Instrumentation of Flow-Through USP IV Dissolution Apparatus to Assess Poorly Soluble Basic Drug Products: a Technical Note.
Supersaturation and precipitation are common limitations encountered especially with poorly soluble basic drugs. The aims of this work were to explore the pattern of dissolution and precipitation of poorly soluble basic drugs using a United States Pharmacopoeia (USP) IV dissolution apparatus and to compare it to the widely used USP II dissolution apparatus. In order to investigate the influence of gastric emptying time on bioavailability, tables of two model drugs (dipyridamole 100 mg and cinnarizine 15 mg) were investigated and pH change from 1.2 to 6.8 were achieved after 10, 20 or 30 min using USP II or USP IV dissolution apparatuses. Using USP II, dipyridamole and cinnarizine concentrations dropped instantly as a result of drug precipitation with drug crystals evident in the dissolution vessel. At pH change times of 10, 20 and 30 min, the total amount of dissolved drug was dependent on pH change time. Using USP IV, at a flow rate of 8 ml/min, it was possible to have comparable release to agitation at 50 rpm using USP II suggesting that comparable hydrodynamic forces are possible. No drop in drug percentage occurs as the dissolved fraction was readily emptied from the flow cell, preventing drug accumulation in the dissolution medium. However, a negligible percentage of drug release took place following pH change. In conclusion, the use of the flow-through cell dissolution provided laminar flow, use of realistic fluid volumes and avoided precipitation of dissolved drug fraction in the gastric phase as it is discharged before pH change.


Glycaemic index and glycaemic load values of commonly consumed foods in the United Arab Emirates.
Glycaemic index (GI) and glycaemic load (GL) values of some commonly consumed foods in the United Arab Emirates were determined with an aim of adding these values to the existing international table of GI and GL values. In all, eighteen test foods categorised into breads (n 5), entrée dishes (n 3), main dishes (n 5) and sweet dishes (n 5) were tested. For each test food, at least fifteen healthy participants consumed 25 or 50 g available carbohydrate portions of a reference food (glucose), which was tested three times, and a test food after an overnight fast, was tested once, on separate occasions. Capillary blood samples were obtained by finger-prick and blood glucose was measured using clinical chemistry analyser. A fasting blood sample was obtained at baseline and before consumption of test foods. Additional blood samples were obtained at 15, 30, 45, 60, 90 and 120 min after the consumption of each test food. The GI value of each test food was calculated as the percentage of the incremental area under the blood glucose curve (IAUC) for the test food of each participant divided by the average IAUC for the reference food of the same participant. The GI values of tested foods ranged from low (55 or less) to high (70 or more). The GI values of various breads and rice-containing dishes were comparable with previously published values. This study provides GI and GL values of previously untested traditional Emirati foods which could provide a useful guide on dietary recommendations for the Emirati population.


M2 differentiation of MonoMac-1 cell line induced by M-CSF and glucocorticoid pathways.
Models of macrophage subtypes require molecular characterization to reliably facilitate their differentiation. Although CD16+ (Fc-gamma III receptor) monocytes that express CD163 (a hemoglobin/haptoglobin receptor) have been implicated in a variety of disease states, the conditions necessary to establish lineages of these cell subtypes remains unknown. The current investigations utilize a cell line derived from acute myelogenous leukemia lineage, MonoMac-1, to interrogate the factors that promote the development of CD16+ macrophages that express CD163. Results implicate the glucocorticoid pathway as well as c-fms signaling based on the action of dexamethasone and macrophage colony-stimulating factor-1 in promoting CD16+ expression, in addition to phorbol myristate acetate and lipopolysaccharides treatment. The ability of glucocorticoid and c-fms receptor antagonists to inhibit CD16+ cell formation further establishes the role of these pathways in CD16 expression in this cell line. In view of the inherent difficulty in working with primary cells as well as donor variation, cell lines may be preferable to primary cells for their consistency. We envision that the process we use to induce CD16 expression in this cell type will be useful for screening and identification of drug candidates potentially useful for the treatment of diseases where the etiology involves the expansion of the CD16+ monocytes subset or the accumulation of CD163+ tissue macrophages.


Impact of iron deficiency anemia on cell-mediated and humoral immunity in children: A case control study.
The precise role of iron in immune regulation especially in children vulnerable to iron deficiency is not fully known. Hence, this study was conducted to evaluate the effects of iron deficiency anemia (IDA) and its treatment with oral iron supplementation on cell-mediated immunity (CMI) and humoral immunity (HMI) in children. A total of 40 children (<15 years) with IDA and 40 age-matched healthy children after satisfying the inclusion criteria were enrolled for this case-control study. Flow cytometric evaluation of absolute and relative numbers of cluster of differentiation 4 (CD4) and CD8 (cluster of differentiation 8) lymphocyte subgroups was carried out to assess the CMI and serum Immunoglobulin G (IgG), Immunoglobulin A (IgA), Immunoglobulin M (IgM) were measured to assess the HMI at baseline and 3 months post oral iron supplementation. Significantly lower levels (P < 0.05) of CD4+ T-cells and decreased CD4:CD8 ratios were observed in the iron deficient children. Iron supplementation significantly improved the CD4+ cell counts and CD4:CD8 ratios. However, immunoglobulin levels weren't different between the two groups. Although IDA did not influence HMI, it significantly impaired CMI, which was improved following iron supplementation for 3 months.


Cognitive and academic outcomes after pediatric liver transplantation: Functional Outcomes Group (FOG) results.
This multicenter study examined prevalence of cognitive and academic delays in children following liver transplant (LT). One hundred and forty-four patients ages 5-7 and 2 years post-LT were recruited through the SPLIT consortium and administered the Wechsler Preschool and Primary Scale of Intelligence, 3rd Edition (WPPSI-III), the Bracken Basic Concept Scale, Revised (BBCS-R), and the Wide Range Achievement Test, 4th edition (WRAT-4). Parents and teachers completed the Behavior Rating Inventory of Executive Function (BRIEF). Participants performed significantly below test norms on intelligence quotient (IQ) and achievement measures (Mean WPPSI-III Full Scale IQ = 94.7 ± 13.5; WRAT-4 Reading = 92.7 ± 17.2; WRAT-4 Math = 93.1 ± 15.4; p < 0001). Twenty-six percent of patients (14% expected) had 'mild to moderate' IQ delays (Full Scale IQ = 71-85) and 4% (2% expected) had 'serious' delays (Full Scale IQ ≤ 70; p < 0.0001). Reading and/or math scores were weaker than IQ in 25%, suggesting learning disability, compared to 7% expected by CDC statistics (p < 0.0001). Executive deficits were noted on the BRIEF, especially by teacher report (Global Executive Composite = 58; p < 0.001). Results suggest a higher prevalence of cognitive and academic delays and learning problems in pediatric LT recipients compared to the normal population.


Argyrophilic nucleolar organizer regions (AgNORs) in mucosal epithelium under experimental denture bases in rats.
The purpose of this study was to evaluate changes in the argyrophilic nucleolar organizer region (AgNOR) counts in mucosal epithelium induced by continuous or intermittent compressive pressure exerted through experimental denture bases and to examine the relationships between the AgNOR count, histopathological changes and the intensity of the pressure under denture bases. Continuous or intermittent compressive pressure exerted through the denture bases was applied to the hard palate of the molar region in rats. A morphometric analysis of AgNORs was performed in denture-supporting tissue 3 days and 1, 2, 4, 8, 12 and 20 weeks after the denture insertion. From the results of this study, it was found that non-pressure contact of the denture bases with palatal tissues did not change the AgNOR count. The AgNOR count was decreased by continuous or intermittent compressive pressure, and then recovered to almost the same level as with the non-pressure contact at 20 weeks following a decrease of the pressure. The AgNOR counts in the epithelium under the denture bases were revealed to be related to the histopathological changes in the denture-supporting tissues and the intensity of the pressure under the denture bases.


In vivo MRI mapping of iron oxide-labeled stem cells transplanted in the heart.
In various stem cell therapy approaches poor cell survival has been recognized as an important factor limiting therapeutic efficacy. Therefore noninvasive monitoring of cell fate is warranted for developing clinically effective stem cell therapy. In this study we investigated the use of voxel-based R₂ mapping as a tool to monitor the fate of iron oxide-labeled cells in the myocardium. Mesenchymal stem cells were transduced with the luciferase gene, labeled with ferumoxide particles and injected in the myocardium of healthy rats. Cell fate was monitored over a period of 8 weeks by bioluminescence and quantitative magnetic resonance imaging. Bioluminescence signal increased during the first week followed by a steep decrease to undetectable levels during the second week. MR imaging showed a sharp increase in R₂ values shortly after injection at the injection site, followed by a very gradual decrease of R₂ over a period of 8 weeks. No difference in the appearances on R₂-weighted images was observed between living and dead cells over the entire time period studied. No significant correlation between the bioluminescence optical data and R₂ values was observed and quantitative R₂ mapping appeared not suitable for the in vivo assessment of stem cell. These results do not follow previous in vitro reports where it was proposed that living cells may be distinguished from dead cells on the basis of the R₂ relaxivities (intracellular and extracellular iron oxides). Cell proliferation, cell migration, cell death, extracellular superparamagnetic iron oxide dispersion and aggregation exhibit different relaxivities. In vivo these processes happen simultaneously, making quantification very complex, if not impossible.


Conformational epitope on gp120 important in CD4 binding and human immunodeficiency virus type 1 neutralization identified by a human monoclonal antibody.
A human monoclonal antibody designated 15e is reactive with the envelope glycoprotein (gp120) of multiple isolates of human immunodeficiency virus type 1 (HIV-1). Antibody 15e also neutralizes HIV-1 with broad specificity and blocks gp120 binding to CD4. Characterization of the 15e epitope shows that it is conformation dependent and is distinct from previously recognized functional domains of gp120, suggesting that this epitope represents a novel site important for HIV-1 neutralization and CD4 binding. These findings have implications for the development of a vaccine for AIDS.


