 100 mg/kg, it inhibited 49.5 or 70.3% of tumor growth, respectively. These results suggest that KRIBB3 is a good drug candidate for cancer therapy.


RME: effects on the nasal septum. A CBCT evaluation.
Three-dimensional images have been evaluated to study the changes that occur on the maxillary complex and the nasal septum, following rapid maxillary expansion (RME) in growing children. The records of 39 consecutively treated patients (20 male, 19 female), aged between 6.2 and 12.3 years (mean 8.6 years, S.D. 1.5), presenting a septal deviation of more than 1 mm, were analysed. Each patient was scanned at two distinct periods. The first image (T0) was obtained before treatment and hence represented the patient's baseline condition prior to expansion. The second time point (T1) was set after removal of the expander. The skeletal modifications induced by RME have been evaluated via cephalometric tracings obtained at T0 and at T1. The time lapse between T0 and T1 was 12 months. The nasal septum has been modified in its length more significantly in its lower rather than its upper tract. The width of the piriform aperture increased of 1.58 mm with an increase in the lateral walls and a downward movement of the nasal floor (consequent to an increase in septal length). There was also a reduction of septal deviation. The control group measurements did not vary during the considered period. Evidences of this study suggest that RME procedures can result in: a potentially positive effect on the nasal septum asymmetry during childhood; an increase of the nasal cavity volume due to a downward and forward movement; a lateral inclination of the nasal cavity reducing air resistance and improving breathing pattern; an increase of the length of the septum in its lower third improving its possible deviations and growth disturbances; favourable effects on the growth of the entire maxillary complex.


Expanded CCUG repeat RNA expression in Drosophila heart and muscle trigger Myotonic Dystrophy type 1-like phenotypes and activate autophagocytosis genes.
Myotonic dystrophies (DM1-2) are neuromuscular genetic disorders caused by the pathological expansion of untranslated microsatellites. DM1 and DM2, are caused by expanded CTG repeats in the 3'UTR of the DMPK gene and CCTG repeats in the first intron of the CNBP gene, respectively. Mutant RNAs containing expanded repeats are retained in the cell nucleus, where they sequester nuclear factors and cause alterations in RNA metabolism. However, for unknown reasons, DM1 is more severe than DM2. To study the differences and similarities in the pathogenesis of DM1 and DM2, we generated model flies by expressing pure expanded CUG ([250]×) or CCUG ([1100]×) repeats, respectively, and compared them with control flies expressing either 20 repeat units or GFP. We observed surprisingly severe muscle reduction and cardiac dysfunction in CCUG-expressing model flies. The muscle and cardiac tissue of both DM1 and DM2 model flies showed DM1-like phenotypes including overexpression of autophagy-related genes, RNA mis-splicing and repeat RNA aggregation in ribonuclear foci along with the Muscleblind protein. These data reveal, for the first time, that expanded non-coding CCUG repeat-RNA has similar in vivo toxicity potential as expanded CUG RNA in muscle and heart tissues and suggests that specific, as yet unknown factors, quench CCUG-repeat toxicity in DM2 patients.


Perioperative Red Blood Cell Transfusion: Harmful or Beneficial to the Patient?
Although the transfusion of red blood cells (RBCs) is safer than ever regarding infections, it is still associated with several adverse reactions and therefore should only be used on the basis of evidence-based triggers. However, prevention of RBC transfusion and subsequent substitution of blood losses with acellular solutions will inevitably result in dilutional anemia. Acute dilutional anemia can be compensated by the body over a wide range of hemoglobin concentrations without a critical restriction of tissue oxygenation. On the other hand, chronic anemia is known to be a potent cause of morbidity and mortality. As a consequence, the impact of perioperative anemia on mortality is difficult to describe, because anemia, as well as the transfusion of RBCs, can influence the clinical outcome. The resulting 'Gordian knot' cannot be cut easily, and this circumstance forces clinical physicians to make a daily trade-off between transfusion-associated and anemia-associated risks. This review focuses on the physiology of oxygen transport, the hazards of acute anemia, the hazards of RBC transfusion, and the literature putting these problems into perspective.


