Complementary detection of prostate-specific antigen using In2O3 nanowires and carbon nanotubes.
We report complementary detection of prostate-specific antigen (PSA) using n-type In2O3 nanowires and p-type carbon nanotubes. Our innovation involves developing an approach to covalently attach antibodies to In2O3 NW surfaces via the onsite surface synthesis of phosphonic acid-succinylimide ester. Electronic measurements under dry conditions revealed complementary response for In2O3 NW and SWNT devices after the binding of PSA. Real-time detection in solution has also been demonstrated for PSA down to 5 ng/mL, a benchmark concentration significant for clinical diagnosis of prostate cancer, which is the most frequently diagnosed cancer.


Asymptomatic peripheral artery disease can limit maximal exercise capacity in chronic obstructive pulmonary disease patients regardless of airflow obstruction and lung hyperinflation.
Background Silent/asymptomatic peripheral artery disease may occur in patients with chronic obstructive pulmonary disease, but it is poorly investigated. The primary aim of this study was to evaluate in chronic obstructive pulmonary disease patients the impact of asymptomatic/silent peripheral artery disease on maximal exercise capacity; the secondary aim was to search for predictors of peripheral artery disease. Methods We prospectively enrolled chronic obstructive pulmonary disease outpatients. Data on anthropometric characteristics, lung function, cardiopulmonary exercise test and ankle-brachial index were recorded. The cut-off of ankle-brachial index used to define patients with peripheral artery disease was ≤0.90. Results We studied 47 patients and found 24 patients (51%) who showed peripheral artery disease. As compared to patients without peripheral artery disease, patients with peripheral artery disease had lower values of peak oxygen uptake, peak workload, energy expenditure (metabolic equivalents) and heart rate recovery, but showed the same degree of airflow obstruction and static and dynamic hyperinflation. In a multivariate linear regression model performed to identify variables predicting metabolic equivalents, ankle-brachial index (β 2.59; 95% confidence interval 0.51-4.67; p = 0.016) was an independent variable. In the search for predictors of peripheral artery disease, heart rate recovery (odds ratio 8.80; 95% confidence interval 1.30-59.35; p = 0.026) increased the risk of peripheral artery disease, whereas metabolic equivalents (odds ratio 0.50; 95% confidence interval 0.26-0.94, p = 0.033) and inhaled corticosteroids+long-acting β2 agonists (odds ratio 0.13; 95% confidence interval 0.02-0.83; p = 0.030) reduced this risk. Conclusions In chronic obstructive pulmonary disease outpatients, asymptomatic/silent peripheral artery disease affects the maximal exercise capacity regardless of airflow obstruction and lung hyperinflation. A delay of heart rate recovery increase the risk of peripheral artery disease, whereas high values of metabolic equivalents and the use of inhaled corticosteroids+long-acting β2 agonists reduces this risk.


Identification of Metabolomic Markers in Frozen or Formalin-Fixed and Paraffin-Embedded Samples of Diffuse Glioma from Adults.
The aim of this study was to identify metabolomic signatures associated with the gliomagenesis pathway (IDH-mutant or IDH-wt) and tumor grade of diffuse gliomas (DGs) according to the 2021 WHO classification on frozen samples and to evaluate the diagnostic performances of these signatures in tumor samples that are formalin-fixed and paraffin-embedded (FFPE). An untargeted metabolomic study was performed using liquid chromatography/mass spectrometry on a cohort of 213 DG samples. Logistic regression with LASSO penalization was used on the frozen samples to build classification models in order to identify IDH-mutant vs. IDH-wildtype DG and high-grade vs low-grade DG samples. 2-Hydroxyglutarate (2HG) was a metabolite of interest to predict IDH mutational status and aminoadipic acid (AAA) and guanidinoacetic acid (GAA) were significantly associated with grade. The diagnostic performances of the models were 82.6% AUC, 70.6% sensitivity and 80.4% specificity for 2HG to predict IDH status and 84.7% AUC, 78.1% sensitivity and 73.4% specificity for AAA and GAA to predict grade from FFPE samples. Thus, this study showed that AAA and GAA are two novel metabolites of interest in DG and that metabolomic data can be useful in the classification of DG, both in frozen and FFPE samples.


On the Power Spectrum of Motor Unit Action Potential Trains Synchronized With Mechanical Vibration.
This study provides a definitive analysis of the spectrum of a motor unit action potential train (MUAPT) elicited by mechanical vibratory stimulation via a detailed and concise mathematical formulation. Experimental studies demonstrated that MUAPs are not exactly synchronized with the vibratory stimulus but show a variable latency jitter, whose effects have not been investigated yet. Synchronized action potential train was represented as a quasi-periodic sequence of a given MU waveform. The latency jitter of action potentials was modeled as a Gaussian stochastic process, in accordance to the previous experimental studies. A mathematical expression for power spectrum of a synchronized MUAPT has been derived. The spectrum comprises a significant continuous component and discrete components at the vibratory frequency and its harmonics. Their relevance is correlated to the level of synchronization: the weaker the synchronization the more relevant is the continuous spectrum. Electromyography (EMG) rectification enhances the discrete components. The derived equations have general validity and well describe the power spectrum of actual EMG recordings during vibratory stimulation. Results are obtained by appropriately setting the level of synchronization and vibration frequency. This paper definitively clarifies the nature of changes in spectrum of raw EMG recordings from muscles undergoing vibratory stimulation. Results confirm the need of motion artifact filtering for raw EMG recordings during stimulation and strongly suggest to avoid EMG rectification that significantly alters the spectrum characteristics.


eRNAbase: a comprehensive database for decoding the regulatory eRNAs in human and mouse.
Enhancer RNAs (eRNAs) transcribed from distal active enhancers serve as key regulators in gene transcriptional regulation. The accumulation of eRNAs from multiple sequencing assays has led to an urgent need to comprehensively collect and process these data to illustrate the regulatory landscape of eRNAs. To address this need, we developed the eRNAbase (http://bio.liclab.net/eRNAbase/index.php) to store the massive available resources of human and mouse eRNAs and provide comprehensive annotation and analyses for eRNAs. The current version of eRNAbase cataloged 10 399 928 eRNAs from 1012 samples, including 858 human samples and 154 mouse samples. These eRNAs were first identified and uniformly processed from 14 eRNA-related experiment types manually collected from GEO/SRA and ENCODE. Importantly, the eRNAbase provides detailed and abundant (epi)genetic annotations in eRNA regions, such as super enhancers, enhancers, common single nucleotide polymorphisms, expression quantitative trait loci, transcription factor binding sites, CRISPR/Cas9 target sites, DNase I hypersensitivity sites, chromatin accessibility regions, methylation sites, chromatin interactions regions, topologically associating domains and RNA spatial interactions. Furthermore, the eRNAbase provides users with three novel analyses including eRNA-mediated pathway regulatory analysis, eRNA-based variation interpretation analysis and eRNA-mediated TF-target gene analysis. Hence, eRNAbase is a powerful platform to query, browse and visualize regulatory cues associated with eRNAs.


Attempts to Understand Oral Mucositis in Head and Neck Cancer Patients through Omics Studies: A Narrative Review.
Oral mucositis (OM) is a common and clinically impactful side effect of cytotoxic cancer treatment, particularly in patients with head and neck squamous cell carcinoma (HNSCC) who undergo radiotherapy with or without concomitant chemotherapy. The etiology and pathogenic mechanisms of OM are complex, multifaceted and elicit both direct and indirect damage to the mucosa. In this narrative review, we describe studies that use various omics methodologies (genomics, transcriptomics, microbiomics and metabolomics) in attempts to elucidate the biological pathways associated with the development or severity of OM. Integrating different omics into multi-omics approaches carries the potential to discover links among host factors (genomics), host responses (transcriptomics, metabolomics), and the local environment (microbiomics).


2,5-Bis{[(-)-(S)-1-(4-bromo-phen-yl)eth-yl]imino-meth-yl}thio-phene.
The title compound, C22H20Br2N2S, was synthesized under solvent-free conditions. The mol-ecule shows crystallographic C 2 symmetry, with the S atom of the central thio-phene ring lying on a twofold rotation axis. Furthermore, as a consequence of the (S,S) stereochemistry, the mol-ecule has a twisted conformation. The dihedral angle between the thio-phene and benzene rings is 72.7 (2)° and that between the two benzene rings is 55.9 (2)°. In the crystal, mol-ecules are arranged in a chevron-like pattern, without any significant inter-molecular inter-actions.


Bayesian Model Averaging for Selection of a Risk Prediction Model for Death within Thirty Days of Discharge: The SILVER-AMI Study.
We describe a selection process for a multivariable risk prediction model of death within 30 days of hospital discharge in the SILVER-AMI study. This large, multi-site observational study included observational data from 2000 persons 75 years and older hospitalized for acute myocardial infarction (AMI) from 94 community and academic hospitals across the United States and featured a large number of candidate variables from demographic, cardiac, and geriatric domains, whose missing values were multiply imputed prior to model selection. Our objective was to demonstrate that Bayesian Model Averaging (BMA) represents a viable model selection approach in this context. BMA was compared to three other backward-selection approaches: Akaike information criterion, Bayesian information criterion, and traditional p-value. Traditional backward-selection was used to choose 20 candidate variables from the initial, larger pool of five imputations. Models were subsequently chosen from those candidates using the four approaches on each of 10 imputations. With average posterior effect probability ≥ 50% as the selection criterion, BMA chose the most parsimonious model with four variables, with average C statistic of 78%, good calibration, optimism of 1.3%, and heuristic shrinkage of 0.93. These findings illustrate the utility and flexibility of using BMA for selecting a multivariable risk prediction model from many candidates over multiply imputed datasets.


