Unique gene expression profiles of donor-matched human retinal and choroidal vascular endothelial cells.
Consistent with clinical observations that posterior uveitis frequently involves the retinal vasculature and recent recognition of vascular heterogeneity, the hypothesis for this study was that retinal vascular endothelium was a cell population of unique molecular phenotype. Donor-matched cultures of primary retinal and choroidal endothelial cells from six human cadavers were incubated with either Toxoplasma gondii tachyzoites (10:1, parasites per cell) or Escherichia coli lipopolysaccharide (100 ng/mL); control cultures were simultaneously incubated with medium. Gene expression profiling of endothelial cells was performed using oligonucleotide arrays containing probes designed to detect 8746 human transcripts. After normalization, differential gene expression was assessed by the significance analysis of microarrays, with the false-discovery rate set at 5%. For selected genes, differences in the level of expression between retinal and choroidal cells were evaluated by real-time RT-PCR. Graphic descriptive analysis demonstrated a strong correlation between gene expression of unstimulated retinal and choroidal endothelial cells, but also highlighted distinctly different patterns of expression that were greater than differences noted between donors or between unstimulated and stimulated cells. Overall, 779 (8.9%) of 8746 transcripts were differentially represented. Of note, the 330 transcripts that were present at higher levels in retinal cells included a larger percentage of transcripts encoding molecules involved in the immune response. Differential gene expression was confirmed for 12 transcripts by RT-PCR. Retinal and choroidal vascular endothelial cells display distinctive gene expression profiles. The findings suggest the possibility of treating posterior uveitis by targeting specific interactions between the retinal endothelial cell and an infiltrating leukocyte.


An extended genotyping framework for Salmonella enterica serovar Typhi, the cause of human typhoid.
The population of Salmonella enterica serovar Typhi (S. Typhi), the causative agent of typhoid fever, exhibits limited DNA sequence variation, which complicates efforts to rationally discriminate individual isolates. Here we utilize data from whole-genome sequences (WGS) of nearly 2,000 isolates sourced from over 60 countries to generate a robust genotyping scheme that is phylogenetically informative and compatible with a range of assays. These data show that, with the exception of the rapidly disseminating H58 subclade (now designated genotype 4.3.1), the global S. Typhi population is highly structured and includes dozens of subclades that display geographical restriction. The genotyping approach presented here can be used to interrogate local S. Typhi populations and help identify recent introductions of S. Typhi into new or previously endemic locations, providing information on their likely geographical source. This approach can be used to classify clinical isolates and provides a universal framework for further experimental investigations.


Traces of Marion B. Sulzberger in the Museum of Wax Moulages in Zurich and their importance for the history of dermatology.
Marion B. Sulzberger was one of the most famous American dermatologists. He had received his training in dermatology in Zurich (Switzerland) from 1926 to 1929. The collection of moulages in Zurich still preserves outstanding wax models of Sulzberger's scientific work. They are impressive examples of the purpose and value of moulages at the beginning of the 20th century being used as teaching aids but also as documents and illustrations in scientific articles and textbooks. Today, the wax models of Sulzberger's work belong to the most important examples of moulages used in medical historical research. They also have value for teaching dermatology to medical students.


Three-dimensional solution structure and conformational plasticity of the N-terminal scavenger receptor cysteine-rich domain of human CD5.
The lymphocyte receptor CD5 influences cell activation by modifying the strength of the intracellular response initiated by antigen engagement. Regulation through CD5 involves the interaction of one or more of its three scavenger receptor cysteine-rich domains present in the extracellular region. Here, we present the 3D solution structure of a non-glycosylated double mutant of the N-terminal domain of human CD5 expressed in Escherichia coli (eCD5d1m), which has enhanced solubility compared to the non-glycosylated wild-type (eCD5d1). In common with a glycosylated form expressed in Pichia pastoris, the [(15)N,(1)H]-correlation spectra of both eCD5d1 and eCD5d1m exhibit non-uniform temperature-dependent signal intensities, indicating extensive conformational fluctuations on the micro-millisecond timescale. Although approximately one half of the signals expected for the domain are absent at 298 K, essentially complete resonance assignments and a solution structure could be obtained at 318 K. Because of the sparse nature of the experimental restraint data and the potentially important contribution of conformational exchange to the nuclear Overhauser effect peak intensity, we applied inferential structure determination to calculate the eCD5d1m structure. The inferential structure determination ensemble has similar features to that obtained by traditional simulated annealing methods, but displays superior definition and structural quality. The eCD5d1m structure is similar to other members of the scavenger receptor cysteine-rich superfamily, but the position of the lone alpha helix differs due to interactions with the unique N-terminal region of the domain. The availability of an experimentally tractable form of CD5d1, together with its 3D structure, provides new tools for further investigation of its function within intact CD5.


Potential for breath test diagnosis of urease positive pathogens in lung infections.
Roles for urease in virulence are accepted for Helicobacter pylori and urinary tract pathogens. However, urease is widely expressed by bacterial and fungal lung pathogens causing emerging and opportunistic lung infections, organisms causing acute exacerbations of chronic bronchitis, mycobacterial lung diseases such as tuberculosis (TB), and ventilator associated pneumonia and health care associated pneumonia. Detection of urease provides a method for rapid in vivo detection of these lung pathogens by inhaled 13C- breath test, and this review discusses the range of lung pathogens that might be amenable to rapid diagnosis.


Syringomyelia: determining risk and protective factors in the conformation of the Cavalier King Charles Spaniel dog.
Syringomyelia (SM) is a painful neurological condition, prevalent in brachycephalic toy breeds including the Cavalier King Charles Spaniel (CKCS). In these breeds, SM is typically secondary to Chiari-like Malformation (CM). There has been much debate in the scientific and veterinary communities to what extent head shape is indicative of either pathology, especially as certain craniosynostosis syndromes in humans (highly associated with CM) have characteristic facial and cranial morphologies. Elucidating a risk morphology would allow for selection away from these traits and proffer further breeding guidelines for the condition. Dogs were measured in multiple countries by means of a standardised bony landmark measuring protocol and photo analysis by blinded, trained researchers. The results found two significant risk factors in the conformation of the CKCS: extent of brachycephaly and distribution of cranium. The study identified a greater amount of cranium distributed caudally (relative to the amount distributed rostrally) to be significantly protective against syrinx development at the levels of three years of age, five years of age and when comparing a sample of SM clear individuals over the age of five to those affected younger than three years of age. A decreased cephalic index (decreasing brachycephaly) was significantly protective at the latter level. Cephalic index and caudal cranium distribution exhibited a negative, linear relationship. Cephalic index demonstrated a positive linear relationship with the amount of doming of the head. This study proposes a risk phenotype of brachycephaly with resulting rostrocaudal doming that is more rostrally distributed and hence sloping caudally. The results of this study may allow for selection against risk aspects of conformation in the CKCS in combination with the British Veterinary Association/Kennel Club CM/SM scheme to enable reduction in CM/SM incidence. Further research comparing this external risk phenotype to the internal presentation upon MRI would determine how these features are indicative of syrinx development. Utilising breeds in which CM free individuals are more available may allow for validation of this risk phenotype for CM or determine alternatives.


