Characterization of N- and O-linked galactosylated oligosaccharides from fission yeast species.
The N- and O-linked oligosaccharides from fission yeast Schizosaccharomyces pombe not only contain large amounts of d-mannose (Man) but also contain large amounts of d-galactose (Gal). Although the galactomannans of S. pombe are mainly composed of α1,2- or α1,3-linked Gals, some of the terminal α1,2-linked Gals are found to be linked to pyruvylated β1,3-linked galactose (PvGal). We have determined the structural characteristics of the N-glycans and O-glycans in three Schizosaccharomyces species (S. japonicus, S. octosporus, and S. cryophilus) using lectin blot, 1H NMR spectroscopy, and size-fractionation high performance liquid chromatography (HPLC), and found that the galactosylation of oligosaccharides was a common feature in fission yeasts. In addition, each of the terminal Galα1,2-, Galβ1,3- and non-substituted Man residues exhibited distinct characteristics. A BLAST search of gene databases in Schizosaccharomyces identified genes homologous to pvg1 encoding pyruvyltransferase of S. pombe. These genes, when expressed in an S. pombe pvg1Δ strains, led to the pyruvylation of non-reducing terminal β-linked Gal, suggesting the biosynthetic pathway of PvGal-containing oligosaccharides is highly conserved in fission yeasts.


Revealing the Pharmacophore of Ipomoeassin F through Molecular Editing.
Ipomoeassin F, the flagship congener of a resin glycoside family exhibited single-digit nanomolar IC50 values against several cancer cell lines. To facilitate drug discovery based on this unique yet underexplored natural product, we performed the most sophisticated SAR studies of ipomoeassin F to date, which not only greatly bettered our understanding of its pharmacophore but also led to the discovery of two new derivatives (3 and 27) with similar potency but improved synthetic profile. The work presented here opens new avenues toward harnessing the medicinal potential of the ipomoeassin family of glycolipids in the future.


Membrane thickness changes ion-selectivity of channel-proteins.
The plasma membrane is a highly dynamic cell-barrier if the nature and distribution of its constituents are considered. Ion channels are embedded in these double lipid bilayers, which modulate their 3D-structures. The structure modulations by the lipid bilayer can assume such a degree that channel activation depends on them, as was shown for the KcsA potassium channel. Here we show that the cation-over-anion selectivity of reconstituted ICln channels can be varied by the thickness of a bilayer build of phosphatidylcholines. The shorter the acyl-chains and therefore the thinner the bilayers of the membrane are, the more potassium selective the channels are. In contrast, the longer the acyl-chains and therefore the thicker the membranes are, the more chloride selective the channels become.


Gender difference on patients' satisfaction and expectation towards mental health care.
Recognizing patient satisfaction and expectation is considered as important components of assessing quality of care. The aim of this study was to determine the gender difference on the patient satisfaction with psychiatrists and explore their expectation from physicians to mental health care needs. This is a prospective cross sectional study conducted during the period from April 2009 to July 2009. Psychiatry hospital of the Hamad Medical Corporation. A total of 1300 psychiatry patients aged 18 to 65 years were approached and 1054 (81.1%) patient's participated in this study. The study sample included only Qatari and other Arab nationals. The study was based on a face to face interview with a designed Patient Doctor Relationship Questionnaire (PDRQ). A standard forward-backward procedure was applied to translate the English version of the PDRQ to Arabic. The main outcome measures of the study were 13-item patient doctor relationship questionnaire (PDRQ) score and other 11 items assessing what patients need from psychiatrists. Also, Socio-demographic data of the patients were collected. PDRQ was administered by qualified nurses among the psychiatry patients. Of the studied patients, 50.9% were males and 49.1% were females. Male patients (55.5%) were more satisfied with the treatment of psychiatrists than females (44.5%). A significant association was observed between male and female patients in terms of marital status (P = 0.02), number of children (P < 0.001), education level (P = 0.001) and monthly household income (P = 0.03). The satisfaction level significantly increased with higher education in both the groups (P = 0.001). The satisfaction level of male patients was significantly higher than female patients in most of the satisfaction areas (P ≤ 0.001). Male patients had significantly high expectations from psychiatrists than females (P < 0.001). Both the groups had different attitude in assessing mental health needs. The study findings revealed that in general, psychiatry patients were quite satisfied with the services provided by the psychiatrists and the study found a gender difference in their satisfaction level with the mental health care. Male patients had significantly greater expectations from psychiatrists than females. The individuals of lower socio-economic status and less education tend to be less satisfied with the mental health care.


Identification and characterization of the Xlsirt cis-acting RNA localization element.
There are many RNAs that are localized to the vegetal cortex of Xenopus laevis oocytes. One family of localized transcripts, Xlsirts (Xenopus laevis short interspersed repeat transcripts), are defined by the presence of non-coding repeat units 79-81 nucleotides long. Endogenous Xlsirt RNAs are localized through the METRO (message transport organizer) pathway that localizes RNAs during stages I and II of oogenesis. Interestingly, exogenous Xlsirt RNAs that are injected into oocytes can utilize both the METRO pathway as well as the Late pathway, which localizes RNAs during the late stages of oogenesis (stages III-VI). In all cases thus far analyzed, the localization process relies on the presence of cis-acting elements on the transcripts that are responsible for directing localization. To better understand the mechanism responsible for the use of the METRO and Late localization pathways, we sought to identify pathway-specific cis-acting localization elements contained in Xlsirts. The results showed that an intact 137 nucleotide element was necessary and sufficient to localize RNAs through the METRO and Late pathways. Further analysis of this element identified putative METRO and Late pathway localization sub-elements. Computer analysis relates the secondary structure of the 137 nt element to its ability to function as a localization element.


Antimicrobial effectiveness of wound matrices containing native extracellular matrix with polyhexamethylene biguanide.
A variety of wound matrix materials that are designed to help heal both acute and chronic wounds are currently available. Because wounds often encounter opportunistic microbes that can delay healing, the effectiveness of these materials is often suboptimal, resulting in delayed or compromised wound healing. The importance of reducing and controlling wound microbes is well recognised and there are several antimicrobial options available to address this unmet clinical need. This study compares the antimicrobial and wound healing capabilities, both in vivo and in vitro against methicillin-resistant Staphylococcus aureus (MRSA) USA 300, for the following compounds: Collagen Wound Matrix-Anti Microbial (CWM-AM); Collagen Wound Matrix-Anti Microbial XT (CWM-AM XT); Antimicrobial Hydrofiber Wound Dressing (AHWD); Dermal Scaffold with Silver (DRSAg); Collagen Extracellular Matrix (CEM); Collagen Wound Matrix (CWM); Matrix Wound Dressing with Silver (MWDAg); Cadexomer Iodine Gel (CIG); Triple Antibiotic Ointment (TAO); and Antimicrobial Wound Gel (AWG). For the in vitro zone of inhibition assay, AWG and CIG had the largest diffused areas, followed by CWM-AM and CWM-AM XT. Furthermore, CWM-AM, CWM-AM XT, AWG, and CIG exhibited a persistent antimicrobial activity for up to 10 days after incubation. However, in the cytotoxicity studies performed using human fibroblasts, CWM-AM and CWM-AM XT had no detrimental effects in cell proliferation and viability, while AWG and CIG were cytotoxic and prohibitive for cell proliferation. Treatments were then assessed for microbiology and wound healing efficacy using an in vivo porcine deep reticular dermal wound model. CWM-AM XT displayed the greatest in vivo antimicrobial activity against MRSA USA300 and expedited the reepithelialisation at a faster rate than other treatment groups. This study shows that a novel collagen matrix containing an antimicrobial agent can reduce the bacterial load and support healing.


Matrix Gla protein polymorphism rs1800801 associates with recurrence of ischemic stroke.
The MGP single nucleotide polymorphism (SNP) rs1800801 has previously been associated with recurrent ischemic stroke in a Spanish cohort. Here, we tested for association of this SNP with ischemic stroke recurrence in a North American Caucasian cohort. Acute ischemic stroke patients admitted between 10/2009 and 12/2016 at three hospitals within a large healthcare system in the northeastern United States that were enrolled in a healthcare system-wide exome sequencing program were retrospectively reviewed. Patients with recurrent stroke within 1 year after index event were compared to those without recurrence. Of 9,348 suspected acute ischemic strokes admitted between 10/2009 and 12/2016, 1,727 (18.5%) enrolled in the exome-sequencing program. Among those, 1,068 patients had exome sequencing completed and were eligible for inclusion. Recurrent stroke within the first year of stroke was observed in 79 patients (7.4%). In multivariable analysis, stroke prior to the index stroke (OR 9.694, 95% CI 5.793-16.224, p ≤ 0.001), pro-coagulant status (OR = 3.563, 95% CI 1.504-8.443, p = 0.004) and the AA genotype of SNP rs1800801 (OR = 2.408, 95% CI 1.079-4.389, p = 0.004) were independently associated with recurrent stroke within the first year. The AA genotype of the MGP SNP rs1800801 is associated with recurrence within the first year after ischemic stroke in North American Caucasians. Study of stroke subtypes and additional populations will be required to determine if incorporation of allelic status at this SNP into current risk scores improves prediction of recurrent ischemic stroke.


Log-odds sequence logos.
DNA and protein patterns are usefully represented by sequence logos. However, the methods for logo generation in common use lack a proper statistical basis, and are non-optimal for recognizing functionally relevant alignment columns. We redefine the information at a logo position as a per-observation multiple alignment log-odds score. Such scores are positive or negative, depending on whether a column's observations are better explained as arising from relatedness or chance. Within this framework, we propose distinct normalized maximum likelihood and Bayesian measures of column information. We illustrate these measures on High Mobility Group B (HMGB) box proteins and a dataset of enzyme alignments. Particularly in the context of protein alignments, our measures improve the discrimination of biologically relevant positions. Our new measures are implemented in an open-source Web-based logo generation program, which is available at http://www.ncbi.nlm.nih.gov/CBBresearch/Yu/logoddslogo/index.html. A stand-alone version of the program is also available from this site. firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Concurrent Infection of the Human Brain with Multiple Borrelia Species.
Lyme disease (LD) spirochetes are well known to be able to disseminate into the tissues of infected hosts, including humans. The diverse strategies used by spirochetes to avoid the host immune system and persist in the host include active immune suppression, induction of immune tolerance, phase and antigenic variation, intracellular seclusion, changing of morphological and physiological state in varying environments, formation of biofilms and persistent forms, and, importantly, incursion into immune-privileged sites such as the brain. Invasion of immune-privileged sites allows the spirochetes to not only escape from the host immune system but can also reduce the efficacy of antibiotic therapy. Here we present a case of the detection of spirochetal DNA in multiple loci in a LD patient's post-mortem brain. The presence of co-infection with Borrelia burgdorferi sensu stricto and Borrelia garinii in this LD patient's brain was confirmed by PCR. Even though both spirochete species were simultaneously present in human brain tissue, the brain regions where the two species were detected were different and non-overlapping. The presence of atypical spirochete morphology was noted by immunohistochemistry of the brain samples. Atypical morphology was also found in the tissues of experimentally infected mice, which were used as a control.