HIV-1 recombinants with multiple parental strains in low-prevalence, remote regions of Cameroon: evolutionary relics?
The HIV pandemic disseminated globally from Central West Africa, beginning in the second half of the twentieth century. To elucidate the virologic origins of the pandemic, a cross-sectional study was conducted of the genetic diversity of HIV-1 strains in villagers in 14 remote locations in Cameroon and in hospitalized and STI patients. DNA extracted from PBMC was PCR amplified from HIV(+) subjects. Partial pol amplicons (N = 164) and nearly full virus genomes (N = 78) were sequenced. Among the 3956 rural villagers studied, the prevalence of HIV infection was 4.9%; among the hospitalized and clinic patients, it was 8.6%. Virus genotypes fell into two distinctive groups. A majority of the genotyped strains (109/164) were the circulating recombinant form (CRF) known to be endemic in West Africa and Central West Africa, CRF02_AG. The second most common genetic form (9/164) was the recently described CRF22_01A1, and the rest were a collection of 4 different subtypes (A2, D, F2, G) and 6 different CRFs (-01, -11, -13, -18, -25, -37). Remarkably, 10.4% of HIV-1 genomes detected (17/164) were heretofore undescribed unique recombinant forms (URF) present in only a single person. Nearly full genome sequencing was completed for 78 of the viruses of interest. HIV genetic diversity was commonplace in rural villages: 12 villages each had at least one newly detected URF, and 9 villages had two or more. These results show that while CRF02_AG dominated the HIV strains in the rural villages, the remainder of the viruses had tremendous genetic diversity. Between the trans-species transmission of SIVcpz and the dispersal of pandemic HIV-1, there was a time when we hypothesize that nascent HIV-1 was spreading, but only to a limited extent, recombining with other local HIV-1, creating a large variety of recombinants. When one of those recombinants began to spread widely (i.e. became epidemic), it was recognized as a subtype. We hypothesize that the viruses in these remote Cameroon villages may represent that pre-epidemic stage of viral evolution.


Clinical update: how to recognize and treat depression in older patients.. Interview by Marc E. Webster.
Geriatrics is pleased to highlight the clinical implications of research topics supported by the American Federation for Aging Research (AFAR). AFAR is a leading private organization supporting research on the aging process and diseases of older populations. More than 900 physicians, scientists, and students have received AFAR grants totaling more than $20 million since AFAR was founded by Irving S. Wright, MD, in 1981. The articles in the New Frontiers series are designed to provide primary care physicians with insight into the pathogenesis, diagnosis, prevention, and treatment of the diseases of aging.


Regional brain stem atrophy in idiopathic Parkinson's disease detected by anatomical MRI.
Idiopathic Parkinson's disease (PD) is a neurodegenerative disorder characterized by the dysfunction of dopaminergic dependent cortico-basal ganglia loops and diagnosed on the basis of motor symptoms (tremors and/or rigidity and bradykinesia). Post-mortem studies tend to show that the destruction of dopaminergic neurons in the substantia nigra constitutes an intermediate step in a broader neurodegenerative process rather than a unique feature of Parkinson's disease, as a consistent pattern of progression would exist, originating from the medulla oblongata/pontine tegmentum. To date, neuroimaging techniques have been unable to characterize the pre-symptomatic stages of PD. However, if such a regular neurodegenerative pattern were to exist, consistent damages would be found in the brain stem, even at early stages of the disease. We recruited 23 PD patients at Hoenn and Yahr stages I to II of the disease and 18 healthy controls (HC) matched for age. T1-weighted anatomical scans were acquired (MPRAGE, 1 mm3 resolution) and analyzed using an optimized VBM protocol to detect white and grey matter volume reduction without spatial a priori. When the HC group was compared to the PD group, a single cluster exhibited statistical difference (p<0.05 corrected for false detection rate, 4287 mm3) in the brain stem, between the pons and the medulla oblongata. The present study provides in-vivo evidence that brain stem damage may be the first identifiable stage of PD neuropathology, and that the identification of this consistent damage along with other factors could help with earlier diagnosis in the future. This damage could also explain some non-motor symptoms in PD that often precede diagnosis, such as autonomic dysfunction and sleep disorders.


Design for an inexpensive but effective cochlear implant.
Widespread application of cochlear implants is limited by cost, especially in developing countries. In this article we present a design for a low-cost but effective cochlear implant system. The system includes a speech processor, four pairs of transmitting and receiving coils, and an electrode array with four monopolar electrodes. All implanted components are passive, reducing to a minimum the complexity of manufacture and allowing high reliability. A four-channel continuous interleaved sampling strategy is used for the speech processor. The processor and transmission link have been evaluated in tests with a subject previously implanted with the Ineraid electrode array and percutaneous connector. A prototype of the link, consisting of four pairs of transmitting and external receiving coils, was used, with the outputs of the receiving coils directed to four intracochlear electrodes through the percutaneous connector. The subject achieved speech reception scores with the prototype system that were equivalent to those achieved with a standard laboratory implementation of a continuous interleaved sampling processor with current-controlled stimuli.


In Vitro Antilithiatic Potential of Kalanchoe pinnata, Emblica officinalis, Bambusa nutans, and Cynodon dactylon.
The study aims at the exploration of calcium oxalate (CaOx) crystal growth inhibition potential of Cynodon dactylon, Emblica officinalis, Kalanchoe pinnata, and Bambusa nutans ethyl acetate fraction rich in polyphenol and flavonoid. Ethyl acetate fraction was separated from the hydromethanolic extract of C. dactylon, E. officinalis, K. pinnata, and B. nutans followed by quantitative analysis for total polyphenol and flavonoid content. Ethyl acetate fraction of all the plants were subjected to in vitro screening for the inhibition of CaO x crystals growth induced by sodium oxalate. The results signify rich presence of polyphenols and flavonoids in K. pinnata and E. officinalis ethyl acetate fractions followed by C. dactylon and B. nutans. Ethyl acetate fractions of B. nutans shoot, E. officinalis fruit, and K. pinnata leaf have excellent in vitro CaO x crystal growth inhibition potential based on both the comparative concentration and the time level to achieve IC50. The study outcome substantiates potential in vitro CaO x crystal dissolution and crystal growth inhibition properties of E. officinalis, B. nutans, C. dactylon, and K. pinnata. Rich presence of caffeic acid, ferulic acid, and luteolin in ethyl acetate fraction of B. nutans leaf, and chebulinic acid, chebulagic acid, gallic acid, ellagic acid, and quercetin of E. officinalis may have produced prominent crystal aggregation inhibition response.


pH titration monitored by quantum cascade laser-based vibrational circular dichroism.
Vibrational circular dichroism (VCD) spectra of aqueous solutions of proline were recorded in the course of titrations from basic to acidic pH using a spectrometer equipped with a quantum cascade laser (QCL) as an infrared light source in the spectral range from 1320 to 1220 cm(-1). The pH-dependent spectra were analyzed by singular value decomposition and global fitting of a two-pK Henderson-Hasselbalch model. The analysis delivered relative fractions of the three different protonation species. Their agreement with the relative fractions obtained from performing the same analysis on pH-dependent Fourier transform infrared (FT-IR) and QCL-IR spectra validates the quantitative results from QCL-VCD. Global fitting of the pH-dependent VCD spectra of L-proline allowed for extraction of pure spectra corresponding to anionic, zwitterionic, and cationic L-proline. From a static experiment, only pure spectra of the zwitterion would be accessible in a straightforward way. A comparison to VCD spectra calculated for all three species led to assignment of vibrational modes that are characteristic for the respective protonation states. The study demonstrates the applicability of QCL-VCD both for quantitative evaluation and for qualitative interpretation of dynamic processes in aqueous solutions.


Game bird carcasses are less persistent than raptor carcasses, but can predict raptor persistence dynamics.
Researchers conduct post-construction fatality monitoring (PCFM) to determine a wind energy facility's direct impacts on wildlife. Results of PCFM can be used to evaluate compliance with permitted take, potentially triggering adaptive management measures or offsetting mitigation; reducing uncertainty in fatality rates benefits wind companies, wildlife agencies, and other stakeholders. As part of PCFM, investigators conduct carcass persistence trials to account for imperfect detection during carcass surveys. In most PCFM studies, pen-raised game birds and other non-raptor surrogates have been used to estimate persistence of all large birds, including raptors. However, there is a growing body of evidence showing carcass persistence varies by bird type; raptor fatality estimates based on game bird carcass persistence may therefore be biased high. We conducted raptor and game bird carcass persistence field trials for 1 year at 6 wind energy facilities. Raptor carcass persistence varied by habitat and season, whereas the best-supported game bird model only included habitat. Raptor persistence probabilities were higher than corresponding game bird persistence probabilities for 13 of the 16 habitat and season combinations. Analysis of a curated large bird persistence meta-dataset showed that raptor carcass persistence varied by season, habitat, and region. The probability of persisting through a 30-day search interval ranged from 0.44 to 0.99 for raptors and from 0.16 to 0.79 for game birds. Raptor persistence was significantly higher than game bird persistence for 95% of the sampled strata. We used these carcass persistence estimates to develop linear mixed-effects models that predict raptor persistence probabilities based on estimated game bird persistence probabilities. Our scaling model provides an important statistical method to address gaps in raptor persistence data at sites in a broad range of landscape contexts in the continental United States and should be used to inform fatality estimation when site-specific raptor persistence data are limited or absent.


Mental health and trauma in asylum seekers landing in Sicily in 2015: a descriptive study of neglected invisible wounds.
In 2015, Italy was the second most common point of entry for asylum seekers into Europe after Greece. The vast majority embarked from war-torn Libya; 80,000 people claimed asylum that year. Their medical conditions were assessed on arrival but their mental health needs were not addressed in any way, despite the likelihood of serious trauma before and during migration. Médecins sans Frontières (MSF), in agreement with the Italian Ministry of Health, provided mental health (MH) assessment and care for recently-landed asylum seekers in Sicily. This study documents mental health conditions, potentially traumatic events and post-migratory living difficulties experienced by asylum seekers in the MSF programme in 2014-15. All asylum seekers transiting the 15 MSF-supported centres were invited to a psycho-educational session. A team of psychologists and cultural mediators then provided assessment and care for those identified with MH conditions. Potentially traumatic events experienced before and during the journey, as well as post-migratory living difficulties, were recorded. All those diagnosed with MH conditions from October 2014 to December 2015 were included in the study. Among 385 individuals who presented themselves for a MH screening during the study period, 193 (50%) were identified and diagnosed with MH conditions. Most were young, West African males who had left their home-countries more than a year prior to arrival. The most common MH conditions were post traumatic stress disorder (31%) and depression (20%). Potentially traumatic events were experienced frequently in the home country (60%) and during migration (89%). Being in a combat situation or at risk of death, having witnessed violence or death and having been in detention were the main traumas. Lack of activities, worries about home, loneliness and fear of being sent home were the main difficulties at the AS centres. MH conditions, potentially traumatic events and post-migratory living difficulties are commonly experienced by recently-arrived ASs, this study suggests that mental health and psychosocial support and improved living circumstances should be integrated into European medical and social services provided by authorities in order to fulfil their humanitarian responsibility and reduce the burden of assimilation on receiving countries.