Langerhans cells regulate cutaneous injury by licensing CD8 effector cells recruited to the skin.
Langerhans cells (LCs) are a distinct population of dendritic cells that form a contiguous network in the epidermis of the skin. Although LCs possess many of the properties of highly proficient dendritic cells, recent studies have indicated that they are not necessary to initiate cutaneous immunity. In this study, we used a tractable model of cutaneous GVHD, induced by topical application of a Toll-like receptor agonist, to explore the role of LCs in the development of tissue injury. By adapting this model to permit inducible and selective depletion of host LCs, we found that GVHD was significantly reduced when LCs were absent. However, LCs were not required either for CD8 T-cell activation within the draining lymph node or subsequent homing of effector cells to the epidermis. Instead, we found that LCs were necessary for inducing transcription of IFN-γ and other key effector molecules by donor CD8 cells in the epidermis, indicating that they license CD8 cells to induce epithelial injury. These data demonstrate a novel regulatory role for epidermal LCs during the effector phase of an inflammatory immune response in the skin.


Strategies to Improve Health Communication: Can Health Professionals be Heroes?
Communicating evidence-based nutrition messages to the public is challenging and is often in conflict with popular opinions, particularly from social media influencers (SMIs). In order to increase engagement with nutrition professionals (NPs) on social media, we aimed to explore young adults' perceptions of the authenticity and trustworthiness of SMIs and NPs Instagram posts. A cross-sectional questionnaire was administered to students (n = 149) from an Australian University. Participants viewed a real-life Instagram profile and one post from both a NP and a SMI. Main outcomes were post authenticity and trustworthiness, and emotional message appeals measured on five-point Likert scales. Regression models were developed to assess whose post (the NP or SMI) was perceived to be more authentic and trustworthy. Participants were young adults (median age (25th, 75th percentiles): 20 (19,21)), with approximately half identifying as female. A high heroic message appeal (+1SD above mean) significantly increased the perceived authenticity of the NPs post only (p = 0.01). Post authenticity enhanced post trustworthiness, but only when a heroic message appeal was used by the NP. When appropriate, NPs should convey positive emotions such as bravery and success to enhance the authenticity and trustworthiness of their posts.


Chronic lactate supplementation does not improve blood buffering capacity and repeated high-intensity exercise.
Since there is conflicting data on the buffering and ergogenic properties of calcium lactate (CL), we investigated the effect of chronic CL supplementation on blood pH, bicarbonate, and high-intensity intermittent exercise performance. Sodium bicarbonate (SB) was used as a positive control. Eighteen athletes participated in this double-blind, placebo-controlled, crossover, fully counterbalanced study. All participants underwent three different treatments: placebo (PL), CL, and SB. The dose was identical in all conditions: 500 mg/kg BM divided into four daily individual doses of 125 mg/kg BM, for five consecutive days, followed by a 2-7-day washout period. On the fifth day of supplementation, individuals undertook four 30-s Wingate bouts for upper body with 3-min recovery between bouts. Total mechanical work (TMW) for the overall protocol and for the initial (1st+2nd) and final (3rd+4th) bouts was determined at each session. Blood pH, bicarbonate, and lactate levels were determined at rest, immediately and 5 min after exercise. CL supplementation did not affect performance (P > 0.05 for the overall TMW as well for initial and final bouts), nor did it affect blood bicarbonate and pH prior to exercise. SB supplementation improved performance by 2.9% for overall TMW (P = 0.02) and 5.9% in the 3rd+4th bouts (P = 0001). Compared to the control session, SB also promoted higher increases in blood bicarbonate than CL and PL (+0.03 ± 0.04 vs +0.009 ± 0.02 and +0.01 ± 0.03, respectively). CL supplementation was not capable of enhancing high-intensity intermittent performance or changing extracellular buffering capacity challenging the notion that this dietary supplement is an effective buffering agent.


Direct measurements of the convective recycling of the upper troposphere.
We present a statistical representation of the aggregate effects of deep convection on the chemistry and dynamics of the upper troposphere (UT) based on direct aircraft observations of the chemical composition of the UT over the eastern United States and Canada during summer. These measurements provide unique observational constraints on the chemistry occurring downwind of convection and the rate at which air in the UT is recycled. These results provide quantitative measures that can be used to evaluate global climate and chemistry models.


A novel cholinergic action of alcohol and the development of tolerance to that effect in Caenorhabditis elegans.
Understanding the genes and mechanisms involved in acute alcohol responses has the potential to allow us to predict an individual's predisposition to developing an alcohol use disorder. To better understand the molecular pathways involved in the activating effects of alcohol and the acute functional tolerance that can develop to such effects, we characterized a novel ethanol-induced hypercontraction response displayed by Caenorhabditis elegans. We compared body size of animals prior to and during ethanol treatment and showed that acute exposure to ethanol produced a concentration-dependent decrease in size followed by recovery to their untreated size by 40 min despite continuous treatment. An increase in cholinergic signaling, leading to muscle hypercontraction, is implicated in this effect because pretreatment with mecamylamine, a nicotinic acetylcholine receptor (nAChR) antagonist, blocked ethanol-induced hypercontraction, as did mutations causing defects in cholinergic signaling (cha-1 and unc-17). Analysis of mutations affecting specific subunits of nAChRs excluded a role for the ACR-2R, the ACR-16R, and the levamisole-sensitive AChR and indicated that this excitation effect is dependent on an uncharacterized nAChR that contains the UNC-63 α-subunit. We performed a forward genetic screen and identified eg200, a mutation that affects a conserved glycine in EAT-6, the α-subunit of the Na(+)/K(+) ATPase. The eat-6(eg200) mutant fails to develop tolerance to ethanol-induced hypercontraction and remains contracted for at least 3 hr of continuous ethanol exposure. These data suggest that cholinergic signaling through a specific α-subunit-containing nAChR is involved in ethanol-induced excitation and that tolerance to this ethanol effect is modulated by Na(+)/K(+) ATPase function.