Occupational exposure to inhalable pathogenic microorganisms in waste sorting.
This study assessed microorganisms in personal inhalable work air samples aiming to identify potential human pathogens, and correlate exposure to adverse health outcomes in waste workers. Full-shift personal exposure was measured in six different waste sorting plants. Microbial concentrations in inhalable air samples were analysed using MALDI-TOF MS for cultivable, and next generation sequencing (NGS) for non-cultivable microorganisms. Concentrations of bacterial and fungal CFUs varied substantially within and between waste sorting plants, ranging from no identifiable organisms to a maximum concentration in the order of 105 CFU/m3. Bacillus and Staphylococcus were among the most abundant bacterial genera, whilst fungal genera were dominated by Aspergillus and Penicillium. Approximately 15% of all identified species were human pathogens classified in risk group 2, whereas 7% belonged to risk group 1. Furthermore, significant correlations between concentrations of fungi in risk group 1 and self-reported adverse symptoms, such as wheezing were identified in exposed workers. The combination of culture-based methods and NGS facilitated the investigation of infectious microbial species with potential pathophysiological properties as well as non-infectious biological agents in inhalable work air samples and thereby contributed to the risk assessment of occupational exposure in waste sorting.


Intramolecular repression of mouse heat shock factor 1.
The pathway leading to transcriptional activation of heat shock genes involves a step of heat shock factor 1 (HSF1) trimerization required for high-affinity binding of this activator protein to heat shock elements (HSEs) in the promoters. Previous studies have shown that in vivo the trimerization is negatively regulated at physiological temperatures by a mechanism that requires multiple hydrophobic heptad repeats (HRs) which may form a coiled coil in the monomer. To investigate the minimal requirements for negative regulation, in this work we have examined mouse HSF1 translated in rabbit reticulocyte lysate or extracted from Escherichia coli after limited expression. We show that under these conditions HSF1 behaves as a monomer which can be induced by increases in temperature to form active HSE-binding trimers and that mutations of either HR region cause activation in both systems. Furthermore, temperature elevations and acidic buffers activate purified HSF1, and mild proteolysis excises fragments which form HSE-binding oligomers. These results suggest that oligomerization can be repressed in the monomer, as previously proposed, and that repression can be relieved in the apparent absence of regulatory proteins. An intramolecular mechanism may be central for the regulation of this transcription factor in mammalian cells, although not necessarily sufficient.


Structural location determines functional roles of the basic amino acids of KR-12, the smallest antimicrobial peptide from human cathelicidin LL-37.
Cationic antimicrobial peptides are recognized templates for developing a new generation of antimicrobials to combat superbugs. Human cathelicidin LL-37 is an essential host defense molecule in human innate immunity. Previously, we identified KR-12 as the smallest antibacterial peptide of LL-37. KR-12 has a narrow activity spectrum since it is active against Gram-negative Escherichia coli but not Gram-positive Staphylococcus aureus. The functional roles of the basic amino acids of KR-12, however, have not yet been elucidated. An alanine scan of cationic amino acids of KR-12 provided evidence for their distinct roles in the activities of the peptides. Bacterial killing and membrane permeation experiments indicate that the R23A and K25A mutants, as well as the lysine-to-arginine mutant, were more potent than KR-12. Another three cationic residues (K18, R19, and R29) of KR-12, which are located in the hydrophilic face of the amphiphathic helix, appeared to be more important in clustering anionic lipids or hemolysis than R23 and K25 in the interfacial region. While the loss of interfacial R23 or K25 reduced peptide helicity, underscoring its important role in membrane binding, the overall increase in peptide activity of KR-12 could be ascribed to the increased peptide hydrophobicity that outweighed the role of basic charge in this case. In contrast, the mutations of interfacial R23 or K25 reduced peptide bactericidal activity of GF-17, an overlapping, more hydrophobic and potent peptide also derived from LL-37. Thus, the hydrophobic context of the peptide determines whether an alanine substitution of an interfacial basic residue increases or decreases membrane permeation and peptide activity.


Apixaban versus warfarin in patients with atrial fibrillation.
Vitamin K antagonists are highly effective in preventing stroke in patients with atrial fibrillation but have several limitations. Apixaban is a novel oral direct factor Xa inhibitor that has been shown to reduce the risk of stroke in a similar population in comparison with aspirin. In this randomized, double-blind trial, we compared apixaban (at a dose of 5 mg twice daily) with warfarin (target international normalized ratio, 2.0 to 3.0) in 18,201 patients with atrial fibrillation and at least one additional risk factor for stroke. The primary outcome was ischemic or hemorrhagic stroke or systemic embolism. The trial was designed to test for noninferiority, with key secondary objectives of testing for superiority with respect to the primary outcome and to the rates of major bleeding and death from any cause. The median duration of follow-up was 1.8 years. The rate of the primary outcome was 1.27% per year in the apixaban group, as compared with 1.60% per year in the warfarin group (hazard ratio with apixaban, 0.79; 95% confidence interval [CI], 0.66 to 0.95; P<0.001 for noninferiority; P=0.01 for superiority). The rate of major bleeding was 2.13% per year in the apixaban group, as compared with 3.09% per year in the warfarin group (hazard ratio, 0.69; 95% CI, 0.60 to 0.80; P<0.001), and the rates of death from any cause were 3.52% and 3.94%, respectively (hazard ratio, 0.89; 95% CI, 0.80 to 0.99; P=0.047). The rate of hemorrhagic stroke was 0.24% per year in the apixaban group, as compared with 0.47% per year in the warfarin group (hazard ratio, 0.51; 95% CI, 0.35 to 0.75; P<0.001), and the rate of ischemic or uncertain type of stroke was 0.97% per year in the apixaban group and 1.05% per year in the warfarin group (hazard ratio, 0.92; 95% CI, 0.74 to 1.13; P=0.42). In patients with atrial fibrillation, apixaban was superior to warfarin in preventing stroke or systemic embolism, caused less bleeding, and resulted in lower mortality. (Funded by Bristol-Myers Squibb and Pfizer; ARISTOTLE ClinicalTrials.gov number, NCT00412984.).


Hepatoprotective effect of Caesalpinia gilliesii and Cajanus cajan proteins against acetoaminophen overdose-induced hepatic damage.
This study aims to evaluate two proteins derived from the seeds of the plants Cajanus cajan (Leguminosae) and Caesalpinia gilliesii (Leguminosae) for their abilities to ameliorate the toxic effects of chronic doses of acetoaminphen (APAP) through the determination of certain biochemical parameters including liver marker enzymes: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and total bilirubin. Also, total protein content and hepatic marker enzyme, lactate dehydrogenase were studied. Moreover, liver antioxidants, glutathione (GSH), nitric oxide, and lipid peroxides were determined in this study. Hepatic adenosine triphosphatase (ATPase), adenylate energy charge (ATP, adenosine diphosphate, adenosine monophosphate, and inorganic phosphate), and phosphate potential, serum interleukin-6, tumor necrosis factor-α, and myeloperoxidase were also examined in the present study. On the other hand, histopathological examination of intoxicated and liver treated with both proteins was taken into consideration. The present results show disturbances in all biochemical parameters and hepatic toxicity signs including mild vascular congestion, moderate inflammatory changes with moderate congested sinusoids, moderate nuclear changes (pyknosis), moderate centrilobular necrosis, fatty changes, nuclear pyknosis vascular congestion, and change in fatty centrilobular necrosis liver. Improvement in all biochemical parameters studied was noticed as a result of treatment intoxicated liver with C. gilliesii and C. cajan proteins either paracetamol with or post paracetamol treatment. These results were documented by the amelioration signs in rat's hepatic architecture. Thus, both plant protein extracts can upregulate and counteract the inflammatory process, minimize damage of the liver, delay disease progression, and reduce its complications.


The prospective validation of the Modified Physiological Triage Tool (MPTT): an evidence-based approach to major incident triage.
Triage is a key principle in the effective management of major incidents. There is limited evidence to support existing triage tools, with a number of studies demonstrating poor performance at predicting the need for a life-saving intervention. The Modified Physiological Triage Tool (MPTT) is a novel triage tool derived using logistic regression, and in retrospective data sets has shown optimum performance at predicting the need for life-saving intervention. Physiological data and interventions were prospectively collected for consecutive adult patients with trauma (>18 years) presenting to the emergency department at Camp Bastion, Afghanistan, between March and September 2011. Patients were considered priority 1 (P1) if they received one or more interventions from a previously defined list. Patients were triaged using existing triage tools and the MPTT. Performance was measured using sensitivity and specificity, and a McNemar test with Bonferroni calculation was applied for tools with similar performance. The study population comprised 357 patients, of whom 214 (59.9%) were classed as P1. The MPTT (sensitivity: 83.6%, 95% CI 78.0% to 88.3%; specificity: 51.0%, 95% CI 42.6% to 59.5%) outperformed all existing triage tools at predicting the need for life-saving intervention, with a 19.6% absolute reduction in undertriage compared with the existing Military Sieve. The improvement in undertriage comes at the expense of overtriage; rates of overtriage were 11.6% higher with the MPTT than the Military Sieve. Using a McNemar test, a statistically significant (p<0.001) improvement in overall performance was demonstrated, supporting the use of the MPTT over the Military Sieve. The MPTT outperforms all existing triage tools at predicting the need for life-saving intervention, with the lowest rates of undertriage while maintaining acceptable levels of overtriage. Having now been validated on both military and civilian cohorts, we recommend that the major incident community consider adopting the MPTT for the purposes of primary triage.


