The cost-effectiveness of prophylaxis with valaciclovir in the management of cytomegalovirus after renal transplantation.
Prophylaxis-based antiviral treatment and intensive monitoring followed by pre-emptive antiviral treatment are both commonly used management strategies to reduce risk of cytomegalovirus (CMV) infection following renal transplantation. This study employed a decision-model approach using published efficacy data and information from a recent survey of French clinical practice to consider the relative costs and outcomes associated with CMV prevention strategies for high-risk patient groups. The cost per case of treating tissue invasive and symptomatic CMV disease was estimated at euro 15,431 and euro 10,852, respectively. In the highest infection-risk patient group (positive donor with no previous CMV history) prophylactic oral valaciclovir was shown to avoid the greatest number of CMV disease cases (35 cases per 100 transplanted patients) and reduced the overall CMV-related costs per transplanted patient by around 14% over a'wait-and-treat' baseline strategy. In contrast, intensive monitoring and preemptive treatment resulted in a much higher cost per transplanted patient. This analysis suggests that prophylactic treatment remains the most cost-effective approach to the management of CMV in renal-transplanted patients. Further comparative studies between prophylactic and pre-emptive treatment would be a valuable addition to the current evidence based on CMV prevention.


Baby steps: pedometer-determined and self-reported leisure-time exercise behaviors of pregnant women.
Research examining women's pregnancy physical activity (PA) behaviors with objective measures is scant. Therefore, 2 studies were conducted to determine the feasibility of pregnant women wearing pedometers and to examine women's self-reported and objectively measured PA behaviors. Participants were pregnant women (Study 1 N=50, Study 2 N=30) who completed the Leisure-Time Exercise Questionnaire (LTEQ) and wore a Yamax pedometer for 3 consecutive days during free living at 20- and 32-weeks gestation. As predicted in Study 1, we found (a) 100% participant agreement in wearing the pedometer and (b) LTEQ min and pedometer-determined indices classified 67% to 86% of the participants as insufficiently active at 20-weeks gestation. In Study 2, as hypothesized, (a) mean steps/d, LTEQ total, strenuous, and mild min of PA were positively associated at 20- and 32-weeks gestation; (b) mean steps/d and LTEQ strenuous min significantly declined from 20- to 32-weeks gestation; and (c) more women were classified as sedentary and low active at 32-weeks (73%) compared with 20-weeks gestation (50%). These findings are consistent with previous epidemiological evidence documenting the decline in women's PA behaviors across the trimesters. They also illustrate that pedometer-determined indices might be a useful tool facilitating PA adoption and maintenance during pregnancy.


Anatomic characteristics of culprit sites in acute coronary syndromes.
A detailed analysis of the anatomic relationships of the site of culprit lesions that have resulted in acute coronary syndromes (ACS) has not been reported. Coronary angiograms of consecutive patients who presented with ACS were analyzed according to multiple anatomic criteria. In left anterior descending artery (LAD) (n = 85), 85% of culprit lesions were located in the first 40 mm from the ostium. The presence of angulation on the lesion increased the risk of an ACS 1.92 times (95% confidence interval [CI] 1.9-3.07), and the presence of bifurcation after the lesion increased the risk 1.65 times (95% CI 1.04-2.62). Angulated lesions located within the first 40 mm from the ostium and before a bifurcation presented an 11-fold increased risk for an ACS. In right coronary artery (RCA) (n = 58), the risk of plaque rupture was almost 2.5 times higher in lesions located between 10 and 50 mm from the ostium compared to those located in 90-130 mm (relative risk [RR] 2.38, 95% CI 1.25-4.56). In left circumflex (LCx) (n = 40), the risk of plaque rupture was almost 4.5 and 5 times higher in the first 20 mm, and between 20 and 40 mm from the ostium, respectively, compared to 60 and 80 mm (relative risk [RR] 4.58, 95% CI 1.01-20.68 for 0-20 mm, and RR 4.95, 95% CI 1.14-21.47 for 20-40 mm) after adjustment for the presence of curve on the lesion. The presence of lesion angulation increased the risk of plaque rupture almost three times (RR 3.22, 95% CI 1.49-6.93). Specific anatomic features of the coronary arteries predispose to development and/or subsequent rupture of vulnerable plaques.


Trace eyeblink conditioning is hippocampally dependent in mice.
The goal of this study was to determine whether trace eyeblink conditioning is a hippocampally dependent associative learning task in the mouse. First, we examined trace intervals of 0, 250, and 500 ms to determine a relatively long trace interval that would support eyeblink conditioning in young adult C57BL/6 mice. Mice rapidly acquired conditioned responses (CRs) with a 0-ms trace interval, acquired CRs with a 250-ms trace interval in approximately 2 days (2 sessions per day), and showed little acquisition with a 500-ms trace interval. Control mice were presented randomly unpaired stimuli and failed to show conditioning. We then determined the effect of lesioning dorsal hippocampal neurons on trace eyeblink conditioning. The hippocampus was injected bilaterally with vehicle (phosphate-buffered saline), 0.1% ibotenic acid, or 1% ibotenic acid. The vehicle group showed >60% CRs. The 0.1% group showed significantly fewer CRs (35-45%). The 1% group showed a level of CRs similar to that of the control mice. All the lesioned mice exhibited >60% CRs when subsequently trained with a 0-ms trace interval. A regression analysis indicated that the volume of area CA1 lesioned was more predictive of the behavioral impairment than the lesion volume of either CA3 or dentate gyrus, or even the total lesion volume. We conclude that dorsal hippocampal neurons play a critical role in eyeblink conditioning when a 250-ms trace interval is used with the C57BL/6 mouse, and that this paradigm will be useful for studying behavior and the in vivo and in vitro electrophysiology of hippocampal neurons in normal and transgenic or knockout mice.


Tracheotomy complications: a retrospective study of 1130 cases.
Tracheotomy is one of the most frequently performed surgical procedures in the critically ill patient. It is frequently performed as an elective therapeutic procedure and only rarely as an emergency procedure. Complications occur in 5% to 40% of tracheotomies depending on study design, patient follow-up, and the definition of the different complications. The mortality rate of tracheotomy is less than 2%. Numerous studies demonstrate a greater complication and mortality rate in emergency situations, in severely ill patients, and in small children. A retrospective study of 1130 consecutive tracheotomies performed during 1 decade (January 1987 through December 1996) is presented. We studied the indications for surgery, the major complications of tracheotomy, and their treatment and outcome. We also noted the overall mortality rate and the specific complications that led to these deaths. In total, 1130 tracheotomies were performed. Major complications occurred in 49 of the cases, and 8 deaths were directly attributed to the tracheotomy. The most common complication was tracheal stenosis, which occurred in 21 cases. Hemorrhage was the second most common complication, which occurred in 9 cases. This is one of the largest series of consecutive tracheotomies compiled. We found a relatively low overall complication and mortality rate compared with other large series. Tracheal stenosis was the most common complication in contrast to other series. Our opinion is that this may reflect tracheal damage originally caused by prolonged intubation before the tracheotomy. We believe that all other complications of tracheotomy may be prevented or minimized by careful surgical technique and postoperative tracheotomy care.


A community planning process for the implementation of evidence-based practice.
Over the past 10 years, increasing attention has been paid to the development and implementation of 'evidence-based' practices or EBPs. In 1998, the National Academy of Sciences' Institute of Medicine began drawing attention to the discrepancy between research findings and practice patterns in the treatment of substance abuse. The community coalition described in this study was developed to create a partnership between substance abuse treatment providers, policy makers, university-based researchers, and consumers to examine treatment advances and evaluate their possible application in local community settings. In the first year the community coalition conducted a needs assessment, with the goal of prioritizing areas for practice change. The needs assessment process included focus groups, key informant interviews, and a survey of substance abuse treatment providers. The questions addressed at each stage of the assessment varied, but collectively all methods formed the basis for consensus-building for a change in practice in substance abuse treatment. The results of the needs assessment were further refined and influenced the development and implementation of two evidence-based practices (EBPs) within multiple agencies. Implications for using a community needs assessment to highlight coalition building and identify and adopting EBPs are discussed.


Nonparametric identification of regulatory interactions from spatial and temporal gene expression data.
The correlation between the expression levels of transcription factors and their target genes can be used to infer interactions within animal regulatory networks, but current methods are limited in their ability to make correct predictions. Here we describe a novel approach which uses nonparametric statistics to generate ordinary differential equation (ODE) models from expression data. Compared to other dynamical methods, our approach requires minimal information about the mathematical structure of the ODE; it does not use qualitative descriptions of interactions within the network; and it employs new statistics to protect against over-fitting. It generates spatio-temporal maps of factor activity, highlighting the times and spatial locations at which different regulators might affect target gene expression levels. We identify an ODE model for eve mRNA pattern formation in the Drosophila melanogaster blastoderm and show that this reproduces the experimental patterns well. Compared to a non-dynamic, spatial-correlation model, our ODE gives 59% better agreement to the experimentally measured pattern. Our model suggests that protein factors frequently have the potential to behave as both an activator and inhibitor for the same cis-regulatory module depending on the factors' concentration, and implies different modes of activation and repression. Our method provides an objective quantification of the regulatory potential of transcription factors in a network, is suitable for both low- and moderate-dimensional gene expression datasets, and includes improvements over existing dynamic and static models.


German diabetes disease management programs are appropriate for restructuring care according to the chronic care model: an evaluation with the patient assessment of chronic illness care instrument.
With the introduction of diabetes disease management programs (DMPs) in Germany, there is a necessity to evaluate whether patients receive care that is congruent to the Chronic Care Model (CCM) and evidence-based behavioral counseling. We examined differences as perceived and experienced by patients with type 2 diabetes between those enrolled in a DMP compared with patients receiving usual care in two federal states of Germany. A random, heterogeneous sample of 3,546 patients (59.3% female) received a mailed questionnaire from their regional health fund, including the German version of the Patient Assessment of Chonic Illness Care (PACIC) instrument, which had additional items for behavioral advice (5A). Two weeks later, a general reminder was sent out. A total of 1,532 questionnaires were returned (response rate 42.2%), and valid data could be obtained for 1,399 patients. Mean age of responders was 70.3 years, of which 53.6% were female. Overall, patients enrolled in a DMP scored significantly higher (3.21 of a possible 5) than patients not enrolled in a DMP (2.86) (P < 0.001). Significant differences in the same direction were found on all five subscales of the PACIC. For the 5A scales, similar differences were found for all five subscales plus the sum score (P < 0.001; mean for DMP = 3.08, mean for non-DMP = 2.78). DMPs, as currently established in primary care in Germany, may impact provided care significantly. The changes in daily practice that have been induced by the DMPs are recognized by patients as care that is more structured and that to a larger extent reflects the core elements of the CCM and evidence-based counseling compared with usual care.