Recent updates on codeine.
Alleviation of pain is a major objective in medicine to increase the quality of life. Analgesics are agents that relieve pain by elevating the pain threshold without disturbing consciousness or altering other sensory modalities. Opium is an isoquinoline alkaloid obtained from poppy plant Papaver somniferum (Papaveraceae). Codeine is an alkaloid prepared from opium or morphine by methylation. Codeine is used as a central analgesic, sedative, hypontic, antinonciceptive, antiperistaltic, and is also recommended in tuberculosis and insomnia due to incessant coughing. The literature information relate mostly to the determination of codeine active components using Gas chromatography (GC), Capillary electrophoresis, Thin layer chromatography, High-performance thin layer chromatography, UV-Vis Spectrophotometry, High-performance liquid chromatography and GC in combination with Mass spectroscopy. This contribution provides a comprehensive review of its analytical and pharmacologic profile of codeine.


The Inhibitory Effect of Extra Virgin Olive Oil and Its Active Compound Oleocanthal on Prostaglandin-Induced Uterine Hypercontraction and Pain-Ex Vivo and In Vivo Study.
Primary dysmenorrhea is a common occurrence in adolescent women and is a type of chronic inflammation. Dysmenorrhea is due to an increase in oxidative stress, which increases cyclooxygenase-2 (COX-2) expression, increases the concentration of prostaglandin F2α (PGF2α), and increases the calcium concentration in uterine smooth muscle, causing excessive uterine contractions and pain. The polyphenolic compound oleocanthal (OC) in extra virgin olive oil (EVOO) has been shown to have an anti-inflammatory and antioxidant effect. This study aimed to investigate the inhibitory effect of extra virgin olive oil and its active ingredient oleocanthal (OC) on prostaglandin-induced uterine hyper-contraction, its antioxidant ability, and related mechanisms. We used force-displacement transducers to calculate uterine contraction in an ex vivo study. To analyze the analgesic effect, in an in vivo study, we used an acetic acid/oxytocin-induced mice writhing model and determined uterus contraction-related signaling protein expression. The active compound OC inhibited calcium/PGF2α-induced uterine hyper-contraction. In the acetic acid and oxytocin-induced mice writhing model, the intervention of the EVOO acetonitrile layer extraction inhibited pain by inhibiting oxidative stress and the phosphorylation of the protein kinase C (PKC)/extracellular signal-regulated kinases (ERK)/ myosin light chain (MLC) signaling pathway. These findings supported the idea that EVOO and its active ingredient, OC, can effectively decrease oxidative stress and PGF2α-induced uterine hyper-contraction, representing a further treatment for dysmenorrhea.


Clinical predictors of mortality in hospitalized patients with infective endocarditis at a tertiary care center in Pakistan.
To evaluate the clinical characteristics and predictors of in-hospital mortality in patients with infective endocarditis at a tertiary care centre in Pakistan. The cross-sectional study was conducted at the Aga Khan University Hospital, Karachi, from January 1, 2002, to December 31, 2006, and comprised 84 consecutive patients hospitalised with infective endocarditis,. All patients underwent verification of the diagnosis according to the Modified Duke criterion. Data was collected through a questionnaire and analysed using Student's t test, Chi square and Fisher's exact test. Univariate and multivariate logistic regression analysis was performed to assess predictors of mortality. Of the total, 53(63%) patients were male and 31(37%) female. The mean age of the patients was 42+17 years. Overall, 34(41%) patients had rheumatic valve disease as a predisposing condition. The most commonly affected valve was mitral in 43(51%) patients, and the most commonly isolated organism was methicillin-resistant staphylococcus aureus in 12(14.3%). Overall in-hospital mortality was 27(32.1%), while 18(21%) patients, developed congestive cardiac failure, 15(18%) developed arrhythmias, 16(19%) developed peripheral embolism and renal failure was present in 38(54%). Besides, 17(20%) patients underwent surgical intervention. The final multivariate model that can be used to predict mortality in this study consisted of the presence of neurological complications (p <0.001) Odds Ratio 7.26, Confidence Interval (2.27-23.18), congestive cardiac heart failure (p <0.023) Odds Ratio 5.39, Confidence Interval (1.26-23.04), and arrhythmias (p <0.034) Odds Ratio 4.21, Confidence Interval (1.11-15.88). Significant predictors of mortality in hospitalised patients with infective endocarditis in our study were the presence of neurological complications, congestive cardiac heart failure, and the presence of arrhythmias.


Effect of carbon to nitrogen ratio on the composition of microbial extracellular polymers in activated sludge.
Effect of carbon to nitrogen ratio (C/N) on the sludge extracellular polymer composition is studied in synthetically fed semi-continuous reactors with 8 days of sludge age. Results show that C/N ratio influences the relative distribution of polymer carbohydrate and protein. At low C/N ratio of 5, polymer extracts have high protein and low carbohydrate content. As the C/N ratio is increased to 17.5 and then to 40, carbohydrate concentration increases sharply and protein concentration decreases.


Syndromic (phenotypic) diarrhoea of infancy/tricho-hepato-enteric syndrome.
Syndromic diarrhoea/tricho-hepato-enteric syndrome (SD/THE) is a rare congenital syndrome. The main features are intractable diarrhoea of infancy, hair abnormalities, facial dysmorphism, intrauterine growth restriction and immune system abnormalities. It has been linked to abnormalities in two components of the putative human ski complex: SKIV2L and TTC37. The long-term outcome of this syndrome is still unknown. We aim to describe the long-term outcome, in the French cohort of patients born since 1992. Review of the clinical and biological features of the 15 patients with SD/THE, followed in France and born between 1992 and 2010. All patients presented typical SD/THE syndrome features, of intractable diarrhoea in infancy requiring parenteral nutrition, a facial dysmorphism with hair abnormalities, and immunological disorders. Half of them also had liver and skin abnormalities. Five children died, among which 3 died due to infections. Probabilities of survival according to the Kaplan-Meier method were 93.3%, 86.7%, 74.3 and 61.9%, respectively at 1 year, 5 years, 10 years and 15 years of age. 3/15 were weaned from parenteral nutrition (PN) with likelihood of weaning being 10% at 5 years and 40% at 10 years. At birth 80% were small for gestational age and the short stature persisted in 60%. Haemophagocytic syndrome was noted in 60% and mild mental retardation was present in 60%. SD/THE is a rare disease with high morbidity and mortality. Management should be focused on nutrition and immunological defects.