[The occurrence of lumbar paralysis in goats and the appearance of Elaphostrongylus cervi in red deer in the canton Ticino].
The aim of the study was to examine the incidence of lumbar paralysis in goats and the possible role of Elaphostrongylus cervi as an agent. For this reason questionnaires concerning the clinical pictures and the incidence of lumbar paralysis were sent to 200 owners of goats and to 9 veterinarians. The study showed that the symptoms of lumbar paralysis had been observed for many years mainly during the winter. Most of the animals suffering from lumbar paralysis originated from the Valley of Leventina and Blenio. The symptoms of lumbar paralysis could be observed from 29 owners of goats. Fecal examination of 36 red deer in the autumn of 1995 was carried out for studying if deer plays a role in spreading the larvae of E. cervi. Twenty-five red deer passed the E. cervi larvae. This could be indicating that deer plays an possible role in the infection of goats with E. cervi.


Two-photon targeted patching (TPTP) in vivo.
Two-photon-excited fluorescence laser-scanning microscopy (2PLSM) has provided a wealth of information about the spatiotemporal properties of biological processes at the single cell and population level. Because such nonlinear optical methods allow for imaging deep within biological tissue, 2PLSM can be combined with patch-clamp techniques to obtain electrophysiological recordings from specific fluorescently labeled cells in vivo. Here a protocol referred to as two-photon targeted patching (TPTP) describes a method that may be used to record from cells in the intact animal labeled by virtually any type of fluorophore. We target neurons that have been optically and genetically identified using green fluorescent protein (GFP) expressed under the control of a specific promoter. TPTP when combined with genetic approaches therefore permits electrophysiological recordings from specified neurons and their compartments, including dendrites. This technique may be repeated in the same preparation many times over the course of several hours and is equally applicable to non-neuronal cell types.


Canadian Network for Mood and Anxiety Treatments (CANMAT) consensus recommendations for functional outcomes in major depressive disorder.
Functional recovery is increasingly recognized as a priority in the treatment of major depressive disorder (MDD), by both clinicians and patients. However, symptom improvement remains the focus of traditional clinical trials for MDD and of the regulatory approval process for new medications and other interventions. Many studies have shown that functional outcomes do not always correspond to symptom-based outcomes. Representatives from clinical practice, professional societies, academia, industry, and government were invited by the Canadian Network for Mood and Anxiety Treatments to develop recommendations for the conceptualization and measurement of functional outcomes in clinical trials of MDD. Definitions and conceptual frameworks to guide assessment of functioning are described, as well as research methodology applicable to the broad spectrum of treatments for MDD. Examples are given for validated instruments, including patient-reported outcome measures. Strategies for knowledge translation and dissemination are suggested and consensus recommendations summarized. As the societal burden and financial costs of MDD continue to escalate, so does the need for evidence-based and cost-effective interventions that demonstrate improvement in functioning. Routine assessment of functional outcomes will benefit not only individuals with MDD but also diverse stakeholders concerned about the efficacy and cost-effectiveness of interventions.


Advanced imaging in acute and chronic deep vein thrombosis.
Deep venous thrombosis (DVT) affecting the extremities is a common clinical problem. Prompt imaging aids in rapid diagnosis and adequate treatment. While ultrasound (US) remains the workhorse of detection of extremity venous thrombosis, CT and MRI are commonly used as the problem-solving tools either to visualize the thrombosis in central veins like superior or inferior vena cava (IVC) or to test for the presence of complications like pulmonary embolism (PE). The cross-sectional modalities also offer improved visualization of venous collaterals. The purpose of this article is to review the established modalities used for characterization and diagnosis of DVT, and further explore promising innovations and recent advances in this field.


Testicular hypofunction and multiple sclerosis risk: a record-linkage study.
The influence of gonadal hormones on multiple sclerosis (MS) is not well characterized and has thus far been investigated primarily in animal models or as a proposed therapeutic approach. We investigated a potential association between testicular hypofunction, as a proxy for low testosterone levels, and MS risk through analysis of linked English national Hospital Episode Statistics from 1999 to 2011. We report a strong positive association between testicular hypofunction and subsequent MS (rate ratio = 4.62, 95% confidence interval = 2.3-8.24, p < 0.0001). Future work should aim more directly to elucidate the relationship between testosterone levels and MS in both males and females.


Further evidence of the developmental origins of osteoarthritis: results from the Hertfordshire Cohort Study.
Investigators have suggested a link between birth weight and both hand and lumbar spine osteoarthritis (OA). In this study, we sought to extend these observations by investigating relationships between growth in early life, and clinical and radiological diagnoses of OA at the hand, knee and hip, among participants from the Hertfordshire Cohort Study. Data were available for 222 men and 222 women. Clinical OA was defined based on American College of Rheumatology criteria. Radiographs were taken of the knees and hips, and graded for the presence of osteophytes and overall Kellgren and Lawrence (KL) score. Lower weight at year one was associated with higher rates of clinical hand OA (OR 1.396, 95% CI 1.05, 1.85, P=0.021). Individuals with lower birth weights were more likely to have hip osteophytes (OR 1.512, 95% CI 1.14, 2.00, P=0.004) and this remained robust after adjustment for confounders. Furthermore, a low weight at one year was also associated with a higher osteophyte number in the lateral compartment of the knee, after adjustment for confounders (OR 1.388, 95% CI 1.01, 1.91, P=0.043). We have found further evidence of a relationship between early life factors and adult OA. These findings accord with previous studies.


Catechol O-methyltransferase mRNA expression in human and rat brain: evidence for a role in cortical neuronal function.
Catechol O-methyltransferase (COMT) is involved in the inactivation of catecholamines, including the neurotransmitter dopamine. A Val(108/158) Met functional polymorphism of the COMT gene has been shown to affect working memory-associated frontal lobe function in humans. In the present study, in situ hybridization histochemistry was employed to determine the mRNA expression profile of COMT in the human prefrontal cortex, striatum and midbrain and in the rat forebrain. In both species, COMT mRNA signals were observed in large pyramidal and smaller neurons in all cortical layers of the prefrontal cortex as well as in medium and large neurons in the striatum. Levels of COMT mRNA were obviously higher in neurons than in glia. The striatum, which receives a dense dopaminergic input, expressed lower levels of COMT mRNA as compared with the prefrontal cortex. Consistent with previous protein expression data, COMT mRNA was abundant in ependymal cells lining the cerebral ventricles. In the midbrain, COMT mRNA was detected in dopaminergic neurons in both species, albeit at low levels. In the rat forebrain, dense labeling was also detected in choroid plexus and hippocampal dentate gyrus and Ammon's horn neurons. Contrary to expectations that COMT would be expressed predominantly in non-neuronal cells, the present study shows that neurons are the main cell populations expressing COMT mRNA in the prefrontal cortex and striatum. Combined with previous data about protein localization, the present results suggest that the membrane-bound isoform of COMT having a high affinity for dopamine is expressed at neuronal dendritic processes in human cortex, consistent with functional evidence that it plays an important role in dopaminergic neurotransmission.


Nucleotide sequence diversity at the alcohol dehydrogenase 1 locus in wild barley (Hordeum vulgare ssp. spontaneum): an evaluation of the background selection hypothesis.
The background selection hypothesis predicts a reduction in nucleotide site diversity and an excess of rare variants, owing to linkage associations with deleterious alleles. This effect is expected to be amplified in species that are predominantly self-fertilizing. To examine the predictions of the background selection hypothesis in self-fertilizing species, we sequenced 1,362 bp of adh1, a gene for alcohol dehydrogenase (Adh; alcohol:NAD+ oxidoreductase, EC 220.127.116.11), in a sample of 45 accessions of wild barley, Hordeum vulgare ssp. spontaneum, drawn from throughout the species range. The region sequenced included 786 bp of exon sequence (part of exon 4, all of exons 5-9, and part of exon 10) and 576 bp of intron sequence (all of introns 4-9). There were 19 sites polymorphic for nucleotide substitutions, 8 in introns, and 11 in exons. Of the 11 nucleotide substitutions in codons, 4 were synonymous and 7 were nonsynonymous, occurring uniquely in the sample. There was no evidence of recombination in the region studied, and the estimated effective population size (Ne) based on synonymous sites was approximately 1.8-4.2 x 10(5). Several tests reveal that the pattern of nonsynonymous substitutions departs significantly from neutral expectations. However, the data do not appear to be consistent with recovery from a population bottleneck, recent population expansion, selective sweep, or strong positive selection. Though several features of the data are consistent with background selection, the distributions of polymorphic synonymous and intron sites are not perturbed toward a significant excess of rare alleles as would be predicted by background selection.


Detection of OXA-48 Gene in Carbapenem-Resistant Escherichia coli and Klebsiella pneumoniae from Urine Samples.
Resistance to carbapenem in Gram-negative bacteria is attributable to their ability to produce carbapenemase enzymes. The main objective of this study was to detect the presence of blaOXA-48 genes in carbapenem-resistant uropathogenic Escherichia coli and Klebsiella pneumoniae isolated from urine samples from patients attending Alka Hospital, Jawalakhel, Lalitpur, Nepal. A total of 1013 mid-stream urine samples were collected from patients with suspected urinary tract infection (UTI) between April and September 2018. The identified isolates underwent antibiotic susceptibility testing using the modified Kirby-Bauer disc-diffusion method. Phenotypic carbapenemase production was confirmed by the modified Hodge test, and the blaOXA-48 gene was detected using conventional polymerase chain reaction. Out of 1013 urine samples, 15.2% (154/1013) had bacterial growth. Among the isolates, 91.5% (141/154) were Gram-negative bacteria, and E. coli was the most common bacterial isolate (62.9%; 97/154), followed by K. pneumoniae 15.6% (24/154). Among 121 bacterial isolates (97 E. coli isolates and 24 K. pneumoniae isolates), 70.3% (52/121) were multidrug-resistant E. coli and 29.7% (22/121) were multidrug-resistant K. pneumoniae. In addition, 9.1% (11/121) were carbapenem resistant (both imipenem and meropenem resistant). Development of multidrug resistance and development of carbapenem resistance were significantly associated (p<0.05). Of the 11 carbapenem-resistant isolates, only seven were carbapenemase producers; of these, 28.6% (2/7) were E. coli, 72.4% (5/7) were K. pneumoniae and 42.8% (3/7) had the blaOXA-48 gene. Of the three bacterial isolates with the blaOXA-48 gene, 33.3% (1/3) were E. coli and 66.7% (2/3) were K. pneumoniae. One in ten isolates of E. coli and K. pneumoniae were carbapenem resistant. Among carbapenem-resistant isolates, one-third of E. coli and two-thirds of K. pneumoniae had the blaOXA-48 gene. OXA-48 serves as a potential agent to map the distribution of resistance among clinical isolates.