[Toxoplasmosis: a survey on meat products in Côte d'Ivoire].
Toxoplasmosis is a worldwide parasitic zoonosis that can cause severe problems under certain circumstances. Before the advent of the last-generation anti-retroviral drugs, estimation predicted that 50% of HIV-infected patients would develop toxoplasmosis (mainly central nervous system forms). It is the first clinical manifestation of AIDS in 20% of patients. This report describes an epidemiological survey on the seroprevalence of Toxoplasma antibodies in bushmeat and pork in the Côte d'Ivoire. The purpose was to determine how the parasite circulates among wild and domestic animals and to evaluate the risk of transmission to humans after ingestion of these meats. Fifteen samples of bushmeat were purchased on markets in 6 different cities. A total of 91 single samples of fresh pork raised at three different modern breeding facilities were collected from a slaughterhouse in Abidjan. Serological testing was performed on muscle fluid using an ELISA test (Pourquier Toxoplasma kit). No bushmeat sample was positive. Global seroprevalence in pork samples was 8.8% [range, 8.2-9.37]. The seroprevalence of toxoplasmosis measured in pork samples produced at modern livestock breeding facilities was lower than values reported in samples produced by traditional breeding in Africa. This finding suggests that the use of modern techniques excluding rodents (good hygiene) can reduce animal contamination. Curing (heat and smoking) may account for the absence of Toxoplasma antibodies in bushmeat. Public information campaigns concerning the risk of consuming meat containing cysts as well as raw vegetables contaminated with oocysts are needed to prevent transmission of toxoplasmosis.


Naturally acquired West Nile virus encephalomyelitis in transplant recipients: clinical, laboratory, diagnostic, and neuropathological features.
In the 2003 West Nile virus (WNV) epidemic, Colorado reported more WNV cases than any other state, including an unprecedented number in organ transplant recipients. Physicians caring for transplant recipients hospitalized with naturally acquired WNV encephalitis provided data to characterize the clinical symptoms, results of diagnostic studies, and outcomes. Eleven transplant recipients were identified (4 kidney, 2 stem cell, 2 liver, 1 lung, and 2 kidney/pancreas). Seven were directly admitted to 1 of the 2 hospitals in the study, and 4 were referred to 1 of these centers from regional hospitals. All but 1 patient had a prodrome typical of WNV encephalitis in nonimmunosuppressed patients. Ten patients developed meningoencephalitis, which in 3 cases was associated with acute flaccid paralysis. One patient developed acute flaccid paralysis without encephalitis. Six patients had significant movement disorders including tremor, myoclonus, or parkinsonism. All patients had cerebrospinal fluid pleocytosis and WNV-specific IgM in the cerebrospinal fluid and/or serum. Cerebrospinal fluid cytologic studies (n = 5) showed atypical lymphocytes, some resembling plasma cells; however, flow cytometry (n = 3) showed that cells were almost exclusively of T-cell (not B-cell or plasma cell) lineage. Magnetic resonance images of the brain were abnormal in 7 of 8 tested patients, and electroencephalograms were abnormal in 7 of 7, with 2 showing periodic lateralized epileptiform discharges. Nine of 11 patients survived infection, but 3 had significant residual deficits. One patient died 17 days after admission, and autopsy findings revealed severe panencephalitic changes with multifocal areas of necrosis in the cerebral deep gray nuclei, brainstem, and spinal cord as well as diffuse macrophage influx in the periventricular white matter. A second patient died of complications of WNV encephalitis 6 months after hospital admission. Naturally acquired WNV encephalitis in transplant recipients shows diagnostic, clinical, and laboratory features similar to those reported in nonimmunocompromised individuals, but neuroimaging, electroencephalography, and autopsy results verify that these patients develop neurological damage at the severe end of the spectrum.


Silencing of RhoC induces macrophage M1 polarization to inhibit migration and invasion in colon cancer via regulating the PTEN/FOXO1 pathway.
Ras homologue family member C (RhoC) is an oncogene in diverse types of human cancers, whereas its regulatory mechanisms involving macrophage polarization is rarely investigated. This study is designed to explore the regulatory role of RhoC in colon cancer and the underlying molecular mechanisms involving macrophage polarization. We detected RhoC expression by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, and analysed the biological function of RhoC knockdown in CC cells by the MTT, wound healing and transwell assay. Macrophage polarization-associated markers, genes associated with migration, phosphatase and tensin homologue (PTEN) and forkhead box O (FOXO) were determined by qRT-PCR and western blot. The xenograft tumour mouse model was used to assess the role of RhoC in vivo. RhoC is highly expressed in CC cells. The cell viability, invasion and migration abilities of CC cells were reduced by knockdown of RhoC. RhoC knockdown promoted M1 polarization, inhibited M2 polarization and decreased levels of genes associated with migration (matrix metalloproteinase-2 and matrix metalloproteinase-9). Silencing of RhoC inhibited tumour growth and expression of genes associated with migration in the xenografted model. In addition, silencing of RhoC promoted PTEN/FOXO1 expression, and PTEN inhibitor (SF1670) reversed the inhibitory effects of RhoC silencing. We demonstrated that silencing of RhoC reduced CC cells invasion and migration, and tumour growth by suppressing M2 macrophage polarization via regulating the PTEN/FOXO1 pathway.


The activation potential of MOF is constrained for dosage compensation.
The H4K16 acetyltransferase MOF plays a crucial role in dosage compensation in Drosophila but has additional, global functions. We compared the molecular context and effect of MOF in male and female flies, combining chromosome-wide mapping and transcriptome studies with analyses of defined reporter loci in transgenic flies. MOF distributes dynamically between two complexes, the dosage compensation complex and a complex containing MBD-R2, a global facilitator of transcription. These different targeting principles define the distribution of MOF between the X chromosome and autosomes and at transcription units with 5' or 3' enrichment. The male X chromosome differs from all other chromosomes in that H4K16 acetylation levels do not correlate with transcription output. The reconstitution of this phenomenon at a model locus revealed that the activation potential of MOF is constrained in male cells in the context of the DCC to arrive at the 2-fold activation of transcription characteristic of dosage compensation.


NSPc1, a novel mammalian Polycomb gene, is expressed in neural crest-derived structures of the peripheral nervous system.
Anterior-posterior (A-P) patterning is a key element in early embryonic development. Polycomb group (PcG) genes act as transcriptional repressors to regulate A-P patterning by either directly or indirectly controlling the coordinated expression of the HOM/Hox homeobox (Curr. Opin. Genet. Dev. 7 (1997) 488; Trends Genet. 13 (1997) 167). We describe the isolation and characterization of a novel mammalian PcG gene, termed Nervous System Polycomb-1 (NSPc1). Human and mouse NSPc1 genes encode proteins with an N-terminal RING finger domain and share homology with Drosophila melanogaster lethal(3)73Ah and the mammalian Mel18 and Bmi1 genes. Transcripts are observed at 10 dpc in the otic vesicle, urogenital bud and dorsal root ganglia. At 11.5 dpc, transcripts are present in a subset of neural crest cell derivatives of the peripheral nervous system, and in the neural tube. NSPc1 expression is ubiquitous in adult tissue.


Effect of Nasal Obstructive Disorders on Sinonasal Symptoms in Children with Different Levels of Bronchial Asthma Control.
Allergic rhinitis (AR) and allergic rhinosinusitis (ARS) are typical upper airway pathologies (UAP) in children with bronchial asthma (BA) frequently accompanied with nasal obstructive diseases (NOD). In order to establish the effect of NOD on correlations between nasal and synonasal symptoms with clinical assessments of asthma control, 82 children, 9.8 [8.9; 10.7] years old, with atopic BA were assessed using ACQ-5 for the BA control level, TNSS for nasal symptoms, and SNOT-20 for synonasal quality of life in combination with rhinovideoendoscopy for NOD. All patients had AR/ARS; in 76.3% (63/82) of children, UAP had a multimorbid character with the presence of NOD. Significant correlations were found between ACQ-5 and TNSS (R=0.40, p < 0.0001) and ACQ-5 and SNOT-20 (R=0.42, p < 0.0001). Correlations between TNSS/ACQ-5 and SNOT-20/ACQ-5 were higher in patients who do not have a combination of AR/ARS with NOD (R=0.67, p=0.0012; R=0.50, p=0.022, resp.) than in patients who have AR/ARS combined with NOD (R=0.30, p=0.015; R=0.26, p=0.04, resp.). Thus, the association of BA control level with the expression of nasal and synonasal symptoms is higher in children who do not have multimorbid UAP.


A comparison of computational methods for detecting bursts in neuronal spike trains and their application to human stem cell-derived neuronal networks.
Accurate identification of bursting activity is an essential element in the characterization of neuronal network activity. Despite this, no one technique for identifying bursts in spike trains has been widely adopted. Instead, many methods have been developed for the analysis of bursting activity, often on an ad hoc basis. Here we provide an unbiased assessment of the effectiveness of eight of these methods at detecting bursts in a range of spike trains. We suggest a list of features that an ideal burst detection technique should possess and use synthetic data to assess each method in regard to these properties. We further employ each of the methods to reanalyze microelectrode array (MEA) recordings from mouse retinal ganglion cells and examine their coherence with bursts detected by a human observer. We show that several common burst detection techniques perform poorly at analyzing spike trains with a variety of properties. We identify four promising burst detection techniques, which are then applied to MEA recordings of networks of human induced pluripotent stem cell-derived neurons and used to describe the ontogeny of bursting activity in these networks over several months of development. We conclude that no current method can provide "perfect" burst detection results across a range of spike trains; however, two burst detection techniques, the MaxInterval and logISI methods, outperform compared with others. We provide recommendations for the robust analysis of bursting activity in experimental recordings using current techniques.