Experience of Shame Mediates the Relationship Between Pathological Narcissism and Suicidal Ideation in Psychiatric Outpatients.
Pathological narcissism, described by 2 dysfunctional phenotypic forms-grandiosity and vulnerability-has often been connected to suicidal tendencies in theoretical and clinical literatures. Furthermore, shame proneness has been implicated as a key mechanism that links these 2 constructs. However, empirical evidence for the presumed relationship between pathological narcissism and suicidal tendencies is sparse, and no prior research has investigated the role of shame proneness in this relationship. The objective of the present research was to investigate the complex relations among pathological narcissism, experience of shame, and suicidal ideation in psychiatric outpatients. A sample of 250 adult psychiatric outpatients (61% female; mean age 39.15 years) were assessed between January and May 2014. The participants filled out the Pathological Narcissism Inventory, the Experience of Shame Scale, and the Suicide Assessment Scale-self-rating. Narcissistic vulnerability was found to have unique positive associations with acute suicidal ideation, whereas narcissistic grandiosity exhibited substantially weaker relations with the same construct. Two dimensions of shame-characterological and bodily shame-mediated the relationship between narcissistic vulnerability and suicidal ideation. The mediating role of behavioral shame was not demonstrated. Narcissistic vulnerability seems to be more strongly related to suicidal tendencies than narcissistic grandiosity, while experience of shame represents one of the underlying mechanisms of this relationship among psychiatric outpatients. The implications of these findings are discussed in relation to the role of pathological narcissism in the psychotherapeutic management of suicidality.


GHB: a life-threatening drug complications and outcome of GHB detoxification treatment-an observational clinical study.
GHB (gammahydroxybutyrate) and its precursors are popular recreational drugs due to their sedative, anxiolytic and sexually stimulating effects. Their use has been steadily increasing in recent years. The detoxification process is complex and prone to high rates of complications while little is known about the pathophysiology. This study aims to elucidate the characteristics of GHB-addicted patients and to evaluate the risks and complications of GHB withdrawal treatment. This observational study describes prospectively the socioeconomic status, clinical history and course of inpatient detoxification treatment of a group of 39 patients suffering from GHB substance use disorder. Detoxification treatment took place in a highly specialized psychiatric inpatient unit for substance use disorders. GHB patients were characterised by being young, well-educated and by living alone. More than 50% of the patients had no regular income. The patients were male and female in equal numbers. Detoxification treatment was complicated, with high rates of delirium (30.8%) and high need for intensive care (20.5%). In our sample, GHB users were young, well-educated people and male and female in equal number. Detoxification proved to be dangerous for GHB-addicted patients. The presence of delirium and the need for transfer to an intensive care unit during detoxification treatment was extraordinarily high, even with appropriate clinical treatment. The reasons for this remain unknown. Therefore an intensive care unit should be available for GHB detoxification treatment. Further studies are needed to evaluate the options for prophylactic treatment of delirium during detoxification.


Primary polycythaemia in a dog.
A 7-year-old, crossbred, desexed bitch had a fluctuating illness of more than 6 months duration, with signs of inappetence, reduced exercise tolerance, restlessness, panting, black diarrhoea, and red eyes and ears. Haematological examination disclosed persistent polycythaemia, reticulocytosis, normoblastaemia , and normal total plasma protein concentration when not dehydrated. Splenomegaly was present, but there was no evidence of significant pulmonary, cardiac or renal disease on physical, radiographic or electrocardiographic examination. Further investigations revealed normal arterial Po2 and oxygen saturation, and increased total erythrocyte volume. Primary polycythaemia was diagnosed. The dog was rendered anaemic by bleeding, then treated with a single injection of radioactive phosphorus. The patient has had no recurrence of signs and remains well 50 months later.