Defining the acute kidney injury and repair transcriptome.
The mammalian kidney has an intrinsic ability to repair after significant injury. However, this process is inefficient: patients are at high risk for the loss of kidney function in later life. No therapy exists to treat established acute kidney injury (AKI) per se: strategies to promote endogenous repair processes and retard associated fibrosis are a high priority. Whole-organ gene expression profiling has been used to identify repair responses initiated with AKI, and factors that may promote the transition from AKI to chronic kidney disease. Transcriptional profiling has shown molecular markers and potential regulatory pathways of renal repair. Activation of a few key developmental pathways has been reported during repair. Whether these are comparable networks with similar target genes with those in earlier nephrogenesis remains unclear. Altered microRNA profiles, persistent tubular injury responses, and distinct late inflammatory responses highlight continuing kidney pathology. Additional insights into injury and repair processes will be gained by study of the repair transcriptome and cell-specific translatome using high-resolution technologies such as RNA sequencing and translational profiling tailored to specific cellular compartments within the kidney. An enhanced understanding holds promise for both the identification of novel therapeutic targets and biomarker-based evaluation of the damage-repair process.


Povidone-iodine ear wash and oral cotrimoxazole for chronic suppurative otitis media in Australian aboriginal children: study protocol for factorial design randomised controlled trial.
Chronic suppurative otitis media (CSOM) is a significant health issue affecting Aboriginal Australians. Long-term hearing loss can cause communication problems, educational disadvantage, and social isolation. Current standard treatment for CSOM in our region is twice daily dry mopping of the pus from the ear canal followed by instillation of ciprofloxacin antibiotic ear drops for up to 16 weeks, or until the discharge resolves for a period of 3 days. The treatment is long, laborious and fails to resolve ear discharge in 70% of cases in remote communities. Bacterial pathogens also persist. Povidone-iodine ear wash is the preferred method of clearing ear discharge in Western Australia. However, evidence of its effectiveness is lacking. In systematic reviews, topical antibiotics (ciprofloxacin) have been shown to be more effective than oral antibiotics or topical antiseptics. Currently, it is unclear whether there are any benefits of combining these treatments. This protocol describes a 2 × 2 factorial randomised controlled trial of two different interventions (povidone-iodine ear wash and oral cotrimoxazole), given as adjunctive therapy to standard treatment for CSOM. 280 children, between 2 months and 17 years of age, Indigenous or non-Indigenous, living in participating Northern Territory (NT) communities are randomised to standard treatment (dry mopping and ciprofloxacin drops) plus one of two topical treatments (dilute povidone-iodine ear wash or no wash) and one of two oral medication treatments (16 weeks of cotrimoxazole or placebo). Current treatment of CSOM in our region shows that eradication of bacterial pathogens from the middle ear space and dry ears is often not achieved. This trial will evaluate the efficacy of adjunctive treatments of antiseptic ear washes and oral antibiotics. Clinical, microbiological and hearing outcomes will be reported. This trial (ACTRN12614000234617) was registered with ANZCTR on 05 April 2014.


Utility of circulating B-RAF DNA mutation in serum for monitoring melanoma patients receiving biochemotherapy.
Somatic B-RAF gene mutation has been identified in many malignancies and detected at a high frequency in cutaneous malignant melanoma. However, the significance of the B-RAF mutation (B-RAFmt) in terms of its prognostic and predictive capabilities for treatment response or disease outcome is not known. We hypothesized that circulating serum B-RAFmt (B-RAFsmt) at V600E, detected in serum, predicts response in melanoma patients receiving concurrent biochemotherapy. A real-time clamp quantitative reverse transcription-PCR assay was designed to assess B-RAFsmt by peptide nucleic acid clamping and a locked nucleic acid hybrid probe. Normal (n = 18) and American Joint Committee on Cancer stage I to IV melanoma patients (n = 103) were evaluated. These included stage IV patients (n = 48) with blood drawn before and after biochemotherapy. Patients were classified as biochemotherapy responders or nonresponders. Responders (n = 24) had a complete or partial response following biochemotherapy; nonresponders (n = 24) developed progressive disease. Of the 103 melanoma patients, 38 (37%) had B-RAFsmt DNA, of which 11 of 34 (32%) were stage I or II, and 27 of 69 (39%) were stage III or IV. Of the 48 biochemotherapy patients, 10 of 24 (42%) patients were positive for the B-RAFsmt in the respective responder and nonresponder groups before treatment. After biochemotherapy, B-RAFsmt was detected in only 1 of 10 patients (10%) in the responder group and 7 of 10 patients (70%) in the nonresponder group. B-RAFsmt is associated with significantly worse (P = 0.039) overall survival in patients receiving biochemotherapy. These studies show the presence and utility of circulating B-RAFsmt DNA in melanoma patients.


Comparison of inhibitory potency by nonpeptide angiotensin II receptor antagonists PD123177 and DuP 753 on proximal nephron and renal transport.
The nonpeptide angiotensin II receptor antagonists, DuP 753 and PD123177, have been recently proposed to efficiently discriminate between two receptor subtypes. Because angiotensin II uses two signaling systems to control transport in the proximal tubule, the hypothesis was entertained that angiotensin-regulated proximal reabsorption might be transduced through two receptor subtypes and, therefore, have two components, sensitive to either DuP 753 or PD123177. Using in vivo microperfusion in the rat, a maximally effective dose of PD123177 (60 or 120 mg/kg, i.v.) had a powerful inhibitory effect on transport in the S1 proximal tubule, significantly (P less than .001) reducing bicarbonate absorption by 50% (360 +/- 4 to 179 +/- 8 peq/mm.min), chloride absorption by 38% (211 +/- 1 to 131 +/- 4 peq/mm.min) and sodium and water absorption by 32% (5.7 +/- 0.1 to 3.9 +/- 0.5 nl/mm.min). These results were only slightly different than previously reported effects on proximal solute and water absorption by DuP 753 (10 mg/kg). The two drugs demonstrated virtually no additivity in their transport inhibitory effects. Although downstream reabsorptive elements compensate for the powerful action of PD123177 in the earliest segment of the nephron, we also showed, using free-flow micropuncture and clearance techniques, that PD123177 induces a substantial diuresis, natriuresis and chloruresis, again similar in magnitude to DuP 753. These results suggest PD123177 has diuretic potency roughly equivalent to that of DuP 753, but do not lend support to the simple thesis that there are two separately acting receptor-signal transduction systems in the proximal tubule for angiotensin II.