On the transition from childhood amnesia to the recall of personal memories.
When adults are asked to report and date personal memories of their pasts, they show childhood amnesia, that is, diminished recall of experiences over the childhood years. This way of demonstrating the phenomenon was supplemented in the present study with a more direct approach: Participants reported events of early childhood that they knew they had experienced (because of family stories, photographs, etc.) but did not actually remember. The resulting cumulative relative frequency distributions produced by the two methods were substantially different, with the median age of remembered events being 6.07 years and of known events, 3.20 years. We suggest that the mean of these two ages, 4.64 years, gives a good indication of when childhood amnesia is eclipsed by personal memories in adults' recall of their personal pasts.


Fully automated and colorimetric foodborne pathogen detection on an integrated centrifugal microfluidic device.
This work describes fully automated and colorimetric foodborne pathogen detection on an integrated centrifugal microfluidic device, which is called a lab-on-a-disc. All the processes for molecular diagnostics including DNA extraction and purification, DNA amplification and amplicon detection were integrated on a single disc. Silica microbeads incorporated in the disc enabled extraction and purification of bacterial genomic DNA from bacteria-contaminated milk samples. We targeted four kinds of foodborne pathogens (Escherichia coli O157:H7, Salmonella typhimurium, Vibrio parahaemolyticus and Listeria monocytogenes) and performed loop-mediated isothermal amplification (LAMP) to amplify the specific genes of the targets. Colorimetric detection mediated by a metal indicator confirmed the results of the LAMP reactions with the colour change of the LAMP mixtures from purple to sky blue. The whole process was conducted in an automated manner using the lab-on-a-disc and a miniaturized rotary instrument equipped with three heating blocks. We demonstrated that a milk sample contaminated with foodborne pathogens can be automatically analysed on the centrifugal disc even at the 10 bacterial cell level in 65 min. The simplicity and portability of the proposed microdevice would provide an advanced platform for point-of-care diagnostics of foodborne pathogens, where prompt confirmation of food quality is needed.


Climate change-informed dietary modeling in Pacific cod: Experimentally-derived effects of temperature and dietary quality on carbon and nitrogen stable isotope trophic discrimination factors.
Stable isotope analysis is a powerful tool for dietary modeling and trophic ecology research. A crucial piece of information for isotopic dietary modeling is the accurate estimation of trophic discrimination factors (TDFs), or the isotopic offset between a consumer's tissue and its diet. In order to parameterize stable isotope dietary models for future climate scenarios, we investigated the effect of water temperature and dietary protein and lipid content on TDFs in juvenile Pacific cod (Gadus macrocephalus). Pacific cod are a commercially and ecologically important species, with stock numbers in the northeast Pacific recently having dropped by more than 70%. We tested four water temperatures (6, 8, 10, and 12°C) and two dietary regimens (low and high lipid content), representing a range of potential ocean temperature and prey quality scenarios, in order to determine carbon and nitrogen TDFs in juvenile Pacific cod. Additionally, we assessed dietary intake and proximate composition of the experimental fish in order to estimate consumption, assimilation, and retention of dietary nutrients. The results of this study suggest that dietary protein catabolism is a primary driver of nitrogen TDF variability in juvenile Pacific cod. Across all temperature treatments from 6 to 12°C, fish reared on the lower quality, lower lipid content diet had higher nitrogen TDFs. The mean TDFs for fish raised on the higher lipid, lower protein diet were +3.40 ‰ for nitrogen (Δ15N) and +0.36 ‰ for lipid-corrected carbon (Δ LC 13C). The mean TDFs for fish raised on the lower lipid, higher protein diet were +4.09 ‰ for nitrogen (Δ15N) and 0.00 ‰ for lipid-corrected carbon (Δ LC 13C). Lipid-corrected carbon isotope data showed that, regardless of temperature, fish consuming the lower lipid diet had essentially no trophic discrimination between diet and bulk tissues. We found no ecologically meaningful differences in TDFs due to water temperature across the 6°experimental range. The results of this experiment demonstrate that dietary quality, and more specifically the use of dietary protein for energetic needs, is a primary driver of trophic discrimination factors. The TDFs determined in this study can be applied to understanding trophic ecology in Pacific cod and closely related species under rapidly changing prey availability and ocean temperature conditions.


Flow-dependency of exhaled nitric oxide in children with asthma and cystic fibrosis.
The concentration of nitric oxide in exhaled air, a marker of airway inflammation, depends critically on the flow of exhalation. Therefore, the aim of this study was to determine the effect of varying the flow on end-expiratory NO concentration and NO output in children with asthma or cystic fibrosis (CF) and in healthy children. Nineteen children with stable asthma, 10 with CF, and 20 healthy children exhaled from TLC while controlling expiratory flow by means of a biofeedback signal at approximately 2, 5, 10 and 20% of their vital capacity per second. NO was measured in exhaled air with a chemiluminescence analyser. Comparisons between the three groups were made by analysing the NO concentration at the endexpiratory plateau and by calculating NO output at different flows. Exhaled NO decreased with increasing flow in all children. Children with asthma had significantly higher NO concentrations than healthy children, but only at the lowest flows. Asthmatics using inhaled steroids (n=13) tended to have lower median exhaled NO than those without steroids. The slope of linearized (log-log transformed) NO/flow plots was significantly steeper in asthmatics than in healthy controls. CF patients had a significantly lower NO concentration and output over the entire flow range studied, compared to asthmatic and control subjects, with a similar NO/flow slope as control subjects. In conclusion, the nitric oxide concentration in exhaled air is highly flow-dependent, and the nitric oxide-flow relationship differs between asthmatics versus cystic fibrosis patients and control subjects. Assessment of the nitric oxide/flow relationship may help in separating asthmatics from normal children.


Significance of p57(Kip2) down-regulation in oncogenesis of bladder carcinoma: an immunohistochemical study.
Cyclin-dependent kinase inhibitors have important roles in the oncogenesis of various tumors including urothelial cancer. The aim of this study was to establish the importance of p57(Kip2), a unique cyclin-dependent kinase inhibitor, in the oncogenesis of bladder carcinoma. This article also focused on another cyclin-dependent kinase inhibitor, p27(Kip1), and telomerase enzyme and examined the relationship between these proteins. Thirty-one patients with urothelial carcinomas of the bladder and 7 cases with normal urinary bladder mucosa were included in the study. Immunohistochemical study was performed by monoclonal antibodies of p27(Kip1), p57(Kip2), and the telomerase subunit (hTERT). All immunohistochemical preparations were evaluated by an immunohistochemical histological score. p57(Kip2) and p27Kip1) expression were seen in all of the cases of normal mucosa. In carcinoma cases, 8 of 31 (25.8%) showed p57(Kip2) nuclear positivity and 20 of 31 (64.5%) expressed nuclear p27(Kip1). HSCOREs of carcinoma cases showed lower scores of nuclear p57(Kip2) and p27(Kip1) than normal mucosa, but only HSCOREs of nuclear p57(Kip2) (P = 0.001) showed statistical significance. Despite unknown significance, cytoplasmic p57(Kip2) and p27(Kip1) were also evaluated. Immunohistochemical analysis showed that carcinomas expressed higher HSCOREs of hTERT than normal mucosa, and there was a significant difference (P = 0.026) between muscle invasive carcinomas and normal mucosa. The data showed that p57(Kip2) down-regulation along with p27(Kip1) is a well-established feature of urothelial carcinoma. Probably, this down-regulation of cyclin-dependent kinase inhibitors supports the proliferation phase of oncogenesis. In the study, we also showed that hTERT expression was up-regulated in higher stages of urothelial carcinoma.


Genetic alterations in astrocytic tumors: hints to a molecular epidemiological interpretation.
General incidence of brain tumors has significantly increased over the past two decades. Although the aetiology of this increase has not been determined, increased life span and improved diagnosis methods can partially be responsible for it. Starting from the epidemiological data on risk factors, we reviewed the molecular events known to be involved in the genesis and progression of the most common brain neoplasm in adult, namely astrocytoma. Alterations in different genes, encoding key regulatory elements in the cell cycle control, were reviewed. In light of the molecular epidemiological notion that carcinogens leave fingerprints, point mutations at the p53 locus from a panel of astrocytic tumor patients from all over the world were analysed. The results of this analysis suggest that the majority of astrocytomas may have a spontaneous origin. In particular, the kind of mutational events observed suggests a major role for mutagenic events occurring at CpG sites. In order to yield valid conclusions on the potential role of environmental mutagenic factors, well designed molecular epidemiological studies on populations clearly showing a higher relative risk of developing brain tumors are needed.


Toxic shock syndrome as the AIDS-defining diagnosis.
Toxic shock syndrome (TSS) has been infrequently reported as a complication of AIDS. We present the case of a 24-year-old man, previously unknown to be positive for the human immunodeficiency virus, presenting in septic shock. The literature on TSS in AIDS is reviewed and the association between the two diseases is presented.


Multilayered structures for p- and s-polarized long-range surface-plasmon-polariton propagation.
A normalized admittance diagram assists in describing and designing multilayered structures to excite long-range surface-plasmon-polariton (LRSPP) waves of either the p- or the s-polarization state. These structures comprise symmetric periodic multilayers on one or both sides of a metal thin film in either the Kretschmann or the Sarid configuration. The normalized admittance diagram even assists in designing structures that can be used to excite LRSPP waves of both polarization states simultaneously.


Longitudinal mediators of achievement in mathematics and reading in typical and atypical development.
Longitudinal studies of neurodevelopmental disorders that are diagnosed at or before birth and are associated with specific learning difficulties at school-age provide one method for investigating developmental precursors of later-emerging academic disabilities. Spina bifida myelomeningocele (SBM) is a neurodevelopmental disorder associated with particular problems in mathematics, in contrast to well-developed word reading. Children with SBM (n=30) and typically developing children (n=35) were used to determine whether cognitive abilities measured at 36 and 60 months of age mediated the effect of group on mathematical and reading achievement outcomes at 8.5 and 9.5 years of age. A series of multiple mediator models showed that: visual-spatial working memory at 36 months and phonological awareness at 60 months partially mediated the effect of group on math calculations, phonological awareness partially mediated the effect of group on small addition and subtraction problems on a test of math fluency, and visual-spatial working memory mediated the effect of group on a test of math problem solving. Groups did not differ on word reading, and phonological awareness was the only mediator for reading fluency and reading comprehension. The findings are discussed with reference to theories of mathematical development and disability and with respect to both common and differing cognitive correlates of math and reading.