Bioinformatics analysis to identify the critical genes, microRNAs and long noncoding RNAs in melanoma.
Melanoma, which is usually induced by ultraviolet light exposure and the following DNA damage, is the most dangerous skin cancer. The purpose of the present study was to screen key molecules involved in melanoma.Microarray data of E-MTAB-1862 were downloaded from the ArrayExpress database, which included 21 primary melanoma samples and 11 benign nevus samples. In addition, the RNASeq version 2 and microRNA (miRNA) sequencing data of cutaneous melanoma were downloaded from The Cancer Genome Atlas database. After identifying the differentially expressed genes (DEGs) using Limma package, enrichment analysis and protein-protein interaction (PPI) network analysis were performed separately for them using DAVID software and Cytoscape software. In addition, survival analysis and regulatory network analysis were further performed by log-rank test and Cytoscape software, respectively. Moreover, real-time reverse transcription polymerase chain reaction (RT-PCR) was performed to further verify the expression patterns of several selected DEGs.A total of 382 DEGs were identified in primary melanoma samples, including 206 upregulated genes and 176 downregulated genes. Functional enrichment analysis showed that COL17A1 was enriched in epidermis development. In the PPI network, CXCL8 (degree = 29) and STAT1 (degree = 28) had higher degrees and could interact with each other. Survival analysis showed that 21 DEGs, 55 long noncoding RNAs (lncRNAs) and 32 miRNAs were found to be associated with prognosis. Furthermore, several regulatory relationships were found in the lncRNA-gene regulatory network (such as RP11-361L15.4 targeting COL17A1) and the miRNA-gene regulatory network (such as hsa-miR-375 targeting CCL27 and hsa-miR-375 targeting insulin-like growth factor 1 receptor [IGF1R]). Real-time RT-PCR results showed that the overall direction of differential expression was consistent except COL17A1.CXCL8 interacted with STAT1, CCL27, and IGF1R targeted by hsa-miR-375, and COL17A1 targeted by RP11-361L15.4 might function in the development and progression of melanoma, which should be verified by more detailed experiments.


The impact of image-directed core biopsy on the practice of breast surgery: a new algorithm for a changing technology.
Open surgical biopsy has long been the standard of care for the diagnosis of breast pathology. As more lesions are diagnosed by mammography, image-directed core biopsy has taken on increasing importance. This study consisted of a consecutive group of 540 patients who underwent breast biopsy for mammographically or sonographically detected lesions. The percentage of those undergoing image-directed biopsy and open biopsy were determined as well as the rate of positive biopsy for each method. Results showed that for patients with nonpalpable lesions, the proportion of image-directed core biopsies rose steadily during the study period, from 0 per cent in early 1993 to 70.1 per cent by early 1996. During this same period, positive biopsy rate rose from 23.9 to 31.0 per cent in the first 18 months of the study to a range of 45.2 to 48.3 per cent in the last 18 months. It was concluded that image-directed core biopsies have begun to impact the practice of breast surgery by replacing, to a great extent, the open surgical biopsy. This study also demonstrates an increased proportion of positive biopsies done by open surgical technique. It was suggested that many of the mammographically benign lesions are diagnosed by image-directed core biopsy, obviating the need for an open technique. An algorithm for managing nonpalpable breast lesions is presented.


When mechanism matters: Bayesian forecasting using models of ecological diffusion.
Ecological diffusion is a theory that can be used to understand and forecast spatio-temporal processes such as dispersal, invasion, and the spread of disease. Hierarchical Bayesian modelling provides a framework to make statistical inference and probabilistic forecasts, using mechanistic ecological models. To illustrate, we show how hierarchical Bayesian models of ecological diffusion can be implemented for large data sets that are distributed densely across space and time. The hierarchical Bayesian approach is used to understand and forecast the growth and geographic spread in the prevalence of chronic wasting disease in white-tailed deer (Odocoileus virginianus). We compare statistical inference and forecasts from our hierarchical Bayesian model to phenomenological regression-based methods that are commonly used to analyse spatial occurrence data. The mechanistic statistical model based on ecological diffusion led to important ecological insights, obviated a commonly ignored type of collinearity, and was the most accurate method for forecasting.


Prostate cancer: horizons in the development of novel anti-cancer strategies.
Prostate cancer is the most common cause of non-cutaneous cancer in men and a leading lethal malignancy with increasing incidence worldwide. Inevitably, all patients will develop androgen-independent disease, which remains the main obstacle to improving survival. Unfortunately, existing first and second line hormonal treatment are condemned to extent survival for only a few years. These sobering data have forced researchers to start exploring in depth the molecular mechanisms implicated in the emergence of androgen independence and of prostate cancer invasion. In this vein, epoch-making discoveries in molecular oncology along with rapid expansion of our knowledge concerning the processes that govern differentiation, apoptosis, immune surveillance, angiogenesis, metastasis, cell cycle and signal transduction control, have unveiled a plethora of specific targets for prostate cancer gene therapy, immunotherapy and anti-invasion therapy, along with a variety of small-molecule compounds which inhibit or stimulate these pathways. These new anti-cancer approaches are in various stages of clinical development, providing exciting perspectives for the future of prostate cancer cure and enriching the current anti-cancer drug quiver with a new spectrum of therapeutic agents. The present review, through extensive literature cross-examination, discusses several novel anti-prostate cancer strategies, emphasizing on approaches that potentially may extend end-stage patient's survival.


Structure of the two transmembrane Cu+ transport sites of the Cu+ -ATPases.
Cu(+)-ATPases drive metal efflux from the cell cytoplasm. Paramount to this function is the binding of Cu(+) within the transmembrane region and its coupled translocation across the permeability barrier. Here, we describe the two transmembrane Cu(+) transport sites present in Archaeoglobus fulgidus CopA. Both sites can be independently loaded with Cu(+). However, their simultaneous occupation is associated with enzyme turnover. Site I is constituted by two Cys in transmembrane segment (TM) 6 and a Tyr in TM7. An Asn in TM7 and Met and Ser in TM8 form Site II. Single site x-ray spectroscopic analysis indicates a trigonal coordination in both sites. This architecture is distinct from that observed in Cu(+)-trafficking chaperones and classical cuproproteins. The high affinity of these sites for Cu(+) (Site I K(a)=1.3 fM(-1), Site II K(a)=1.1 fM(-1)), in conjunction with reversible direct Cu(+) transfer from chaperones, points to a transport mechanism where backward release of free Cu(+) to the cytoplasm is largely prevented.


The disconnected: COVID-19 and disparities in access to quality broadband for higher education students.
The COVID-19 pandemic forced many higher education institutions (HEIs) across the world to cancel face-to-face teaching, close campus facilities, and displace staff and students to work and learn from home. Given the persistent nature of the pandemic, many HEIs have continued to deliver courses online and/or use a blended learning approach. However, there are concerns around differences in student access to digital learning resources while at home, including high quality broadband connectivity. This is important, since variation in connectivity may impact the type of online/blended model that faculty can deliver or constrain student engagement with online content. In this context, this paper combines national data on the domiciles of students enrolled in Irish HEIs with detailed spatial data on broadband coverage to estimate the number of higher education students 'at risk' of poor access to high quality internet connectivity. Overall it finds that one-in-six students come from areas with poor broadband coverage, with large disparities by geography and by HEI. It also finds that students from the poorest broadband coverage areas are more likely to be socioeconomically disadvantaged. As a result, this paper recommends that HEIs use their detailed registration data to help identify and support at-risk students. In particular, the results suggest that some HEIs may need to prioritise access to campus facilities and services to less well-off students living in poor broadband coverage areas. The online version contains supplementary material available at 10.1186/s41239-021-00262-1.


Consensus-based policy recommendations for geriatric emergency care.
The purpose of this paper is to establish policy recommendations to address service and care delivery challenges facing hospital emergency departments (EDs) responding to the needs of increasing numbers of older adults. The consensus development process used an international expert interdisciplinary panel, convened at an international conference. Following a round table discussion and think-tank session, a nominal group method with constant comparative analysis and coding techniques was used to identify policy recommendations. Two rounds of electronic input followed the face-to-face meeting to reach consensus on priority ranking of the policy recommendations. Findings underwent an external review by four independent experts. A total of seven categories of policy recommendations were developed: education, integration and coordination of care, resources, ED physical environment, evidence-based practice, research and evaluation, and advocacy. The consensus development process did not include a systematic literature review on the topic. However, participants included experts in their disciplines. The recommendations may assist administrators, policy makers, clinicians, and researchers on future directions for improving emergency care and service delivery for older adults. The paper describes the process and results of a consensus development activity for ED care and services of older adults.


Spinal cord ischemia/infarct after cauda equina syndrome from disc herniation - A case study and literature review.
Spinal cord infarction is rare and occurs in 12/100,000; it represents 0.3%-2% of central nervous system infarcts. Here, we present a patient who developed recurrent bilateral lower extremity paraplegia secondary to spinal cord infarction 1 day after a successful L4-5 microdiscectomy in a patient who originally presented with a cauda equina syndrome. A 56-year-old patient presented with an acute cauda equina syndrome characterized by severe lower back pain, a right foot drop, saddle anesthesia, and acute urinary retention. When the lumbar magnetic resonance imaging (MRI) revealed a large right paracentral lumbar disc herniation at the L4-L5 level, the patient underwent an emergency minimally invasive right-sided L4-5 discectomy. Immediately, postoperatively, the patient regained normal function. However, 1 day later, while having a bowel movement, he immediately developed the recurrent paraplegia. The new lumbar MRI revealed acute ischemia and an infarct involving the distal conus medullaris. Further, workup was negative for a spinal cord vascular malformation, thus leaving an inflammatory postsurgical vasculitis as the primary etiology of delayed the conus medullaris infarction. Acute neurologic deterioration after spinal surgery which does not neurologically correlate with the operative level or procedure performed should prompt the performance of follow-up MR studies of the neuraxis to rule out other etiologies, including vascular lesions versus infarctions, as causes of new neurological deficits.


Antitubercular inhaled therapy: opportunities, progress and challenges.
Pulmonary tuberculosis remains the commonest form of this disease and the development of methods for delivering antitubercular drugs directly to the lungs via the respiratory route is a rational therapeutic goal. The obvious advantages of inhaled therapy include direct drug delivery to the diseased organ, targeting to alveolar macrophages harbouring the mycobacteria, reduced risk of systemic toxicity and improved patient compliance. Research efforts have demonstrated the feasibility of various drug delivery systems employing liposomes, polymeric microparticles and nanoparticles to serve as inhalable antitubercular drug carriers. In particular, nanoparticles have emerged as a remarkably useful tool for this purpose. While some researchers have preferred dry powder inhalers, others have emphasized nebulization. Beginning with the respiratory delivery of a single antitubercular drug, it is now possible to deliver multiple drugs simultaneously with a greater therapeutic efficacy. More experience and expertise have been observed with synthetic polymers, nevertheless, the possibility of using natural polymers for inhaled therapy has yet to be explored. Several key issues such as patient education, cost of treatment, stability and large scale production of drug formulations, etc. need to be addressed before antitubercular inhaled therapy finds its way from theory to clinical reality.