Regulation of coenzyme A biosynthesis by glucagon and glucocorticoid in adult rat liver parenchymal cells.
We studied the effects of glucagon, dibutyryl cyclic AMP and dexamethasone on the rate of [(14)C]pantothenate conversion to CoA in adult rat liver parenchymal cells in primary culture. The presence of 30nm-glucagon increased the rate by about 1.5-fold relative to control cultures (range 1.4-2.3) and 2.4-fold relative to cultures containing 1-3m-i.u. of insulin/ml. The half-maximal effect was obtained at 3nm-glucagon. Dibutyryl cyclic AMP plus theophylline also enhanced the rate by about 1.5-fold. Dexamethasone acted synergistically with glucagon; glucagon at 0.3nm had no effect when added alone, but resulted in a 1.7-fold enhancement when added in the presence of dexamethasone (maximum effect at 50nm). The 1.4-fold enhancement caused by the addition of saturating glucagon concentrations was increased to a 3-fold overall enhancement by the addition of dexamethasone. However, dexamethasone added alone over the range 5nm to 5mum had no effect on the rate of [(14)C]pantothenate conversion to CoA. The stimulatory effect of dibutyryl cyclic AMP plus theophylline was also enhanced by the addition of dexamethasone. Changes in intracellular pantothenate concentration or radioactivity could not account for the stimulatory effects of glucagon, dibutyryl cyclic AMP or dexamethasone. Addition of 18mum-cycloheximide, an inhibitor of protein synthesis, decreased the rate of incorporation of [(14)C]pantothenate into CoA and the enhancement of this rate by glucagon and dibutyryl cyclic AMP plus theophylline in a reversible manner. These results demonstrate an influence of glucagon, dibutyryl cyclic AMP and glucocorticoids on the intracellular mechanism regulating total CoA concentrations in the liver.


The interpretation of leukoplakia in laryngeal pathology.
Leukoplakia is only a descriptive clinical term designating a white patch or plaque of the mucosa and must be complemented by histology. On the other hand, keratosis is an exclusively histological term denoting pathological production and accumulation of keratin on the surface of the laryngeal epithelium. Leukoplakia is usually keratosis, but not always. Keratosis can mask various epithelial changes, from simple hyperplasia to invasive squamous carcinoma and is only the superficially visible manifestation of an underlying pathological process. Keratosis means total replacement of superficial epithelial cells by keratin filaments, and dissolution of the nuclei. When nuclei are retained in keratinized cells, the process is termed parakeratosis. Therefore, keratosis can be classified as a separate entity only when histopathological examination reveals superficial keratotic changes accompanying a normal squamous epithelium. To identify the presence of keratosis in various benign laryngeal entities divided according to severity of epithelial abnormalities, and to determine whether keratosis has any prognostic value, we performed a retrospective analysis on bioptic material on 4,291 tissue specimens over a period of 15 years. Our results suggest that keratosis must be considered as only one sign of the disorder within the complex of other pathological changes and not as a distinct pathological entity. For this very reason, keratinization of the epithelial surface was not included among significant parameters used for the grading of epithelial changes into the particular group according to Kambic-Lenart classification.


Complex human disorders and molecular system engineering: historical perspective and potential impacts.
The challenging nature of complex human disorders has taught us that we can not untangle a disorder unless we understand how the "engine" of molecular systems works. After learning the basic physiology of different organs in the human body, a "molecular revolution" occurred, which has now generated a huge amount of information regarding the function of individual molecules in human cells. The difficult task, however, is to understand how thousands of molecules communicate and work together to deliver a specific function, and more importantly, what goes wrong when the system fails and causes different diseases. The emerging field of systems biology is now opening the door for engineers, to join molecular biologists and enter the era of molecular biomedical engineering.


Primary cutaneous coccidioidomycosis.
Primary cutaneous coccidioidomycosis is usually secondary to laboratory or autopsy inoculation, but can result from trauma in endemic areas. We describe an unusual case of primary inoculation cutaneous coccidioidomycosis occurring in an immunocompetent woman in a nonendemic area. The clinical features, diagnostic tests, and therapy of this extremely rare disease are reviewed.


Genetics redraws pelagic biogeography of Calanus.
Planktonic copepods of the genus Calanus play a central role in North Atlantic/Arctic marine food webs. Here, using molecular markers, we redrew the distributional ranges of Calanus species inhabiting the North Atlantic and Arctic Oceans and revealed much wider and more broadly overlapping distributions than previously described. The Arctic shelf species, C. glacialis, dominated the zooplankton assemblage of many Norwegian fjords, where only C. finmarchicus has been reported previously. In these fjords, high occurrences of the Arctic species C. hyperboreus were also found. Molecular markers revealed that the most common method of species identification, prosome length, cannot reliably discriminate the species in Norwegian fjords. Differences in degree of genetic differentiation among fjord populations of the two species suggested that C. glacialis is a more permanent resident of the fjords than C. finmarchicus We found no evidence of hybridization between the species. Our results indicate a critical need for the wider use of molecular markers to reliably identify and discriminate these morphologically similar copepod species, which serve as important indicators of climate responses.


The "Patient-centered coordination by a care team" questionnaire achieves satisfactory validity and reliability.
Increasing prevalence of chronic conditions and multimorbidity challenges health care systems and calls for patient-centered coordination of care. Implementation and evaluation of health policies focusing on the development of patient-centered coordination of care needs valid instruments measuring this dimension of care. The aim of this validation study was to assess the psychometric properties of the French version of the 14-item Patient-Centered Coordination by a Care Team (PCCCT) questionnaire in a primary care setting. PCCCT provides a total score from 0 (worst coordination) to 42 (best coordination). 165 adult patients consulting in primary care with one or more chronic condition(s) completed questionnaires (including PCCCT) at recruitment. After three weeks, participants completed PCCCT again, either by mail (group A) or during a telephone interview (group B). At recruitment, the mean (SD) PCCCT score was 33.3 (7.7). Exploratory factor analysis revealed a 2-dimension structure, 8 items relating to patient involvement and 6 items relating to coordination (factors loadings ranging from 0.34 to 0.88). PCCCT score correlated significantly with subscales of Haggerty's continuity questionnaire, Spearman correlation coefficients (95% confidence interval) ranging from 0.40 (0.22-0.57) to 0.52 (0.38-0.63). Internal consistency was excellent: Cronbach alpha 0.90 (0.79-0.92). Reliability was good, with an intraclass correlation coefficient of 0.68 (0.55-0.78) for test-retest reliability (group A) and of 0.65 (0.46-0.79) for reliability between the self-administered and the interviewer-administered versions of the questionnaire (group B,). The PCCCT questionnaire presents satisfactory validity and reliability; it can be used for the evaluation of health organizations involving team work in primary care.


Elevated asymmetric dimethylarginine in young adult survivors of childhood acute lymphoblastic leukemia: a preliminary report.
Adult survivors of childhood malignancy are predisposed to late cardiovascular (CV) complications. Our aim was to estimate plasma levels of the endogenous nitric oxide formation inhibitor asymmetric dimethylarginine (ADMA), in long-term survivors of childhood acute lymphoblastic leukemia (ALL) treated with only chemotherapy. ADMA and its isomer symmetric dimethylarginine (SDMA) were measured in 25 former ALL patients (aged 18-28 years) who had survived without recurrent disease ≥ 5 years from completing chemotherapy without cranial irradiation, and in 20 healthy controls (aged 20-31 years). Characteristics of the both groups were similar, except for lower high-density lipoproteins-cholesterol (HDL-C) in ALL survivors. Compared to controls, the former ALL patients exhibited significant, albeit small, rises in levels of ADMA (0.63 ± 0.09 [SD] vs. 0.57 ± 0.07 μmol/L; p=0.016), but not SDMA, with a consequently increased ADMA to SDMA ratio (1.08 ± 0.22 vs. 0.91 ± 0.16; p=0.004). The effect of former ALL on ADMA was attenuated (intergroup p=0.10 [ANCOVA]) upon adjustment for HDL-C (ADMA vs. HDL-C regression coefficient: -0.065 ± 0.030 [SEM]; p=0.03). ADMA is elevated in adult childhood ALL survivors, which can reflect late detrimental chemotherapy effects, partially related to minor lipid profile changes. Whether these subtle ADMA elevations might herald future CV morbidity, remains to be elucidated.


Contrast-enhanced MR of the facial nerve in patients with posttraumatic peripheral facial nerve palsy.
To estimate the value of noncontrast and contrast-enhanced T1-weighted MR imaging in detecting the underlying mechanisms of injury and regeneration in immediate- or delayed-onset posttraumatic peripheral facial nerve palsy. Twenty-four patients with posttraumatic peripheral facial nerve palsy were examined on a 1.5-T MR imaging unit with precontrast and postcontrast T1-weighted spin-echo and gradient-echo sequences. Abnormal enhancement of the distal intrameatal nerve segment was visible in 92% of the patients up to 2 years after their initial trauma. A hematoma within the geniculate ganglion was seen in 33% of the patients with a longitudinal fracture. The greater superficial petrosal nerve (in 32% of patients) and the geniculate ganglion (in 48% of patients) were thick and intensely enhancing. Hematoma within the cochlea/vestibule or enhancement of the cochlea/vestibule and the vestibulocochlear (eighth) nerve was observed in transverse fractures. MR images can show long-lasting abnormal nerve enhancement, especially in the distal intrameatal nerve segment, related to the long-lasting breakdown of the blood/peripheral nerve barrier associated with nerve degeneration and regeneration after traumatic stretching of the greater superficial petrosal nerve. Additionally, intraoperatively observed perineural and intraneural scar formation leads to thickening and intense enhancement of the affected nerve segments on MR images. A hematoma in the region of the geniculate ganglion can be seen in some but not all patients. Associated damage of the inner ear structures in patients with transverse fractures is also visible on MR images.


[Decontamination of cell cultures by removing mycoplasmas: a comparison of 3 methods].
Three different methods of decontamination of cell cultures from mycoplasma were compared: by macrophages in the presence of antibiotics, by ultraviolet radiation in the presence of 5-bromuridin and Hoechst-33258, by cloning of cells in the presence of antibiotics metacycline and doxicycline. Only the latter method was effective to decontaminate the cell cultures from mycoplasma. Twelve cell lines were decontaminated and tested for the presence of mycoplasma by microbiological method, by staining with the dye Hoechst-33258 and autoradiography. All the cell lines were free of mycoplasma.