Community-level HIV/STI interventions and their impact on alcohol use in urban poor populations in India.
This paper describes an Indo-US, research and intervention project for HIV/STI prevention and sexual risk reduction in urban poor communities in Mumbai, India in which formative research established the importance of reduction in alcohol use as one of the central features of the intervention. As a part of formative research, in-depth interviews with married women and men indicated that alcohol had a direct negative effect on marital relationships, violence, household economics and men's involvement in extramarital sex. The project utilized diverse community intervention mechanisms over the course of a three year intervention effort. Comparison of pre-post intervention, cross-sectional samples showed a significant drop in overall use of alcohol in the study communities. Analysis of a longitudinal panel sample identified sub-groups of married men based on their demographic, behavioral and attitudinal characteristics at baseline who stopped drinking during the intervention period. Results also demonstrated that a reduction in men's alcohol use during the intervention period was associated with a reduction in sexual risk behavior and related variables.


Systemic restoration of UBA1 ameliorates disease in spinal muscular atrophy.
The autosomal recessive neuromuscular disease spinal muscular atrophy (SMA) is caused by loss of survival motor neuron (SMN) protein. Molecular pathways that are disrupted downstream of SMN therefore represent potentially attractive therapeutic targets for SMA. Here, we demonstrate that therapeutic targeting of ubiquitin pathways disrupted as a consequence of SMN depletion, by increasing levels of one key ubiquitination enzyme (ubiquitin-like modifier activating enzyme 1 [UBA1]), represents a viable approach for treating SMA. Loss of UBA1 was a conserved response across mouse and zebrafish models of SMA as well as in patient induced pluripotent stem cell-derive motor neurons. Restoration of UBA1 was sufficient to rescue motor axon pathology and restore motor performance in SMA zebrafish. Adeno-associated virus serotype 9-UBA1 (AAV9-UBA1) gene therapy delivered systemic increases in UBA1 protein levels that were well tolerated over a prolonged period in healthy control mice. Systemic restoration of UBA1 in SMA mice ameliorated weight loss, increased survival and motor performance, and improved neuromuscular and organ pathology. AAV9-UBA1 therapy was also sufficient to reverse the widespread molecular perturbations in ubiquitin homeostasis that occur during SMA. We conclude that UBA1 represents a safe and effective therapeutic target for the treatment of both neuromuscular and systemic aspects of SMA.


Crystal structure of the calcium release-activated calcium channel Orai.
The plasma membrane protein Orai forms the pore of the calcium release-activated calcium (CRAC) channel and generates sustained cytosolic calcium signals when triggered by depletion of calcium from the endoplasmic reticulum. The crystal structure of Orai from Drosophila melanogaster, determined at 3.35 angstrom resolution, reveals that the calcium channel is composed of a hexameric assembly of Orai subunits arranged around a central ion pore. The pore traverses the membrane and extends into the cytosol. A ring of glutamate residues on its extracellular side forms the selectivity filter. A basic region near the intracellular side can bind anions that may stabilize the closed state. The architecture of the channel differs markedly from other ion channels and gives insight into the principles of selective calcium permeation and gating.


Structure of the complex between pyridoxal 5'-phosphate and the tyrosine 225 to phenylalanine mutant of Escherichia coli aspartate aminotransferase determined by isotope-edited classical Raman difference spectroscopy.
The azomethine (Schiff base) linkage between the epsilon-amino group of active-site lysine 258 and the carbonyl moiety of enzyme-bound pyridoxal 5'-phosphate (PLP) normally exhibits absorbance maxima at ca. 360 (high-pH form) or ca. 430 nm (low-pH form). However, the absorbance maximum is shifted from 358 to 386 nm, a value which is similar to that of free PLP (lambda max = 388 nm), in a mutant form of Escherichia coli aspartate aminotransferase (AATase) in which tyrosine 225, which normally donates a hydrogen bond to the phenolate function of PLP, has been replaced with phenylalanine (Y225F). This spectral shift suggested that PLP binds to Y225F as the free aldehyde. The following evidence from isotope-edited classical Raman spectroscopy proves conclusively that the near-UV spectrum is anomalous and that PLP is bound to Y225F as a Schiff base: (1) A strong cofactor peak at 1630 cm-1 in the holoenzyme-minus-apoenzyme difference spectrum of the unprotonated form of Y225F is red-shifted by 18 cm-1 in enzyme labeled with 15N at lysine 258 and other positions. (2) This isotope-induced red shift is similar to that observed in the unprotonated form of the model Schiff base, PLP-valine. (3) The Raman spectrum of Y225F is unchanged in H(2)18O, while peaks at ca. 1670 cm-1 in the spectrum of free PLP or in that of a mutant of AATase in which Lys-258 is replaced with Ala, are red-shifted by ca. 30 cm-1 in H(2)18O.(ABSTRACT TRUNCATED AT 250 WORDS)


Dissociating the effects of similarity, salience, and top-down processes in search for linearly separable size targets.
In two experiments, we explored the role of foreknowledge on visual search for targets defined along the size continuum. Targets were of large, medium, or small size and of high or low similarity relative to the distractors. In Experiment 1, we compared search for known and unknown singleton feature targets as a function of their size and similarity to the distractors. When distractor similarity was high, target foreknowledge benefited targets at the end of the size continuum (i.e., large and small) relatively more than targets of medium size. In Experiment 2, participants were given foreknowledge of what the target was not The beneficial effect of foreknowledge for endpoint targets was reduced. The data indicate the role of top-down templates in search, which can be "tuned" more effectively for targets at the ends of feature dimensions.