Can MMP-9 be a Prognosticator Marker for Oral Squamous Cell Carcinoma?
Invasion and metastasis of malignant tumours severely endanger the life of cancer patients. Squamous cell carcinoma is one of the commonly found malignancies in the oral cavity and its survival rate has not improved from past few decades. Since an important risk factor for oral squamous cell carcinoma is the presence of epithelial dysplasia, it is necessary to check the presence of a prognosticator marker in both of them. As matrix metalloproteinase's (MMP's) are involved in degradation of type IV collagen, which are one of the important components of extracellular matrix components which play a relevant role in several steps of tumour progression such as invasion and metastasis. We have studied MMP-9 expression to evaluate its prognostic potential in oral cancers as well as oral epithelial dysplasia along with tissues of normal oral epithelium. The expression was examined using immunohistochemistry procedure with MMP-9 in 100 samples including cases of epithelium from normal oral mucosa, oral dysplastic lesions and oral squamous cell carcinoma. One set of formalin fixed, paraffin embedded sections of the three categories were stained by haematoxylin and eosin. The sections were then evaluated under microscope. Data was examined for statistical significance using SPSS 13.0 by Mann-Whitney Test and Kruskal-Wallis Test. With MMP-9 gain of expression was noted from Control group to oral squamous cell carcinoma. Cytoplasmic staining was seen with MMP-9. Statistically highly significant differences were seen between oral epithelial dysplasia and oral squamous cell carcinoma and statistically significant differences were found between the control group and the oral squamous cell carcinoma group. This study suggested that oral squamous cell carcinoma shows higher MMP-9 expression as compared to oral epithelial dysplasia followed by epithelium from normal oral mucosa. However, no correlation was found among the histological grades of oral squamous cell carcinoma.


A 10-Minute "Mix and Read" Antibody Assay for SARS-CoV-2.
Accurate and rapid diagnostic tools are needed for management of the ongoing coronavirus disease 2019 (COVID-19) pandemic. Antibody tests enable detection of individuals past the initial phase of infection and help examine vaccine responses. The major targets of human antibody response in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are the spike glycoprotein (SP) and nucleocapsid protein (NP). We have developed a rapid homogenous approach for antibody detection termed LFRET (protein L-based time-resolved Förster resonance energy transfer immunoassay). In LFRET, fluorophore-labeled protein L and antigen are brought to close proximity by antigen-specific patient immunoglobulins of any isotype, resulting in TR-FRET signal. We set up LFRET assays for antibodies against SP and NP and evaluated their diagnostic performance using a panel of 77 serum/plasma samples from 44 individuals with COVID-19 and 52 negative controls. Moreover, using a previously described SP and a novel NP construct, we set up enzyme linked immunosorbent assays (ELISAs) for antibodies against SARS-CoV-2 SP and NP. We then compared the LFRET assays with these ELISAs and with a SARS-CoV-2 microneutralization test (MNT). We found the LFRET assays to parallel ELISAs in sensitivity (90-95% vs. 90-100%) and specificity (100% vs. 94-100%). In identifying individuals with or without a detectable neutralizing antibody response, LFRET outperformed ELISA in specificity (91-96% vs. 82-87%), while demonstrating an equal sensitivity (98%). In conclusion, this study demonstrates the applicability of LFRET, a 10-min "mix and read" assay, to detection of SARS-CoV-2 antibodies.


A narrow-band injection-seeded pulsed titanium:sapphire oscillator-amplifier system with on-line chirp analysis for high-resolution spectroscopy.
A narrow-band tunable injection-seeded pulsed titanium:sapphire laser system has been developed for application in high-resolution spectroscopic studies at the fundamental wavelengths in the near infrared as well as in the ultraviolet, deep ultraviolet, and extreme ultraviolet after upconversion. Special focus is on the quantitative assessment of the frequency characteristics of the oscillator-amplifier system on a pulse-to-pulse basis. Frequency offsets between continuous-wave seed light and the pulsed output are measured as well as linear chirps attributed mainly to mode pulling effects in the oscillator cavity. Operational conditions of the laser are found in which these offset and chirp effects are minimal. Absolute frequency calibration at the megahertz level of accuracy is demonstrated on various atomic and molecular resonance lines.


Planning prompts to promote uptake of HIV services among men: a randomised trial in rural Uganda.
Interventions informed by behavioural economics, such as planning prompts, have the potential to increase HIV testing at minimal or no cost. Planning prompts have not been previously evaluated for HIV testing uptake. We conducted a randomised clinical trial to evaluate the effectiveness of low-cost planning prompts to promote HIV testing among men. We randomised adult men in rural Ugandan parishes to receive a calendar planning prompt that gave them the opportunity to make a plan to get tested for HIV at health campaigns held in their communities. Participants received either a calendar showing the dates when the community health campaign would be held (control group) or a calendar showing the dates and prompting them to select a date and time when they planned to attend (planning prompt group). Participants were not required to select a date and time or to share their selection with study staff. The primary outcome was HIV testing uptake at the community health campaign. Among 2362 participants, 1796 (76%) participants tested for HIV. Men who received a planning prompt were 2.2 percentage points more likely to test than the control group, although the difference was not statistically significant (77.1% vs 74.9%; 95% CI -1.2 to 5.7 percentage points, p=0.20). The planning prompt was more effective among men enrolled ≤40 days before the campaigns (3.6 percentage-point increase in testing; 95% CI -2.9 to 10.1, p=0.27) than among men enrolled >40 days before the campaigns (1.8 percentage-point increase; 95% CI -2.3 to 5.8, p=0.39), although the effects within the subgroups were not significant. These findings suggest that planning prompts may be an effective behavioural intervention to promote HIV testing at minimal or no cost. Large-scale studies should further assess the impact and cost-effectiveness of such interventions.


Learning the dynamical system behind sensory data.
This letter looks at the physics behind sensory data by identifying the parameters that govern the physical system and estimating them from sensory observations. We extend Takens's delay-embedding theorem to the dynamical systems controlled by parameters. An embedding of the product space of the phase and the parameter spaces of the dynamical system can be obtained by the delay-embedding map, provided that the parameter of the dynamical system changes slowly. The reconstruction error is bounded for slowly varying parameters. A manifold learning technique is applied to the embedding obtained to extract a low-dimensional global coordinate system representing the product space. The phase space of the deterministic dynamics can be contracted by using the adjacency relationship in time, which enables recovery of only the parameter space. As examples, the manifolds of synthetic and real-world vowels with time-varying fundamental frequency (F(0)) are analyzed, and the F(0) contours are extracted by an unsupervised algorithm. Experimental results show that the proposed method leads to robust performance under various noise conditions and rapid changes of F(0) compared with the current state-of-the-art F(0) estimation algorithms.


Bacterial and archaeal diversity in an iron-rich coastal hydrothermal field in Yamagawa, Kagoshima, Japan.
Physicochemical characteristics and archaeal and bacterial community structures in an iron-rich coastal hydrothermal field, where the temperature of the most active hot spot reaches above 100 °C, were investigated to obtain fundamental information on microbes inhabiting a coastal hydrothermal field. The environmental settings of the coastal hydrothermal field were similar in some degree to those of deep-sea hydrothermal environments because of its emission of H2, CO2, and sulfide from the bottom of the hot spot. The results of clone analyses based on the 16S rRNA gene led us to speculate the presence of a chemo-synthetic microbial ecosystem, where chemolithoautotrophic thermophiles, primarily the bacterial order Aquificales, function as primary producers using H2 or sulfur compounds as their energy source and CO2 as their carbon source, and the organic compounds synthesized by them support the growth of chemoheterotrophic thermophiles, such as members of the order Thermales and the family Desulfurococcaceae. In addition, the dominance of members of the bacterial genus Herbaspirillum in the high temperature bottom layer led us to speculate the temporal formation of mesophilic zones where they can also function as primary producing or nitrogen-fixing bacteria.


Quality assessment of mental health care by people with severe mental disorders: a participatory research project.
This study assessed the perceived quality of care by consumers with severe mental disorders. A questionnaire investigating service quality was developed by a consumer focus group and filled by 204 consumers. In five areas the negative evaluations exceeded or closely approximated the positive ones: choice of professionals, waiting times, information about illness and medications. All five do not refer to the outcomes of care, but to the concept of responsiveness. The results confirmed that people with severe mental disorders can give value judgments on various aspects of care. However, even in a service strongly oriented towards community care, the consumers' needs in sensitive areas concerning choices, respect and autonomy are not met. The application of the concept of responsiveness to quality improvement may help services to meet consumers' expectations.


In-silico screening of small molecule inhibitors against Lactate Dehydrogenase (LDH) of Cryptosporidium parvum.
Cryptosporidium parvum is a protozoan parasite which causes waterborne diseases known as Cryptosporidiosis. It is an acute enteric diarrheal disease being severe in the case of immunocompromised individuals and children. C. parvum mainly depends on the glycolysis process for energy production and LDH (Lactate Dehydrogenase) is a key controller of this process. In this study from different in-silico approaches such as structure-based, ligand-based and de novo drug design; a total of 40 compounds were selected for docking studies against LDH. The study reported a compound CHEMBL1784973 from Pathogen Box as the best inhibitor in terms of docking score and pharmacophoric features. Furthermore, the binding mode of the best-reported inhibitor was validated through molecular dynamics simulation for a time interval of 70 ns in water environment. The findings resulted in the stable conformation of the inhibitor in the active site of the protein. This study will be helpful for experimental validation.