Leniency and halo effects in marking undergraduate short research projects.
Supervisors are often involved in the assessment of projects they have supervised themselves. Previous research suggests that detailed marking sheets may alleviate leniency and halo effects. We set out to determine if, despite using such a marking schedule, leniency and halo effects were evident in the supervisors' marking of undergraduate short research projects (special study modules (SSM)). Review of grades awarded by supervisors, second markers and control markers to the written reports of 4th year medical students who had participated in an SSM during two full academic years (n = 399). Paired t-tests were used to compare mean marks, Pearson correlation to look at agreement between marks and multiple linear regression to test the prediction of one mark from several others adjusted for one another. There was a highly significant difference of approximately half a grade between supervisors and second markers with supervisors marking higher. (t = 3.12, p < 0.01, difference in grade score = 0.42, 95% CI for mean difference 0.18-0.80). There was a high correlation between the two marks awarded for performance of the project and the written report by the supervisor (r = 0.75), but a low-modest correlation between supervisor and second marker (r = 0.28). Linear regression analysis of the influence of the supervisors' mark for performance on their mark for the report gave a non-significant result. This suggests a leniency effect but no halo effect. This study shows that with the use of structured marking sheet for assessment of undergraduate medical students, supervisors marks are not associated with a halo effect, but leniency does occur. As supervisor assessment is becoming more common in both under graduate and postgraduate teaching new ways to improve objectivity in marking and to address the leniency of supervisors should be sought.


Impaired expression of ciliary neurotrophic factor in Charcot-Marie-Tooth type 1A neuropathy.
We investigated the contribution of Schwann cell-derived ciliary neurotrophic factor (CNTF) to the pathogenesis of Charcot-Marie-Tooth disease type 1A (CMT1A) and addressed the question as to whether it plays a role in the development of axonal damage observed in the disease, with aging. Ciliary neurotrophic factor was underexpressed in experimental CMT1A but not in other models of hereditary neuropathies. Sciatic nerve crush experiments and dosage of CNTF at different time points showed that expression of this trophic factor remained significantly lower in CMT1A rats than in normal controls; moreover, in uninjured CMT1A sciatic nerves CNTF levels further decreased with ageing, thus paralleling the molecular signs of axonal impairment, that is increased expression of non-phosphorylated neurofilaments and amyloid precursor protein. Administration of CNTF to dorsal root ganglia cultures reduced dephosphorylation of neurofilaments in CMT1A cultures, without improving demyelination. Taken together, these results provide further evidence that the production of CNTF by Schwann cells is markedly reduced in CMT1A. Moreover, the observations suggest that trophic support to the axon is impaired in CMT1A and that further studies on the therapeutic use of trophic factors or their derivatives in experimental and human CMT1A are warranted.


Influence of acetaminophen and ibuprofen on skeletal muscle adaptations to resistance exercise in older adults.
Evidence suggests that consumption of over-the-counter cyclooxygenase (COX) inhibitors may interfere with the positive effects that resistance exercise training has on reversing sarcopenia in older adults. This study examined the influence of acetaminophen or ibuprofen consumption on muscle mass and strength during 12 wk of knee extensor progressive resistance exercise training in older adults. Thirty-six individuals were randomly assigned to one of three groups and consumed the COX-inhibiting drugs in double-blind placebo-controlled fashion: placebo (67 ± 2 yr; n = 12), acetaminophen (64 ± 1 yr; n = 11; 4 g/day), and ibuprofen (64 ± 1 yr; n = 13; 1.2 g/day). Compliance with the resistance training program (100%) and drug consumption (via digital video observation, 94%), and resistance training intensity were similar (P > 0.05) for all three groups. Drug consumption unexpectedly increased muscle volume (acetaminophen: 109 ± 14 cm(3), 12.5%; ibuprofen: 84 ± 10 cm(3), 10.9%) and muscle strength (acetaminophen: 19 ± 2 kg; ibuprofen: 19 ± 2 kg) to a greater extent (P < 0.05) than placebo (muscle volume: 69 ± 12 cm(3), 8.6%; muscle strength: 15 ± 2 kg), when controlling for initial muscle size and strength. Follow-up analysis of muscle biopsies taken from the vastus lateralis before and after training showed muscle protein content, muscle water content, and myosin heavy chain distribution were not influenced (P > 0.05) by drug consumption. Similarly, muscle content of the two known enzymes potentially targeted by the drugs, COX-1 and -2, was not influenced (P > 0.05) by drug consumption, although resistance training did result in a drug-independent increase in COX-1 (32 ± 8%; P < 0.05). Drug consumption did not influence the size of the nonresistance-trained hamstring muscles (P > 0.05). Over-the-counter doses of acetaminophen or ibuprofen, when consumed in combination with resistance training, do not inhibit and appear to enhance muscle hypertrophy and strength gains in older adults. The present findings coupled with previous short-term exercise studies provide convincing evidence that the COX pathway(s) are involved in the regulation of muscle protein turnover and muscle mass in humans.


Differentiated membrane specializations and myofibrillar breakdown and recovery in cultured adult cardiac myocytes treated with TPA and diacylglycerol.
Cultured adult rat ventricular cardiac muscle cells were treated with either 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or diacylglycerol (DAG) and observed by in situ transmission electron microscopy. Membrane specializations present in untreated cells (intercalated discs, transverse tubules, plasmalemmal couplings) were also present after TPA and DAG treatment. In the case of the transverse tubular system, there was morphological evidence for active growth. Our studies showed that myofilaments began to become disorganized after 12-24 h of TPA treatment and that after 2 days of exposure to TPA the breakdown of sarcomeres was essentially complete. Myocytes that were treated with TPA for 2 days and then allowed to recover in control medium for 5 days contained sarcomeres in various stages of reassembly. These data indicate that TPA-treated cardiac myocytes retain several membrane specializations, suggesting that there are separate controls for myofilament organization and the maintenance of these differentiated plasmalemmal regions. Furthermore, the ability of the myocytes to recover from TPA treatment may provide investigators with a useful model with which to study myofibrillogenesis.


Investigation into the effect of humidity on drug-drug interactions using the atomic force microscope.
The atomic force microscope (AFM) has been used to characterize the cohesive nature of a micronized pharmaceutical powder used for inhalation therapy. Salbutamol sulfate (also referred to as albuterol sulfate), a therapeutic drug commonly delivered from dry powder inhalers (DPI), was chosen as a model system because the cohesion and subsequent de-agglomeration during inhalation are critical aspects to the efficacy of such a delivery system. Salbutamol sulfate drug particulates were mounted on V-shaped AFM cantilevers using a novel micromanipulation technique. Force-distance curves obtained from the measurements between cantilever drug probes and model compacts of salbutamol sulfate were integrated to determine separation energies. The effect of humidity (15-75% RH) on the energy required to separate a drug particle from model drug surface was determined using a custom-built perfusion apparatus attached to the AFM. Separation energy measurements over 10 x 10-microm areas of the compact surface (n = 4096) exhibited log normal distributions (apparent linear regression, R(2) >or= 0.97). Significant increases in the median separation energies (p < 0.05) between the salbutamol sulfate drug probes and salbutamol sulfate model surfaces were observed as humidity was increased. This result is most likely attributed to capillary interactions becoming more dominant at higher humidities. This investigation has shown the AFM to be a powerful technique for quantification of the separation energies between micronized drug particulates, highlighting the potential of the AFM as a rapid preformulation tool.


Characterization of proliferin-related protein.
Proliferin-related protein (mPRP) is a member of the PRL/GH family in the mouse. We have generated an antiserum against mPRP expressed as a bacterial fusion protein; this antiserum detects mPRP in the conditioned media of placental tissue cultures as a heterogeneous population of glycoproteins. We have also expressed mPRP in mammalian tissue culture cells and purified the secreted protein. N-terminal sequence analysis of the purified protein reveals that it is secreted as a 214 amino acid protein after removal of a 30 amino acid signal polypeptide. An antiserum raised against the purified protein detects high levels of mPRP in maternal serum during gestation. The site of synthesis of this protein has been localized by in situ hybridization to the basal zone of the day-10 mouse placenta, which is distinct from the site of synthesis of other placental proteins in this family.


Profile measurements in the plasma edge of mega amp spherical tokamak using a ball pen probe.
The ball pen probe (BPP) technique is used successfully to make profile measurements of plasma potential, electron temperature, and radial electric field on the Mega Amp Spherical Tokamak. The potential profile measured by the BPP is shown to significantly differ from the floating potential both in polarity and profile shape. By combining the BPP potential and the floating potential, the electron temperature can be measured, which is compared with the Thomson scattering (TS) diagnostic. Excellent agreement between the two diagnostics is obtained when secondary electron emission is accounted for in the floating potential. From the BPP profile, an estimate of the radial electric field is extracted which is shown to be of the order ∼1 kV/m and increases with plasma current. Corrections to the BPP measurement, constrained by the TS comparison, introduce uncertainty into the ER measurements. The uncertainty is most significant in the electric field well inside the separatrix. The electric field is used to estimate toroidal and poloidal rotation velocities from E × B motion. This paper further demonstrates the ability of the ball pen probe to make valuable and important measurements in the boundary plasma of a tokamak.


Liver transplantation and pancreatic resection: a single-center experience and a review of the literature.
Liver transplantation may occasionally be indicated in patients with unique clinical scenarios. Little is known regarding the outcomes of patients who have had a pancreatic resection prior to, in combination with, or after liver transplantation. A retrospective review of all patients undergoing liver transplantation from March 1998 to March 2008 identified 17 patients who also underwent pancreatic resection. An additional literature review was performed. Five underwent pancreatic resection prior to liver transplantation (1.7, 3.6, 3.8, 6.8, and 8.1 years), another 9 underwent pancreatic resection together with liver transplantation, and 3 underwent pancreatic resection after liver transplantation (2.2, 2.6, and 3.8 years). Indications for pancreatic resection included cholangiocarcinoma (n = 6), neuroendocrine tumor (n = 5), pancreatic cancer (n = 2), gastrointestinal stromal tumor (n = 1), periampullary adenocarcinoma (n = 1), duodenal adenomas (n = 1), and benign pancreatic mass (n = 1). Indications for liver transplantation were metastatic neuroendocrine tumor disease (n = 5), primary sclerosing cholangitis (n = 5), hepatitis C virus (n = 2), metastatic gastrointestinal stromal tumor (n = 1), Klatskin tumor (n = 1), alcohol cirrhosis (n = 1), alpha-1 antitrypsin deficiency (n = 1), and chemotherapy-induced cirrhosis (n = 1). One patient died intraoperatively, 7 patients died of tumor recurrence, 2 patients died from transplant complications, and 7 patients are still alive. Pancreatic resection-related complications included 4 pancreatic fistulas. A literature review confirmed liver transplantation/pancreatic resection-related complications. In conclusion, liver transplantation and pancreatic resection remain uncommon, and a good outcome can be achieved. Recurrence of malignant disease is the main factor limiting survival, and specific morbidity may be related to pancreatic resection and liver transplantation.