Genotype X diet interactions in mice predisposed to mammary cancer: II. Tumors and metastasis.
High dietary fat intake and obesity may increase the risk of susceptibility to certain forms of cancer. To study the interactions of dietary fat, obesity, and metastatic mammary cancer, we created a population of F(2) mice cosegregating obesity QTL and the MMTV-PyMT transgene. We fed the F(2) mice either a very high-fat or a matched-control-fat diet, and we measured growth, body composition, age at mammary tumor onset, tumor number and severity, and formation of pulmonary metastases. SNP genotyping across the genome facilitated analyses of QTL and QTL x diet interaction effects. Here we describe effects of diet on mammary tumor and metastases phenotypes, mapping of tumor/metastasis modifier genes, and the interaction between dietary fat levels and effects of cancer modifiers. Results demonstrate that animals fed a high-fat diet are not only more likely to experience decreased mammary cancer latency but increased tumor growth and pulmonary metastases occurrence over an equivalent time. We identified 25 modifier loci for mammary cancer and pulmonary metastasis, likely representing 13 unique loci after accounting for pleiotropy, and novel QTL x diet interactions at a majority of these loci. These findings highlight the importance of accurately modeling not only the human cancer characteristics in mice but also the environmental exposures of human populations.


Binding of an organo-osmium(II) anticancer complex to guanine and cytosine on DNA revealed by electron-based dissociations in high resolution Top-Down FT-ICR mass spectrometry.
The Os(II) arene anticancer complex [(η(6)-bip)Os(en)Cl](+) (Os1-Cl; where bip = biphenyl, and en = ethylenediamine) binds strongly to DNA. Here we investigate reactions between Os1-Cl and the self-complementary 12-mer oligonucleotide 5'-TAGTAATTACTA-3' (DNA12) using ultra high resolution Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS). Identification of the specific sites of DNA osmiation with {(η(6)-bip)Os(en)}(2+) was made possible by the use of Electron Detachment Dissociation (EDD) which produced a wide range of assignable osmiated MS/MS fragments. In contrast, the more commonly used CAD and IRMPD techniques produced fragments which lose the bound osmium. These studies reveal that not only is guanine G3 a strong binding site for {(η(6)-bip)Os(en)}(2+) but, unexpectedly, so too is cytosine C10. Interestingly, the G3/C10 di-osmiated adduct of DNA12 also formed readily but did not undergo such facile fragmentation by EDD, perhaps due to folding induced by van der Waal's interactions of the bound osmium arene species. These new insights into osmium arene DNA adducts should prove valuable for the design of new organometallic drugs and contribute to understanding the lack of cross resistance of this organometallic anticancer complex with cisplatin.


Bacterial aetiology of otitis media in children in Pakistan aged 0-59 months; laboratory surveillance data from 2004 to 2013: comparison between before and after the introduction of Hib vaccination.
Otitis media (OM) is a leading cause of childhood illness. In Pakistan, the estimated incidence of OM-associated hearing impairment is >40/10,000 population and OM-associated mortality is 50-79·9/10×10(6) population. No OM microbiology data are available from Pakistan since 2004. To describe the microbiology of OM in children aged 0-59 months in Pakistan. Laboratory data on ear pus specimens taken from children seen between 2004 and 2013 were retrieved from the Laboratory Information Systems of the Aga Khan University and entered into Microsoft Excel and SPSS version 16.0. Bacterial culture results from 277 specimens were analysed. Staphylococcus aureus and Pseudomonas aeruginosa were the organisms most commonly isolated, followed by Streptococcus pneumoniae and Haemophilus influenzae. Polymicrobial cultures significantly increased in the post-Hib vaccination period from 19·5% to 32·7% (P = 0·038). H. influenzae also increased significantly from 16·8% to 24·5% (P = 0·038). An increase in H. influenzae may reflect non-b capsular types (not determined in the study), or even capsular types from areas with low vaccine coverage. Increases in polymicrobial cultures and H. influenzae warrant further study.


Improvement of drug delivery with a breath actuated pressurised aerosol for patients with poor inhaler technique.
BACKGROUND The metered dose inhaler is difficult to use correctly, synchronising actuation with inhalation being the most important problem. A breath actuated pressurised inhaler, designed to help patients with poor inhaler technique, was compared with a conventional metered dose inhaler in terms of aerosol deposition and bronchodilator response. METHODS Radioaerosol deposition and bronchodilator response to 100 micrograms salbutamol were measured in 18 asthmatic patients, who inhaled from a conventional metered dose inhaler by their own chosen metered dose inhaler technique, from a conventional metered dose inhaler by a taught metered dose inhaler technique, and from a breath actuated pressured inhaler (Autohaler). RESULTS In the 10 patients who could coordinate actuation and inhalation of the inhaler on their own deposition of aerosol in the lungs and bronchodilator response were equivalent on the three study days. By contrast, in the eight patients who could not coordinate the mean (SEM) percentage of the dose deposited in the lungs with their own inhaler technique (7.2% (3.4%] was substantial lower than those attained by the taught metered dose inhaler technique (22.8% (2.5%] and by Autohaler (20.8% (1.7%]. CONCLUSION Although of little additional benefit to asthmatic patients with good coordination, the Autohaler is potentially a valuable aid to those with poor coordination, and should be considered in preference to a conventional metered dose inhaler in any patient whose inhaler technique is not known to be satisfactory.


Chemotaxis in microfluidic devices--a study of flow effects.
The use of microfluidic devices has become increasingly popular in the study of chemotaxis due to the exceptional control of flow properties and concentration profiles on the length scale of individual cells. In these applications, it is often neglected that cells, attached to the inner surfaces of the microfluidic chamber, are three-dimensional objects that perturb and distort the flow field in their vicinity. Depending on the interplay of flow speed and geometry with the diffusive time scale of the chemoattractant in the flow, the concentration distribution across the cell membrane may differ strongly from the optimal gradient in a perfectly smooth channel. We analyze the underlying physics in a two-dimensional approximation and perform systematic numerical finite element simulations to characterize the three-dimensional case and to identify optimal flow conditions.


Evidence for Heterodimerization and Functional Interaction of the Angiotensin Type 2 Receptor and the Receptor MAS.
The angiotensin type 2 receptor (AT2R) and the receptor MAS are receptors of the protective arm of the renin-angiotensin system. They mediate strikingly similar actions. Moreover, in various studies, AT2R antagonists blocked the effects of MAS agonists and vice versa. Such cross-inhibition may indicate heterodimerization of these receptors. Therefore, this study investigated the molecular and functional interplay between MAS and the AT2R. Molecular interactions were assessed by fluorescence resonance energy transfer and by cross correlation spectroscopy in human embryonic kidney-293 cells transfected with vectors encoding fluorophore-tagged MAS or AT2R. Functional interaction of AT2R and MAS was studied in astrocytes with CX3C chemokine receptor-1 messenger RNA expression as readout. Coexpression of fluorophore-tagged AT2R and MAS resulted in a fluorescence resonance energy transfer efficiency of 10.8 ± 0.8%, indicating that AT2R and MAS are capable to form heterodimers. Heterodimerization was verified by competition experiments using untagged AT2R and MAS. Specificity of dimerization of AT2R and MAS was supported by lack of dimerization with the transient receptor potential cation channel, subfamily C-member 6. Dimerization of the AT2R was abolished when it was mutated at cysteine residue 35. AT2R and MAS stimulation with the respective agonists, Compound 21 or angiotensin-(1-7), significantly induced CX3C chemokine receptor-1 messenger RNA expression. Effects of each agonist were blocked by an AT2R antagonist (PD123319) and also by a MAS antagonist (A-779). Knockout of a single of these receptors made astrocytes unresponsive for both agonists. Our results suggest that MAS and the AT2R form heterodimers and that-at least in astrocytes-both receptors functionally depend on each other.


Electrochemical detection of chloride levels in sweat using silver nanoparticles: a basis for the preliminary screening for cystic fibrosis.
Cystic fibrosis is a common disease which has an associated characteristic symptom of high sweat chloride content. Thus, chloride ion quantification in sweat is important towards the screening of cystic fibrosis. Electrochemical methods, being cost effective and convenient, can be exploited for this. The electrochemical oxidation of silver nanoparticles in the absence of chloride ions gives one voltammetric signal related to the oxidation of silver to silver ions. The presence of chloride ions in the solution causes the appearance of an additional signal at a lower potential which is related to the oxidation of silver to silver chloride. This signal has a peak height which correlates linearly with the concentration of chloride ions from 2 mM to 40 mM when the electrochemical experiments are performed on silver nanoparticle modified screen printed electrodes. Thus, reliable quantification was found to be achievable. Furthermore, chloride ion levels of diluted synthetic sweat samples are measured accurately with the modified electrodes. Thus, the detection of the chloride ion concentration with a silver nanoparticle modified electrode provides a proof-of-concept for a point-of-care system for preliminary screening of cystic fibrosis.


Article Effect and Mechanism of Ganoderma lucidum Polysaccharides on Human Fibroblasts and Skin Wound Healing in Mice.
To investigate the effects of Ganoderma lucidum polysaccharides (GL-PS) on human fibroblasts and skin wound healing in Kunming male mice and to explore the putative molecular mechanism. Primary human skin fibroblasts were cultured. The viability of fibroblasts treated with 0, 10, 20, 40, 80, and 160 μg/mL of GL-PS, respectively were detected by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-Htetrazolium bromide (MTT). The migration ability of fibroblasts treated with 0, 10, 20, and 40 μg/mL of GL-PS were measured by transwell assay. The secretion of the C-terminal peptide of procollagen type I (CICP) and transforming growth factor-β1 (TGF-β1) in the cell supernatant was tested by enzyme-linked immunosorbent assay. The expression of β-catenin was detected by Western blot. Furthermore, the Kunming mouse model with full-layer skin resection trauma was established, and was treated with 10, 20, and 40 mg/mL of GL-PS, respectively as external use. The size of the wound was measured daily, complete healing time in each group was recorded and the percentage of wound contraction was calculated. Compared with the control group, 10, 20, and 40 μg/mL of GL-PS significantly increased the viability of fibroblasts, promoted the migration ability of fibroblasts, and up-regulated the expressions of CICP and TGF-β1 in fibroblasts (Plt;0.05 or Plt;0.01). The expression of β-catenin in fibroblasts treated with 20 and 40 μg/mL of GL-PS was significantly higher than that of the control group (Plt;0.01). Furthermore, after external use of 10, 20, and 40 mg/mL of GL-PS, the rates of wound healing in mice were significantly higher and the wound healing time was significantly less than the control group (Plt;0.05 or Plt;0.01). A certain concentration of GL-PS may promote wound healing via activation of the Wnt/β-catenin signaling pathway and up-regulation of TGF-β1, which might serve as a promising source of skin wound healing.