Selective quantification of viable Escherichia coli bacteria in biosolids by quantitative PCR with propidium monoazide modification.
Quantitative differentiation of live cells in biosolids samples, without the use of culturing-based approaches, is highly critical from a public health risk perspective, as recent studies have shown significant regrowth and reactivation of indicator organisms. Persistence of DNA in the environment after cell death in the range of days to weeks limits the application of DNA-based approaches as a measure of live cell density. Using selective nucleic acid intercalating dyes like ethidium monoazide (EMA) and propidium monoazide (PMA) is one of the alternative approaches to detecting and quantifying viable cells by quantitative PCR. These compounds have the ability to penetrate only into dead cells with compromised membrane integrity and intercalate with DNA via their photoinducible azide groups and in turn inhibit DNA amplification during PCRs. PMA has been successfully used in different studies and microorganisms, but it has not been evaluated sufficiently for complex environmental samples such as biosolids. In this study, experiments were performed with Escherichia coli ATCC 25922 as the model organism and the uidA gene as the target sequence using real-time PCR via the absolute quantification method. Experiments with the known quantities of live and dead cell mixtures showed that PMA treatment inhibits PCR amplification from dead cells with over 99% efficiency. The results also indicated that PMA-modified quantitative PCR could be successfully applied to biosolids when the total suspended solids (TSS) concentration is at or below 2,000 mg·liter(-1).


Oral iloprost as a treatment for Raynaud's syndrome: a double blind multicentre placebo controlled study.
To compare the efficacy, tolerance and safety of 50-150 micrograms orally administered iloprost given twice a day versus placebo in patients with Raynaud's syndrome. The study was multicentre (n = 3), double blind and placebo controlled. Sixty three patients who had eight or more vasospastic attacks per week were enrolled. After a one week run-in period, all patients received either iloprost or placebo treatment to a maximum tolerated dose of 150 micrograms twice a day for 10 days. Diary cards assessed the duration and severity of the vasospastic attacks. Side effects were monitored by direct questioning. A global assessment of treatment efficacy was made by the patient at the end of treatment and two weeks later. Patient opinion tended to favour iloprost at the end of the 10 day treatment phase (p = 0.09) and this was significant at day 24 (the follow up visit) (p = 0.011). Although the duration and severity of attacks tended to decrease in the iloprost treated group, these results tended not to reach statistical significance (for severity p = 0.06 at end of treatment, p = 0.09 on day 24). Iloprost administered intravenously has been shown to be of benefit in the treatment of the Raynaud's syndrome associated with systemic sclerosis, but this route of administration is inconvenient. This study evaluated the use of iloprost administered orally to patients with Raynaud's syndrome. Patient documented improvement was significantly improved by iloprost. Diary card analysis showed a trend in favour of iloprost, but these results did not reach statistical significance.


New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression in Retinal Ganglion Cells of Mouse and Primate Retina.
Retinal gene therapy is leading the neurological gene therapy field, with 32 ongoing clinical trials of recombinant adeno-associated virus (rAAV)-based therapies. Importantly, over 50% of those trials are using restricted promoters from human genes. Promoters that restrict expression have demonstrated increased efficacy and can limit the therapeutic to the target cells thereby reducing unwanted off-target effects. Retinal ganglion cells are a critical target in ocular gene therapy; they are involved in common diseases such as glaucoma, rare diseases such as Leber's hereditary optic neuropathy, and in revolutionary optogenetic treatments. Here, we used computational biology and mined the human genome for the best genes from which to develop a novel minimal promoter element(s) designed for expression in restricted cell types (MiniPromoter) to improve the safety and efficacy of retinal ganglion cell gene therapy. Gene selection included the use of the first available droplet-based single-cell RNA sequencing (Drop-seq) dataset, and promoter design was bioinformatically driven and informed by a wide range of genomics datasets. We tested seven promoter designs from four genes in rAAV for specificity and quantified expression strength in retinal ganglion cells in mouse, and then the single best in nonhuman primate retina. Thus, we developed a new human-DNA MiniPromoter, Ple345 (NEFL), which in combination with intravitreal delivery in rAAV9 showed specific and robust expression in the retinal ganglion cells of the nonhuman-primate rhesus macaque retina. In mouse, we also developed MiniPromoters expressing in retinal ganglion cells, the hippocampus of the brain, a pan neuronal pattern in the brain, and peripheral nerves. As single-cell transcriptomics such as Drop-seq become available for other cell types, many new opportunities for additional novel restricted MiniPromoters will present.


Randomized controlled trial to evaluate a novel two-catheter technique for urethral catheterization in anesthetized healthy female cats and small dogs.
To evaluate a novel 2-catheter technique for urethral catheterization in female cats and small dogs and compare the time required for and success rates achieved by use of the novel technique versus traditional methods (blind technique in cats and digital palpation in dogs) as performed by personnel (catheter placers [CPs]) with different levels of experience in urinary catheter placement. 39 healthy sexually intact female animals (24 cats and 15 dogs weighing < 10 kg). 2 CPs were board certified in veterinary surgery, 1 of whom had experience with the novel technique, and the other did not. The third CP was a veterinary surgical intern who was unfamiliar with the novel technique. For each animal enrolled in the study, 1 CP performed catheterization with the novel technique and traditional methods. Data recorded included the time required for successful catheterization and whether a successful catheterization was achieved within a 3-minute time limit. The overall success rates were 79.5% (31/39 animals) with the novel technique and 43.6% (17/39 animals) with traditional methods. Median times for successful catheter placement were 48 seconds for the novel technique and 41 seconds for traditional methods. Among CPs, success rates or times to successful catheter placement did not differ significantly. Study results suggested that the novel 2-catheter technique for urethral catheterization may be a more efficient option than traditional methods for gaining access to the urinary bladder in cats and small dogs, particularly when patient size limits use of instrumentation or digital palpation.


Associations between lamb survival and prion protein genotype: analysis of data for ten sheep breeds in Great Britain.
Selective breeding programmes, based on prion protein (PrP) genotype, have been introduced throughout the European Union to reduce the risk of sheep transmissible spongiform encephalopathies (TSEs). These programmes could have negative consequences on other important traits, such as fitness and production traits, if the PrP gene has pleiotropic effects or is in linkage disequilibrium with genes affecting these traits. This paper presents the results of an investigation into associations between lamb survival and PrP genotype in ten mainstream sheep breeds in Great Britain (GB). In addition, the reasons for lamb deaths were examined in order to identify any associations between these and PrP genotype. Survival times from birth to weaning were analysed for over 38000 lambs (2427 dead and 36096 live lambs) from 128 flocks using Cox proportional hazard models for each breed, including additive animal genetic effects. No significant associations between PrP genotype and lamb survival were identified, except in the Charollais breed for which there was a higher risk of mortality in lambs of the ARR/VRQ genotype compared with those of the ARR/ARR genotype. Significant effects of birth weight, litter size, sex, age of dam and year of birth on survival were also identified. For all breeds the reasons for death changed significantly with age; however, no significant associations between reason for death and PrP genotype were found for any of the breeds. This study found no evidence to suggest that a selective breeding programme based on PrP genotype will have a detrimental effect on lamb survival. The only significant effect of PrP genotype identified was likely to be of little consequence because an increased risk of mortality was associated with a genotype that is selected against in current breeding strategies.