Application of a new combined model including radiological indicators to predict difficult airway in patients undergoing surgery for cervical spondylosis.
Airway management is crucial in clinical anesthesia. Many complications associated with airway management result from unexpected difficult airway, but predicting a difficult airway is a major challenge. We investigated the efficacy of a new combined model including radiological indicators to predict difficult airway in patients undergoing surgery for cervical spondylosis, a population with a high incidence of difficult airway. We randomly enrolled 303 patients scheduled for elective surgery for cervical spondylosis at Peking University Third Hospital between August 2012 and March 2013. Preoperatively, patients were evaluated for difficult airway according to a clinical index and parameters on lateral cervical radiographs and magnetic resonance images. Difficult airway was defined as Cormack-Lehane grades III-IV. Logistic regression was used to identify a combined (clinical and radiological) model for difficult airway. A receiver operating characteristic (ROC) curve was used to describe the effectiveness of prediction. We identified three clinical predictive factors using the ROC curve: mouth opening, sternomental distance, and neck mobility. We created a clinical model using three factors: gender, age, and mouth opening, with odds ratios (OR) of 0.370, 1.034, and 0.358, respectively. Using the clinical and radiological parameters, we formulated a combined model with five risk factors: gender, mouth opening, atlanto-occipital gap, the angle from the second to sixth cervical vertebraes in the neutral position, and the angle difference of d (the angle between the laryngeal axis and the epiglottic axis) from the neutral position to extension (OR: 0.107, 0.355, 0.846, 1.057, and 0.952, respectively). The sensitivity and specificity of the combined model were 80.0% and 65.7%, respectively, and the ROC curve confirmed that the combined model was better than any single clinical predictor and the clinical model. The efficacy of the combined model including both clinical and radiological indicators was better than any single clinical predictor or the clinical model in patients undergoing elective surgery for cervical spondylosis.


The Medical Data Index (MDI) dependency module: a shared database to assist discharge planning and audit.
As part of the development of integrated, patient-based hospital information technology (IT) systems in the South-Western Region of England, a module has been developed which will hold core data pertaining to the functional status and current resources of elderly or disabled patients. Its purpose is to assist early identification of unmet needs and facilitate prompt transfer to community care. The module provides a shared database, which is completed or updated as necessary on admission and is then available to all appropriate users of the hospital system, avoiding duplication of data collection. In addition to details of home circumstances and support, it includes brief, standardized assessment scales for activities of daily living and mental state, which will identify the need for specialist referral. A summary is provided for easy communication with other care agencies.


Leg ulceration in sickle cell disease: medieval medicine in a modern world.
Leg ulceration is now recognized as an important complication of sickle cell disease, especially of the SS genotype. Since there is no convincing evidence of delayed healing of operation scars or of wounds elsewhere in the body, it must be concluded that factors specific to the lower leg render patients prone to delayed healing at this site. Many lesions are traumatic in origin and since there is considerable variation in healing rates among the normal population, it is useful to define chronic leg ulceration on the basis of a minimal duration, which in Jamaican studies has required at least 3 months and sometimes 6 months before healing. This minimal duration avoids the difficulties of interpreting the significance of briefer lesions since the moment of final healing may be poorly defined (patients may conclude that a scab represents healing whereas small lesions persist beneath) and often goes undocumented as patients may not report and medical attendants may not enquire, the date of final healing.


The effects of naloxone on human breast cancer progression: in vitro and in vivo studies on MDA.MB231 cells.
Naloxone is viewed as a specific competitive opioid antagonist acting at the level of opioid receptors (μ, δ, and κ) with blended agonist-adversary or agonist action. The role of naloxone in tumor cell growth has been poorly studied in human cancer cell lines. In the present study, we report findings from in vitro and in vivo experiments performed to evaluate the effects of naloxone on human breast cancer cell growth and progression. In vitro assays were conducted on estrogen receptor-negative human breast carcinoma cells, MDA.MB231, treated with naloxone at different concentrations (10-100 μM). In vivo experiments were performed on a mouse model of human triple-negative breast cancer generated by using MDA.MB231 injected subcutaneously in mice. Naloxone was daily intraperitoneally injected in mice at 0.357 mg/kg for 2 weeks and at 0.714 mg/kg for the next 2 weeks. Microvessels formation was detected by fluorescein isothiocyanate-dextran (100 μL) injected into the tail vein of mice and confirmed by immunohistochemistry with CD31 on mice tumor sections. In vitro tests showed that the cell proliferation of MDA.MB231 was inhibited by naloxone in a dose-dependent manner, whereas the cell death was increased. In vivo studies demonstrated that tumors of mice treated with naloxone were significantly smaller than those observed in the control groups, as long as naloxone was administered. Finally, naloxone was not able to impair the microvessel formation in tumors of treated mice. Our data showed, for the first time, that naloxone reduced breast cancer progression without affecting angiogenesis.