Overexpression of AtPCS1 in tobacco increases arsenic and arsenic plus cadmium accumulation and detoxification.
The heterologous expression of AtPCS1 in tobacco plants exposed to arsenic plus cadmium enhances phytochelatin levels, root As/Cd accumulation and pollutants detoxification, but does not prevent root cyto-histological damages. High phytochelatin (PC) levels may be involved in accumulation and detoxification of both cadmium (Cd) and arsenic (As) in numerous plants. Although polluted environments are frequently characterized by As and Cd coexistence, how increased PC levels affect the adaptation of the entire plant and the response of its cells/tissues to a combined contamination by As and Cd needs investigation. Consequently, we analyzed tobacco seedlings overexpressing Arabidopsis phytochelatin synthase1 gene (AtPCS1) exposed to As and/or Cd, to evaluate the levels of PCs and As/Cd, the cyto-histological modifications of the roots and the Cd/As leaf extrusion ability. When exposed to As and/or Cd the plants overexpressing AtPCS1 showed higher PC levels, As plus Cd root accumulation, and detoxification ability than the non-overexpressing plants, but a blocked Cd-extrusion from the leaf trichomes. In all genotypes, As, and Cd in particular, damaged lateral root apices, enhancing cell-vacuolization, causing thinning and stretching of endodermis initial cells. Alterations also occurred in the primary structure region of the lateral roots, i.e., cell wall lignification in the external cortex, cell hypertrophy in the inner cortex, crushing of endodermis and stele, and nuclear hypertrophy. Altogether, As and/or Cd caused damage to the lateral roots (and not to the primary one), with such damage not counteracted by AtPCS1 overexpression. The latter, however, positively affected accumulation and detoxification to both pollutants, highlighting that Cd/As accumulation and detoxification due to PCS1 activity do not reduce the cyto-histological damage.


Injection pressure as a marker of intraneural injection in procedures of peripheral nerves blockade.
The blockade of peripheral nerves carries a certain risk of unwanted complications, which can follow after unintentional intraneural injection of a local anesthetic. Up till today, the research of measuring injection pressure has been based on animal models, even though the histological structure of periphery nerve is different for animal and human population, so the application pressure which presages intraneural injection in human population is still unknown. As material in performing this study there have been used 12 Wistar rats and 12 delivered stillborns. After bilateral access to the median nerve, we applied 3 ml of 2% lidocaine with epinephrine, with the help of automatic syringe charger. The needle was at first placed perineural on one side, and then intraneural on the other side of both examination groups. During every application the pressure values were monitored using the manometer, and then they were analyzed by special software program BioBench. All perineural injections resulted with the pressure < or = 27.92 kPa, while the majority of intraneural injections were combined with the injection pressure > or = 69.8 kPa. The difference between intraneural and perineural injection pressures for the two different examination groups (rats and delivered stillborns) was not statistically significant (P>0.05). As prevention from intraneural injections today are in use two methods: the method of causing paresthesia or the method of using the peripheral nerve stimulator. However the nerve injury can still occur, independent from the technique used. If our results are used in clinical practice on human population, than the high injection pressure could be the marker of intraneural lodging of a needle.


Hematopoietic stem cell transplantation for aplastic anemia and paroxysmal nocturnal hemoglobinuria: current evidence and recommendations.
Aplastic anemia (AA) is a bone marrow failure syndrome characterized by pancytopenia and an empty bone marrow. Standard treatments for AA include immunosuppressive therapy and bone marrow transplantation (BMT). BMT is the preferred option for young AA patients with a sibling donor, whereas in older patients or in those to be grafted from an unrelated donor BMT is exploited as second-line treatment. Current results of BMT for AA demonstrate cure rates up to 80 and 70% in BMT from HLA-matched siblings and unrelated donor, respectively, with age and stem cell source largely affecting the outcome. BMT is also a potential treatment option for paroxysmal nocturnal hemoglobinuria, a rare hematological disorder characterized by complement-mediated intravascular hemolytic anemia, thrombophilia and bone marrow failure.


Activity and asymmetry index of masticatory muscles in women with and without dysfunction temporomandibular.
[corrected] Compare the symmetry of the activity and masticatory muscles in individuals with TMD and asymptomatic. The study included 50 women, while 31 had temporomandibular disorders (TMD) and 19 were asymptomatic (control group), aged between 19 and 40 years. The volunteers were subjected to clinical examination of the diagnostic criteria in research with TMD (RDC/TMD) with the aim of diagnosing volunteers with or without TMD, and evaluate the electromyographic activity of the right temporalis muscle (TR) and left (TL), right masseter (MR) and left (ML), in situations of rest, isometric contraction of the muscles of the jaw elevators. We obtained the rates of activity and asymmetry in each situation and for the collection and TMD control groups. For the index of activity for the rest there was significant difference (p = 0.0008) between the control group and the TMD group, with predominance of temporal muscle, was not observed difference between groups for the index of activity during the isometric contraction (p = 0.1069). For the index of asymmetry no difference between groups during rest, for the masseter muscles (p = 0.4182) and the temporal (p = 0.7614), and also during the isometry for both masseter muscles (p = 0.8691) and for time (p = 0.6643). The control group showed prevalence of TMD and temporal muscle during rest, which did not occur in the isometry, and no difference for the index of asymmetry between the groups for the masseter and temporal muscles.


Signalling in viral entry.
Viral infections are serious battles between pathogens and hosts. They can result in cell death, elimination of the virus or latent infection keeping both cells and pathogens alive. The outcome of an infection is often determined by cell signalling. Viruses deliver genomes and proteins with signalling potential into target cells and thereby alter the metabolism of the host. Virus interactions with cell surface receptors can elicit two types of signals, conformational changes of viral particles, and intracellular signals triggering specific cellular reactions. Responses by cells include stimulation of innate and adaptive immunity, growth, proliferation, survival and apoptosis. In addition, virus-activated cell signalling boosts viral entry and gene delivery, as recently shon for adenoviruses and adeno-associated viruses. This review illustrates that multiple activation of host cells during viral entry profoundly impacts the elaborate relationship between hosts and viral pathogens.


Genome and metabolic network of "Candidatus Phaeomarinobacter ectocarpi" Ec32, a new candidate genus of Alphaproteobacteria frequently associated with brown algae.
Rhizobiales and related orders of Alphaproteobacteria comprise several genera of nodule-inducing symbiotic bacteria associated with plant roots. Here we describe the genome and the metabolic network of "Candidatus Phaeomarinobacter ectocarpi" Ec32, a member of a new candidate genus closely related to Rhizobiales and found in association with cultures of the filamentous brown algal model Ectocarpus. The "Ca. P. ectocarpi" genome encodes numerous metabolic pathways that may be relevant for this bacterium to interact with algae. Notably, it possesses a large set of glycoside hydrolases and transporters, which may serve to process and assimilate algal metabolites. It also harbors several proteins likely to be involved in the synthesis of algal hormones such as auxins and cytokinins, as well as the vitamins pyridoxine, biotin, and thiamine. As of today, "Ca. P. ectocarpi" has not been successfully cultured, and identical 16S rDNA sequences have been found exclusively associated with Ectocarpus. However, related sequences (≥97% identity) have also been detected free-living and in a Fucus vesiculosus microbiome barcoding project, indicating that the candidate genus "Phaeomarinobacter" may comprise several species, which may colonize different niches.


The virtual patient as a learning tool: a mixed quantitative qualitative study.
The use of simulation in medical education has been widely accepted. There are different types of medical simulators that vary in both accuracy to emulate the real world (fidelity) and cost of development or acquisition. There is significant evidence that supports the use of high-fidelity simulators (i.e. mannequins or dummies) to prepare students for clinical environments, less attention has been given to low-fidelity simulators. This article aims to present evidence regarding the effectiveness of a low-fidelity simulator: Virtual Patient (VP), which develops several interactive computer-based clinical scenarios, seeking to promote an alternative learning environment and the development of necessary medical skills such as clinical reasoning in students of medicine. A quasi-experimental study was designed to investigate the results on the development of history taking and clinical reasoning skills in a group of undergraduate medical students, in a course devised under the concepts of constructivism in education, which used the Virtual Patient as the fundamental teaching tool. Results were measured through a mixed, quantitative and qualitative study, triangulating the results of the students' skills evaluation when facing a clinical case represented by an actor patient before and after the course. Additionally, the description of the students' and tool's performance was measured by way of a qualitative study. The comparison of the students' skills on the evaluation matrix before-and-after the course evidenced a statistically significant advance (p < 0.01) in all aspects (interview, physical exam, clinical judgment, relevance of medical exams, and presentation of case). Students described the VP as an easy-to-use and motivating tool for learning without stress, especially at the beginning of their career. VP allowed them to create logical and structured processes, to be wrong without consequences, and to review and reassess information available. From the professor perspective, it allowed a better follow-up of the students' learning process and favored reflections on the teaching-learning process. VP proved to be a valuable and useful tool for the development of clinical reasoning and history taking skills in medical students, as part of a constructivist learning course.


Regulation of CTLA-4 expression during T cell activation.
T cell activation requires at least two distinct signals, including signaling via the Ag-specific TCR and a costimulatory pathway. The best characterized costimulatory pathway involves the CD28 molecule, which is expressed constitutively on T cells and binds the family of B7 counter-receptors on APCs. Inhibition of this costimulatory pathway prevents T cell activation and can lead to long-term T cell unresponsiveness or anergy. In contrast, CTLA4, which is homologous to CD28, has been shown to be a negative regulator of T cell activation. The CTLA4 molecule is not expressed on resting T cells, but is induced after the initial steps of T cell activation. To address the regulation of CTLA4 expression, we have analyzed CTLA4 at the level of cell surface expression, mRNA, rate of transcription, and rate of decay of message. Nuclear runoff results show an increase in the rate of transcription following T cell activation. Our analyses of non-T cells, including B cells, mastocytoma, and fibroblasts, by Northern blot analysis detect only T cell expression of CTLA4. Reporter gene analysis indicates that 335 bp of upstream CTLA4 sequence are sufficient to control inducibility. We have identified important regulatory regions that control inducible and cell-specific CTLA4 expression. These results also suggest that both positive and negative response elements modulate the transcriptional regulation of CTLA4 gene expression. Understanding the regulation of CTLA4 should provide insight into the regulation of T cell activation at the molecular level.