Photosensitized killing of cultured fibroblasts from patients with peroxisomal disorders due to pyrene fatty acid-mediated ultraviolet damage.
The influence of pyrene-fatty acids on the resistance of cells to ultraviolet (UV) radiation was investigated in cultured fibroblasts from patients with five types of peroxisomal disorders. All showed reduced survival compared to control. The effect varied with the biochemical defect involved and the chain length of the pyrene fatty acid. Reduced survival was observed in cells deficient in plasmalogens (rhizomelic chondrodysplasia punctata) and in cells deficient in peroxisomal fatty acid oxidation (bifunctional enzyme deficiency), which accumulated pyrene-fatty acids. X-linked adrenoleukodystrophy fibroblasts accumulated pyrene-fatty acids and showed increased UV sensitivity only when exposed to longer-chain pyrene fatty acids. UV radiation resistance was lowest in cells with combined impairment of plasmalogen synthesis and fatty acid oxidation (Zellweger syndrome, neonatal adrenoleukodystrophy), suggesting that UV sensitivity correlates inversely with the ratio of plasmalogens to radical producing substances. Fibroblasts deficient in plasmalogens gained normal UV resistance when their plasmalogen levels were normalized by hexadecylglycerol. UV resistance increased when Zellweger cells were fused with X-linked adrenoleukodystrophy cells, and also when Zellweger cells belonging to different complementation groups were fused. The results provide leads to the pathogenesis of the multiple malformations associated with peroxisomal disorders and a method for the selection of cells in which the metabolic defect has been corrected.


[Enterovirus non-poliomyelitis infections in Krasnodar region].
Evaluation of epidemic situation by non-poliomyelitis enterovirus infections in Krasnodar region in multi-year dynamics and characterization of clinical course of enterovirus serous meningitis in hospitalize patients. Retrospective analysis of non-poliomyelitis enterovirus infection epidemi process manifestations during 2002-2012 in Krasnodar region territory based on data of Center of Hygien and Epidemiology in Krasnodar Region. Clinical-epidemiologic characteristics of enterovirus infections in Krasnodar region are presented. Landscape ofenteroviruses isolated from the environment of some territories of the region and from the biological material of patients with various diseases is demonstrated. Clinical features ofenterovirus meningitis course are characterized. Enterovirus transmission b contact route was established to be the most frequent. A lack of pathognomonic symptoms and awareness o physicians of various specialties regarding diagnostics of this infection are the clinical problems of non-po liomyelitis enterovirus diseases.


Shifting ranges and conservation challenges for lemurs in the face of climate change.
Geospatial modeling is one of the most powerful tools available to conservation biologists for estimating current species ranges of Earth's biodiversity. Now, with the advantage of predictive climate models, these methods can be deployed for understanding future impacts on threatened biota. Here, we employ predictive modeling under a conservative estimate of future climate change to examine impacts on the future abundance and geographic distributions of Malagasy lemurs. Using distribution data from the primary literature, we employed ensemble species distribution models and geospatial analyses to predict future changes in species distributions. Current species distribution models (SDMs) were created within the BIOMOD2 framework that capitalizes on ten widely used modeling techniques. Future and current SDMs were then subtracted from each other, and areas of contraction, expansion, and stability were calculated. Model overprediction is a common issue associated Malagasy taxa. Accordingly, we introduce novel methods for incorporating biological data on dispersal potential to better inform the selection of pseudo-absence points. This study predicts that 60% of the 57 species examined will experience a considerable range of reductions in the next seventy years entirely due to future climate change. Of these species, range sizes are predicted to decrease by an average of 59.6%. Nine lemur species (16%) are predicted to expand their ranges, and 13 species (22.8%) distribution sizes were predicted to be stable through time. Species ranges will experience severe shifts, typically contractions, and for the majority of lemur species, geographic distributions will be considerably altered. We identify three areas in dire need of protection, concluding that strategically managed forest corridors must be a key component of lemur and other biodiversity conservation strategies. This recommendation is all the more urgent given that the results presented here do not take into account patterns of ongoing habitat destruction relating to human activities.


[The Global Initiative for Asthma and familial medicine-resident physicians. Impact of an educational strategy].
The Global Initiative for Asthma (GINA) provides effective medical care to asthmatic patients. Knowledge and appropriate handling of GINAS' management guidelines for medicine residents significantly reduce asthma morbidity and mortality. To evaluate the level of knowledge about asthma on Global initiative for Asthma program before and after an educational strategy in residents. Forty-four residents of first, second, and third grade of UMF No. 21 and No. 28 took part in this project A magisterial lecture and a workshop about clinical cases on asthma were carried out. The lecture lasted three hours and asthma surveys, before and after the educational strategy; were applied. Descriptive statistics as well as t test in SPSS were used for statistical analysis. Average grade of the 44 residents before the workshop was of 4.24 +/- 1.6, and after the workshop of 6.00 +/- 1.71. Therewere no significant differences among groups peryear of medical residency onthe initial assessment; however, residents of first year showed an increment on asthma knowledge (p < 0.05) in the subsequent test. There were no differences among groups. Asthma educational workshops increase residents' awareness of asthma, mainly first-year medical students.


Personalized therapy for pancreatic cancer: Do we need better targets, arrows, or both?
Pancreatic cancer is predicted by the Pancreatic Cancer Action Network to soon become the 2nd leading cause of cancer related deaths in this country. This cannot be attributed to a lack of resources focused on interrogating the genomic landscape of pancreatic cancer genomes. Additionally, a large number of researchers and federal dollars have been directed towards inhibiting the most frequent genetic lesion in pancreatic cancer, Kras. Even with these advances, cytotoxic chemotherapy is currently the best option for patients with metastatic disease. Besides developing better early detection strategies, translating our knowledge of the molecular aspects of pancreatic cancer to the clinic via a targeted, personalized medicine approach may be the best way to dramatically improve patient outcomes. Herein, we briefly describe the scope of the problem in targeting pancreatic cancer on both the cellular and molecular levels. We also outline some promising, ongoing efforts to develop better therapeutic interventions for this deadly disease. For instance, we discuss novel strategies to target molecules and pathways that go beyond targeting genetic mutations and the level of transcriptional gene regulation. Finally, we summarize a clinical trial that is designed to answer the question whether with the available arsenal of drugs and molecular targets, can we now "personalize" therapy for pancreatic cancer patients?


Effective estimation of correct platelet counts in pseudothrombocytopenia using an alternative anticoagulant based on magnesium salt.
Pseudothrombocytopenia remains a challenge in the haematological laboratory. The pre-analytical problem that platelets tend to easily aggregate in vitro, giving rise to lower platelet counts, has been known since ethylenediamine-tetra acetic acid EDTA and automated platelet counting procedures were introduced in the haematological laboratory. Different approaches to avoid the time and temperature dependent in vitro aggregation of platelets in the presence of EDTA were tested, but none of them proved optimal for routine purposes. Patients with unexpectedly low platelet counts or flagged for suspected aggregates, were selected and smears were examined for platelet aggregates. In these cases patients were asked to consent to the drawing of an additional sample of blood anti-coagulated with a magnesium additive. Magnesium was used in the beginning of the last century as anticoagulant for microscopic platelet counts. Using this approach, we documented 44 patients with pseudothrombocytopenia. In all cases, platelet counts were markedly higher in samples anti-coagulated with the magnesium containing anticoagulant when compared to EDTA-anticoagulated blood samples. We conclude that in patients with known or suspected pseudothrombocytopenia the magnesium-anticoagulant blood samples may be recommended for platelet counting.


Subgingival microbiota in squirrel monkeys with naturally occurring periodontal diseases.
The squirrel monkey (Saimiri sciureus) has been proposed as an in vivo model for the study of subgingival colonization by suspected periodontopathogens, such as black-pigmented porphyromonads and prevotellas (BP/P). However, the indigenous microbiota of the squirrel monkey has not been well described. Therefore, in order to more fully characterize the oral microbiota of these animals, we studied two groups of squirrel monkeys from widely different sources. Group I consisted of 50 breeding colony monkeys ranging in age from 9 months to over 6 years which had been raised in captivity; group II consisted of 16 young sexually mature monkeys recently captured in the wild in Guyana. Group I animals in captivity had developed moderate to severe gingivitis, with a mean gingival index (GI) of 2.6; 52% of the sites bled, 26% had detectable calculus, and 83% had detectable BP/P. A group I subset (six animals), for which predominant cultivable microbiota was described, had a mean GI of 2.4. Colony morphology enumeration revealed that five of the six subset animals were detectably colonized with BP/P (range, 0 to 16.9%) and Actinobacillus actinomycetemcomitans (range, 0 to 3.9%); all subset animals were colonized with Fusobacterium species (range, 0.8 to 3.6%), Actinomyces species (range, 2.3 to 11%), and gram-positive cocci (range, 1.4 to 21.4%). Predominant cultivable microbiota results revealed the presence of many bacterial species commonly found in the human gingival sulcus. At baseline, group II animals were clinically healthy and had a mean GI of 1.4; 67% of the sites bled and 2.1% had calculus, and none of the animals had detectable BP/P. Neisseriae were very common in noninflamed sites. Subsequently, when inflamed sites were compared with noninflamed sites in group II animals after they had been maintained in captivity for 6 months, inflamed sites exhibited a more complex microbiota and increased proportions of gram-negative rods and asaccharolytic bacteria.


Nanocarriers and nonviral methods for delivering antiangiogenic factors for glioblastoma therapy: the story so far.
Angiogenesis, the formation of new blood vessels, is an essential component of glioblastoma (GB) progression. The development of angiogenesis inhibitor therapy, including treatments targeting vascular endothelial growth factor (VEGF) in particular, raised new hopes for the treatment of GB, but no Phase III clinical trial to date has reported survival benefits relative to standard treatment. There are several possible reasons for this limited efficacy, including VEGF-independent angiogenesis, induction of tumor invasion, and inefficient antiangiogenic factor delivery to the tumor. Efforts have been made to overcome these limitations by identifying new angiogenesis inhibitors that target angiogenesis through different mechanisms of action without inducing tumor invasion, and through the development of viral and nonviral delivery methods to improve antiangiogenic activity. Herein, we describe the nonviral methods, including convection-enhanced delivery devices, implantable polymer devices, nanocarriers, and cellular vehicles, to deliver antiangiogenic factors. We focus on those evaluated in intracranial (orthotopic) animal models of GB, the most relevant models of this disease, as they reproduce the clinical scenario of tumor progression and therapy response encountered in GB patients.