Aqueous extract of betel nut-induced adducts on pMTa4 DNA acquires stability in the presence of Na+ and K+ ions.
Betel nut (BN), a natural carcinogen to humans, is used as a masticator across the globe by a large segment of the human population. The primary carcinogens of BN are alkaloids, in particular arecoline. Upon nitrosation, arecoline can potentially interact with DNA, forming adducts and initiating carcinogenesis. Though considerable evidence exists in support of the carcinogenicity of BN, the molecular mechanism of its induction of carcinogenesis is unknown. This investigation was undertaken to directly demonstrate adduct formation on DNA and to study its characteristics, such as its frequency of formation and stability. A plasmid DNA construct, pMTa4, was chosen to determine the stoichiometry and dynamics of adduct formation. This construct was exposed to aqueous extract of betel nut (AEBN) in vitro and in vivo and analyzed. Spectrophotometric analysis revealed a significant red shift in the pMTa4 DNA spectrum. The gel electrophoretic mobility of pMTa4 DNA was also retarded in an AEBN dose- and exposure time-dependent manner, indicating BN-specific adduct formation on the DNA. These results conclusively demonstrate that adducts are formed on DNA by BN extract, and suggest that one AEBN-induced adduct was formed every 3 NT on pMTa4 DNA under the experimental conditions. Trace amounts of monovalent cations, such as Na+ or K+ ion (≥0.5 mmol), conferred stability to the adducts on DNA, which were otherwise unstable beyond 24 h.


Developing a Role Activity Performance Scale.
This article describes the Role Activity Performance Scale (RAPS) and presents data from a study of its reliability and validity. The RAPS evaluates an individual's functional level, using 12 subscales that represent a range of life roles. It was developed as an instrument for evaluating the impact of occupational therapy and other treatment modalities on the functioning of psychiatric patients. It is also useful in the diagnostic, treatment, and discharge planning processes. Validity and interrater reliability testing involved a sample of 30 patients in two major diagnostic groups. Interrater correlations for subscales and total scores were above .80 and statistically significant. A comparison of the RAPS with other accepted psychosocial rating scales showed strong correlations in scores. The psychometric evidence from this preliminary study supports the continued development and use of the RAPS for both clinical and research purposes.


Parasitological procedures, skills, and areas of knowledge used by small-animal practitioners in North America.
We designed a study to assess veterinarians' competency regarding parasitological procedures, skills, and areas of knowledge currently used in small-animal practice. The outcome will help us refine our curriculum on the basis of the parasitological working knowledge and skill sets that small-animal practitioners are using today. A questionnaire was developed and sent to small-animal practitioners. Their responses provided general information on practice characteristics, parasitological procedures used, and client education. Parasitological procedures included those to diagnose helminths, protozoa, and ectoparasites. We focused on three questions: "Do you perform or request this procedure?" "Where is this procedure performed?" and "What is your frequency?" The respondents were 478 small-animal practitioners. We performed descriptive analyses of practice characteristics along with bivariate and multivariate analyses. These analyses revealed the clinical competence of parasitological diagnoses performed or requested by small-animal practitioners. The results showed that more involved or time-consuming methods such as fecal flotation using centrifugation and the Baermann test are more often sent to a diagnostic laboratory and are requested more often by veterinarians in larger practices (i.e., those that employ more veterinarians). The outcomes also suggest that the main diagnostician may not fully understand the tests available at the diagnostic laboratory, which has an impact on decision making for management, treatment, and prevention of parasites and ultimately client education. In addition, small-animal practitioners who have been in practice longer and practices that employ five veterinarians or fewer (i.e., smaller practices) offer more client education.


Anterior Stafne bone defect mimicking a residual cyst: a case report.
Stafne bone defects (SBDs) are asymptomatic lingual bone depressions of the lower jaw that are frequently caused by soft tissue inclusion. The common variant of SBDs exists at the third molar region of the mandible below the inferior dental canal and has been mostly diagnosed incidentally during routine radiographic examination. The anterior variant of a SBD (ASBD) is relatively uncommon and is located in the premolar region of the mandible. Sublingual salivary glands are thought to be responsible for ASBDs. However, other structures such as lymphoid or vascular tissues might be associated with ASBDs. In the present report, an ASBD which was mimicking a residual cyst was diagnosed with the aid of a three-dimensional CT scan. ASBDs might be confused with other odontogenic or non-odontogenic pathologies because of their location and lower occurrence rate. Advanced imaging modalities, especially CT scans, are useful to assess such lesions in order to avoid unnecessary surgery.


Reduced ABA Accumulation in the Root System is Caused by ABA Exudation in Upland Rice (Oryza sativa L. var. Gaoshan1) and this Enhanced Drought Adaptation.
Lowland rice (Nipponbare) and upland rice (Gaoshan 1) that are comparable under normal and moderate drought conditions showed dramatic differences in severe drought conditions, both naturally occurring long-term drought and simulated rapid water deficits. We focused on their root response and found that enhanced tolerance of upland rice to severe drought conditions was mainly due to the lower level of ABA in its roots than in those of the lowland rice. We first excluded the effect of ABA biosynthesis and catabolism on root-accumulated ABA levels in both types of rice by monitoring the expression of four OsNCED genes and two OsABA8ox genes. Next, we excluded the impact of the aerial parts on roots by suppressing leaf-biosynthesized ABA with fluridone and NDGA (nordihydroguaiaretic acid), and measuring the ABA level in detached roots. Instead, we proved that upland rice had the ability to export considerably more root-sourced ABA than lowland rice under severe drought, which improved ABA-dependent drought adaptation. The investigation of apoplastic pH in root cells and root anatomy showed that ABA leakage in the root system of upland rice was related to high apoplastic pH and the absence of Casparian bands in the sclerenchyma layer. Finally, taking some genes as examples, we predicted that different ABA levels in rice roots stimulated distinct ABA perception and signaling cascades, which influenced its response to water stress.


Implementation of data security and data privacy provisions will bring sweeping changes to laboratory service providers.
The Health Insurance Portability and Accountability Act included substantial changes involving handling of health information by establishing national standards for electronic transactions, data privacy, and data security. The first final rule for electronic transaction standards was published August 17, 2000. The remaining final rules are expected to be published in Winter 2000. Providers, such as clinical laboratories, will have 26 months from the data of publication to comply. The civil monetary fines for noncompliance are substantial. This article will review the key provisions of the data security and data privacy proposed rules. These provisions will touch virtually every aspect of electronic claims submissions, electronic data transactions, and the electronic storage of medical information. The proposed rules will require a coordinated approach by providers to develop the policies and procedures, and the technical and physical infrastructure to protect health information. Moreover, providers will need to identify a privacy officer, to review existing privacy policies to compare the proposed rule with any existing state laws to determine which may be more stringent, and to develop new policies to address the particular requirements of the final rule.


Enhanced and coordinated in vivo expression of inflammatory cytokines and nitric oxide synthase by chondrocytes from patients with osteoarthritis.
To evaluate the sites of expression of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), and inducible nitric oxide synthase (iNOS) in patients with inflammatory and degenerative joint diseases. Cytokines and iNOS were detected by immunohistochemistry analysis of synovial and cartilage biopsy specimens obtained at knee arthroscopy in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), osteoarthritis (OA), and traumatic knee arthritis. Cytokine and iNOS expression was quantified using computerized image analysis. IL-1beta, TNFalpha, and iNOS were highly expressed by synovial cells (lining layer cells, infiltrating leukocytes, endothelial cells) from patients with inflammatory arthritides and significantly less by synovial cells from patients with OA and traumatic arthritis. In contrast, the latter patients showed high chondrocyte expression of cytokines and iNOS while RA and PsA patients had only minor chondrocyte positivity. In both joint compartments, IL-1beta expression, TNFalpha expression, and iNOS expression were strongly correlated. The enhanced and coordinated expression of IL-1beta, TNFalpha, and iNOS by chondrocytes strongly supports the hypothesis that chondrocytes are the major site of production of mediators of inflammation in human OA, thus playing a primary role in the pathogenesis of this disease.


Habitual exercise and arterial aging.
Aging affects the function and structure of arteries and increases the risk of cardiovascular diseases (CVD). In healthy sedentary adults, aging is associated with increased stiffness (reduced compliance) of large elastic arteries; impaired vascular endothelial function, including reductions in endothelium-dependent dilation (EDD), release of tissue-type plasminogen activator (fibrinolytic capacity) and endothelial progenitor cell number and function; increased intima-media wall thickness (IMT); and peripheral vasoconstriction (decreased basal leg blood flow). Habitual physical activity/increased aerobic exercise capacity is associated with reduced risk of CVD. Compared with their sedentary peers, adults who regularly perform aerobic exercise demonstrate smaller or no age-associated increases in large elastic artery stiffness, reductions in vascular endothelial function, and increases in femoral artery IMT. A short-term, moderate-intensity aerobic exercise intervention (brisk daily walking for 12 wk) improves carotid artery compliance and can restore vascular endothelial function in previously sedentary middle-aged and older adults. Reduced oxidative stress may be an important mechanism contributing to these effects. Habitual resistance exercise increases (high-intensity) or does not affect (moderate-intensity) large elastic artery stiffness, and prevents/restores the age-associated reduction in basal leg blood flow independent of changes in leg fat-free mass. Habitual exercise favorably modulates several expressions of arterial aging, thus preserving vascular function and possibly reducing the risk of CVD.