Europium bis(dimethylsilyl)amides including mixed-valent Eu3[N(SiHMe2)2]6[μ-N(SiHMe2)2]2.
Trivalent Eu[N(SiHMe2)2]3(THF)2 can easily be synthesized by applying a routine salt metathesis protocol (EuCl3(THF)2 and 3 equiv. of Li[N(SiHMe2)2] in n-hexane) which crystallizes isotypically to its analogues of the rare-earth metal series (space group P21/c). Transsilylamination of Eu[N(SiMe3)2]2(THF)2 with a slight excess of HN(SiHMe2)2 in n-hexane-THF yields the divalent trinuclear compound Eu{[μ-N(SiHMe2)2]2Eu[N(SiHMe2)2](THF)}2, the solid-state structure of which differs significantly from the samarium and ytterbium analogues by showing three unique molecules in the asymmetric unit of which one is related to the two others by an inversion. Using crude Eu[N(SiMe3)2]3 in transsilylamination reactions with HN(SiHMe2)2 in n-hexane afforded n-hexane-insoluble trivalent ate complexes {MEu[N(SiHMe2)2]4}n (M = Na, K) depending on the synthesis conditions of Eu[N(SiMe3)2]3. Performing the transsilylamination of Eu[N(SiMe3)2]3 with a large excess of HN(SiHMe2)2 at elevated temperatures gave reproducibly the donor-free, mixed-valent, trinuclear compound Eu(II){[μ-N(SiHMe2)2]Eu(III)[N(SiHMe2)2]3}2 in good yield.


The multiple antibiotic resistance (mar) locus and its significance.
Chromosomally encoded systems involved in low level resistance of bacteria to different classes of antibiotics (mainly beta-lactams, chloramphenicol, quinolones and tetracycline), disinfectants and in resistance to organic solvents have been the focus of considerable interest in recent years. The multiple antibiotic resistance (mar) locus of Escherichia coli and Salmonella is perhaps the best described system involved in this type of resistance which is induced by MarA, the activator protein encoded by the marRAB locus. The mar -locus is reported to mediate resistance primarily by up-regulating efflux of some antibiotics, disinfectants and organic solvents via the AcrAB-TolC efflux pump and down regulating influx through Outer Membrane Protein F (OmpF). Whilst the level of antibiotic resistance conferred by marRAB is only low level, there are increasing data to suggest that marRAB and related systems are important in clinical antibiotic resistance, possibly as a 'stepping stone' to higher levels of resistance. Other related systems include up-regulation of RobA, SoxS and AcrAB which give rise to a similar resistance phenotype to that conferred by up-regulation of MarA. The aim of this paper is to review the function and significance of the mar -locus and related systems with a particular focus on its implications in veterinary medicine.


Dissection and immunohistochemistry of larval, pupal and adult Drosophila retinas.
The compound eye of Drosophila melanogaster consists of about 750 ommatidia (unit eyes). Each ommatidium is composed of about 20 cells, including lens-secreting cone cells, pigment cells, a bristle cell and eight photoreceptors (PRs) R1-R8. The PRs have specialized microvillar structures, the rhabdomeres, which contain light-sensitive pigments, the Rhodopsins (Rhs). The rhabdomeres of six PRs (R1-R6) form a trapezoid and contain Rh1. The rhabdomeres of R7 and R8 are positioned in tandem in the center of the trapezoid and share the same path of light. R7 and R8 PRs stochastically express different combinations of Rhs in two main subtypes: In the 'p' subtype, Rh3 in pR7s is coupled with Rh5 in pR8s, whereas in the 'y' subtype, Rh4 in yR7s is associated with Rh6 in yR8s. Early specification of PRs and development of ommatidia begins in the larval eye-antennal imaginal disc, a monolayer of epithelial cells. A wave of differentiation sweeps across the disc and initiates the assembly of undifferentiated cells into ommatidia. The 'founder cell' R8 is specified first and recruits R1-6 and then R7. Subsequently, during pupal development, PR differentiation leads to extensive morphological changes, including rhabdomere formation, synaptogenesis and eventually rh expression. In this protocol, we describe methods for retinal dissections and immunohistochemistry at three defined periods of retina development, which can be applied to address a variety of questions concerning retinal formation and developmental pathways. Here, we use these methods to visualize the stepwise PR differentiation at the single-cell level in whole mount larval, midpupal and adult retinas (Figure 1).


A longitudinal study of the effects of age and time to death on hospital costs.
Recent studies indicate that approaching death, rather than age, may be the main demographic driver of health care costs. Using a 29-year longitudinal English dataset, this paper uses more robust methods to examine the effects of age and proximity to death on hospital costs. A random effects panel data two-part model shows that approaching death affects costs up to 15 years prior to death. The large tenfold increase in costs from 5 years prior to death to the last year of life overshadows the 30% increase in costs from age 65 to 85. Hence, expenditure projections must consider remaining life expectancy in the populations.


Genotype-covariate interaction effects and the heritability of adult body mass index.
Obesity is a worldwide epidemic, with major health and economic costs. Here we estimate heritability for body mass index (BMI) in 172,000 sibling pairs and 150,832 unrelated individuals and explore the contribution of genotype-covariate interaction effects at common SNP loci. We find evidence for genotype-age interaction (likelihood ratio test (LRT) = 73.58, degrees of freedom (df) = 1, P = 4.83 × 10-18), which contributed 8.1% (1.4% s.e.) to BMI variation. Across eight self-reported lifestyle factors, including diet and exercise, we find genotype-environment interaction only for smoking behavior (LRT = 19.70, P = 5.03 × 10-5 and LRT = 30.80, P = 1.42 × 10-8), which contributed 4.0% (0.8% s.e.) to BMI variation. Bayesian association analysis suggests that BMI is highly polygenic, with 75% of the SNP heritability attributable to loci that each explain <0.01% of the phenotypic variance. Our findings imply that substantially larger sample sizes across ages and lifestyles are required to understand the full genetic architecture of BMI.


Analysis of the Microbial Community in an Acidic Hollow-Fiber Membrane Biofilm Reactor (Hf-MBfR) Used for the Biological Conversion of Carbon Dioxide to Methane.
Hydrogenotrophic methanogens can use gaseous substrates, such as H2 and CO2, in CH4 production. H2 gas is used to reduce CO2. We have successfully operated a hollow-fiber membrane biofilm reactor (Hf-MBfR) for stable and continuous CH4 production from CO2 and H2. CO2 and H2 were diffused into the culture medium through the membrane without bubble formation in the Hf-MBfR, which was operated at pH 4.5-5.5 over 70 days. Focusing on the presence of hydrogenotrophic methanogens, we analyzed the structure of the microbial community in the reactor. Denaturing gradient gel electrophoresis (DGGE) was conducted with bacterial and archaeal 16S rDNA primers. Real-time qPCR was used to track changes in the community composition of methanogens over the course of operation. Finally, the microbial community and its diversity at the time of maximum CH4 production were analyzed by pyrosequencing methods. Genus Methanobacterium, related to hydrogenotrophic methanogens, dominated the microbial community, but acetate consumption by bacteria, such as unclassified Clostridium sp., restricted the development of acetoclastic methanogens in the acidic CH4 production process. The results show that acidic operation of a CH4 production reactor without any pH adjustment inhibited acetogenic growth and enriched the hydrogenotrophic methanogens, decreasing the growth of acetoclastic methanogens.


Sleep duration and sleep-related problems in different occupations in the Hordaland Health Study.
The aim of this study was to examine the relationship between occupation and sleep duration, sleepiness, insufficient sleep, and insomnia in day and shift workers (including night work and watches). The study was population-based and cross-sectional, and relied on self-administered questionnaires. It was conducted as part of the 1997-1999 Hordaland Health Study in collaboration with the Norwegian National Health Screening Service. Aged 40-45 years, 7782 participants answered a sleep questionnaire, reporting their occupation and whether or not they were employed in shift work. Our study found differences in sleep duration during the working week between occupational groups; in both shift and day workers. Craft workers, plant operators, and drivers slept less than leaders, and non-personal and personal service workers. Within some occupations (leaders, personal service workers, and plant operators), shift workers slept less than day workers. The mean sleep duration of shift workers was 15 minutes shorter than that of day workers. Rise times, but not bedtimes, were earlier in craft-and construction workers, plant operators, and drivers than in leaders and non-personal and personal service workers, particularly day workers. When adjusted for shift work and working hours - compared to leaders - craft workers, plant operators, and drivers had an increased risk of daytime sleepiness (odds ratio 1.5, 1.8, and 1.8 respectively) and of falling asleep at work (odds ratio 1.6, 2.1 and 2.0 respectively). Shift workers had an increased risk of falling asleep at work and insomnia. Occupation has separate effects on sleep duration and sleep-related problems, independent of the effects of shift work.


Quercetin lowers plasma uric acid in pre-hyperuricaemic males: a randomised, double-blinded, placebo-controlled, cross-over trial.
Elevated plasma uric acid concentration is a risk factor for gout, insulin resistance and type 2 diabetes. Quercetin, a flavonoid found in high levels in onions, tea and apples, inhibits xanthine oxidoreductase in vitro, the final step in intracellular uric acid production, indicating that quercetin might be able to lower blood uric acid in humans. We determined the effects of 4 weeks of oral supplementation of quercetin on plasma uric acid, blood pressure and fasting glucose. This randomised, double-blinded, placebo-controlled, cross-over trial recruited twenty-two healthy males (19-60 years) with baseline plasma uric acid concentration in the higher, but still considered healthy, range (339 (SD 51) µmol/l). The intervention included one tablet containing 500 mg quercetin daily for 4 weeks, compared with placebo, with a 4-week washout period between treatments. The primary outcome was change in concentrations of plasma uric acid after 2 and 4 weeks; secondary outcome measures were changes in fasting plasma glucose, 24-h urinary excretion of uric acid and resting blood pressure. After quercetin treatment, plasma uric acid concentrations were significantly lowered by -26·5 µmol/l (95% CI, -7·6, -45·5; P=0·008), without affecting fasting glucose, urinary excretion of uric acid or blood pressure. Daily supplementation of 500 mg quercetin, containing the bioavailable amount of quercetin as present in approximately 100 g red onions, for 4 weeks, significantly reduces elevated plasma uric acid concentrations in healthy males.