Mendelian disease research in the Plain populations of Lancaster County, Pennsylvania.
Founder populations have long contributed to our knowledge of rare disease genes and phenotypes. From the pioneering work of Dr. Victor McKusick to today, research in these groups has shed light on rare recessive phenotypes, expanded the clinical spectrum of disease, and facilitated disease gene identification. Current clinical and research studies in these special groups augment the wealth of knowledge already gained, provide new insights into emerging problems such as variant interpretation and reduced penetrance, and contribute to the development of novel therapies for rare genetic diseases. Clinical developments over the past 30 years have altered the fundamental relationship with the Lancaster Plain communities: research has become more collaborative, and the knowledge imparted by these studies is now being harnessed to provide cutting-edge translational medicine to the very community of vulnerable individuals who need it most.


The mechanisms of propofol-induced vascular relaxation and modulation by perivascular adipose tissue and endothelium.
Propofol causes hypotension due to relaxation of vascular smooth muscle cells through its direct or indirect vasodilator effects. Perivascular adipose tissue (PVAT) and endothelium attenuate vascular contraction, and the function of PVAT is altered in hypertension and diabetes. Whether PVAT affects the action of anesthetics on vascular function is unknown. We studied the mechanisms of propofol-induced relaxation in relation to the involvement of PVAT and endothelium. Thoracic aortic rings from Wistar rats were prepared with or without PVAT (PVAT+ and PVAT-), intact endothelium (E+), or both, or with the endothelium removed (E-) for functional studies. In phenylephrine precontracted vessels, propofol-induced relaxation was highest with both PVAT and E+ and lowest in vessels denuded of both PVAT and endothelium. Propofol-induced relaxation occurred via both endothelium-dependent and -independent mechanisms. The relaxation response induced by propofol was significantly reduced by nitric oxide synthase inhibitor (l-NNA), K(+) channel blockers (tetraethylammonium and glibenclamide) in E+ and E- vessels, and by soluble guanylyl cyclase inhibitor 1H-(1,2,4) oxadiazolo (4,3-A) quinazoline-1-one and hydrogen peroxide scavenger (catalase) in E- vessels. The presence of PVAT significantly enhanced the relaxation response induced by propofol. In contrast to phenylephrine precontracted vessels in which the presence of PVAT or endothelium had an effect, in vessels precontracted with KCl, propofol-induced relaxation was similar among the 4 types of vessel preparation. PVAT enhances the relaxation effect induced by propofol in rat aorta through both endothelium-dependent and endothelium-independent pathways thus highlighting the clinical importance of PVAT.


Identification and treatment of osteochondritis dissecans of the distal sagittal ridge of the third metacarpal bone.
Osteochondritis dissecans (OCD) of the distal sagittal ridge of the third metacarpal bone (McIII) has not previously been described in the literature. To describe the clinical, radiological and arthroscopic features of OCD of the distal sagittal ridge of the McIII and to report outcome in a series of cases. Retrospective case series. Case records and images of horses with OCD of the distal sagittal ridge of the McIII admitted to a single referral centre between February 2006 and February 2013 were reviewed. Follow-up information was obtained by telephone questionnaire and/or racing performance. Osteochondritis dissecans of the distal sagittal ridge of the McIII was found in 16 lame horses; 9 unilaterally and 7 bilaterally. Lesions were consistently identified with flexed lateromedial radiographs and in 21 joints in dorsopalmar projections also. These were arthroscopically accessible from the dorsal compartment with metacarpophalangeal joints maximally flexed. This permitted fragment removal and lesion debridement. Thirteen of 14 horses with follow-up of ≥12 months post surgery performed athletically including 11 of 12 Thoroughbreds that raced. Unlike more proximal lesions, OCD of the distal sagittal ridge appears to affect the McIII preferentially. Lesions were identified on dorsopalmar and flexed lateromedial radiographs only and are amenable to arthroscopic surgery. Clinicians should be aware of the potential for OCD to affect the distal sagittal ridge of the McIII together with the radiographic projections needed to identify its presence. A modified arthroscopic approach to the dorsal metacarpophalangeal joint is necessary in order to access lesions.


Clinical decision-making regarding intravenous hydration in normal labor in a birth center setting.
Intravenous hydration (IVH) in normal labor is used routinely in most hospital settings despite the lack of support from recent research. However, it is not used routinely at one in-hospital, Alternative Birthing Center (ABC), in the Midwest. The purpose of this study was to explore and describe the criteria and perspectives of the certified nurse-midwives (CNMs) and nurses (RNs) in their decision-making process toward nonroutine use of IVH. In-depth interviews with five CNMs and four RNs that work in the ABC provided insight into how they balanced patient preference, research-based knowledge, and clinical judgments, in making decisions about IVH. The RNs were slightly more in favor of IVH than the CNMs, but not because of philosophic differences. Rather, the different roles and responsibilities of CNMs and RNs seemed to be most important. CNMs had a stronger commitment to client preferences, whereas RNs were more concerned about their responsibility for efficiently starting IVs prior to an emergency situation. Understanding professional differences in the decision-making process for IVH can enhance clinical functioning of the caregiving team by helping CNMs anticipate and address potential conflicts.


Alcohol use and prior alcohol-related convictions as predictors of probation officer perceptions and sentencing.
Presentence reports for 740 offenders were content analyzed. Regression techniques were used to evaluate a number of predictors of sentencing, including the effects of number of prior alcohol-related convictions and whether the offender was intoxicated while committing the offense he was charged with. The two most prominent variables affecting the severity of sentence were the probation officer's assessment of the offender's probable success on probation and the legal seriousness of the offense. While failing to have any significant direct effects on sentencing, our measures of alcohol use had significant indirect effects which were mediated by the probation officer's assessment of success on probation and legal seriousness. An extensive criminal record of prior alcohol-related convictions resulted in a poorer prognosis for success on probation this, in turn, led to more severe sentences for these offenders. Intoxication while committing an offense was related to the commission of minor crimes which, subsequently, yielded more lenient treatment for alcohol-users compared to nonusers. Finally, native offenders were more likely than nonnatives to have many prior alcohol-related convictions and to have been intoxicated while committing an offense.


First record of insect-plant interaction in Late Cretaceous fossils from Nelson Island (South Shetland Islands Archipelago), Antarctica.
Despite the enormous paleobotanical record on different islands of the Antarctic Peninsula, the evidence of insect activity associated with fossilized plants is scarce. Here we report the first evidence of insect-plant interaction from Cretaceous deposits, more precisely from a new locality at the Rip Point area, Nelson Island (Antarctic Peninsula). The macrofossil assemblage includes isolated Nothofagus sp. leaf impressions, a common component of the Antarctic paleoflora. Two hundred leaves were examined, of which 15 showed evidence of insect activity, displaying variations in size, shape, and preservation. Two types of interaction damage, galls and mines, were identified. A single specimen retained a circular scar recognized as galling scar, while meandering tracks were considered mines. These traces of herbivore insect activity, correspond to the oldest known record of this type of interaction of West Antarctica and the oldest record of insect-plant interaction in Nothofagus sp. reported so far.


Maternal anxiety from pregnancy to 2 years postpartum: transactional patterns of maternal early adversity and child temperament.
The aims of this study were to examine the anxiety trajectories of women from pregnancy to 2 years postpartum and to assess the influence of their early life experiences and the temperament of the child on these trajectories. We evaluated state anxiety (State-Trait Anxiety Inventory) at pregnancy and 3, 6, 12, 18, and 24 months postpartum and determined its course as a function of self-reported early adverse experiences (Childhood Trauma Questionnaire) and the temperament of the child at 18 months (Early Child Behavior Questionnaire). Based on growth curve modeling, we found that anxiety followed a general U-shape pattern from gestation to 2 years postpartum, which was modified by early life experience of women. Greater early adversity was associated with higher gestational anxiety, followed by a marked decrease once the baby was born, and subsequent increase during the later postpartum period. The temperament of the child also modulated anxiety trajectories. Thus, mothers of children high in negative affectivity and who also experienced greater early adversity had elevated and flat anxiety trajectories, while child extraversion was associated with increasing anxiety courses approaching 2 years postpartum. These results show that maternal anxiety dynamically changes through the postpartum period with a course that is affected by previous and current experiences.


Association Between Different Forms of Elder Mistreatment and Cognitive Change.
Objectives: Elder mistreatment (EM) is associated with worse physical health and psychological well-being, but little is known regarding its cognitive consequences. Methods: Data were derived from the T1-T2 PINE (n = 2713). EM was measured by psychological, physical, and sexual mistreatment; financial exploitation; and caregiver neglect. Cognitive function was evaluated by global cognition, episodic memory, working memory, processing speed, and Mini-Mental State Examination. Linear regression was used. Results: At baseline, physical mistreatment and caregiver neglect were associated with lower global cognition and cognitive domains, while psychological mistreatment was associated with higher cognitive function. Older adults with incident psychological mistreatment, physical mistreatment, and caregiver neglect had a faster decrease in global cognition and cognitive domains over 2 years. Discussion: This study is among the first to examine the association between different forms of EM and cognitive change. These findings provide a basis for engaging ethnic minorities with EM to maintain cognitive health.


Physiological sources of reductant for nitrogen fixation activity in Nostoc sp. strain UCD 7801 in symbiotic association with Anthoceros punctatus.
Pure cultures of the symbiotic cyanobacterium-bryophyte association with Anthoceros punctatus were reconstituted by using Nostoc sp. strain UCD 7801 or its 3-(3,4-dichlorophenol)-1,1-dimethylurea (DCMU)-resistant mutant strain, UCD 218. The cultures were grown under high light intensity with CO2 as the sole carbon source and then incubated in the dark to deplete endogenous reductant pools before measurements of nitrogenase activities (acetylene reduction). High rates of light-dependent acetylene reduction were obtained both before starvation in the dark and after recovery from starvation, regardless of which of the two Nostoc strains was reconstituted in the association. Rates of acetylene reduction by symbiotic tissue with the wild-type Nostoc strain decreased 99 and 96% after 28 h of incubation in the dark and after reexposure to light in the presence of 5 microM DCMU, respectively. Supplementation of the medium with glucose restored nitrogenase activity in the dark to a rate that was 64% of the illuminated rate. In the light and in the presence of 5 microM DCMU, acetylene reduction could be restored to 91% of the uninhibited rate by the exogenous presence of various carbohydrates. The rate of acetylene reduction in the presence of DCMU was 34% of the uninhibited rate of tissue in association with the DCMU-resistant strain UCD 218. This result implies that photosynthates produced immediately by the cyanobacterium can supply at least one-third of the reductant required for nitrogenase activity on a short-term basis in the symbiotic association. However, high steady-state rates of nitrogenase activity by symbiotic Nostoc strains appear to depend on endogenous carbohydrate reserves, which are presumably supplied as photosynthate from both A. punctatus tissue and the Nostoc strain.