Engineering hydrogels with affinity-bound laminin as 3D neural stem cell culture systems.
Laminin incorporation into biological or synthetic hydrogels has been explored to recapitulate the dynamic nature and biological complexity of neural stem cell (NSC) niches. However, the strategies currently explored for laminin immobilization within three-dimensional (3D) matrices do not address a critical aspect influencing cell-matrix interactions, which is the control over laminin conformation and orientation upon immobilization. This is a key feature for the preservation of the protein bioactivity. In this work, we explored an affinity-based approach to mediate the site-selective immobilization of laminin into a degradable synthetic hydrogel. Specifically, a four-arm maleimide terminated poly(ethylene glycol) (PEG-4MAL) macromer was functionalized with a mono-PEGylated recombinant human N-terminal agrin (NtA) domain, to promote high affinity binding of laminin. Different NtA concentrations (10, 50 and 100 μM) were used to investigate the impact of NtA density on laminin incorporation, hydrogel biophysical properties, and biological outcome. Laminin was efficiently incorporated for all the conditions tested (laminin incorporation >95%), and the developed hydrogels revealed mechanical properties (average storage modulus (G') ranging from 187 to 256 Pa) within the values preferred for NSC proliferation and neurite branching and extension. Affinity-bound laminin PEG-4MAL hydrogels better preserve laminin bioactivity, compared to unmodified hydrogels and hydrogels containing physically entrapped laminin, this effect being dependent on NtA concentration. This was evidenced by the 10 μM NtA-functionalized PEG-4MAL gels incorporating laminin that support enhanced human NSC proliferation and neurite extension, compared to the latter. Overall, this work highlights the potential of the proposed engineered matrices to be used as defined 3D platforms for the establishment of artificial NSC niches and as extracellular matrix-mimetic microenvironments to support human NSC transplantation.


Serum concentrations of ionized calcium, vitamin D3, and parathyroid hormone in captive thick-billed parrots (Rhynchopsitta pachyrhyncha).
Serum collected from 68 thick-billed parrots (Rhynchopsitta pachyrhyncha) from 15 institutions was analyzed for ionized Ca (iCa), total Ca (tCa), P, total protein (TP), albumin (Alb), parathyroid hormone (PTH), and vitamin D3. Values were not distributed normally; 95% frequency intervals were as follows: iCa (0.82-1.3 mmol/L), tCa (1.37-2.09 mmol/L,), P (0.35-1.75 mmol/L), TP (21-39 g/L), Alb (9-13 g/L), PTH (0-65.68 pmol/L), and vitamin D3 (5.2-51 nmol/L). Sixty percent (+/-7.5%) of tCa was ionized. Female thick-billed parrots had significantly higher mean iCa (1.11 mmol/L, n = 22) than male thick-billed parrots (1.05 mmol/L, n = 32). tCa and iCa values in thick-billed parrots were lower than the reported values for other psittacine species. A significant positive linear relationship existed between Alb-TP and iCa-tCa ratios. A significant inverse linear relationship was also identified between the tCa-P ratio and PTH. These findings are consistent with known domestic avian Ca physiology.


Self-Reported Physical Activity Among American Indian Adults From Two Diverse Regions.
Physical activity may be a protective factor against the disproportionate rates of chronic diseases faced by American Indians. Nevertheless, few studies report any cultural adoptions made to capture physical activity behaviors among this hard-to-reach population. Existing studies reporting the prevalence of physical activity among American Indians are often aggregated and tend to obscure regional, local, and tribal-level variations. This study examines the prevalence of physical activity and inactivity levels, along with associated factors, among rural dwelling American Indian adults from 2 distinct regions. Baseline self-reported data were collected using a culturally modified version of the International Physical Activity Questionnaire (IPAQ) short form during the Obesity Research Prevention and Evaluation of Intervention Effectiveness in Native North Americans trial (OPREVENT) among rural American Indian adults (aged 18-75 years) from 5 tribal communities in Michigan and New Mexico. Most participants were classified as moderately physically active (43.5%), and the majority reported access to physical activity facilities (83.5%). Michigan participants reported engaging in more moderate and total physical activity than those in New Mexico (P < .001) and reported spending less time sitting (P < .001). Differences in physical activity among the American Indian communities may be due to regional variations in occupations, climate, and tribal and community support and infrastructure. The unexpected high level of activity evokes uncertainty in the accuracy and appropriateness of the data collection instrument. Research is needed to understand culturally appropriate approaches to measure physical activity and inactivity among rural American Indians.


Dual roles for ubiquitination in the processing of sperm organelles after fertilization.
The process of fertilization involves a cell fusion event between the sperm and oocyte. Although sperm contain mitochondria when they fuse with the oocyte, paternal mitochondrial genomes do not persist in offspring and, thus, mitochondrial inheritance is maternal in most animals. Recent evidence suggests that paternal mitochondria may be eliminated via autophagy after fertilization. In C. elegans, sperm-specific organelles called membraneous organelles (MO) cluster together with paternal mitochondria immediately after fertilization. These MOs but not the mitochondria become polyubiquitinated and associated with proteasomes. The current model for the elimination of paternal mitochondria in C. elegans is that ubiquitination of the MOs induces the formation of autophagosomes which also capture the mitochondria and cause their degradation. Sperm-derived mitochondria and MOs show a sharp decrease in number during the time between sperm-oocyte fusion and the onset of mitosis. During this time, paternal mitochondria remain closely clustered with the MOs. Two types of polyubiquitin chains are observed on the MOs: K48-linked ubiquitin chains which are known to lead to proteasomal degradation and K63-linked ubiquitin chains which have been linked to autophagy. K48-linked ubiquitin chains and proteasomes show up on MOs very soon after sperm-oocyte fusion. These are present on MOs for only a short period of time. Maternal proteasomes localize to MOs and sperm proteasomes localize to structures that are at the periphery of the MO cluster suggesting that these two proteasome populations may have different roles in degrading paternal material. K63-linked ubiquitin chains appear on MOs early and remain throughout the first several cell divisions. Since there are two different types of polyubiquitin chains associated with sperm organelles and their timing differs, it suggests that ubiquitin has two or more roles in the processing of sperm components after fertilization. The K63 chains potentially provide a signal for autophagy of paternal organelles, whereas the K48 chains and proteasomes may be involved in degradation of specific proteins.


What is stable pain control? A prospective longitudinal study to assess the clinical value of a personalized pain goal.
A universal consensus regarding standardized pain outcomes does not exist. The personalized pain goal has been suggested as a clinically relevant outcome measure. To assess the feasibility of obtaining a personalized pain goal and to compare a clinically based personalized pain goal definition versus a research-based study definition for stable pain. Prospective longitudinal descriptive study. The attending physician completed routine assessments, including a personalized pain goal and the Edmonton Classification System for Cancer Pain, and followed patients daily until stable pain control, death, or discharge. Stable pain for cognitively intact patients was defined as pain intensity less than or equal to desired pain intensity goal (personalized pain goal definition) or pain intensity ⩽3 (Edmonton Classification System for Cancer Pain study definition) for three consecutive days with <3 breakthroughs per day. A total of 300 consecutive advanced cancer patients were recruited from two acute care hospitals and a tertiary palliative care unit. In all, 231/300 patients (77%) had a pain syndrome; 169/231 (73%) provided a personalized pain goal, with 113/169 (67%) reporting a personalized pain goal ⩽3 (median = 3, range = 0-10). Using the personalized pain goal definition as the gold standard, sensitivity and specificity of the Edmonton Classification System for Cancer Pain definition were 71.3% and 98.5%, respectively. For mild (0-3), moderate (4-6), and severe (7-10) pain, the highest sensitivity was for moderate pain (90.5%), with high specificity across all three categories (95%-100%). The personalized pain goal is a feasible outcome measure for cognitively intact patients. The Edmonton Classification System for Cancer Pain definition closely resembles patient-reported personalized pain goals for stable pain and would be appropriate for research purposes. For clinical pain management, it would be important to include the personalized pain goal as standard practice.


A Review of Commonly Used Intrauterine Contraceptive Devices.
The radiographic appearances of the most commonly used intrauterine contraceptives devices are reviewed, and methods are suggested to determine the extrauterine location of these devices. Complications resulting from the use of these devices are also discussed.


Pharmacological characterization of protein phosphatase activities in preparations from failing human hearts.
beta-Adrenoceptor stimulation acts in the heart in part by increasing the phosphorylation state of phospholamban and phospholemman. There is evidence that the beta-adrenoceptor-mediated increase in phospholamban phosphorylation is in part due to inhibition of type 1 phosphatases. The aim of the present study was to elucidate which phosphatases dephosphorylate phospholamban and phospholemman in the human heart. In the past, cardiac serine/threonine phosphatases have been studied using phosphorylase a as substrate. Here, type 1 and type 2A phosphatase activities were studied in preparations from failing human hearts using phosphorylated phospholamban and phospholemman as substrates. Phospholamban and phospholemman phosphatase activity was detectable in human cardiac homogenates. Moreover, using a heparin-Sepharose column, the catalytic subunits of type 1 and type 2A phosphatases could be separated from human ventricles. Okadaic acid and cantharidin inhibited phosphatase activities dephosphorylating phospholamban, phospholemman, and phosphorylase a in homogenates in a concentration-dependent manner. However, okadaic acid was more potent. Cantharidin inhibited type 2A and type 1 activities against all substrates studied with IC50 values <15 nM and >290 nM, respectively. Okadaic acid inhibited type 1 and type 2A phosphatase activities as effectively but 10-30 times more potently than cantharidin. This work provides evidence that in the human heart, type 1 and 2A phosphatases are involved in the dephosphorylation of phospholamban and phospholemman and could play a role in the effects of beta-adrenergic stimulation in the heart.


Fur regulates the expression of iron-stress genes in the cyanobacterium Synechococcus sp. strain PCC 7942.
A homologue of the 'ferric uptake regulation' gene (fur) was isolated from the cyanobacterium Synechococcus sp. strain PCC 7942 by an Escherichia coli-based 'in vivo repression assay'. The assay uses a reporter-gene construct containing the promoter region of the iron-regulated cyanobacterial gene isiA, fused to the coding region for chloramphenicol acetyltransferase. The isolated gene codes for a protein that has 41% sequence similarity (36% identity) to Fur from E. coli and contains the putative iron-binding motif found in the Fur proteins of purple bacteria. No significant similarity was found to the DxtR repressor that regulates the expression of toxin and siderophore production in Gram-positive bacteria. Insertional mutagenesis of the cloned cyanobacterial fur gene led to the creation of heteroallelic mutants that showed iron-deficiency symptoms in iron-replete medium, including the constitutive production of flavodoxin and of hydroxamate siderophores. Failure to eliminate wild-type copies of the fur gene from the polyploid genome of Synechococcus 7942 implies that in this cyanobacterium Fur may have essential functions in addition to the regulation of genes involved in iron scavenging or photosynthetic electron transport.