A Practical Guide to the Measurement and Analysis of DNA Methylation.
DNA methylation represents a fundamental epigenetic mark that is associated with transcriptional repression during development, maintenance of homeostasis, and disease. In addition to methylation-sensitive PCR and targeted deep-amplicon bisulfite sequencing to measure DNA methylation at defined genomic loci, numerous unsupervised techniques exist to quantify DNA methylation on a genome-wide scale, including affinity enrichment strategies and methods involving bisulfite conversion. Both affinity-enriched and bisulfite-converted DNA can serve as input material for array hybridization or sequencing using next-generation technologies. In this practical guide to the measurement and analysis of DNA methylation, the goal is to convey basic concepts in DNA methylation biology and explore genome-scale bisulfite sequencing as the current gold standard for assessment of DNA methylation. Bisulfite conversion chemistry and library preparation are discussed in addition to a bioinformatics approach to quality assessment, trimming, alignment, and methylation calling of individual cytosine residues. Bisulfite-converted DNA presents challenges for standard next-generation sequencing library preparation protocols and data-processing pipelines, but these challenges can be met with elegant solutions that leverage the power of high-performance computing systems. Quantification of DNA methylation, data visualization, statistical approaches to compare DNA methylation between sample groups, and examples of integrating DNA methylation data with other -omics data sets are also discussed. The reader is encouraged to use this article as a foundation to pursue advanced topics in DNA methylation measurement and data analysis, particularly the application of bioinformatics and computational biology principles to generate a deeper understanding of mechanisms linking DNA methylation to cellular function.


Transvaginal sonographic evaluation of endometrial polyps.
The purpose of this study was to correlate the sonographic and histologic findings of focal lesions of the endometrium as depicted by transvaginal sonography. Sixteen focal endometrial masses were encountered during an 8 month period. The transvaginal sonographic appearance was correlated with findings at surgery or hysteroscopy. All uniformly hyperechoic endometrial masses were benign endometrial polyps. Five heterogeneous lesions were complicated endometrial polyps; one of these showed a focus of endometrial carcinoma. Four small and uniformly hypoechoic lesions were all prolapsed submucosal leiomyomata. Our results showed that a well-defined, uniformly hyperechoic mass within the endometrial cavity is most suggestive of a benign endometrial polyp. Larger masses with a heterogeneous appearance are nonspecific and warrant further evaluation and biopsy.


Development and verification of a mouse model for Roux-en-Y gastric bypass surgery with a small gastric pouch.
Existing mouse models of Roux-en-Y gastric bypass (RYGB) surgery are not comparable to human RYGB in gastric pouch volume for a large or absent gastric volume. The aim of this study was to develop and characterize a mouse RYGB model that closely replicates gastric pouch size of human RYGB surgery of about 5% of total gastric volume. We established this model in diet-induced obese (DIO) mice of C57BL/6J. This surgery resulted in a sustained 30% weight loss, entirely accounted for by decreased fat mass but not lean mass, compared to sham-operated mice on the high fat diet. Compared to sham-operated mice, energy expenditure corrected for total body weight was significantly increased by about 25%, and substrate utilization was shifted toward higher carbohydrate utilization at 8 weeks after RYGB when body weight had stabilized at the lower level. The energy expenditure persisted and carbohydrate utilization was even more pronounced when the mice were fed chow diet. Although significantly increased during daytime, overall locomotor activity was not significantly different. In response to cold exposure, RYGB mice exhibited an improved capacity to maintain the body temperature. In insulin tolerance test, exogenous insulin-induced suppression of plasma glucose levels was significantly greater in RYGB mice at 4 weeks after surgery. Paradoxically, food intake measured at 5 weeks after surgery was significantly increased, possibly in compensation for increased fecal energy loss and energy expenditure. In conclusion, this new model is a viable alternative to existing murine RYGB models and the model matches human RYGB surgery in anatomy. This model will be useful for studying molecular mechanisms involved in the beneficial effects of RYGB on body weight and glucose homeostasis.