Ophthalmologic findings in acquired immune deficiency syndrome (AIDS).
Forty-one homosexually active men had ophthalmologic and immunologic evaluations. Four of eight with acquired immune deficiency syndrome (AIDS) had abnormal ocular findings that included cotton-wool spots, retinal hemorrhages, cytomegalovirus, retinitis and conjunctivitis due to cytomegalovirus, and keratoconjunctivitis sicca. The other four patients with AIDS and 33 homosexual male controls had normal ocular examinations. Patients with AIDS and abnormal eye findings had a notably lower total leukocyte count, absolute lymphocyte count, percentage T-helper lymphocytes, helper-suppressor lymphocyte ratio, hematocrit level, and platelet count than patients with AIDS and normal results on eye examination or controls. All patients with AIDS and abnormal eye examination results died; the four other patients with AIDS and normal eye findings remain alive. These observations suggest that ophthalmologic abnormalities are common in patients with AIDS, are associated with severe immunoregulatory abnormalities, and carry a poor prognosis.


Systematic review of the incidence of discitis after cervical discography.
Cervical discography is not uniformly used in part because of the fear of discitis. Studies report widely varying rates of this life-threatening infection. The aim of this study was to estimate the incidence of discitis after cervical discography, delineate the consequences of discitis, and identify factors that may influence this complication. Meta-analysis. Studies pertaining to cervical discography were identified by a literature review and bibliographic search. These were screened for inclusion into the meta-analysis by two reviewers. Data were collected on a wide range of clinical and demographic variables including age, gender, morbidities, number of patients, number of discograms, use of prophylactic antibiotics, type of surgical prep, number of needles used, and the number of patients and discs infected. Primary data were used to calculate the incidence of discitis per patient and per disc. Fourteen studies were included in the analysis. Both procedural details and demographic information on patients were missing from eight studies. The mean age of patients ranged from 41 to 47 years, and gender distribution varied greatly. Antibiotics use was reported in three studies. Cervical discography was complicated by postprocedural discitis in 22 of 14,133 disc injections (0.15%) and 21 of 4,804 patients (0.44%). Only one patient suffered from an infection at more than one spinal level. The rate of discitis after cervical discography is relatively low. This can perhaps be further decreased by the use of prophylactic intradiscal antibiotics. Should the ability of cervical discography to improve surgical outcomes be proven, the fear of discitis should not preclude performance of disc provocation.


The regenerative plasticity of isolated urodele myofibers and its dependence on MSX1.
The conversion of multinucleate postmitotic muscle fibers to dividing mononucleate progeny cells (cellularisation) occurs during limb regeneration in salamanders, but the cellular events and molecular regulation underlying this remarkable process are not understood. The homeobox gene Msx1 has been studied as an antagonist of muscle differentiation, and its expression in cultured mouse myotubes induces about 5% of the cells to undergo cellularisation and viable fragmentation, but its relevance for the endogenous programme of salamander regeneration is unknown. We dissociated muscle fibers from the limb of larval salamanders and plated them in culture. Most of the fibers were activated by dissociation to mobilise their nuclei and undergo cellularisation or breakage into viable multinucleate fragments. This was followed by microinjection of a lineage tracer into single fibers and analysis of the labelled progeny cells, as well as by time-lapse microscopy. The fibers showing morphological plasticity selectively expressed Msx1 mRNA and protein. The uptake of morpholino antisense oligonucleotides directed to Msx1 led to a specific decrease in expression of Msx1 protein in myonuclei and marked inhibition of cellularisation and fragmentation. Myofibers of the salamander respond to dissociation by activation of an endogenous programme of cellularisation and fragmentation. Lineage tracing demonstrates that cycling mononucleate progeny cells are derived from a single myofiber. The induction of Msx1 expression is required to activate this programme. Our understanding of the regulation of plasticity in postmitotic salamander cells should inform strategies to promote regeneration in other contexts.


Determination of heme in microorganisms using HPLC-MS/MS and cobalt(III) protoporphyrin IX inhibition of heme acquisition in Escherichia coli.
One of the main threats to the achievements in modern medicine is antimicrobial resistance. Molecular targeting of bacterial acquisition mechanisms of heme has been suggested to be an alternative to antibiotics. In the present study, HPLC-MS/MS combined with a simple clean-up based on liquid-liquid extraction has been developed and evaluated for simultaneous determination of heme and porphyrin heme precursors in microorganisms. Experimental design was used to optimize the extraction parameters, to obtain a method with high recovery, low matrix effects, and high precision. The effects of additives in the culture medium on the biosynthesis of heme were studied using Escherichia coli as a model microorganism. 5-Aminolaevulinic acid and hemin increased the heme concentration in E. coli by a factor of 1.5 and 4.5, respectively. Addition of 5-aminolaevulinic acid bypassed the E. coli negative feedback control of heme biosynthesis, which led to high amounts of intracellular porphyrins. The high heme concentration obtained when hemin was used as a culture additive shows that E. coli has an uptake of heme from its surroundings. In contrast, addition of cobalt protoporphyrin IX to the growth medium reduced the amount of heme in E. coli, demonstrating this compound's ability to mimic real heme and inhibit the heme acquisition mechanisms.


Non-destructive vacuum decay method for pre-filled syringe closure integrity testing compared with dye ingress testing and high-voltage leak detection.
In reaction to the limitations of the traditional sterility test methods, in 2008, the U.S. Food and Drug Administration issued the guidance "Container and Closure System Integrity Testing in Lieu of Sterility Testing as a Component of the Stability Protocol for Sterile Products" encouraging sterile drug manufacturers to use properly validated physical methods, apart from conventional microbial challenge testing, to confirm container closure integrity as part of the stability protocol. The case study presented in this article investigated the capability of four container closure integrity testing methods to detect simulated defects of different sizes and types on glass syringes, prefilled both with drug product intended for parenteral administration and sterile water. The drug product was a flu vaccine (Agrippal, Novartis Vaccines, Siena, Italy). Vacuum decay, pharmacopoeial dye ingress test, Novartis specific dye ingress test, and high-voltage leak detection were, in succession, the methods involved in the comparative studies. The case study execution was preceded by the preparation of two independent sets of reference prefilled syringes, classified, respectively, as examples of conforming to closure integrity requirements (negative controls) and as defective (positive controls). Positive controls were, in turn, split in six groups, three of with holes laser-drilled through the prefilled syringe glass barrel, while the other three with capillary tubes embedded in the prefilled syringe plunger. These reference populations were then investigated by means of validated equipment used for container closure integrity testing of prefilled syringe commercial production; data were collected and analyzed to determine the detection rate and the percentage of false results. Results showed that the vacuum decay method had the highest performance in terms of detection sensitivity and also ensured the best reliability and repeatability of measurements. An innovative technical solution, preventing possible prefilled syringe plunger movement during container closure integrity testing execution, is presented as well. The growing need to meet sterile drug products' regulatory, quality, and safety expectations has progressively driven new developments and improvements both in container closure integrity testing methods and in the respective equipment, over the last years. Indeed, container closure integrity testing establishes the container closure system capability to provide required protection to the drug product and to demonstrate maintenance of product sterility over its shelf life. This article describes the development of four container closure integrity testing approaches for the evaluation of glass prefilled syringe closure integrity, including two destructive (pharmacopoeial and Novartis specific dye ingress test) and two non-destructive (vacuum decay and high-voltage leak detection) methods. The important finding from the validation of comparative studies was that the vacuum decay method resulted in the most effective, reliable and repeatable detection of defective samples, whether the defect was exposed to sterile water, to drug product, or to air. Complete sets of known defects were created for this purpose (5 μm, 10 μm, 20 μm certified leakages by laser drilled holes and capillary tubes). All investigations and studies were conducted at Bonfiglioli Engineering S.r.l. (Vigarano Pieve, Ferrara, Italy) and at Novartis Vaccines (Sovicille, Siena, Italy).


Interactive analysis of systems biology molecular expression data.
Systems biology aims to understand biological systems on a comprehensive scale, such that the components that make up the whole are connected to one another and work through dependent interactions. Molecular correlations and comparative studies of molecular expression are crucial to establishing interdependent connections in systems biology. The existing software packages provide limited data mining capability. The user must first generate visualization data with a preferred data mining algorithm and then upload the resulting data into the visualization package for graphic visualization of molecular relations. Presented is a novel interactive visual data mining application, SysNet that provides an interactive environment for the analysis of high data volume molecular expression information of most any type from biological systems. It integrates interactive graphic visualization and statistical data mining into a single package. SysNet interactively presents intermolecular correlation information with circular and heatmap layouts. It is also applicable to comparative analysis of molecular expression data, such as time course data. The SysNet program has been utilized to analyze elemental profile changes in response to an increasing concentration of iron (Fe) in growth media (an ionomics dataset). This study case demonstrates that the SysNet software is an effective platform for interactive analysis of molecular expression information in systems biology.


[Inequity in health affects the development in Peru].
Unfair and avoidable differences in exercising the right to health coexist with the Universal Declaration of Human Rights since 1948. Some causes of the persistent inequality inferred after Alma-Ata are insufficient funding, development of national health systems without adequate prioritization, and since the 80s with economic consensus to introduce the health market model. Health in Peru is still an area of little progress, ostensible inequality and limited participation in development. Policies in the 20th century are still insufficient; and missing opportunities like increasing the value of exports over a decade ago is a recurring issue. For a health reform to be successful in terms of equality and development, it is necessary to agree on a state policy, establish a modern and equitable social security funding system and eradicate the inconsistencies in the national health system.