GeP3: A Small Indirect Band Gap 2D Crystal with High Carrier Mobility and Strong Interlayer Quantum Confinement.
We propose a two-dimensional crystal that possesses low indirect band gaps of 0.55 eV (monolayer) and 0.43 eV (bilayer) and high carrier mobilities similar to those of phosphorene, GeP3. GeP3 has a stable three-dimensional layered bulk counterpart, which is metallic and known from experiment since 1970. GeP3 monolayer has a calculated cleavage energy of 1.14 J m-2, which suggests exfoliation of bulk material as viable means for the preparation of mono- and few-layer materials. The material shows strong interlayer quantum confinement effects, resulting in a band gap reduction from mono- to bilayer, and then to a semiconductor-metal transition between bi- and triple layer. Under biaxial strain, the indirect band gap can be turned into a direct one. Pronounced light absorption in the spectral range from ∼600 to 1400 nm is predicted for monolayer and bilayer and promises applications in photovoltaics.


An evaluation of population pharmacokinetics in therapeutic trials. Part I. Comparison of methodologies.
NONMEM, a computer program that uses the method of extended least-squares analysis, has been advocated as a means of obtaining estimates of population pharmacokinetic parameters when only fragmentary information can be obtained from subjects. To assess the performance of this program, we compared NONMEM with traditional methods for the estimation of population pharmacokinetic parameters with data collected during a phase III clinical trial of alprazolam. NONMEM estimates of the population mean clearance and its coefficient of variation were identical to the estimates obtained with traditional pharmacokinetic techniques. Moreover, NONMEM estimates of these parameters remained stable even when a few as three data points were available per subject. NONMEM estimates of the mean volume of distribution and its coefficient of variation appear to be overestimated, apparently because of the sampling scheme used to generate data for the NONMEM analysis. Suggestions for the effective use of NONMEM in clinical trials, to maximize the benefits of this approach, are provided. Our results lend further support for the use of NONMEM to estimate population pharmacokinetic parameters of a drug from data generated during phase III clinical trials.


Elevated expression of SATB1 is involved in pancreatic tumorigenesis and is associated with poor patient survival.
Special AT‑rich sequence‑binding protein 1 (SATB1) is a master chromatin organizer which has been reported to be implicated in tumor progression in breast and lung cancer. However, its functions in pancreatic tumorigenesis have yet to be elucidated. In the present study, the involvement of SATB1 in pancreatic cancer development was investigated in human BxPC‑3 pancreatic adenocarcinoma cells. Short hairpin (sh)RNA was used to stably downregulate SATB1 expression, and functional assays, including cell proliferation, colony formation, soft agar and migration assays, were performed in vitro. In addition, a mouse pancreatic cancer xenograft model was created to examine the tumor‑promoting properties of SATB1 in vivo. The present findings demonstrated that stable knockdown of SATB1 expression inhibited the proliferation, colony formation, anchorage‑independent growth and suppressed the migratory capabilities of BxPC‑3 cells in vitro. In addition, SATB1 downregulation significantly inhibited tumor growth in xenografted mice in vivo. Furthermore, SATB1 was revealed to be upregulated in human pancreatic cancer tissue samples compared with matched non‑cancerous adjacent tissues, and high SATB1 expression was associated with poor patient survival. Overall, the present study demonstrated that SATB1 promoted the proliferation of pancreatic cancer cells in vitro. In addition, SATB1 expression was revealed to be upregulated in human pancreatic cancer tissues and its upregulation was associated with poor patient survival. Therefore, SATB1 may have potential as a novel prognostic biomarker and therapeutic target for the treatment of patients with pancreatic cancer.


Symmetric Projection Attractor Reconstruction Analysis as a Method to Assess Seismocardiogram Quality in a Healthy Population.
Signal quality significantly affects the processing, analysis, and interpretation of biomedical signals. There are many procedures for assessing signal quality that use averaged numerical values, thresholding, analysis in the time or frequency domain, or nonlinear approaches. An interesting approach to the assessment of signal quality is using symmetric projection attractor reconstruction (SPAR) analysis, which transforms an entire signal into a two-dimensional plot that reflects the waveform morphology. In this study, we present an application of SPAR to evaluate the quality of seismocardiograms (SCG signals) from the CEBS database, a publicly available seismocardiogram signal database. Visual inspection of symmetric projection attractors suggests that high-quality (clean) seismocardiogram projections resemble six-pointed asterisks (*), and any deviation from this shape suggests the influence of noise and artifacts.Clinical relevance- SPAR analysis enables quick identification of noise and artifacts that can affect the reliability of the diagnosis of cardiovascular diseases based on SCG signals.


Renal dysfunction after oral acyclovir treatment in a geriatric woman: a case report.
Oral acylovir treatment rarely results in renal dysfunction. In reported cases concerning renal failure due to oral acyclovir, baseline creatinine levels were unknown or impaired or a predisposing factor for nephrotoxicity was present. We report a 78-y-old female with documented normal baseline renal function and no contributing possible nephrotoxic factor, who developed irreversible renal dysfunction after oral acyclovir treatment. We suggest that in geriatric age groups chronic renal insufficiency should be taken into consideration during oral acyclovir therapy.


Treatment of canine blastomycosis with amphotericin B and ketoconazole.
The treatment of 62 dogs with blastomycosis was reviewed to identify prognostic factors and response to various treatment regimens. Severity of lung involvement, as determined by radiography, and the number of nonsegmented neutrophils were useful prognostic factors. Females survived treatment better than did males, but females were more prone to relapse. Ketoconazole treatment at a dose of 10 mg/kg daily for 60 days was not as effective as amphotericin B. The most notable adverse effect of amphotericin B treatment was nephrotoxicosis . Treatment with amphotericin B followed by ketoconazole was as effective as amphotericin B alone and resulted in less nephrotoxicosis . Most dogs that had relapses were retreated effectively with amphotericin B and/or ketoconazole. Canine blastomycosis was shown to be a treatable disease, with a cure rate of approximately 75%.


Virological and serological characterization of vaccinated and non-vaccinated piglet subpopulations coming from vaccinated and non-vaccinated sows.
The present study describes the virological and serological profiles of PCV2 vaccinated (V) and non-vaccinated (NV) piglet subpopulations coming from V and NV sows in a PCV2 subclinically infected farm. Four hundred seventy-six piglets born from V or NV sows were further subdivided in a total of four groups: NV sows-NV pigs (NV-NV), NV sows-V pigs (NV-V); V sows-NV pigs (V-NV) and V sows-V pigs (V-V). Seventy-five pigs were randomly selected at the beginning of the trial from each group and they were bled at 4, 8, 12, 16, 21 and 25 weeks of age. All animals included in the trial were weighed at 4 and 25 weeks of age and their average daily weight gain (ADWG) was calculated. Serum samples obtained at different time points were used to assess PCV2 infection (viremia) and the level of antibodies by means of immunoperoxidase monolayer assay (IPMA) against this pathogen. IPMA titers (classified in high, medium or low) and PCR results (positive or negative) were analyzed using a multiple correspondence and K-means cluster analysis. According to these tests, animals included in the study were classified into the following four clusters: (1) 93 piglets that were viremic mainly from 12 to 25 weeks of age and with PCV2 antibody titers increasing over time; (2) 75 piglets with late PCV2 infection and seroconversion (later than 16 weeks of age); (3) 26 piglets with high but decreasing PCV2 antibody titers and low percentages of PCV2 PCR positive serum samples; and (4) 105 piglets with medium and high IPMA titers throughout the trial and sporadic PCR positive samples. The defined subpopulations of piglets were observed in all experimental groups (NV-NV, NV-V, V-NV and V-V) although in variable percentages. Thus, animals from clusters 1 and 2 belonged mainly to the NV-NV and V-NV groups and animals from clusters 3 and 4 were distributed mainly into the NV-V and V-V groups. Finally, the ADWG of pigs belonging to clusters 3 and 4 was significantly higher (p=0.02) than that of pigs belonging to clusters 1 and 2. Within each cluster, no statistically significant differences were found in ADWG between treatment groups.


Prediction of Losartan-Active Carboxylic Acid Metabolite Exposure Following Losartan Administration Using Static and Physiologically Based Pharmacokinetic Models.
The aim of this study was to evaluate a strategy based on static and dynamic physiologically based pharmacokinetic (PBPK) modeling for the prediction of metabolite and parent drug area under the time-concentration curve ratio (AUCm/AUCp) and their PK profiles in humans using in vitro data when active transport processes are involved in disposition. The strategy was applied to losartan and its pharmacologically active metabolite carboxylosartan as test compounds. Hepatobiliary transport including transport-mediated uptake, canilicular and basolateral efflux, and metabolic clearance estimates were obtained from in vitro studies using human liver microsomes and sandwich-cultured hepatocytes. Human renal clearance of carboxylosartan was estimated from dog renal clearance using allometric scaling approach. All clearance mechanisms were mechanistically incorporated in a static model to predict the relative exposure of carboxylosartan versus losartan (AUCm/AUCp). The predicted AUCm/AUCp were consistent with the observed data following intravenous and oral administration of losartan. Moreover, the in vitro parameters were used as initial parameters in PBPK permeability-limited disposition models to predict the concentration-time profiles for both parent and its active metabolite after oral administration of losartan. The PBPK model was able to recover the plasma profiles of both losartan and carboxylosartan, further substantiating the validity of this approach.


Intramolecular Pauson-Khand reactions of alpha,beta-unsaturated esters and related electron-deficient olefins.
The intramolecular Pauson-Khand (PK) reaction of a variety of electron-poor enynes having an ester, cyano, or phosphonate group at the olefin terminus is described. Depending on the reaction conditions and substitution at the enyne, their dicobalthexacarbonyl complexes led preferentially to the exocyclic 1,3-diene or to the PK cyclopentenone product. In general, the 1,3-diene was obtained as the major product under N-oxide-promoted conditions, while the PK product was selectively formed in refluxing acetonitrile.