Improved music perception with explicit pitch coding in cochlear implants.
Music perception and appraisal is very poor in cochlear implant (CI) subjects partly because (musical) pitch is inadequately transmitted by the current clinically used sound processors. A new sound processing scheme (F0mod) was designed to optimize pitch perception, and its performance for music and pitch perception was compared in four different experiments to that of the current clinically used sound processing scheme (ACE) in six Nucleus CI24 subjects. In the F0mod scheme, slowly varying channel envelopes are explicitly modulated sinusoidally at the fundamental frequency (F0) of the input signal, with 100% modulation depth and in phase across channels to maximize temporal envelope pitch cues. The results of the four experiments show that: (1) F0 discrimination of single-formant stimuli was not significantly different for the two schemes, (2) F0 discrimination of musical notes of five instruments was three times better with the F0mod scheme for F0 up to 250 Hz, (3) melody recognition of familiar Flemish songs (with all rhythm cues removed) was improved with the F0mod scheme, and (4) estimates of musical pitch intervals, obtained in a musically trained CI subject, matched more closely the presented intervals with the F0mod scheme. These results indicate that explicit F0 modulation of the channel envelopes improves music perception in CI subjects.


The YlmG protein has a conserved function related to the distribution of nucleoids in chloroplasts and cyanobacteria.
Reminiscent of their free-living cyanobacterial ancestor, chloroplasts proliferate by division coupled with the partition of nucleoids (DNA-protein complexes). Division of the chloroplast envelope membrane is performed by constriction of the ring structures at the division site. During division, nucleoids also change their shape and are distributed essentially equally to the daughter chloroplasts. Although several components of the envelope division machinery have been identified and characterized, little is known about the molecular components/mechanisms underlying the change of the nucleoid structure. In order to identify new factors that are involved in the chloroplast division, we isolated Arabidopsis thaliana chloroplast division mutants from a pool of random cDNA-overexpressed lines. We found that the overexpression of a previously uncharacterized gene (AtYLMG1-1) of cyanobacterial origin results in the formation of an irregular network of chloroplast nucleoids, along with a defect in chloroplast division. In contrast, knockdown of AtYLMG1-1 resulted in a concentration of the nucleoids into a few large structures, but did not affect chloroplast division. Immunofluorescence microscopy showed that AtYLMG1-1 localizes in small puncta on thylakoid membranes, to which a subset of nucleoids colocalize. In addition, in the cyanobacterium Synechococcus elongates, overexpression and deletion of ylmG also displayed defects in nucleoid structure and cell division. These results suggest that the proper distribution of nucleoids requires the YlmG protein, and the mechanism is conserved between cyanobacteria and chloroplasts. Given that ylmG exists in a cell division gene cluster downstream of ftsZ in gram-positive bacteria and that ylmG overexpression impaired the chloroplast division, the nucleoid partitioning by YlmG might be related to chloroplast and cyanobacterial division processes.


Targeting epidermal growth factor-overexpressing triple-negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor.
Traditional cancer therapy and regular immunotherapy are ineffective for treating triple-negative breast cancer (TNBC) patients. Recently, chimeric antigen receptor-engineered natural killer cells (CAR NK) have been applied to target several hormone receptors on different cancer cells to improve the efficacy of immunotherapy. Furthermore, epidermal growth factor receptor (EGFR) is a potential therapeutic target for TNBC. Here, we demonstrated that EGFR-specific CAR NK cells (EGFR-CAR NK cells) could be potentially used to treat patients with TNBC exhibiting enhanced EGFR expression. We investigated the cytotoxic effects of EGFR-CAR NK cells against TNBC cells in vitro and in vivo. The two types of EGFR-CAR NK cells were generated by transducing lentiviral vectors containing DNA sequences encoding the single-chain variable fragment (scFv) regions of the two anti-EGFR antibodies. The cytotoxic and anti-tumor effects of the two cell types were examined by performing cytokine release and cytotoxicity assays in vitro, and tumor growth assays in breast cancer cell line-derived xenograft (CLDX) and patient-derived xenograft (PDX) mouse models. Both EGFR-CAR NK cell types were activated by TNBC cells exhibiting upregulated EGFR expression and specifically triggered the lysis of the TNBC cells in vitro. Furthermore, the two EGFR-CAR NK cell types inhibited CLDX and PDX tumors in mice. This study suggested that treatment with EGFR-CAR NK cells could be a promising strategy for TNBC patients.