Transposon tagging using Ty elements in yeast.
We have used the ability to induce high levels of Ty transposition to develop a method for transposon mutagenesis in Saccharomyces cerevisiae. To facilitate genetic and molecular analysis, we have constructed GAL1-promoted TyH3 or Ty917 elements that contain unique cloning sites, and marked these elements with selectable genes. These genes include the yeast HIS3 gene, and the plasmid PiAN7 containing the Tn903 NEO gene. The marked Ty elements retain their ability to transpose, to mutate the LYS2, LYS5, or STE2 genes, and to activate the promoterless his3 delta 4 target gene. Ty elements containing selectable genes are also useful in strain construction, in chromosomal mapping, and in gene cloning strategies.


Physical Exhaustion Induced Variations in Event-Related Potentials and Cognitive Task Performance in Young Adults.
Physical exhaustion is not always peripheral, and it is the brain that causes the sensation of fatigue either due to decrease of metabolic resources or due to central activation process that regulates attention and performance. This study was undertaken to observe the variations in event-related potentials (ERPs) and cognitive performance after an exhausting physical exercise. A total of 60 healthy young adult subjects were included in the study. The study was conducted in 2 phases with at least a week gap between the phases. The participants answered a Multidimensional Fatigue Inventory (MFI-20) questionnaire before and after trials in each phase to measure the induced physical exhaustion. In phase I (control trial), the ERP data were processed using P300, Standard auditory "oddball paradigm," on computerized evoked potential recorder (RMS EMG MK-2) using 10/20 system to know the engagement of attention after which participants were given to perform cognitive tasks such as "Stroop Test, Trial Making Test and Mini Mental State Examination." In Phase II (exercise trial), the participants were instructed to cycle as hard as they could, till they could not continue anymore, which was followed by recording of P300-evoked potentials and performance of cognitive tasks as in Phase I. Paired t test was used to compare between any dependent variables. Fatigue-related subjective measures (MFI-20) showed that both mental and physical exhaustion were significantly greater in the exercise-involved cognitive trial than in the control trial. Lower P300 latencies reflect faster reaction time; however, their response accuracies were poorer resulting in poorer cognitive performances. Participants subjected to control trial performed better in terms of higher percentage accuracy but with slow reaction time. The participants experienced more fatigue physically and mentally during the exercise that involved cognitive tasks. An apparent decrease in attention based on decreased percentage accuracy of response was evident, implying that fatigue, performance, and attention are interdependent.


Stereotactic Radiosurgery for Benign (World Health Organization Grade I) Cavernous Sinus Meningiomas-International Stereotactic Radiosurgery Society (ISRS) Practice Guideline: A Systematic Review.
Stereotactic radiosurgery (SRS) has become popular as a standard treatment for cavernous sinus (CS) meningiomas. To summarize the published literature specific to the treatment of CS meningioma with SRS found through a systematic review, and to create recommendations on behalf of the International Stereotactic Radiosurgery Society. Articles published from January 1963 to December 2014 were systemically reviewed. Three electronic databases, PubMed, EMBASE, and The Cochrane Central Register of Controlled Trials, were searched. Publications in English with at least 10 patients (each arm) were included. Of 569 screened abstracts, a total of 49 full-text articles were included in the analysis. All studies were retrospective. Most of the reports had favorable outcomes with 5-yr progression-free survival (PFS) rates ranging from 86% to 99%, and 10-yr PFS rates ranging from 69% to 97%. The post-SRS neurological preservation rate ranged from 80% to 100%. Resection can be considered for the treatment of larger (>3 cm in diameter) and symptomatic CS meningioma in patients both receptive to and medically eligible for open surgery. Adjuvant or salvage SRS for residual or recurrent tumor can be utilized depending on factors such as tumor volume and proximity to adjacent critical organs at risk. The literature is limited to level III evidence with respect to outcomes of SRS in patients with CS meningioma. Based on the observed results, SRS offers a favorable benefit to risk profile for patients with CS meningioma.


Platelet activation in type 2 diabetes mellitus.
The abnormal metabolic state that accompanies diabetes renders arteries susceptible to atherosclerosis, being capable of altering the functional properties of multiple cell types, including endothelium and platelets. In particular, an altered platelet metabolism and changes in intraplatelet signaling pathways may contribute to the pathogenesis of atherothrombotic complications of diabetes. A variety of mechanisms may be responsible for enhanced platelet aggregation. Among them, hyperglycemia may represent a causal factor for in vivo platelet activation, and may be responsible for nonenzymatic glycation of platelet glycoproteins, causing changes in their structure and conformation, as well as alterations of membrane lipid dynamics. Furthermore, hyperglycemia-induced oxidative stress is responsible for enhanced peroxidation of arachidonic acid to form biologically active isoprostanes, which represents an important biochemical link between impaired glycemic control and persistent platelet activation. Finally, increased oxidative stress is responsible for activation of transcription factors and expression of redox-sensitive genes leading to a phenotypic switch of endothelium toward an adhesive, pro-thrombotic condition, initial platelet activation, adhesion and subsequent platelet aggregate formation. All this evidence is strengthened by the results of clinical trials documenting the beneficial effects of metabolic control on platelet function, and by the finding that aspirin treatment may even be more beneficial in diabetic than in high-risk non-diabetic patients. Attention to appropriate medical management of diabetic patients will have great impact on long-term outcome in this high-risk population.


A systematic review comparing antiretroviral adherence descriptive and intervention studies conducted in the USA.
We examined the extent to which studies aimed at testing interventions to improve antiretroviral adherence have targeted the facilitators of and barriers known to affect adherence. Of the 88 reports reviewed, 41 were reports of descriptive studies conducted with US HIV-positive women and 47 were reports of intervention studies conducted with US HIV-positive persons. We extracted from the descriptive studies all findings addressing any factor linked to antiretroviral adherence and from the intervention studies, information on the nature of the intervention, the adherence problem targeted, the persons targeted for the intervention, and the intervention outcomes desired. We discerned congruence between the prominence of substance abuse as a factor identified in the descriptive studies as a barrier to adherence and its prominence as the problem most addressed in those reports of intervention studies that specified the problems targeted for intervention. We also discerned congruence between the prominence of family and provider support as factors identified in the descriptive studies as facilitators of adherence and the presence of social support as an intervention component and outcome variable. Less discernible in the reports of intervention studies was specific attention to other factors prominent in the descriptive studies, which may be due to the complex nature of the problem, individualistic and rationalist slant of interventions, or simply the ways interventions were presented. Our review raises issues about niche standardization and intervention tailoring, targeting, and fidelity.


Lack of choice in caregiving decision and caregiver risk of stress, North Carolina, 2005.
An aspect of caregiving that has received little attention is the degree to which the choice to provide care affects a caregiver's emotional well-being. We compared a population-based sample of informal caregivers who reported having a choice in caring with caregivers who did not have a choice in caring to determine the extent to which choice affects caregivers' self-reported stress. We identified 341 informal caregivers who completed a caregiving module appended to the 2005 North Carolina Behavioral Risk Factor Surveillance System survey. We determined participants' self-reported stress by using a 5-point scale that was dichotomized and used adjusted binomial logistic regression to assess the risk of stress given lack of choice in caregiving. In the fully adjusted model, caregivers without a choice in caring were more than 3 times as likely to report stress as caregivers with a choice in caring. High level of burden also increased stress. Caregivers with no choice in caring were most commonly the primary caregiver of a parent. Caregivers who do not have a choice in caregiving were at increased risk of stress, which may predispose them to poor health outcomes. Further investigation is needed to determine whether interventions that target caregivers without a choice in caring can reduce their levels of stress.


Rare potential complications of thyroid fine needle biopsy.
Thyroid fine needle biopsy (FNB) is the procedure of choice for the management of thyroid nodules. Serious complications after FNB are rare, but there is also an underestimation of complication risk because of record, selection and publication biases. Apart from the well-documented post-FNB complications, we hypothesized that there are potential complications following FNB, albeit supported by limited evidence in the literature. According to our hypothesis, there may be five distinct expected rare complications: 1) cyst fluid leakage; 2) anaphylactic reaction; 3) pneumothorax; 4) thromboembolism and 5) needle tract seeding of medullary thyroid carcinoma (MTC) or thyroid lymphoma. Cyst fluid leakage and pneumothorax may be of minimal clinical significance. Needle tract seeding of MTC or thyroid lymphoma may not have significant clinical consequences, if someone considers the easiness and effectiveness of surgical removal of needle tract seeding in cases of differentiated thyroid carcinoma. On the contrary, anaphylactic reaction or thromboembolism may be life-threatening. The performers of thyroid FNB are hereby encouraged to publish these complications, if they ever occur, because awareness of them could render FNB even safer.


Conceptual model and experimental framework to determine the contributions of direct and indirect photoreactions to the solar disinfection of MS2, phiX174, and adenovirus.
Sunlight inactivates waterborne viruses via direct (absorption of sunlight by the virus) and indirect processes (adsorption of sunlight by external chromophores, which subsequently generate reactive species). While the mechanisms underlying these processes are understood, their relative importance remains unclear. This study establishes an experimental framework to determine the kinetic parameters associated with a virus' susceptibility to solar disinfection and proposes a model to estimate disinfection rates and to apportion the contributions of different inactivation processes. Quantum yields of direct inactivation were determined for three viruses (MS2, phiX174, and adenovirus), and second-order rate constants associated with indirect inactivation by four reactive species ((1)O2, OH(•), CO3(•-), and triplet states) were established. PhiX174 exhibited the greatest quantum yield (1.4 × 10(-2)), indicating that it is more susceptible to direct inactivation than MS2 (2.9 × 10(-3)) or adenovirus (2.5 × 10(-4)). Second-order rate constants ranged from 1.7 × 10(7) to 7.0 × 10(9) M(-1) s(-1) and followed the sequence MS2 > adenovirus > phiX174. A predictive model based on these parameters accurately estimated solar disinfection of MS2 and phiX174 in a natural water sample and approximated that of adenovirus within a factor of 6. Inactivation mostly occurred by direct processes, though indirect inactivation by (1)O2 also contributed to the disinfection of MS2 and adenovirus.