Knowledge translation in developing countries.
There is increasing evidence that the application of knowledge in developing countries is failing. One reason is the woeful shortage of health workers, but as this is redressed, it is also crucial that we have an evidence base of what works to minimize the "know-do gap." The World Health Organization and other international organizations are actively building momentum to promote research to determine effective strategies for knowledge translation (KT). At this time, the evidence base for the effectiveness of those strategies is not definitive in developed countries and is relatively sparse in developing countries. It appears, however, that the effectiveness of these strategies is highly variable and dependent on the setting, and success hinges on whether the strategies have been tailored. A useful framework to provide direction for tailoring interventions is the Ottawa Model of Research Use (OMRU). Underlying OMRU is the principle that success rests with tailoring KT strategies to the salient barriers and supports found within the setting. The model recommends that barriers and supports found in the practice environment or as characteristics of potential adopters and the evidence-based innovation or research evidence be assessed and then the KT strategy tailored and executed. The model also recommends that whether the research has been applied and has resulted in improved health outcomes should be measured. Studies in developing countries, although few, illustrate that the OMRU approach may be a valid method of tackling the challenges of KT strategies to improve health care in developing countries.


[Latest news and perspectives in cardiogenetics].
In recent years, cardiogenetics is emerging as a major discipline for the study of many pathologies, with immediate clinical effects for patients who were previously managed by the cardiologist alone. Recent acquisitions have allowed significant improvements in terms of diagnostic characterization, prognostic stratification and guidance for treatment for both patients with genetic disease and their family members. At present, cardiogenetics has an important role for the clinical management of patients with cardiomyopathies and channelopathies. We present an updated review of the literature and a proposal of organizational model.


Using a human visual system model to optimize soft-copy mammography display: influence of MTF compensation.
The investigators developed an efficient method for optimizing cathode ray tube (CRT) monitor performance for digital mammography, based on the correlation between the performance of human observers and the performance of a mathematical computer model of the human visual system. The investigators examined observer performance on soft-copy display of mammographic images that were either unprocessed or processed to compensate for modulation transfer function (MTF) deficiencies in the CRT display. The results were used to validate the human visual system model. Six radiologists viewed a series of 250 mammographic images with microcalcification clusters with different contrast levels on a CRT monitor. The images were viewed twice: once without image processing and once with processing designed to compensate for MTF deficiencies in the CRT monitor. The images were analyzed with the JNDmetrix Visual Discrimination Model, which is based on the principles of just-noticeable difference measurement and frequency-channel vision modeling. Receiver operating characteristic (ROC) curves were generated for the human observers and compared statistically with the model observers' performance. Both human and model performance was better overall with the MTF-compensated images, especially for microcalcifications in the midlevel contrast range. There was a very high correlation between human and model observers. The use of image-processing methods to compensate for limitations in the MTF of CRT monitors can improve the detection performance of radiologists searching for microcalcifications in mammographic images, and a model based on characteristics of the human visual system can be used to predict human observer results accurately.


Motivational attributions about intimate partner violence among male and female perpetrators.
Given the widely acknowledged negative impact and pervasiveness of intimate partner violence (IPV), many have sought to discover why people perpetrate IPV. To this end, IPV risk factors have been extensively studied and attributions have recently received more attention. Evidence suggests males and females have similar attributions and both engage in IPV. Yet, IPV tactics and attribution rates appear to differ by perpetrator sex. This study explores whether males and females tend to attribute different reasons to their IPV, and whether these attributions help to distinguish among physical and psychological IPV tactics. The study tests three hypotheses: that IPV tactics and attributions will differ between females and males, that females and males will have similar latent constructs associated with types of attributions made about IPV, and that males and females will have different relationships among attribution factors and IPV tactics. This study is based on a cross-sectional survey of 5,035 18- to 25-year-old undergraduate university students in the Southeastern United States. Analyses used SPSS 20 to conduct basic and bivariate analyses, multiway frequency analysis, reliability analysis, and logistic regression, and MPlus 6.12 to conduct exploratory and confirmatory factor analysis. Overall, the results provide support for the three study hypotheses, although there was inconsistency in findings for the second hypothesis. The results suggest important practice and research implications, which are discussed. The study fills a gap in the limited literature on IPV perpetrator motivational attributions by presenting a nuanced analysis of an early measure of IPV attributions.


Inhibition of lipopolysaccharide-associated endotoxin activities in vitro and in vivo by the human anti-lipid A monoclonal antibody SdJ5-1.17.15.
The present study evaluated the effect of a novel anti-lipid A monoclonal antibody, termed SdJ5, on the in vitro production of tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta by endotoxin- or lipopolysaccharide (LPS)-challenged human peripheral blood mononuclear cells (hPBMC). In addition, the present study determined whether SdJ5 could neutralize the in vivo toxicity of LPS. SdJ5, at a concentration equal to or greater than 3 micrograms/ml, specifically inhibited TNF-alpha and interleukin-1 beta production by hPBMC stimulated with every type of LPS and lipid A assessed. SdJ5 also showed a significantly greater inhibition of cytokine production than a nonrelevant human immunoglobulin M myeloma control. The SdJ5-mediated inhibition of TNF-alpha production was rapid, as the simultaneous addition of the SdJ5 and LPS still resulted in a marked decrease in hPBMC cytokine synthesis. The ability of SdJ5 to neutralize in vivo toxicity was also determined by using LPS from four different strains of gram-negative bacteria. LPS, when preincubated with SdJ5, resulted in a significant decrease in the 24-h mortality rate compared with that for the control. These studies show that the anti-lipid A monoclonal antibody SdJ5 can modulate LPS-induced cytokine production in vitro and increase the survival rate of rats challenged with lethal doses of LPS.


The path of nasal reconstruction: from ancient India to the present.
Nasal reconstruction, first described over 2600 years ago in India, has undergone several modifications in its development. However, there is a misconception that this surgery began with the forehead flap repair, and the history of the subsequent two and a half millennia is poorly represented in most surgical texts. This article presents a review of the historical significance of nasal injuries in antiquity and the various technical developments along the path to modern nasal reconstruction, from ancient India through medieval Europe to modern England and America. Although written texts from 6th-century BC India through 16th-century AD Europe described pedicle flap repair of nasal defects, it was not until the late 18th century that the first written description of the forehead flap appeared. Forehead flap repair developed on an alternate pathway, being transmitted via an oral tradition, typically within families of craftsmen, at least as early as the 14th century. These ancient authors recognized the need for accurate flap design and sizing, donor site repair, precise tissue apposition, protection of the flap pedicle, hemostasis, deepithelialization of the wound, and replacement of intranasal lining. By appreciating ancient surgical techniques and examining the particular issues these procedures attempted to address, the surgeon will gain an additional and more detailed perspective of nasal reconstruction that will aid in providing excellent care to his patients.


Alterations in pattern of rapid eye movement activity during REM sleep in depression.
The aim of the study was to evaluate REM sleep parameters, especially the temporal characteristics of rapid eye movement activity, in depressed patients, and to compare three different methods for scoring of REM density. The sleep of 15 nonmedicated depressed patients and 13 healthy controls was recorded during two consecutive nights. Sleep recordings were scored by raters blinded to the diagnosis. In comparison to healthy controls depressed patients showed an increased REM density and increased REM activity. Both groups differed also regarding the pattern of REM density changes between REM sleep periods (REMPs). Whereas in healthy controls REM density in the first REMP was significantly lower than in the successive REMPs, no such difference was found in depressed patients. )n visual inspection we failed to find any significant differences in the time course of REM activity within the first REMP in depressed patients. All applied methods for scoring of REM density distinguished depressed patients from healthy controls with comparable accuracy.


Photocrosslinkable polysaccharide hydrogel composites based on dextran or pullulan-amylose blends with cytokines for a human co-culture model of human osteoblasts and endothelial cells.
Although a large body of research has been devoted to biomaterial development for bone tissue engineering and related medical disciplines in the last few years, novel and optimized materials especially for bone fractures of critical sizes demand continued development. In this respect, polysaccharide-based hydrogels demonstrate beneficial properties and fulfill the main requirements for a bone tissue scaffold as they are hydrophilic, biocompatible, and biodegradable. The aim of the present study was the development of a natural polysaccharide-based scaffold material that can integrate with the host tissue and support bone regeneration. For this purpose, we prepared and investigated two polymer hydrogel composites of photocrosslinkable derivatives of either pure dextran or a mixture of amylose and pullulan with varying composition. In order to increase their biological activity the swollen hydrogel matrices were compounded with stromal derived growth factor (SDF-1) and bone morphogenic protein (BMP-2). As skeletal development is known to depend on angiogenesis, these hydrogel systems were subjected to mono- and co-culture models of human primary osteoblasts (hOBs) with human endothelial cells (HUVEC - as precursors for blood vessel development). The effect of cytokines on hydrogel-dependent cell behavior was analyzed in the presence and absence of the growth factors SDF-1 and BMP-2. Both the employed cell types grew on all cytokine-modified hydrogel composites, which was assessed by optical microscopy and proliferation assays. Migration assays indicated enhanced HUVEC migration under the influence of SDF-1 and real-time PCR demonstrated an enhanced expression of cell-specific markers for growth factor-modified hydrogels, thus demonstrating their functional bioactivity. Our results demonstrate the fundamental potential of such multi-component polysaccharide hydrogel composites as biomaterials for bone regeneration.


Effects of dopamine D1 receptor full agonists in rats trained to discriminate SKF 38393.
Although the dopaminergic pharmacology of the D1 receptor full agonists, dinapsoline, dihydrexidine and the prodrug ABT-431 have been studied, no information is available on the ability of these agonists to substitute for the D1 agonist SKF 38393 in rats trained to discriminate this compound from vehicle. The present study was designed to characterize the potential D1 discriminative stimulus effects of these compounds. The selective dopamine D1-receptor agonists dihydrexidine [(+/-)-trans-10,11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a] phenanthridine hydrochloride], ABT-431 [(-)-trans-9,10-diacetyloxy-2-propyl-4,5,5a,6,7,11b-hexahydro-3-thia-5-azacyclopent-1-ena[c]phenanthrene hydrochloride], the diacetyl prodrug derivative of A-86929, and dinapsoline [9-dihydroxy-2,3,7,11b-tetrahydro-1H-naph[1,2,3-de]isoquinoline] were studied in rats trained to discriminate racemic SKF 38393 [(+/-)-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol], a selective D1 receptor partial agonist from vehicle. All of the agonists substituted fully for the discriminative stimulus effects of SKF 38393. The rank order of potency for substitution was ABT-431 > dinapsoline > dihydrexidine > SKF 38393. The D1 receptor antagonist, SCH 23390, blocked the discriminative stimulus effects of SKF 38393. The D3/D2-receptor agonist PD 128,907 [S(+)-(4aR,10bR)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]-benzopyrano[4,3-b]-1,4-oxazin-9-ol] did not substitute up to doses that produced profound rate-suppressant effects. Thus, consistent with their D1 receptor pharmacology, the full D1-receptor agonists substituted completely for the discriminative stimulus of SKF 38393.