Biocontrol of aflatoxigenic Aspergillus parasiticus by native Debaryomyces hansenii in dry-cured meat products.
Dry-cured meat products, such as dry-cured ham or dry-fermented sausages, are characterized by their particular ripening process, where a mould population grows on their surface. Some of these moulds are hazardous to the consumers because of their ability to produce mycotoxins including aflatoxins (AFs). The use of native yeasts could be considered a potential strategy for controlling the presence of AFs in dry-cured meat products. The aim of this work was to evaluate the antagonistic activity of two native Debaryomyces hansenii strains on the relative growth rate and the AFs production in Aspergillus parasiticus. Both D. hansenii strains significantly reduced the growth rates of A. parasiticus when grown in a meat-model system at different water activity (aw) conditions. The presence of D. hansenii strains caused a stimulation of AFs production by A. parasiticus at 0.99 aw. However, at 0.92 aw the yeasts significantly reduced the AFs concentration in the meat-model system. The relative expression levels of the aflR and aflS genes involved in the AFs biosynthetic pathway were also repressed at 0.92 aw in the presence of both D. hansenii strains. These satisfactory results were confirmed in dry-cured ham and dry-fermented sausage slices inoculated with A. parasiticus, since both D. hansenii strains significantly reduced AFs amounts in these matrices. Therefore, both tested D. hansenii strains could be proposed as biocontrol agents within a HACCP framework to minimize the hazard associated with the presence of AFs in dry-cured meat products.


Expression of herpes simplex virus ICP47 and human cytomegalovirus US11 prevents recognition of transgene products by CD8(+) cytotoxic T lymphocytes.
The in vivo persistence of gene-modified cells may be limited by the development of a host immune response to vector-encoded proteins. Herpesviruses evade cytotoxic T-lymphocyte (CTL) recognition by expressing genes which interfere selectively with presentation of viral antigens by class I major histocompatibility complex (MHC) molecules. Here, we studied the use of retroviral vectors encoding herpes simplex virus ICP47, human cytomegalovirus (HCMV) US3, or HCMV US11 to decrease presentation of viral proteins and transgene products to CD8(+) CTL. Human fibroblasts and T cells transduced to express the ICP47, US3, or US11 genes alone exhibited a decrease in cell surface class I MHC expression. The combination of ICP47 and US11 rendered fibroblasts negative for surface class I MHC and allowed a class I MHC-low population of T cells to be sorted by flow cytometry. Fibroblasts and T cells expressing both ICP47 and US11 were protected from CTL-mediated lysis and failed to stimulate specific memory T-cell responses to transgene products in vitro. Our findings suggest that expression of immunoregulatory viral gene products could be a potential strategy to prolong transgene expression in vivo.


Transfer of peanut allergy from donor to recipient after liver transplant.
31 years old female with a history of contact dermatitis, eczema, allergic rhinitis, pernicious anemia, alopecia areata and latent tuberculosis was treated concurrently with methotrexate along with isoniazid and pyridoxine. Five months into the therapy she developed acute onset jaundice progressing into fulminant liver failure with altered mentation and worsening liver function tests. Extensive workup including serological and histopathological evaluation revealed drug-induced liver injury as the etiology of her liver failure and she underwent a successful orthotropic liver transplant. On post-transplant follow-up at four months, she was noted to have an allergic reaction consisting of a perioral rash and swelling (without anaphylaxis) after receiving a kiss from her significant other who had just eaten a peanut butter chocolate. She denied any history of allergic reaction to peanuts prior to the transplant. Percutaneous skin testing revealed immediate hypersensitivity to peanut, hazelnut, and pecan believed to be acquired newly post-transplant. Further investigation revealed that the organ donor had a documented history of systemic anaphylaxis from the peanut allergy and a positive peanut-specific IgE level. Also, another parallel solid organ recipient (lung transplant) from the same organ donor experienced a serious anaphylactic reaction after peanut exposure. This is a case of food (peanut) allergy transfer from the donor to the recipient after the liver transplant. This case highlights the importance of incorporating known donor allergies as a part of pre-transplant screening, given the potentially serious consequences from the transfer of allergies to a previously anergic recipient.


Searching for new pharmacological targets for the treatment of Alzheimer's disease in Down syndrome.
Individuals with Down syndrome are at increased risk of developing Alzheimer's disease due to increase gene dosage resulting from chromosome 21 triplication. Although virtually all adults with Down syndrome will exhibit the major neuropathological hallmarks that define Alzheimer's disease, not all of them will develop the clinical symptoms associated with this disorder (i.e. dementia). Therefore, a good understanding of the pathophysiology of Alzheimer's disease in Down syndrome will be crucial for the identification of novel pharmacological targets to develop disease-modifying therapies for the benefit of Down syndrome individuals and for Alzheimer's sufferers alike. The study of biomarkers will also be essential for the development of better screening tools to identify dementia at its incipient stages. This review discusses the best-validated pharmacological targets for the treatment of cognitive impairment and Alzheimer's disease in Down syndrome. We further examine the relevance of newly discovered biological markers for earlier dementia diagnosis in this population.


Retrospective analysis of natural products provides insights for future discovery trends.
Understanding of the capacity of the natural world to produce secondary metabolites is important to a broad range of fields, including drug discovery, ecology, biosynthesis, and chemical biology, among others. Both the absolute number and the rate of discovery of natural products have increased significantly in recent years. However, there is a perception and concern that the fundamental novelty of these discoveries is decreasing relative to previously known natural products. This study presents a quantitative examination of the field from the perspective of both number of compounds and compound novelty using a dataset of all published microbial and marine-derived natural products. This analysis aimed to explore a number of key questions, such as how the rate of discovery of new natural products has changed over the past decades, how the average natural product structural novelty has changed as a function of time, whether exploring novel taxonomic space affords an advantage in terms of novel compound discovery, and whether it is possible to estimate how close we are to having described all of the chemical space covered by natural products. Our analyses demonstrate that most natural products being published today bear structural similarity to previously published compounds, and that the range of scaffolds readily accessible from nature is limited. However, the analysis also shows that the field continues to discover appreciable numbers of natural products with no structural precedent. Together, these results suggest that the development of innovative discovery methods will continue to yield compounds with unique structural and biological properties.


An International Survey of Approaches to Chair Restraint of Nonhuman Primates.
Specifically designed restraint chairs are the preferred method of restraint for research studies that require NHP to sit in place for sustained periods of time. In light of increasing emphasis on refinement of restraint to improve animal wellbeing, it is important to have a better understanding of this potentially stressful procedure. Although chair restraint is used internationally, very little published information is available on this subject. We developed a survey to obtain an overview of equipment, procedures, and plans for improvement regarding chair restraint. We received 101 responses from people working in academic, government, contract research, and pharmaceutical laboratories within the Americas, Europe and Asia. Findings indicate that the majority of laboratories using restraint chairs work with macaque species. Restraint chairs are used for a wide range of procedures, including cognitive testing, recording neuronal activity, functional MRI, intravenous infusion, and blood sampling. Approximately 2/3 of laboratories use an enclosed 'box chair,' which the animal is trained to enter and then to extend its head through an opening on the top of the chair; the remaining one third of laboratories use an 'open chair' design, in which manual handling or the pole-and-collar system is used to transfer and secure the animal into the chair. Respondents reported that when selecting the type of chair to use, they considered comfort for the animal, ease of use, and the ability to adjust fit between animals of different sizes. Various training methods and timeframes are used to prepare macaques for restraint chair procedures. Several laboratories are incorporating greater use of positive reinforcement training. The community that uses these restraint procedures needs to work together to define best practice; our survey results can help in that effort.


Total antioxidant response in patients with schizophrenia.
There is a large amount of convincing data demonstrating that reactive oxygen species (ROS) are involved in initiation and development of many different forms of neuropsychiatric disorders. The levels of oxidants and antioxidants in schizophrenia have been evaluated. However, measurements of total antioxidant response (TAR) were not evaluated up to now. Therefore, the objectives of this study are to investigate plasma TAR levels in schizophrenia subtypes. A total of 76 patients with schizophrenia and 25 healthy volunteers were included in the study. Positive and Negative Syndrome Scale (SANS and SAPS, respectively) were applied to patients. TAR values were determined in the plasma of normal healthy controls and patients with schizophrenia. Plasma TAR levels of each schizophrenia subtype were significantly lower than healthy controls (P < 0.01 for disorganized, residual and undifferentiated subtypes and P < 0.01 for paranoid subtype). When intragroup comparisons were performed, paranoid subtype had higher plasma TAR levels compared to other subtypes (P < 0.01). Accordingly, as a whole group, patients with schizophrenia had lower plasma TAR levels compared to controls. Plasma TAR levels were significantly and negatively correlated with SANS scores, and duration of illness was evaluated but not related to other parameters. Consequently, the present study further emphasizes the growing consideration that free radical damage may have an important etiopathogenetic role on the development of schizophrenia and suggests that decreased plasma total antioxidant levels may be related to the progression of illness.


Views on neurodegeneration as a basis for neuroprotective strategies.
Evidence is presented to demonstrate neurodegenerative processes in Parkinson's disease which are interconnected and may be synergistic in a way that they self-perpetuate progression. Free iron plays a predominant role, because it may be continuously and unlimitedly taken up through a disturbed blood-brain-barrier. Iron's toxic action is at both neuronal and glial sites. Loss of tyrosine hydroxylase protein and activity and fibrillation of alpha-synuclein connected with disturbed proteasomal protein breakdown contribute to cell death, as are changes in neuromelanin concentration and binding affinity, e.g. for iron. The interplay of genetic disturbances and neuronal and glial pathological processes involving the functioning of the blood-brain barrier, eventually initiated via an ascending toxic process, is the key for attacking vulnerable catecholaminergic neurons such as those in the substantia nigra and locus coeruleus. Neuroprotective therapeutic strategies are difficult to achieve because of the immanent complexity of cell death cascades.