Treatment of Non-Small-Cell Lung Cancer with Erlotinib following Gefitinib-Induced Hepatotoxicity: Review of 8 Clinical Cases.
Objective. Gefitinib often induces liver damage. A few reports have described that the subsequent administration of erlotinib was associated with less hepatotoxicity, but the safety and efficacy of this treatment are still not fully investigated. Therefore, we evaluated retrospectively the patients with erlotinib following gefitinib-induced hepatotoxicity. Methods and Patients. We retrospectively reviewed the medical records between December 2007 and March 2010. The patients were evaluated including the following information: age, gender, histology of lung cancer, performance status, smoking status, epidermal growth factor receptor (EGFR) mutation status, liver metastasis, viral hepatitis, alcoholic liver injury, clinical response, and hepatotoxicity due to EGFR tyrosine kinase inhibitors. Results. We identified 8 patients with erlotinib following gefitinib-induced hepatotoxicity. All achieved disease control by gefitinib. Hepatotoxicity was grades 2 and 3 in 3 and 5 patients, respectively. The median duration of treatment with gefitinib was 112.5 days and the median time to gefitinib-induced hepatotoxicity was 51.5 days. The median duration of treatment with erlotinib was 171.5 days. Grade 1 and 2 erlotinib-induced hepatotoxicity was observed in 2 and 1 patient, respectively. Conclusions. Erlotinib administration with careful monitoring is thought to be a good alternative strategy for patients who respond well to gefitinib treatment but experience hepatotoxicity.


Thyroid dose estimates for a cohort of Belarusian children exposed to radiation from the Chernobyl accident.
The U.S. National Cancer Institute, in collaboration with the Belarusian Ministry of Health, is conducting a study of thyroid cancer and other thyroid diseases in a cohort of about 12,000 persons who were exposed to fallout from the Chernobyl accident in April 1986. The study subjects were 18 years old or younger at the time of exposure and resided in Belarus in the most contaminated areas of the Gomel and Mogilev Oblasts, as well as in the city of Minsk. All cohort members had at least one direct thyroid measurement made in April-June 1986. Individual data on residential history, consumption of milk, milk products and leafy vegetables as well as administration of stable iodine were collected for all cohort members by means of personal interviews conducted between 1996 and 2007. Based on the estimated (131)I activities in the thyroids, which were derived from the direct thyroid measurements, and on the responses to the questionnaires, individual thyroid doses from intakes of (131)I were reconstructed for all cohort members. In addition, radiation doses to the thyroid were estimated for the following minor exposure pathways: (a) intake of short-lived (132)I, (133)I and (132)Te by inhalation and ingestion; (b) external irradiation from radionuclides deposited on the ground; and (c) ingestion intake of (134)Cs and (137)Cs. Intake of (131)I was the major pathway for thyroid exposure; its mean contribution to the thyroid dose was 92%. The thyroid doses from (131)I intakes varied from 0.5 mGy to almost 33 Gy; the mean was estimated to be 0.58 Gy, while the median was 0.23 Gy. The reconstructed doses are being used to evaluate the risk of thyroid cancer and other thyroid diseases in the cohort.


Large wind shift on the great plains during the medieval warm period.
Spring-summer winds from the south move moist air from the Gulf of Mexico to the Great Plains. Rainfall in the growing season sustains prairie grasses that keep large dunes in the Nebraska Sand Hills immobile. Longitudinal dunes built during the Medieval Warm Period (800 to 1000 years before the present) record the last major period of sand mobility. These dunes are oriented NW-SE and are composed of cross-strata with bipolar dip directions. The trend and structure of the dunes record a drought that was initiated and sustained by a historically unprecedented shift of spring-summer atmospheric circulation over the Plains: Moist southerly flow was replaced by dry southwesterly flow.


How Flow Reduction Influences the Intracranial Aneurysm Occlusion: A Prospective 4D Phase-Contrast MRI Study.
Flow-diverter stents are widely used for the treatment of wide-neck intracranial aneurysms. Various parameters may influence intracranial aneurysm thrombosis, including the flow reduction induced by flow-diverter stent implantation, which is assumed to play a leading role. However, its actual impact remains unclear due to the lack of detailed intra-aneurysmal flow measurements. This study aimed to clarify this relationship by quantitatively measuring the intra-aneurysmal flow using 4D phase-contrast MR imaging. We acquired prospective pre- and post-stent implantation 4D phase-contrast MR imaging data of a consecutive series of 23 patients treated with flow-diverter stents. Velocity field data were combined with the intraprocedural 3D angiogram vessel geometries for precise intracranial aneurysm extraction and partial volume correction. Intra-aneurysmal hemodynamic modifications were compared with occlusion outcomes at 6 and 12 months. The averaged velocities at systole were lower after flow-diverter stent implantation for all patients and ranged from 21.7 ± 7.1 cm/s before to 7.2 ± 2.9 cm/s after stent placement. The velocity reduction was more important for the group of patients with aneurysm thrombosis at 6 months (68.8%) and decreased gradually from 66.2% to 55% for 12-month thrombosis and no thrombosis, respectively (P = .08). We propose an innovative approach to measure intracranial flow changes after flow-diverter stent implantation. We identified a trend between flow reduction and thrombosis outcome that brings a new insight into current understanding of the flow-diversion treatment response.


Towards a caring practice: reflections on the processes and components of arts-health practice.
Background: Many arts and health studies have focused on evaluating the impact of participatory arts activities, but few have considered the processes and components shaping the wellbeing outcomes of the participants. This paper uses a relational approach to health to explore the elements at play in participatory arts.Methods: The study uses an action-research case study of a 12-week visual art programme in a nursing home with 10 participants. Data collected were analysed and interpreted using grounded theory to obtain general themes and to facilitate focused analysis.Results: The study identifies the participants, wellbeing outcomes, the environment and the quality of activities as key concerns of arts-health practice. It also reflects on the link between caring and arts-health practice to highlight several caring attributes that promote a caring arts-health practice.Conclusions: The study findings provide a practice framework that can guide decision making and action to foster a caring arts-health practice.


Large-Area Graphene Films as Target Surfaces for Highly Reproducible Matrix-Assisted Laser Desorption Ionization Suitable for Quantitative Mass Spectrometry.
Due to the known sweet-spot issues that intrinsically arise from inhomogeneous formation of matrix-analyte crystals utilized as samples in matrix-assisted laser desorption ionization (MALDI) mass spectrometry, its reproducibility and thus its applications for quantification have been somewhat limited. In this paper, we report a simple strategy to improve the uniformity of matrix-analyte crystal spots, which we realized by adapting large-area graphene films, i.e., non-inert, interacting surfaces, as target surfaces. In this example, the graphitic surfaces of the graphene films interact with excess matrix molecules during the sample drying process, which induces spontaneous formation of optically uniform MALDI sample crystal layers on the film surfaces. Further, mass spectrometric imaging reveals that the visible uniformity is indeed accompanied by reproducible MALDI ionization over an entire sample spot, which greatly suppresses the appearance of sweet spots. The results of this study confirm that the proposed method achieves good linear responses of ion intensity to the analyte concentration (R2 > 0.99) with small relative standard deviations (σ < 10%), which is a range applicable for quantitative measurements using MALDI mass spectrometry. Graphical Abstract .


Vitamin D threshold to prevent aromatase inhibitor-related bone loss: the B-ABLE prospective cohort study.
Aromatase inhibitor (AI)-related bone loss is associated with increased fracture rates. Vitamin D might play a role in minimising this effect. We hypothesised that 25-hydroxy-vitamin D concentrations [25(OH)D] after 3 months supplementation might relate to bone loss after 1 year on AI therapy. We conducted a prospective cohort study from January 2006 to December 2011 of a consecutive sample of women initiating AI for early breast cancer who were ineligible for bisphosphonate therapy and stayed on treatment for 1 year (N = 232). Serum 25(OH)D was measured at baseline and 3 months, and lumbar spine (LS) bone mineral density at baseline and 1 year. Subjects were supplemented with daily calcium (1 g) and vitamin D(3) (800 IU) and additional oral 16,000 IU every 2 weeks if baseline 25(OH)D was <30 ng/ml. Linear regression models were fitted to adjust for potential confounders. After 1 year on AI therapy, 232 participants experienced a significant 1.68 % [95 % CI 1.15-2.20 %] bone loss at LS (0.017 g/cm(2) [0.012-0.024], P < 0.0001). Higher 25(OH)D at 3 months protected against LS bone loss (-0.5 % per 10 ng/ml [95 % CI -0.7 to -0.3 %], adjusted P = 0.0001), and those who reached levels ≥40 ng/ml had reduced bone loss by 1.70 % [95 % CI 0.4-3.0 %; adjusted P = 0.005] compared to those with low 25(OH)D levels (<30 ng/ml). We conclude that improved vitamin D status using supplementation is associated with attenuation of AI-associated bone loss. For this population, the current Institute of Medicine target recommendation of 20 ng/ml might be too low to ensure good bone health.


Madagascar corals track sea surface temperature variability in the Agulhas Current core region over the past 334 years.
The Agulhas Current (AC) is the strongest western boundary current in the Southern Hemisphere and is key for weather and climate patterns, both regionally and globally. Its heat transfer into both the midlatitude South Indian Ocean and South Atlantic is of global significance. A new composite coral record (Ifaty and Tulear massive Porites corals), is linked to historical AC sea surface temperature (SST) instrumental data, showing robust correlations. The composite coral SST data start in 1660 and comprise 200 years more than the AC instrumental record. Numerical modelling exhibits that this new coral derived SST record is representative for the wider core region of the AC. AC SSTs variabilities show distinct cooling through the Little Ice Age and warming during the late 18(th), 19th and 20th century, with significant decadal variability superimposed. Furthermore, the AC SSTs are teleconnected with the broad southern Indian and Atlantic Oceans, showing that the AC system is pivotal for inter-ocean heat exchange south of Africa.