KatharoSeq Enables High-Throughput Microbiome Analysis from Low-Biomass Samples.
Microbiome analyses of low-biomass samples are challenging because of contamination and inefficiencies, leading many investigators to employ low-throughput methods with minimal controls. We developed a new automated protocol, KatharoSeq (from the Greek katharos [clean]), that outperforms single-tube extractions while processing at least five times as fast. KatharoSeq incorporates positive and negative controls to reveal the whole bacterial community from inputs of as few as 50 cells and correctly identifies 90.6% (standard error, 0.013%) of the reads from 500 cells. To demonstrate the broad utility of KatharoSeq, we performed 16S rRNA amplicon and shotgun metagenome analyses of the Jet Propulsion Laboratory spacecraft assembly facility (SAF; n = 192, 96), 52 rooms of a neonatal intensive care unit (NICU; n = 388, 337), and an endangered-abalone-rearing facility (n = 192, 123), obtaining spatially resolved, unique microbiomes reproducible across hundreds of samples. The SAF, our primary focus, contains 32 sOTUs (sub-OTUs, defined as exact sequence matches) and their inferred variants identified by the deblur algorithm, with four (Acinetobacter lwoffii, Paracoccus marcusii, Mycobacterium sp., and Novosphingobium) being present in >75% of the samples. According to microbial spatial topography, the most abundant cleanroom contaminant, A. lwoffii, is related to human foot traffic exposure. In the NICU, we have been able to discriminate environmental exposure related to patient infectious disease, and in the abalone facility, we show that microbial communities reflect the marine environment rather than human input. Consequently, we demonstrate the feasibility and utility of large-scale, low-biomass metagenomic analyses using the KatharoSeq protocol. IMPORTANCE Various indoor, outdoor, and host-associated environments contain small quantities of microbial biomass and represent a niche that is often understudied because of technical constraints. Many studies that attempt to evaluate these low-biomass microbiome samples are riddled with erroneous results that are typically false positive signals obtained during the sampling process. We have investigated various low-biomass kits and methods to determine the limit of detection of these pipelines. Here we present KatharoSeq, a high-throughput protocol combining laboratory and bioinformatic methods that can differentiate a true positive signal in samples with as few as 50 to 500 cells. We demonstrate the application of this method in three unique low-biomass environments, including a SAF, a hospital NICU, and an abalone-rearing facility.


Modelling approaches to dose estimation in children.
Most of the drugs on the market are originally developed for adults and dosage selection is based on an optimal balance between clinical efficacy and safety. The aphorism 'children are not small adults' not only holds true for the selection of suitable drugs and dosages for use in children but also their susceptibility to adverse drug reactions. Since children may not be subject to dose escalation studies similar to those carried out in the adult population, some initial estimation of dose in paediatrics should be obtained via extrapolation approaches. However, following such an exercise, well-conducted PK-PD or PK studies will still be needed to determine the most appropriate doses for neonates, infants, children and adolescents.


Composition and energy harvesting capacity of the gut microbiota: relationship to diet, obesity and time in mouse models.
Increased efficiency of energy harvest, due to alterations in the gut microbiota (increased Firmicutes and decreased Bacteroidetes), has been implicated in obesity in mice and humans. However, a causal relationship is unproven and contributory variables include diet, genetics and age. Therefore, we explored the effect of a high-fat (HF) diet and genetically determined obesity (ob/ob) for changes in microbiota and energy harvesting capacity over time. Seven-week-old male ob/ob mice were fed a low-fat diet and wild-type mice were fed either a low-fat diet or a HF-diet for 8 weeks (n=8/group). They were assessed at 7, 11 and 15 weeks of age for: fat and lean body mass (by NMR); faecal and caecal short-chain fatty acids (SCFA, by gas chromatography); faecal energy content (by bomb calorimetry) and microbial composition (by metagenomic pyrosequencing). A progressive increase in Firmicutes was confirmed in both HF-fed and ob/ob mice reaching statistical significance in the former, but this phylum was unchanged over time in the lean controls. Reductions in Bacteroidetes were also found in ob/ob mice. However, changes in the microbiota were dissociated from markers of energy harvest. Thus, although the faecal energy in the ob/ob mice was significantly decreased at 7 weeks, and caecal SCFA increased, these did not persist and faecal acetate diminished over time in both ob/ob and HF-fed mice, but not in lean controls. Furthermore, the proportion of the major phyla did not correlate with energy harvest markers. The relationship between the microbial composition and energy harvesting capacity is more complex than previously considered. While compositional changes in the faecal microbiota were confirmed, this was primarily a feature of high-fat feeding rather than genetically induced obesity. In addition, changes in the proportions of the major phyla were unrelated to markers of energy harvest which changed over time. The possibility of microbial adaptation to diet and time should be considered in future studies.


Editorial: Making clinical trials smarter (and more interesting).
At its heart, the field of child psychology and psychiatry is geared towards using science to develop interventions that promote children's healthy development and treat behavioural and emotional difficulties when they arise. While there have been some successes (e.g. stimulant medication for ADHD, parent training for child conduct problems, Fonagy et al., ) serious challenges lie ahead if we are to achieve reliable and lasting improvements for a larger number of children, and for a broader spectrum of problems.


Potentially inappropriate medication use by elderly Mexican Americans.
Use of inappropriate medications by the elderly is a public health concern with potentially serious health consequences. Research indicates relatively high rates of inappropriate prescription drug usage for older whites and African Americans. However, rates for older Mexican Americans are unknown. To examine the prevalence and predictors of inappropriate prescription medication use by older Mexican Americans. A cross-sectional study of 3050 Mexican Americans aged > or =65 years living in the southwestern US was conducted. In-home interviews in 1993 and 1994 assessed prescription medication use. Descriptive statistics and logistic regression models were used to estimate prevalence and risk of inappropriate prescription drug use. Approximately 12% (n = 365) of the sample had used at least 1 of 32 potentially inappropriate prescription medications within 2 weeks of the baseline assessment. Four drugs, chlorpropamide, propoxyphene, amitriptyline, and dipyridamole, accounted for 54% of all inappropriate prescribing. Unmarried subjects, those with >/=1 chronic diseases, high depressive symptoms, frequent physician visits, and combined Medicaid and Medicare insurance were more likely to have used at least 1 of the 32 potentially inappropriate drugs. The prevalence of inappropriate medication use is lower in older Mexican Americans than in similar white and African American populations. More studies are needed on the pattern of inappropriate prescribing over time and on intervention programs to reduce potentially adverse health outcomes in older Mexican Americans most at risk.