Fabrication of a Molecularly Imprinted Nano-Interface-Based Electrochemical Biosensor for the Detection of CagA Virulence Factors of H. pylori.
H. pylori is responsible for several stomach-related diseases including gastric cancer. The main virulence factor responsible for its establishment in human gastric cells is known as CagA. Therefore, in this study, we have fabricated a highly sensitive MIP-based electrochemical biosensor for the detection of CagA. For this, an rGO and gold-coated, screen-printed electrode sensing platform was designed to provide a surface for the immobilization of a CagA-specific, molecularly imprinted polymer; then it was characterized electrochemically. Interestingly, molecular dynamics simulations were studied to optimize the MIP prepolymerization system, resulting in a well-matched, optimized molar ratio within the experiment. A low binding energy upon template removal indicates the capability of MIP to recognize the CagA antigen through a strong binding affinity. Under the optimized electrochemical experimental conditions, the fabricated CagA-MIP/Au/rGO@SPE sensor exhibited high sensitivity (0.275 µA ng-1 mL-1) and a very low limit of detection (0.05 ng mL-1) in a linear range of 0.05-50 ng mL-1. The influence of other possible interferents in analytical response has also been observed with the successful determination of the CagA antigen.


Proliferative and clonogenic heterogeneity of cells from EMT6/Ro multicellular spheroids induced by the glucose and oxygen supply.
The culture of EMT6/Ro multicellular tumor spheroids in a 20-fold range of glucose concentrations and a 4-fold range of oxygen tensions had profound effects on the cell subpopulations which developed in the spheroids. As the spheroids increased in diameter the rate of accumulation of cells with a G1-phase DNA content was much greater for those spheroids cultured in the lower oxygen and glucose concentrations. Growth fractions of 600- to 800-micron-diameter spheroids, calculated from either DNA content or [3H]thymidine labeling data, showed a direct correlation with the glucose concentrations in the culture medium. The cells from spheroids cultured in low glucose concentrations also showed a more rapid loss in clonogenicity as a function of increasing spheroid diameter. Lowering the oxygen tension enhanced these effects of reduced glucose concentrations. Selective dissociation of 600- to 800-micron-diameter spheroids demonstrated that both the proliferation index and the clonogenic capacity of the spheroid cells decreased with increasing depth into the spheroid cell rim. The rates of decrease of both of these parameters as a function of depth into the spheroid were greater for spheroids cultured in the lower oxygen and glucose concentrations. These results indicate that both the glucose and oxygen supply are critical in the development of nonproliferating and nonclonogenic cell subpopulations in spheroids.


Mimicking multivalent protein-carbohydrate interactions for monitoring the glucosamine level in biological fluids and pharmaceutical tablets.
An easily synthesizable probe has been employed for dual mode sensing of glucosamine in pure water. The method was also applied for glucosamine estimation in blood serum samples and pharmaceutical tablets. Further, selective detection of glucosamine was also achieved using portable color strips.


A qualitative exploration of health workers' and clients' perceptions of barriers to completing four antenatal care visits in Morogoro Region, Tanzania.
Antenatal care (ANC) remains an important contact point on the continuum of care for mothers and children in low- and middle-income countries. In Tanzania, the proportion of pregnant women completing at least four ANC visits (ANC-4) dropped from 70% to 43% between 1999 and 2010. To identify potential causes of the decline in the number of ANC visits, we conducted qualitative research at 18 health centres in Morogoro Region, exploring providers' communication about ANC visits and clients' and providers' perceptions of changes in ANC services and barriers to completing four visits. We also observed counselling messages delivered during 203 ANC consultations. Our results indicate that provider communication about ANC visit recommendations is inadequate, and confusion exists among clients about when and how often they should attend. Participants highlighted how the scale up of Prevention of Mother-to-Child Transmission, with routine human immunodeficiency virus testing for women and their male partners, presents additional barriers for some women. Changes to the timing and content of ANC services following the adoption of the Focused ANC model was described by participants as changing women's perceptions and decisions in how they utilize ANC services. In particular, condensed delivery of technical interventions fostered a sense among clients that multiple visits are unnecessary. Other barriers that may contribute to declining ANC-4 include changing norms about family planning and birth spacing, out-of-pocket costs for clients and informal practices adopted by health facilities and providers such as turning women away who attend early in pregnancy or are not accompanied by male partners. Further research is needed to determine the role and extent that these barriers may be contributing to declining ANC-4. Issues of poor communication, supply inadequacies and informal practices, deserve immediate attention from the health system.


Desiccation survival in an Antarctic nematode: molecular analysis using expressed sequenced tags.
Nematodes are the dominant soil animals in Antarctic Dry Valleys and are capable of surviving desiccation and freezing in an anhydrobiotic state. Genes induced by desiccation stress have been successfully enumerated in nematodes; however we have little knowledge of gene regulation by Antarctic nematodes which can survive multiple environmental stresses. To address this problem we investigated the genetic responses of a nematode species, Plectus murrayi, that is capable of tolerating Antarctic environmental extremes, in particular desiccation and freezing. In this study, we provide the first insight into the desiccation induced transcriptome of an Antarctic nematode through cDNA library construction and suppressive subtractive hybridization. We obtained 2,486 expressed sequence tags (ESTs) from 2,586 clones derived from the cDNA library of desiccated P. murrayi. The 2,486 ESTs formed 1,387 putative unique transcripts of which 523 (38%) had matches in the model-nematode Caenorhabditis elegans, 107 (7%) in nematodes other than C. elegans, 153 (11%) in non-nematode organisms and 605 (44%) had no significant match to any sequences in the current databases. The 1,387 unique transcripts were functionally classified by using Gene Ontology (GO) hierarchy and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The results indicate that the transcriptome contains a group of transcripts from diverse functional areas. The subtractive library of desiccated nematodes showed 80 transcripts differentially expressed during desiccation stress, of which 28% were metabolism related, 19% were involved in environmental information processing, 28% involved in genetic information processing and 21% were novel transcripts. Expression profiling of 14 selected genes by quantitative Real-time PCR showed 9 genes significantly up-regulated, 3 down-regulated and 2 continuously expressed in response to desiccation. The establishment of a desiccation EST collection for Plectus murrayi, a useful model in assessing the structural, physiological, biochemical and genetic aspects of multiple stress tolerance, is an important step in understanding the genome level response of this nematode to desiccation stress. The type of transcript analysis performed in this study sets the foundation for more detailed functional and genome level analyses of the genes involved in desiccation tolerance in nematodes.


Increased EPO Levels Are Associated With Bone Loss in Mice Lacking PHD2 in EPO-Producing Cells.
The main oxygen sensor hypoxia inducible factor (HIF) prolyl hydroxylase 2 (PHD2) is a critical regulator of tissue homeostasis during erythropoiesis, hematopoietic stem cell maintenance, and wound healing. Recent studies point toward a role for the PHD2-erythropoietin (EPO) axis in the modulation of bone remodeling, even though the studies produced conflicting results. Here, we used a number of mouse strains deficient of PHD2 in different cell types to address the role of PHD2 and its downstream targets HIF-1α and HIF-2α in bone remodeling. Mice deficient for PHD2 in several cell lineages, including EPO-producing cells, osteoblasts, and hematopoietic cells (CD68:cre-PHD2f/f ) displayed a severe reduction of bone density at the distal femur as well as the vertebral body due to impaired bone formation but not bone resorption. Importantly, using osteoblast-specific (Osx:cre-PHD2f/f ) and osteoclast-specific PHD2 knock-out mice (Vav:cre- PHD2f/f ), we show that this effect is independent of the loss of PHD2 in osteoblast and osteoclasts. Using different in vivo and in vitro approaches, we show here that this bone phenotype, including the suppression of bone formation, is directly linked to the stabilization of the α-subunit of HIF-2, and possibly to the subsequent moderate induction of serum EPO, which directly influenced the differentiation and mineralization of osteoblast progenitors resulting in lower bone density. Taken together, our data identify the PHD2:HIF-2α:EPO axis as a so far unknown regulator of osteohematology by controlling bone homeostasis. Further, these data suggest that patients treated with PHD inhibitors or EPO should be monitored with respect to their bone status. © 2016 American Society for Bone and Mineral Research.


Cocarcinogenic effects of intrahepatic bile acid accumulation in cholangiocarcinoma development.
Bile acid accumulation in liver with cholangiolar neoplastic lesions may occur before cholestasis is clinically detected. Whether this favors intrahepatic cholangiocarcinoma development has been investigated in this study. The E. coli RecA gene promoter was cloned upstream from Luc2 to detect in vitro direct genotoxic ability by activation of SOS genes. This assay demonstrated that bile acids were not able to induce DNA damage. The genotoxic effect of the DNA-damaging agent cisplatin was neither enhanced nor hindered by the hepatotoxic and hepatoprotective glycochenodeoxycholic and glycoursodeoxycholic acids, respectively. In contrast, thioacetamide metabolites, but not thioacetamide itself, induced DNA damage. Thus, thioacetamide was used to induce liver cancer in rats, which resulted in visible tumors after 30 weeks. The effect of bile acid accumulation on initial carcinogenesis phase (8 weeks) was investigated in bile duct ligated (BDL) animals. Serum bile acid measurement and determination of liver-specific healthy and tumor markers revealed that early thioacetamide treatment induced hypercholanemia together with upregulation of the tumor marker Neu in bile ducts, which were enhanced by BDL. Bile acid accumulation was associated with increased expression of interleukin (IL)-6 and downregulation of farnesoid X receptor (FXR). Bile duct proliferation and apoptosis activation, with inverse pattern (BDL > thioacetamide + BDL >> thioacetamide vs. thioacetamide > thioacetamide + BDL > BDL), were observed. In conclusion, intrahepatic accumulation of bile acids does not induce carcinogenesis directly but facilitates a cocarcinogenic effect due to stimulation of bile duct proliferation, enhanced inflammation, and reduction in FXR-dependent chemoprotection. This study reveals that bile acids foster cocarcinogenic events that impact cholangiocarcinoma.