Effects of worksite health promotion interventions on employee diets: a systematic review.
Public health strategies place increasing emphasis on opportunities to promote healthy behaviours within the workplace setting. Previous research has suggested worksite health promotion programmes have positive effects on physical activity and weight loss, yet little is known regarding their effects on dietary behaviour. The aim of this review was to assess the effects of worksite interventions on employee diets. Electronic databases (MEDLINE, The Cochrane Library, PsycINFO, EMBASE, LexisNexis) were searched for relevant articles published between 1995 and April 2009. Studies were eligible for inclusion if they were peer-reviewed English language publications describing a worksite-based health promotion intervention with minimum study duration of eight weeks. All study designs were eligible. Studies had to report one or more diet-related outcome (energy, fat, fruit, or vegetable intakes). Methodological quality was assessed using a checklist that included randomisation methods, use of a control group, and study attrition rates. Sixteen studies were included in the review. Eight programmes focussed on employee education, and the remainder targeted change to the worksite environment, either alone or in combination with education. Study methodological quality was moderate. In general, worksite interventions led to positive changes in fruit, vegetable and total fat intake. However, reliance on self-reported methods of dietary assessment means there is a significant risk of bias. No study measured more robust outcomes such as absenteeism, productivity, or healthcare utilisation. The findings of this review suggest that worksite health promotion programmes are associated with moderate improvement in dietary intake. The quality of studies to date has been frequently sub-optimal and further, well designed studies are needed in order to reliably determine effectiveness and cost-effectiveness. Future programmes to improve employee dietary habits should move beyond individual education and aim to intervene at multiple levels of the worksite environment.


Temporal Rac1 - HIF-1 crosstalk modulates hypoxic survival of aged neurons.
Neurodegenerative diseases are frequently associated with hypoxic conditions. During hypoxia the neuronal cytoskeleton is rapidly reorganized and such abnormalities are directly linked to adverse outcomes. Besides their roles as master regulators of the cytoskeleton, the Rho GTPases are also involved in cellular processes stimulated by hypoxic stress. We investigated the contribution of Rac1-mediated signaling to hypoxic responses of mature neurons using primary cortical cells cultured for 17 days in vitro. We show Rac1 is both upregulated and activated during hypoxia. Pharmacological inhibition of Rac1, but not RhoA, completely abrogated hypoxic HIF-1α stabilization and expression of the HIF-1 targets VEGF and GLUT1. Furthermore activity of JNK and GSK3β were also highly dependent on Rac1 activity and biphasic effects were observed after 6 and 24h of exposure. Notably, inhibition of either pathway suppressed HIF-1α accumulation. Although inhibition of Rac1 did not affect neuronal viability during acute exposure cell death was strongly induced after 24h revealing a time-dependent effect of Rac1 signaling on survival. Thus hypoxia-activated Rac1 is critical for neuronal HIF-1α stabilization and survival during oxygen deprivation via integration of complex signaling cascades.


Loss of Cited2 affects trophoblast formation and vascularization of the mouse placenta.
Cited2 is widely expressed in the developing embryo and in extraembryonic tissues including the placenta. Gene expression can be induced by a number of factors; most notably by the hypoxia inducible transcription factor, HIF1, under low oxygen conditions. Cited2 encodes for a transcriptional co-factor that in vitro can act as both a positive and negative regulator of transcription. This function is due to its interaction with CBP/p300 and appears to depend on whether Cited2 enables CBP/p300 to interact with the basic transcriptional machinery, or if its binding prevents such an interaction from occurring. Here, we report a novel function for Cited2 in placenta formation, following gene deletion in mouse. In the absence of Cited2 the placenta and embryo are significantly small from 12.5 and 14.5 dpc respectively, and death occurs in utero. Cited2 null placentas have fewer differentiated trophoblast cell types; specifically there is a reduction in trophoblast giant cells, spongiotrophoblasts and glycogen cells. In addition, the fetal vasculature of the placenta is disorganised and there are fewer anastomosing capillaries. Given that Cited2 is expressed in both trophoblasts and the fetal vasculature, the observed defects fit well with the sites of gene expression. We conclude that Cited2 is required for normal placental development and vascularisation, and hence for embryo viability.


New ecological aspects of hantavirus infection: a change of a paradigm and a challenge of prevention--a review.
In the last decades a significant number of so far unknown or underestimated pathogens have emerged as fundamental health hazards of the human population despite intensive research and exceptional efforts of modern medicine to embank and eradicate infectious diseases. Almost all incidents caused by such emerging pathogens could be ascribed to agents that are zoonotic or expanded their host range and crossed species barriers. Many different factors influence the status of a pathogen to remain unnoticed or evolves into a worldwide threat. The ability of an infectious agent to adapt to changing environmental conditions and variations in human behavior, population development, nutrition, education, social, and health status are relevant factors affecting the correlation between pathogen and host. Hantaviruses belong to the emerging pathogens having gained more and more attention in the last decades. These viruses are members of the family Bunyaviridae and are grouped into a separate genus known as Hantavirus. The serotypes Hantaan (HTN), Seoul (SEO), Puumala (PUU), and Dobrava (DOB) virus predominantly cause hemorrhagic fever with renal syndrome (HFRS), a disease characterized by renal failure, hemorrhages, and shock. In the recent past, many hantavirus isolates have been identified and classified in hitherto unaffected geographic regions in the New World (North, Middle, and South America) with characteristic features affecting the lungs of infected individuals and causing an acute pulmonary syndrome. Hantavirus outbreaks in the United States of America at the beginning of the 10th decade of the last century fundamentally changed our knowledge about the appearance of the hantavirus specific clinical picture, mortality, origin, and transmission route in human beings. The hantavirus pulmonary syndrome (HPS) was first recognized in 1993 in the Four Corners Region of the United States and had a lethality of more than 50%. Although the causative virus was first termed in connection with the geographic name of its outbreak region the analysis of the individual viruses indicate that the causing virus of HPS was a genetically distinct hantavirus and consequently termed as Sin Nombre virus. Hantaviruses are distributed worldwide and are assumed to share a long time period of co-evolution with specific rodent species as their natural reservoir. The degree of relatedness between virus serotypes normally coincides with the relatedness between their respective hosts. There are no known diseases that are associated with hantavirus infections in rodents underlining the amicable relationship between virus and host developed by mutual interaction in hundreds of thousands of years. Although rodents are the major reservoir, antibodies against hantaviruses are also present in domestic and wild animals like cats, dogs, pigs, cattle, and deer. Domestic animals and rodents live jointly in a similar habitat. Therefore the transmission of hantaviruses from rodents to domestic animals seems to be possible, if the target organs, tissues, and cell parenchyma of the co-habitat domestic animals possess adequate virus receptors and are suitable for hantavirus entry and replication. The most likely incidental infection of species other than rodents as for example humans turns hantaviruses from harmless to life-threatening pathogenic agents focusing the attention on this virus group, their ecology and evolution in order to prevent the human population from a serious health risk. Much more studies on the influence of non-natural hosts on the ecology of hantaviruses are needed to understand the directions that the hantavirus evolution could pursue. At least, domestic animals that share their environmental habitat with rodents and humans particularly in areas known as high endemic hantavirus regions have to be copiously screened. Each transfer of hantaviruses from their original natural hosts to other often incidental hosts is accompanied by a change of ecology, a change of environment, a modulation of numerous factors probably influencing the pathogenicity and virulence of the virus. The new environment exerts a modified evolutionary pressure on the virus forcing it to adapt and probably to adopt a form that is much more dangerous for other host species compared to the original one.


Anti-HER2 antibody and heregulin suppress growth of HER2-overexpressing human breast cancer cells through different mechanisms.
Previous reports have shown that certain anti-HER2 antibodies and heregulin can inhibit clonogenic growth of breast and ovarian cancers that overexpress HER2. Anti-HER2 antibodies bind to HER2 directly, whereas heregulin does not bind to HER2 alone, but rather interacts with HER2 through the formation of heterodimers with HER3 or HER4. The purpose of the present study was to elucidate the mechanisms by which anti-HER2 antibody and heregulin inhibit tumor growth. The anti-HER2 monoclonal antibody (mAb) ID5 was found to block G1-S progression of the cell cycle, whereas heregulin inhibited passage through G2-M. Compatible with the effects on the cell cycle, treatment with mAb ID5 decreased levels of cyclin-dependent kinase (CDK) 2, cyclin E, and CDK6 proteins and reduced cyclin E-CDK2-associated kinase activity; mAb HD5-treated cells had increased p27Kip1 expression and an increased association of p27Kip1 with CDK2. In contrast, treatment with heregulin increased protein levels of CDK2, CDK6, CDC2, and cyclin B1. More Retinoblastoma protein was found in the hypophosphorylated state in the cells treated with mAb ID5, whereas more retinoblastoma protein was in the hyperphosphorylated state in heregulin-treated cells. Heregulin was able to induce cell differentiation as assessed by Oil Red O staining and apoptosis as assessed by sub-G1 peak on flow cytometry and the presence of DNA fragmentation in ApopTag histochemistry staining. Neither differentiation nor apoptosis was observed in the cells treated with mAb ID5. We conclude that anti-HER-2 mAb ID5 and heregulin exert growth inhibition through different mechanisms. In mammary cells overexpressing HER2, anti-HER2 mAb ID5 induces G1 arrest, whereas heregulin induces G2-M arrest, cell differentiation, and apoptosis.


Dissolution enhancement of gliclazide using pH change approach in presence of twelve stabilizers with various physico-chemical properties.
The micronization using milling process to enhance dissolution rate is extremely inefficient due to a high energy input, and disruptions in the crystal lattice which can cause physical or chemical instability. Therefore, the aim of the present study is to use in situ micronization process through pH change method to produce micron-size gliclazide particles for fast dissolution hence better bioavailability. Gliclazide was recrystallized in presence of 12 different stabilizers and the effects of each stabilizer on micromeritic behaviors, morphology of microcrystals, dissolution rate and solid state of recrystallized drug particles were investigated. The results showed that recrystallized samples showed faster dissolution rate than untreated gliclazide particles and the fastest dissolution rate was observed for the samples recrystallized in presence of PEG 1500. Some of the recrystallized drug samples in presence of stabilizers dissolved 100% within the first 5 min showing at least 10 times greater dissolution rate than the dissolution rate of untreated gliclazide powders. Micromeritic studies showed that in situ micronization technique via pH change method is able to produce smaller particle size with a high surface area. The results also showed that the type of stabilizer had significant impact on morphology of recrystallized drug particles. The untreated gliclazide is rod or rectangular shape, whereas the crystals produced in presence of stabilizers, depending on the type of stabilizer, were very fine particles with irregular, cubic, rectangular, granular and spherical/modular shape. The results showed that crystallization of gliclazide in presence of stabilizers reduced the crystallinity of the samples as confirmed by XRPD and DSC results. In situ micronization of gliclazide through pH change method can successfully be used to produce micron-sized drug particles to enhance dissolution rate.