Claudin-19 mediates the effects of NO on the paracellular pathway in thick ascending limbs.
Claudins are a family of tight junction proteins that provide size and charge selectivity to solutes traversing the paracellular space. Thick ascending limbs (TALs) express numerous claudins, including claudin-19. Nitric oxide (NO), via cGMP, reduces dilution potentials in perfused TALs, a measure of paracellular permeability, but the role of claudin-19 is unknown. We hypothesized that claudin-19 mediates the effects of NO/cGMP on the paracellular pathway in TALs via increases in plasma membrane expression of this protein. We measured the effect of the NO donor spermine NONOate (SPM) on dilution potentials with and without blocking antibodies and plasma membrane expression of claudin-19. During the control period, the dilution potential was -18.2 ± 1.8 mV. After treatment with 200 μmol/l SPM, it was -14.7 ± 2.0 mV (P < 0.04). In the presence of claudin-19 antibody, the dilution potential was -12.7 ± 2.1 mV. After SPM, it was -12.9 ± 2.4 mV, not significantly different. Claudin-19 antibody alone had no effect on dilution potentials. In the presence of Tamm-Horsfall protein antibody, SPM reduced the dilution potential from -9.7 ± 1.0 to -6.3 ± 1.1 mV (P < 0.006). Dibutyryl-cGMP (500 µmol/l) reduced the dilution potential from -19.6 ± 2.6 to -17.2 ± 2.3 mV (P < 0.002). Dibutyryl-cGMP increased expression of claudin-19 in the plasma membrane from 29.9 ± 3.8% to 65.9 ± 10.1% of total (P < 0.011) but did not change total expression. We conclude that claudin-19 mediates the effects of the NO/cGMP signaling cascade on the paracellular pathway.


[Antibodies to the structural elements of the neuron in patients with amyotrophic lateral sclerosis and in their healthy relatives].
The results of determining antibodies to the structural elements of neuron in the serum and CSF of 100 patients with lateral amyotrophic sclerosis (LAS), in the serum of their relatives (n-67), and in biological fluids of 50 patients suffering from other neurologic diseases allowed the following conclusions to be made. The predominance of antibody demonstration in the CSF of LAS patients over that in the serum may attest to the immune response on the part of the CNS. The presence of antibodies in the relatives (not-blood ones for the most part) of LAS patients suggests an infectious nature of the disease.


CCK1R agonists: a promising target for the pharmacological treatment of obesity.
Almost 30 years have passed since Gibbs, Young, and Smith demonstrated the ability of exogenously administered cholecystokinin (CCK) to inhibit food intake in rats. This observation was the beginning of very extensive studies into the role CCK plays in the regulation of food intake in mammals. CCK is a brain-gut peptide, which exists in multiple forms. CCK peptides exert their action on two distinct receptor subtypes: CCK-A (Alimentary) now called the CCK1R, mostly expressed peripherally; and CCK-B (Brain), renamed the CCK2R, which is primarily present in the brain. Through the use of subtype-selective agonists and antagonists for the CCK receptor, it was determined that the effect of CCK on feeding was dependent on agonist induced activation of peripheral CCK1 receptors. This discovery was followed by intense research with the goal of identifying small molecule agonists on the CCK1 receptor as potentially useful agents for the treatment of obesity. This review will attempt to summarize the results of this research.


Agronomic importance of first development of chickpea (Cicer arietinum L.) under semi-arid conditions: II. Seed imbibition.
Due to the slowness growth and weakness of the first developments of chickpea (Cicer arietinum L.), it could not combated with weeds and easily caught up by Ascochyta blight (Ascochyta rabiei (Pass) Labr.) disease. Additionally, due to biotic and abiotic stress factors, esp. at the late sowing, important seed yield losses could be happened. To be able to avoid from them is only possible to accelerate of its first development as possible as. So, one of the best solutions to is to use chemical compounds such as Humic Acid (HA) known soil regulator under the semi-arid conditions. With this aim this research was performed in a Randomized Complete Block Design (RCBD) with four replications under semi-arid field conditions during (2008/2009) and (2009/2010) in Turkiye. Two cultivars (V1 = Gokce and V2 = Ispanyol) and four seed imbibition methods (A0 = 0, A1 = Tap Water, A2 = 1/2 Tap Water + 1/2 Humic acid (HA), A3 = Full HA, as w/w) and seven yield components Plant Height (PH), Number of Branches per Plant (NBP), Number of Pods per Plant (NPP), First Pod Height (NFP), Number of Seeds per Pod (NSP), Seed Weight per Plant (SWP) and 100-Seed weight (HSW) were investigated. The PH and FPH were affected the A0, the NBP, NPP and NSP were affected the A2 and the SWP and HSW were given the varied but not clear responses according to varieties for all the parameters in A1. The A0 and A1 were encouraged the germination and top soil of the plant but, the A2 to A3 were encouraged root system's development. It was concluded that the A2 is a promising method which makes the maximum and positive effect to the first development of the chickpea agronomy under the semi-arid conditions.


Organization of the gymnotiform fish pallium in relation to learning and memory: II. Extrinsic connections.
This study describes the extrinsic connections of the dorsal telencephalon (pallium) of gymnotiform fish. We show that the afferents to the dorsolateral and dorsomedial pallial subdivisions of gymnotiform fish arise from the preglomerular complex. The preglomerular complex receives input from four clearly distinct regions: (1) descending input from the pallium itself (dorsomedial and dorsocentral subdivisions and nucleus taenia); (2) other diencephalic nuclei (centroposterior, glomerular, and anterior tuberal nuclei and nucleus of the posterior tuberculum); (3) mesencephalic sensory structures (optic tectum, dorsal and ventral torus semicircularis); and (4) basal forebrain, preoptic area, and hypothalamic nuclei. Previous studies have implicated the majority of the diencephalic and mesencephalic nuclei in electrosensory, visual, and acousticolateral functions. Here we discuss the implications of preglomerular/pallial electrosensory-associated afferents with respect to a major functional dichotomy of the electric sense. The results allow us to hypothesize that a functional distinction between electrocommunication vs. electrolocation is maintained within the input and output pathways of the gymnotiform pallium. Electrocommunication information is conveyed to the pallium through complex indirect pathways that originate in the nucleus electrosensorius, whereas electrolocation processing follows a conservative pathway inherent to all vertebrates, through the optic tectum. We hypothesize that cells responsive to communication signals do not converge onto the same targets in the preglomerular complex as cells responsive to moving objects. We also hypothesize that efferents from the dorsocentral (DC) telencephalon project to the dorsal torus semicircularis to regulate processing of electrocommunication signals, whereas DC efferents to the tectum modulate sensory control of movement.


Obesity, diabetes, and preoperative hyperglycemia as predictors of periprosthetic joint infection: a single-center analysis of 7181 primary hip and knee replacements for osteoarthritis.
Diabetes and obesity are common in patients undergoing joint replacement. Studies analyzing the effects of diabetes and obesity on the occurrence of periprosthetic joint infection have yielded contradictory results, and the combined effects of these conditions are not known. The one-year incidence of periprosthetic joint infections was analyzed in a single-center series of 7181 primary hip and knee replacements (unilateral and simultaneous bilateral) performed between 2002 and 2008 to treat osteoarthritis. The data regarding periprosthetic joint infection (defined according to Centers for Disease Control and Prevention criteria) were collected from the hospital infection register and were based on prospective, active surveillance. Patients diagnosed with diabetes were identified from the registers of the Social Insurance Institution of Finland. The odds ratios (ORs) for infection and the accompanying 95% confidence intervals (CIs) were calculated with use of binary logistic regression with adjustment for age, sex, American Society of Anesthesiologists risk score, arthroplasty site, body mass index, and diabetic status. Fifty-two periprosthetic joint infections occurred during the first postoperative year (0.72%; 95% CI, 0.55% to 0.95%). The infection rate increased from 0.37% (95% CI, 0.15% to 0.96%) in patients with a normal body mass index to 4.66% (95% CI, 2.47% to 8.62%) in the morbidly obese group (adjusted OR, 6.4; 95% CI, 1.7 to 24.6). Diabetes more than doubled the periprosthetic joint infection risk independent of obesity (adjusted OR, 2.3; 95% CI, 1.1 to 4.7). The infection rate was highest in morbidly obese patients with diabetes; this group contained fifty-one patients and periprosthetic infection developed in five (9.8%; 95% CI, 4.26% to 20.98%). In patients without a diagnosis of diabetes at the time of the surgery, there was a trend toward a higher infection rate in association with a preoperative glucose level of ≥6.9 mmol/L (124 mg/dL) compared with <6.9 mmol/L. The infection rate was 1.15% (95% CI, 0.56% to 2.35%) in the former group compared with 0.28% (95% CI, 0.15% to 0.53%) in the latter, and the adjusted OR was 3.3 (95% CI, 0.96 to 11.0). The type of diabetes medication was not associated with the infection rate. Diabetes and morbid obesity increased the risk of periprosthetic joint infection following primary hip and knee replacement. The benefits of joint replacement should be carefully weighed against the incidence of postoperative infection, especially in morbidly obese patients. Prognostic Level II. See Instructions for Authors for a complete description of levels of evidence.


Stigma/style cell cycle inhibitor 1 (SCI1), a tissue-specific cell cycle regulator that controls upper pistil development.
A cDNA encoding a small lysine-rich protein of unknown function was identified in a tobacco (Nicotiana tabacum) stigma/style suppression subtractive hybridization cDNA library. After its characterization, the corresponding gene was designated stigma/style cell cycle inhibitor 1 (SCI1). Fluorescence microscopy with an SCI1-GFP protein fusion demonstrated its nuclear localization, which was confined to the interchromatic region. Real-time RT-PCR and in situ hybridization experiments showed that SCI1 is stigma/style-specific and developmentally regulated. SCI1 RNAi knockdown and overexpression plants had stigmas/styles with remarkably enlarged and reduced areas, respectively, which was attributable to differences in cell numbers. These results indicate that SCI1 is a tissue-specific negative cell cycle regulator. The differences in cell division had an effect on the timing of the differentiation of the stigmatic papillar cells, suggesting that their differentiation is coupled to stigma cell divisions. This is consistent with a role for SCI1 in triggering differentiation through cell proliferation control. Our results revealed that SCI1 is a novel tissue-specific gene that controls cell proliferation/differentiation, probably as a component of a developmental signal transduction pathway.


[Benign tumors of the uterus and ovaries in women with chronic obstructive bronchopulmonary diseases].
Some aspects were studied of origination of uterine myoma and ovarian cyst in 121 women younger than 35 years of age presenting with chronic obstructive bronchopulmonary pathology. Uterine and ovarian benign tumors are more likely to develop against the background of chronic obstructive pulmonary pathology in the presence of respiratory failure, pulmonary emphysema, bronchiectatic disease, grave progress of the illness, development of lung disease at age, less than 10, with the above disease being more than ten years in duration.