Bilateral adrenalectomy of lambs in utero: effects on maternal hormone levels at induced parturition.
Progesterone, 17 alpha, 20 alpha-dihydroxypregn-4-en-3-one, androstenedione, total unconjugated oestrogen and oestrone sulphate have been measured by radioimmunoassays in maternal utero-ovarian venous, maternal peripheral venous and/or foetal posterior vena caval plasma from six sheep bearing bilaterally adrenalectomized lambs, in which premature parturition was induced by administration of glucocorticoid. Three of the ewes were overiectomized, and in one of these three animals the foetal testes were also excised, at the time of foetal adrenalectomy. Adrenalectomy was judged to be complete on the basis of plasma cortisol levels in the neonatal lambs, and by examination of the site of ablation at necropsy. In all cases foetal administration of glucocorticoid led to the onset of labour, and lambing, and in all animals the hormonal changes preceding parturition were indistinguishable (either qualitatively or quantitatively) from the changes observed in animals carrying intact lambs. Since therapy with glucocorticoid alone successfully compensates for ablation of the foetal adrenal cortex, it is suggested that glucocorticoid is the only adrenal product required to cause parturition, and that foetal adrenal secretion of androgens may be unnecessary.


Energizing Richard Wolf E-Line stricture blades can result in electrosurgical burns.
Stricture blades sold by Richard Wolf for use with its E-Line resectoscopes can be mistaken for electrodes and used for electrosurgical applications, potentially resulting in alternate-site electrosurgical burns. Users of these products--and similar products that may be marketed by other endoscopy suppliers--must be adequately trained on the proper identification and use of these instruments.


Oxidative Stability of Sardine Oil Embedded in Spray-dried Egg White Powder and Its Use for n-3 Unsaturated Fatty Acid Fortification of Cookies.
Fine droplets of sardine oil either embedded in or covered with egg white powder, which had been prepared from an oil-in-water emulsion containing 10% protein-oil (9:1) mixture by spray-drying and freeze-drying, were examined for their oxidative stability during storage under moderate conditions (RH = 45-55%, 40°C). The antioxidative effect was estimated by measuring the peroxide value as well as the residual unsaturated fatty acid (mainly eicosapentaenoic and docosahexaenoic acids). Microencapsulation of the oil droplets with albumen by freeze-drying was not so effective in stabilizing against oxidation as that by spray-drying, probably because of the porosity of a protein coating on oil or leakage of oil through crevices. As a practical application of powdered sardine oil (i.e., entrapped in spray-dried albumen particles), plain cookies and those enriched with the oil were baked, and any difference in taste between them was evaluated by a paired preference test assessed by 32 amateur panelists. Sardine oil fortification of the cookies was judged not to affect their quality from the results of the sensory test. Spray-dried egg white powder inclusive of sardine oil was stable during prolonged storage, so that its use would be favorable for supplying an n-3 series of polyunsaturated fatty acids.


Distal Pancreatectomy with Celiac Axis Resection (Modified Appleby Procedure) and Arterial Reconstruction for Locally Advanced Pancreatic Adenocarcinoma After FOLFIRINOX Chemotherapy and Chemoradiation Therapy.
Resectability of pancreatic carcinoma (PC) is directly linked to vascular extension (Tempero MA et al. in J Natl Compr Canc Netw 15(8):1028-1061, 2017. https://doi.org/10.6004/jnccn.2017.0131 ; Isaji S et al. in Pancreatology 18(1):2-11, 2018. https://doi.org/10.1016/j.pan.2017.11.011 ). Involvement of the celiac axis (CA) is typically a contraindication to surgery. High postoperative morbidity and subsequent poor prognosis have been observed in this case, especially for contact > 180° requiring arterial resection (Tempero MA et al. 2017). Recent medical advances in PC treatment, such as FOLFIRINOX-based chemotherapy eventually followed by chemoradiation therapy, offer the potential to select tumour for surgery and to obtain a negative-margin resection even in case of unresectable PC at diagnosis (Suker M et al. in Lancet Oncol 17(6):801-10, 2016. https://doi.org/10.1016/s1470-2045(16)00172-8 ; Pietrasz D et al. in Ann Surg Oncol 26(1):109-117, 2019. https://doi.org/10.1245/s10434-018-6931-6 ). A major pathologic response has been observed in more than 20% of patients after this treatment and is associated with an improved survival (Suker M et al. 2016; Pietrasz D et al. 2019). This evolution allows aggressive surgical strategies with the possibility of long-term disease control for patients showing a good response to induction treatment. This video presents the case of a 66-year-old man diagnosed with a locally advanced ductal adenocarcinoma of the pancreatic body with a 360° involvement of the CA and the hepatic artery. After eight courses of FOLFIRINOX chemotherapy and a capecitabin-based chemoradiation, a surgical exploration was planned for potential resection. The key steps of the procedure are presented, i.e. surgical exposition, assessment of resectability with frozen sections of peri-arterial tissues, en bloc resection (Strasberg SM et al. in Surgery 133(5):521-527, 2003. https://doi.org/10.1067/msy.2003.146 ), and primary end-to-end arterial reconstruction. A modified Appleby operation for locally advanced PC is a technically challenging but feasible procedure in experienced teams. It offers the possibility of en bloc R0 resection of a locally advanced PC with the potential of long-term disease local control. This video may help surgeons to perform this complex intervention.