On-chip density-based sorting of supercooled droplets and frozen droplets in continuous flow.
The freezing of supercooled water to ice and the materials which catalyse this process are of fundamental interest to a wide range of fields. At present, our ability to control, predict or monitor ice formation processes is poor. The isolation and characterisation of frozen droplets from supercooled liquid droplets would provide a means of improving our understanding and control of these processes. Here, we have developed a microfluidic platform for the continuous flow separation of frozen from unfrozen picolitre droplets based on differences in their density, thus allowing the sorting of ice crystals and supercooled water droplets into different outlet channels with 94 ± 2% efficiency. This will, in future, facilitate downstream or off-chip processing of the frozen and unfrozen populations, which could include the analysis and characterisation of ice-active materials or the selection of droplets with a particular ice-nucleating activity.


Periodontal disease increases risk for chronic obstructive pulmonary disease.
The aim of this study was to explore whether a periodontal disease could be a risk indicator for a chronic obstructive pulmonary disease (COPD). The examined group comprised 93 patients with COPD (mean age 65.8 years). The control group comprised 43 systemically healthy individuals (mean age 62.1 years). Respiratory and periodontal conditions were examined in both groups. COPB subjects had significantly worse periodontal conditions than controls (p < 0.05) with regard to each parameter of periodontal condition, except for gingival inflammation. COPD patients had higher Plaque Index than control patients (82.84 +/- 22.81 vs. 57.15 +/- 26.96; p < 0.001), higher periodontal depth (3.02 +/- 0.92 vs. 2.57 +/- 0.79 mm; p = 0.007), higher gingival recession (1.97 +/- 1.09 vs. 0.91 +/- 0.79 mm; p < 0.001), and higher mean clinical attachment loss (CAL) (4.12 +/- 1.74 vs. 2.91 +/- 1.27 mm; p < 0.001). Multiple logistic regression model, after controlling for other risk indicators, showed that periodontal disease, presented as CAL > or = 4 mm at > or = 60% sites, was associated with odds ratio of 3.2 (95% CI 1.0-9.8) for the COPB group. Data suggest that periodontal disease could be a risk indicator for COPD.


Saponins from Sanguisorba officinalis Improve Hematopoiesis by Promoting Survival through FAK and Erk1/2 Activation and Modulating Cytokine Production in Bone Marrow.
Radix Sanguisorbae, the root of Sanguisorba officinalis L. is used as traditional Chinese medicine. In recent decades, it has been reported to be clinically effective against myelosuppression induced by chemotherapy and/ or radiotherapy. However, the underlining mechanism has not been well studied. In this work, we evaluated the hematopoietic effect of total saponins from S. officinalis L. on myelosuppressive mice induced by cyclophosphamide and by60Co-γ-irradiation and confirmed the therapeutic effect. Then, we found total saponins and their characteristic constituents Ziyuglycoside I and Ziyuglycoside II can inhibit apoptosis of TF-1 cells caused by cytokine deprivation, and promote survival of mouse bone marrow nuclear cells through focal adhesion kinase (FAK) and extracellular signal-regulated kinase 1/2 (Erk1/2) activation in vitro. In addition, they can down-regulate macrophage inflammatory protein 2 (MIP-2), platelet factor 4 (PF4) and P-selectin secretion, which are reported to be suppressive to hematopoiesis, both in vitro and in vivo. These results suggest that promotion of survival through FAK and Erk1/2 activation and inhibition of suppressive cytokines in the bone marrow is likely to be the pharmacological mechanism underlying the hematopoietic effect of saponins from S. officinalis L.


Determination of the effective refractive index of nanoparticulate ITO layers.
Nanoparticles of transparent conducting oxides, such as indium tin oxide, can be used in printing techniques to generate functional layers for various optoelectronic devices. Since these deposition methods do not create fully consolidated films, the optical properties of such layers are expected to be notably different from those of the bulk material and should be characterized on their own. In this work we present a way to measure the effective refractive index of a particulate ITO layer by refraction of light. The obtained data points are used to identify an accurate layer model for spectroscopic ellipsometry. In this way the complex refractive index of the particle layer is determined in a wide spectral range from ultra violet to near infrared.


Role of salivary transcriptomics as potential biomarkers in oral cancer: A systematic review.
Transcriptomes in saliva can be used as potential biomarkers for both diagnostic and response to treatment in oral squamous cell carcinoma (OSCC). In this review, we explored their application in this increasingly common disease. PubMed, EMBASE, Scopus, Web of Science and grey literature from January 1990 to May 2017 were searched. Two independent reviewers performed the study selection according to eligibility criteria. A total of nine studies were included. Three studies showed increased expression of DUSP1, IL8, IL1B, OAZ1, SAT1, S100P and two showed increased expression of miRNA-31 among study groups compared to normal healthy controls. The sensitivity ranged from 14% to 100%, while the specificity ranged from 38% to 100%. miRNA-27b had the highest AUC (write in full) of 0.9643 and DUSP1 had the minimum AUC of 0.41. Salivary transcriptomics may play an effective role as a robust and non-invasive biomarker sighting tool for the diagnosis and management of OSCC.


Metabolism of D-galactose is dispensable for the induction of the beta-galactosidase (bgaD) and lactose permease (lacpA) genes in Aspergillus nidulans.
In this study, we analyze the expression of the Aspergillus nidulans bgaD-lacpA gene couple (encoding an intracellular beta-galactosidase and a lactose permease) in the presence of D-galactose. This monosaccharide can be catabolized via alternative, independent pathways in this model organism. The inductive capabilities of intermediates of the two alternative routes of D-galactose utilization were addressed in loss-of-function mutants defective in a defined step in one of the two pathways. In a galactokinase (galE9) mutant, the cluster is strongly induced by D-galactose, suggesting that formation of Leloir pathway intermediates is not required. The expression profiles of bgaD and lacpA were similar in wild type, L-arabinitol dehydrogenase (araA1), and hexokinase (hxkA1) negative backgrounds, indicating that intermediates of the oxido-reductive pathway downstream of galactitol are not necessary either. Furthermore, bgaD-lacpA transcription was not induced in any of the tested strains when galactitol was provided as the growth substrate. An hxkA1/galE9 double mutant cannot grow on d-galactose at all, but still produced bgaD and lacpA transcripts upon transfer to d-galactose. We therefore concluded that the physiological inducer of the bgaD-lacpA gene cluster upon growth on D-galactose is the nonmetabolized sugar itself.


Transcriptional organization of pneumococcal psrP-secY2A2 and impact of GtfA and GtfB deletion on PsrP-associated virulence properties.
Pneumococcal serine-rich repeat protein (PsrP) is a glycoprotein that mediates Streptococcus pneumoniae attachment to lung cells and promotes biofilm formation. Herein, we investigated the transcriptional organization of psrP-secY2A2, the 37-kbp pathogenicity island encoding PsrP and its accessory genes. PCR amplification of cDNA and RNA-seq analysis found psrP-secY2A2 to be minimally composed of three operons: psrP-glyA, glyB, and glyC-asp5. Transcription of all three operons was greatest during biofilm growth and immunoblot analyses confirmed increased PsrP production by biofilm pneumococci. Using gas chromatography-mass spectrometry we identified monomeric N-acetylglucosamine as the primary glycoconjugate present on a recombinant intracellular version of PsrP, i.e. PsrP1-734. This finding was validated by immunoblot using lectins with known carbohydrate specificities. We subsequently deleted gtfA and gtfB, the GTFs thought to be responsible for addition of O-linked N-acetylglucosamine, and tested for PsrP and its associated virulence properties. These deletions negatively affected our ability to detect PsrP1-734 in bacterial whole cell lysates. Moreover, S. pneumoniae mutants lacking these genes pheno-copied the psrP mutant and were attenuated for: biofilm formation, adhesion to lung epithelial cells, and pneumonia in mice. Our studies identify the transcriptional organization of psrP-secY2A2 and show the indispensable role of GtfA and GtfB on PsrP-mediated pneumococcal virulence.


Effect of miR‑146a‑5p on tumor growth in NSCLC using chick chorioallantoic membrane assay and bioinformatics investigation.
Our previous study demonstrated that the expression of miR‑146a‑5p was downregulated in non‑small cell lung cancer (NSCLC) tissue, which affected the progression and prognosis of patients with NSCLC. Thus, the present study was conducted to investigate the functional mechanism of miR‑146a‑5p in tumorigenesis and angiogenesis in NSCLC. Following the construction of a H460 NSCLC cell line in which miR‑146a‑5p was overexpressed via lentivirus transduction, the NSCLC chick embryo chorioallantoic membrane (CAM) model was established by transplanting miR‑146a‑5p‑overexpressing NSCLC cells into the CAM. Then, the size of the neoplasms within the CAM was measured, the vessel ratio was calculated, and the cellular morphology, metastasis and inflammation of tumor cell was observed using hematoxylin and eosin staining. The target genes of miR‑146a‑5p were predicted by 12 online software programs; these genes were then subjected to Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway annotations using the Database for Annotation, Visualization and Integrated Discovery 6.7 as well as constructed into a protein interaction network using protein‑protein interaction from Search Tool for the Retrieval of Interacting Genes/Proteins. The xenograft tumor size and angiogenesis conditions of the miR‑146a‑5p‑overexpressing group (volume 6.340±0.066 mm3, vessel ratio 9.326±0.083) was obviously restricted (P<0.001) when compared with the low expression group (volume 30.13±0.06 mm3, vessel ratio 16.94±0.11). In addition, marked necrosis along with inflammatory cell infiltration was observed with the HE‑stained slices from the miR‑146a‑5p low expression group. Regarding the results of the target gene prediction, cancer and toll‑like receptor signaling were the two most significant pathways represented among the target genes, while JUN, EGFR and RAC1 were the most relevant proteins among the selected potential targets of miR‑146a‑5p. In a CAM xenograft tumor model, overexpression of miR‑146a‑5p inhibited the tumorigenesis and angiogenesis of an NSCLC cell line. miR‑146a‑5p may act as a tumor suppressor gene in NSCLC and have moderate prognostic value in lung cancer.