Amphiphilic gold nanoparticles perturb phase separation in multidomain lipid membranes.
Amphiphilic gold nanoparticles with diameters in the 2-4 nm range are promising as theranostic agents thanks to their spontaneous translocation through cell membranes. This study addresses the effects that these nanoparticles may have on a distinct feature of plasma membranes: lipid lateral phase separation. Atomic force microscopy, quartz crystal microbalance, and molecular dynamics are combined to study the interaction between model neuronal membranes, which spontaneously form ordered and disordered lipid domains, and amphiphilic gold nanoparticles having negatively charged surface functionalization. Nanoparticles are found to interact with the bilayer and form bilayer-embedded ordered aggregates. Nanoparticles also suppress lipid phase separation, in a concentration-dependent fashion. A general, yet simple thermodynamic model is developed to show that the change of lipid-lipid enthalpy is the dominant driving force towards the nanoparticle-induced destabilization of phase separation.


Roles of the Picornaviral 3C Proteinase in the Viral Life Cycle and Host Cells.
The Picornaviridae family comprises a large group of non-enveloped viruses that have a major impact on human and veterinary health. The viral genome contains one open reading frame encoding a single polyprotein that can be processed by viral proteinases. The crucial 3C proteinases (3C(pro)s) of picornaviruses share similar spatial structures and it is becoming apparent that 3C(pro) plays a significant role in the viral life cycle and virus host interaction. Importantly, the proteinase and RNA-binding activity of 3C(pro) are involved in viral polyprotein processing and the initiation of viral RNA synthesis. In addition, 3C(pro) can induce the cleavage of certain cellular factors required for transcription, translation and nucleocytoplasmic trafficking to modulate cell physiology for viral replication. Due to interactions between 3C(pro) and these essential factors, 3C(pro) is also involved in viral pathogenesis to support efficient infection. Furthermore, based on the structural conservation, the development of irreversible inhibitors and discovery of non-covalent inhibitors for 3C(pro) are ongoing and a better understanding of the roles played by 3C(pro) may provide insights into the development of potential antiviral treatments. In this review, the current knowledge regarding the structural features, multiple functions in the viral life cycle, pathogen host interaction, and development of antiviral compounds for 3C(pro) is summarized.


T-cell chronic lymphocytic leukemia or small-cell variant of T-cell prolymphocytic leukemia: a historical perspective and search for consensus.
There is a rich history behind the extinct entity 'T-cell chronic lymphocytic leukemia (T-CLL)' and the now-established replacement, small-cell variant of T-cell prolymphocytic leukemia (T-PLL-sv). Herein, we review the history of the events, observations, and discussions that led to this replacement. We also provide a systematic analysis of all previously reported cases of T-PLL-sv as well as our four new additional cases. Despite the higher frequency of a normal karyotype and perhaps an overrepresented CD4(-) CD8(-) immunophenotype among these patients (compared to T-PLL in general) as well as bland morphology (that makes them superficially appear more similar to B-CLL), we argue that the current World Health Organization (WHO)-based classification as T-PLL-sv is adequate and should continue for the time being. Morphologically, T-PLL-sv represents approximately one-fifth of all T-PLL cases. However, morphology alone does not determine the clinical course and should not be the basis for clinical decision making and prognostication. We propose a clonal evolution model in which mature T-cell leukemias classified in the past as T-CLL are perhaps T-PLL diagnosed early in the course of the disease. Future research using next-generation sequencing, comparative genomic hybridization, and molecular array studies, including serial analyses of individual cases over time, is needed to better identify this rarely diagnosed, inherently controversial form of T-cell leukemia.


Epidemiologic data on the relationships of caloric intake, energy balance, and weight gain over the life span with longevity and morbidity.
Animal experiments have shown that calorically restricted (CR) animals weigh less and live longer than their ad libitum-fed peers. Are these observations applicable to human beings? This is an important question because the prevalence of obesity in America has increased markedly over recent years. We examine whether there are physiologic effects that occur with CR in humans that could plausibly explain the observed longevity of laboratory animals associated with CR. We also review epidemiologic data from observational and interventional studies on the relationships of caloric intake, energy balance, and weight gain with age-related diseases and longevity. Additionally, data on whether long-term, sustained maintenance of weight loss is feasible, as well as the degree of CR achieved in clinical trials, are summarized. Finally, we provide recommendations regarding further epidemiologic research that will help clarify unanswered questions in these areas.


Association of longer telomeres with better health in centenarians.
Prior animal model studies have demonstrated an association between telomere length and longevity. Our study examines telomere length in centenarians in good health versus poor health. Using DNA from blood lymphocytes, telomere length was measured by quantitative polymerase chain reaction in 38 sex- and age-matched centenarians (ages 97-108). "Healthy" centenarians (n = 19) with physical function in the independent range and the absence of hypertension, congestive heart failure, myocardial infarction, peripheral vascular disease, dementia, cancer, stroke, chronic obstructive pulmonary disease, and diabetes were compared to centenarians with physical function limitations and > or =2 of the above conditions (n = 19). Healthy centenarians had significantly longer telomeres than did unhealthy centenarians (p =.0475). Our study demonstrated that investigations of the association between telomere length and exceptional longevity must take into account the health status of the individuals. This raises the possibility that perhaps it is not exceptional longevity but one's function and health that may be associated with telomere length.


Prediction of type 1 diabetes postpartum in patients with gestational diabetes mellitus by combined islet cell autoantibody screening: a prospective multicenter study.
Women with gestational diabetes mellitus (GDM) have a considerable risk of developing diabetes later in life. To determine the predictive value of autoantibody markers in gestational diabetic pregnancy for the development of type 1 diabetes postpartum, we tested 437 patients with GDM (289 women treated with diet only [GDM-A] and 148 requiring insulin treatment during pregnancy [GDM-B]) for antibodies to islet cells (ICAs), GAD (GADAs), and tyrosine phosphatase ICA512/IA-2 (IA2As). We prospectively followed them with repeated oral glucose tolerance tests and antibody determinations for up to 7 years postpartum (mean, 1.6 years; range, 0-7.2 years). The cumulative risk of diabetes up to 5 years postpartum was 17% (95% CI 12-22%). The risk of type 1 diabetes was 3% (2-5%) by 9 months and 7% (4-9%) 2 years after delivery. At delivery, 8.5% of all patients were ICA+, 9.5% were GADA+, 6.2% were IA2A+, and 18.1% were positive for at least one antibody (12.6% for GDM-A vs. 30.4% for GDM-B, P < 0.0001). During follow-up, GADAs persisted in 75%, ICAs in 35%, and IA2As in 30% of the subjects positive for the respective marker at delivery. By 2 years postpartum, 29% (19-39%) of patients positive for at least one antibody developed type 1 diabetes, compared with 2% (1-4%) of antibody-negative patients (P < 0.0001). Thereby, the risk for type 1 diabetes 2 years postpartum increased with the number of antibodies present at delivery from 17% (6-28%) for one antibody, to 61% (30-91%) for two antibodies, and to 84% (55-100%) for 3 antibodies. Risk of progression to type 1 diabetes postpartum was also associated with the status of parity. Women with one or more pregnancies before the index pregnancy had a higher risk for type 1 diabetes 2 years after delivery (14.7% [4.9.-24.5%]) than women having their first (i.e., index) pregnancy (5% [2.9-7.1%]) (P < 0.006). A comparison of different prediction strategies showed that single antibody screening with GADA yielded the highest sensitivity of 63% (45-75%), compared with ICA (48% [31-65%]) and IA2A (34% [13-47%]). Combined screening with two autoantibodies increased sensitivity to 74% (58-90%) and 75% (60-92%) when using GADA plus ICA or GADA plus IA2A, respectively. Screening with all three markers improved sensitivity further to 82% (67-100%). Beta-cell autoantibodies determined at delivery in women with GDM are highly predictive for the development of type 1 diabetes postpartum. Autoantibody screening in pregnant women with GDM from populations at high risk for type 1 diabetes should therefore be considered to allow early diagnosis and appropriate therapy.


Reconstruction of the eyelids after mohs surgery.
A variety of defects of the upper and lower eyelids can be encountered after Mohs surgery. The goals of eyelid reconstruction are to provide structural and functional restoration with an acceptable aesthetic result. A thorough knowledge of the intricate anatomy of the eyelids combined with familiarity of the wide variety of reconstructive options is required to achieve these goals. In this article, we review the anatomy of the eyelids, and we present commonly used techniques for reconstruction of the upper and lower eyelids and the medial and lateral canthal regions after Mohs surgery.


A novel brief treatment for methamphetamine use disorders in South Africa: a randomised feasibility trial.
Effective brief treatments for methamphetamine use disorders (MAUD) are urgently needed to complement longer more intensive treatments in low and middle income countries, including South Africa. To address this gap, the purpose of this randomised feasibility trial was to determine the feasibility of delivering a six-session blended imaginal desensitisation, plus motivational interviewing (IDMI) intervention for adults with a MAUD. We enrolled 60 adults with a MAUD and randomly assigned them 1:1 to the IDMI intervention delivered by clinical psychologists and a control group who we referred to usual care. Feasibility measures, such as rates of recruitment, consent to participate in the trial and retention, were calculated. Follow-up interviews were conducted at 6 weeks and 3 months post-enrollment. Over 9 months, 278 potential particiants initiated contact. Following initial screening 78 (28%) met inclusion criteria, and 60 (77%) were randomised. Thirteen of the 30 participants assigned to the treatment group completed the intervention. Both psychologists were highly adherent to the intervention, obtaining a fidelity rating of 91%. In total, 39 (65%) participants completed the 6-week follow-up and 40 (67%) completed the 3-month follow-up. The intervention shows potential effectiveness in the intention-to-treat analysis where frequency of methamphetamine use was significantly lower in the treatment than in the control group at both the 6 week and 3-month endpoints. No adverse outcomes were reported. This feasibility trial suggests that the locally adapted IDMI intervention is an acceptable and safe intervention as a brief treatment for MAUD in South Africa. Modifications to the study design should be considered in a fully powered, definitive controlled trial to assess this potentially effective intervention. Trial registration The trial is registered with the Pan African Clinical Trials Registry (Trial ID: PACTR201310000589295).


Is vitamin D deficiency a confounder in alcoholic skeletal muscle myopathy?
Excessive intake of alcohol is often associated with low or subnormal levels of vitamin D even in the absence of active liver disease. As vitamin D deficiency is a well-recognized cause of myopathy, alcoholic myopathy might be related to vitamin D deficiency. Chronic alcoholic myopathy affects approximately half of chronic alcoholics and is characterized by the insidious development of muscular weakness and wasting. Although alcohol or its metabolites may have a direct toxic effect on muscles, the relationship between alcoholic myopathy and vitamin D deficiency has not been examined extensively. We reviewed the literature on alcoholic myopathy and hypovitaminosis D myopathy and compared the pathophysiological findings to designate possible mechanisms of vitamin D action in alcohol-related myopathy. Given the strong interdependency of suboptimal levels of vitamin D, phosphate, and magnesium in chronic alcohol abuse, we hypothesize that combined deficiencies interfere with membrane and intracellular metabolic processes in chronic alcohol-related myopathy; however, it is not yet possible to define exact mechanisms of interaction.