Light Regulation of LoCOP1 and Its Role in Floral Scent Biosynthesis in Lilium 'Siberia'.
Light is an important environmental signal that governs plant growth, development, and metabolism. Constitutive photomorphogenic 1 (COP1) is a light signaling component that plays a vital role in plant light responses. We isolated the COP1 gene (LoCOP1) from the petals of Lilium 'Siberia' and investigated its function. The LoCOP1 protein was found to be the most similar to Apostasia shenzhenica COP1. LoCOP1 was found to be an important factor located in the nucleus and played a negative regulatory role in floral scent production and emission using the virus-induced gene silencing (VIGS) approach. The yeast two-hybrid, β-galactosidase, and bimolecular fluorescence complementation (BiFC) assays revealed that LoCOP1 interacts with LoMYB1 and LoMYB3. Furthermore, light modified both the subcellular distribution of LoCOP1 and its interactions with LoMYB1 and MYB3 in onion cells. The findings highlighted an important regulatory mechanism in the light signaling system that governs scent emission in Lilium 'Siberia' by the ubiquitination and degradation of transcription factors via the proteasome pathway.


[Monophasic Salmonella Typhimurium outbreak due to the consumption of roast pork meat].
This report presents an outbreak of monophasic Salmonella enteric serovar Typhimurium fagotipe 4, 5, 12: i:-, in a motorcycle concentration in Valladolid. Information was collected to one hundred and twelve affected from seven Spanish Autonomous Communities. The epidemiological investigation associated the outbreak with the consumption of roast pork with sauce sandwiches sold at a street market in that event.


Spreading vasodilatation in the murine microcirculation: attenuation by oxidative stress-induced change in electromechanical coupling.
Regulation of blood flow in microcirculatory networks depends on spread of local vasodilatation to encompass upstream arteries; a process mediated by endothelial conduction of hyperpolarization. Given that endothelial coupling is reduced in hypertension, we used hypertensive Cx40ko mice, in which endothelial coupling is attenuated, to investigate the contribution of the renin-angiotensin system and reduced endothelial cell coupling to conducted vasodilatation of cremaster arterioles in vivo. When the endothelium was disrupted by light dye treatment, conducted vasodilatation, following ionophoresis of acetylcholine, was abolished beyond the site of endothelial damage. In the absence of Cx40, sparse immunohistochemical staining was found for Cx37 in the endothelium, and endothelial, myoendothelial and smooth muscle gap junctions were identified by electron microscopy. Hyperpolarization decayed more rapidly in arterioles from Cx40ko than wild-type mice. This was accompanied by a shift in the threshold potential defining the linear relationship between voltage and diameter, increased T-type calcium channel expression and increased contribution of T-type (3 μmol l(-1) NNC 55-0396), relative to L-type (1 μmol l(-1) nifedipine), channels to vascular tone. The change in electromechanical coupling was reversed by inhibition of the renin-angiotensin system (candesartan, 1.0 mg kg(-1) day(-1) for 2 weeks) or by acute treatment with the superoxide scavenger tempol (1 mmol l(-1)). Candesartan and tempol treatments also significantly improved conducted vasodilatation. We conclude that conducted vasodilatation in Cx40ko mice requires the endothelium, and attenuation results from both a reduction in endothelial coupling and an angiotensin II-induced increase in oxidative stress. We suggest that during cardiovascular disease, the ability of microvascular networks to maintain tissue integrity may be compromised due to oxidative stress-induced changes in electromechanical coupling.