Cones, pringles, and grain boundary landscapes in graphene topology.
A polycrystalline graphene consists of perfect domains tilted at angle alpha to each other and separated by the grain boundaries (GB). These nearly one-dimensional regions consist in turn of elementary topological defects, 5-pentagons and 7-heptagons, often paired up into 5-7 dislocations. Energy G(alpha) of GB computed for all range 0 <or= alpha <or= pi/3, shows a slightly asymmetric behavior, reaching approximately 5 eV/nm in the middle, where the 5's and 7's qualitatively reorganize in transition from nearly armchair to zigzag interfaces. Analysis shows that two-dimensional (2D) nature permits the off-plane relaxation, unavailable in three-dimensional (3D) materials, qualitatively reducing the energy of defects on one hand while forming stable 3D landscapes on the other. Interestingly, while the GB display small off-plane elevation, the random distributions of 5's and 7's create roughness that scales inversely with defect concentration, h approximately n(-1/2).


Anti-inflammatory and metabolic actions of FXR: insights into molecular mechanisms.
The farnesoid X receptor (FXR) is a ligand-activated transcription factor belonging to the nuclear receptor (NR) superfamily. FXR plays an important role in positively regulating genes (transactivation) involved in bile acid homeostasis, fat and glucose metabolism. Recently, it has become clear that an additional important role for FXR consists of downregulating genes involved in inflammation. Because of this broad spectrum of regulated genes, therapeutically targeting FXR with full agonists will likely result in adverse side effects, in line with what is described for other NRs. It may therefore be necessary to develop selective FXR modulators. However, the molecular mechanisms that distinguish between FXR-mediated transactivation and transrepression are currently unknown. For other NRs, post-translational modifications such as SUMOylation and phosphorylation have been reported to be unique to either transactivation or transrepression. Here, we review current knowledge on post-translational regulation of FXR with respect to transactivation and transrepression. Ultimately, increased understanding of the different mechanisms of transactivation and transrepression of nuclear receptors will aid in the development of NR drugs with fewer side effects.


Efficacy and safety of intracavernosal alprostadil in men with erectile dysfunction. The Alprostadil Study Group.
Erectile dysfunction is a common medical problem affecting many men. Although several intracavernosal therapies are available, their efficacy and safety have not been studied systematically. We investigated the efficacy and safety of alprostadil formulated for intracavernosal treatment in three separate multi-institutional, prospective studies in men with erectile dysfunction of vasculogenic, neurogenic, psychogenic, and mixed causes. Clinical and laboratory evaluations of erection and the feasibility of satisfactoriness of sexual activity as assessed both by the men and by their partners were the primary measures of efficacy. In a dose-response study of 296 men, all doses of alprostadil were superior to placebo and there was a significant dose-response relation (P < / = 0.001), resulting in higher response rates with increasing doses of alprostadil (from 2.5 to 20 microg). In a dose-finding study of 201 men, the minimal effective dose was < / = 2 microg in 23, 20, 38 and 23 percent of men with erectile dysfunction of neurogenic, vasculogenic, psychogenic, or mixed causes, respectively. In a six-month self-injection study in 683 men, the participants reported being able to have sexual activity after 94 percent of the injections. The men and their partners rated the sexual activity as satisfactory after 87 and 86 percent of the injections, respectively. Penile pain, usually mild, occurred in 50 percent of the men at some time but after only 11 percent of the injections. Prolonged erections occurred in 5 percent of the men, priapism in 1 percent, penile fibrotic complications in 2 percent, and hematoma or ecchymosis in 8 percent. In men with erectile dysfunction, intracavernosal injection of alprostadil is an effective therapy with tolerable side effects.


Situation of onchocerciasis transmission in 2020 in the Cascades region of Burkina Faso.
Faced with the focal resurgence of onchocerciasis reported since 2004 in the South-West of Burkina Faso, the Neglected Tropical Diseases Control Program adopted a resumption of biannual community-directed treatment with ivermectin, since 2011 in the Cascades region and since 2013 in the South West region. The objective of this study was to assess the situation of onchocerciasis transmission in the Cascades region, nine years after the resumption of mass drug administration. This cross-sectional and descriptive survey concerned people over 5. The traditional method of skin snip on both iliac crests was performed for the parasitological diagnosis of onchocerciasis. The Ov-16 serological test was carried out in children aged 5 to 9 years. In 22 surveyed villages, the overall prevalence of microfilariae was 0.11% and below the tolerable threshold of 5%. It was less than 5% in all the villages (n = 22), less than 1% in 21 villages (99%) and zero in 19 villages (86.36%). The community microfilarial loads varied from 0.01 to 0.05 mf/b. Out of 946 children tested for OV-16, only one 9-year-old was positive and whose skin snip examination was negative. All the positive cases came from endemical areas in Côte d'Ivoire. Population migration is a risk factor for introducing the parasite into Burkina Faso; it also is risk factor for the effective elimination of onchocerciasis which requires the joint development of a control strategy between neighboring countries.


Cationic lipid polymerization as a novel approach for constructing new DNA delivery agents.
In vivo gene delivery mediated by cationic lipids is often compromised by aggregation due to complexation with proteins in the blood. To improve the stability of cationic lipid-DNA complexes, the present study aimed to develop a novel approach in which a poly(cationic lipid) (PCL) is utilized to form stable cationic polyplexes for gene transfection. Hydrogenation of the acrylamide analogue of betaAE-DMRI, the polymerizable precursor of PCL, provided a monomeric lipid derivative (MHL) which was used for direct comparison of corresponding lipoplex stability, toxicity, and transfection activity. Various formulations of cationic liposomes, such as MHL, MHL-cholesterol (Chol), PCL, PCL-Chol, DOTAP-Chol, and commercially available lipofectamine were generated and examined in this study. The new poly(cationic lipid) did not display any significant toxicity to rat hepatocytes or Hep G2 cells as indicated by an LDH leakage assay. Furthermore, PCL was significantly less toxic than MHL, DOTAP-Chol or lipofectamine. Suspensions of PCL were resistant to aggregation even after 24 h of exposure to solutions containing 50 and 100% fetal bovine serum (FBS). In contrast, suspensions of lipofectamine extensively aggregated after 24 h of exposure to 50% FBS. To examine the influence of lipid polymerization on gene transfer activity, liposome-mediated transfections of a luciferase vector (pGL3) were performed in Hep G2 and Alexander cell lines. The luciferase activity of the PCL formulations in Hep G2 cells were similar to those of the MHL, DOTAP-Chol and lipofectamine formulations, demonstrating that lipid polymerization does not compromise transfection activity. In comparison to the monomeric precursor MHL and to the industry transfection standards DOTAP and lipofectamine, the novel poly(cationic lipid) exhibited the lowest cytotoxicity, was the most resistant to serum-induced aggregation and had comparable transfection activity when coformulated with cholesterol. This novel polymerization approach for the development of stable and active polyplexes may prove a valuable alternative for in vivo gene delivery.


Non-target time trend screening: a data reduction strategy for detecting emerging contaminants in biological samples.
Non-targeted mass spectrometry-based approaches for detecting novel xenobiotics in biological samples are hampered by the occurrence of naturally fluctuating endogenous substances, which are difficult to distinguish from environmental contaminants. Here, we investigate a data reduction strategy for datasets derived from a biological time series. The objective is to flag reoccurring peaks in the time series based on increasing peak intensities, thereby reducing peak lists to only those which may be associated with emerging bioaccumulative contaminants. As a result, compounds with increasing concentrations are flagged while compounds displaying random, decreasing, or steady-state time trends are removed. As an initial proof of concept, we created artificial time trends by fortifying human whole blood samples with isotopically labelled standards. Different scenarios were investigated: eight model compounds had a continuously increasing trend in the last two to nine time points, and four model compounds had a trend that reached steady state after an initial increase. Each time series was investigated at three fortification levels and one unfortified series. Following extraction, analysis by ultra performance liquid chromatography high-resolution mass spectrometry, and data processing, a total of 21,700 aligned peaks were obtained. Peaks displaying an increasing trend were filtered from randomly fluctuating peaks using time trend ratios and Spearman's rank correlation coefficients. The first approach was successful in flagging model compounds spiked at only two to three time points, while the latter approach resulted in all model compounds ranking in the top 11 % of the peak lists. Compared to initial peak lists, a combination of both approaches reduced the size of datasets by 80-85 %. Overall, non-target time trend screening represents a promising data reduction strategy for identifying emerging bioaccumulative contaminants in biological samples. Graphical abstract Using time trends to filter out emerging contaminants from large peak lists.


Cell origin, volume and arrangement are drivers of articular cartilage formation, morphogenesis and response to injury in mouse limbs.
Limb synovial joints are composed of distinct tissues, but it is unclear which progenitors produce those tissues and how articular cartilage acquires its functional postnatal organization characterized by chondrocyte columns, zone-specific cell volumes and anisotropic matrix. Using novel Gdf5CreERT2 (Gdf5-CE), Prg4-CE and Dkk3-CE mice mated to R26-Confetti or single-color reporters, we found that knee joint progenitors produced small non-migratory progenies and distinct local tissues over prenatal and postnatal time. Stereological imaging and quantification indicated that the columns present in juvenile-adult tibial articular cartilage consisted of non-daughter, partially overlapping lineage cells, likely reflecting cell rearrangement and stacking. Zone-specific increases in cell volume were major drivers of tissue thickening, while cell proliferation or death played minor roles. Second harmonic generation with 2-photon microscopy showed that the collagen matrix went from being isotropic and scattered at young stages to being anisotropic and aligned along the cell stacks in adults. Progenitor tracing at prenatal or juvenile stages showed that joint injury provoked a massive and rapid increase in synovial Prg4+ and CD44+/P75+ cells some of which filling the injury site, while neighboring chondrocytes appeared unresponsive. Our data indicate that local cell populations produce distinct joint tissues and that articular cartilage growth and zonal organization are mainly brought about by cell volume expansion and topographical cell rearrangement. Synovial Prg4+ lineage progenitors are exquisitely responsive to acute injury and may represent pioneers in joint tissue repair.