Completeness of cancer registry data in a small Iranian province: A capture-recapture approach.
The incidence of cancer is rising in Iran, and hence it is important to assess the accuracy of the Iranian cancer registry dataset. In this study, the completeness of the cancer registry in the Kohgiluyeh and Boyer-Ahmad (K&B) province is evaluated. The data of registered cases of cancer of people who were living in the K&B province at the time of diagnosis were obtained from the provincial cancer registry offices in K&B, Fars and all other neighbouring provinces. A capture-recapture method along with log-linear statistical modelling were used for analysis. The results indicated that of 2029 known cases of cancer, only 1400 (31%) were registered by the K&B cancer registry office. Age-adjusted incidence rates for all common types of cancer rose from 307.0 per 100,000 (95% confidence interval (CI); 293.8, 320.3, based on observed cases) to 376.4 per 100,000 (95% CI; 361.7, 391.1, based on expected number of cases estimated by capture-recapture analysis) ( p < 0.01). The completeness of cancer registry data varied significantly for different types of cancer. Results suggest that the provincial cancer dataset, which is a part of the national cancer registry programme, is neither complete nor representative. A major improvement in case finding, registry procedures and effective data sharing by provincial cancer registry offices is needed in order to provide valid data for epidemiology of cancer in Iran.


Gestational Lyme borreliosis. Implications for the fetus.
Great diversity of clinical expression of signs and symptoms of gestational Lyme borreliosis parallels the diversity of prenatal syphilis. It is documented that transplacental transmission of the spirochete from mother to fetus is possible. Further research is necessary to investigate possible teratogenic effects that might occur if the spirochete reaches the fetus during the period of organogenesis. Autopsy and clinical studies have associated gestational Lyme borreliosis with various medical problems including fetal death, hydrocephalus, cardiovascular anomalies, neonatal respiratory distress, hyperbilirubinemia, intrauterine growth retardation, cortical blindness, sudden infant death syndrome, and maternal toxemia of pregnancy. Whether any or all of these associations are coincidentally or causally related remains to be clarified by further investigation. It is my expectation that the spectrum of gestational Lyme borreliosis will expand into many of the clinical domains of prenatal syphilis.


Independent external validation of the QRISK3 cardiovascular disease risk prediction model using UK Biobank.
To externally evaluate the performance of QRISK3 for predicting 10 year risk of cardiovascular disease (CVD) in the UK Biobank cohort. We used data from the UK Biobank, a large-scale prospective cohort study of 403 370 participants aged 40-69 years recruited between 2006 and 2010 in the UK. We included participants with no previous history of CVD or statin treatment and defined the outcome to be the first occurrence of coronary heart disease, ischaemic stroke or transient ischaemic attack, derived from linked hospital inpatient records and death registrations. Our study population included 233 233 women and 170 137 men, with 9295 and 13 028 incident CVD events, respectively. Overall, QRISK3 had moderate discrimination for UK Biobank participants (Harrell's C-statistic 0.722 in women and 0.697 in men) and discrimination declined by age (<0.62 in all participants aged 65 years or older). QRISK3 systematically overpredicted CVD risk in UK Biobank, particularly in older participants, by as much as 20%. QRISK3 had moderate overall discrimination in UK Biobank, which was best in younger participants. The observed CVD risk for UK Biobank participants was lower than that predicted by QRISK3, particularly for older participants. It may be necessary to recalibrate QRISK3 or use an alternate model in studies that require accurate CVD risk prediction in UK Biobank.


Patterns and Outcomes in Hepatocellular Carcinoma Patients with Portal Vein Invasion: A Multicenter Prospective Cohort Study.
Sorafenib is a proven first-line treatment recommended for hepatocellular carcinoma (HCC) patients with portal vein invasion (PVI). However, multiple treatment modalities are used in clinical practice as a first-line option. This study is a prospective, observational, multicenter, cohort study evaluating patterns of treatment modalities and outcomes for HCC patients with PVI. The baseline characteristics, treatment modalities, and outcomes were prospectively collected for 287 newly diagnosed HCC patients with PVI between August 2015 and July 2016 from 16 sites in Korea. During a median 7.8 months of follow-up (range 0.3-24.6 months), mortality was observed in 123 (42.9%) patients. Decision tree analysis classified patients into five subgroups with different outcomes. The patterns of treatment were very heterogeneous, and there was no dominant treatment modality. The most commonly used treatment modality was transarterial chemoembolization (TACE) (20.2%) followed by TACE plus external beam radiation therapy (17.8%) and sorafenib (12.5%). When stratified according to the extent of PVI, sorafenib treatment showed comparable outcomes when the PVI extent was lobal or main/bilateral, yet showed worse outcomes when the PVI extent was limited to the segmental level compared to those who received treatment other than sorafenib. HCC patients with PVI comprise a heterogeneous population and are treated with various treatment modalities with diverse clinical outcomes in clinical practice. Subclassification of HCC patients with PVI is required to minimize heterogeneity and should be considered for the selection of treatment modalities and future clinical trials.


Prediction of selective fetal growth restriction and twin-to-twin transfusion syndrome in monochorionic twins.
To study the correlation of discrepancy between crown-rump length (CRL) and nuchal translucency (NT) in monochorionic twins at 11-14 weeks of gestation and subsequent development of twin-to-twin transfusion syndrome (TTTS) and selective fetal growth restriction (sFGR). Retrospective cohort study. Tertiary-care Fetal Medicine Unit, London. Monochorionic twin pregnancies with known outcome. Inter-twin discrepancy was calculated as a percentage of the larger CRL and smaller NT and compared among those developing TTTS, those with sFGR and those with normal outcome. Receiver operating characteristic (ROC) curves were constructed to evaluate the performance of inter-twin discrepancy in prediction of sFGR and TTTS. Development of TTTS and sFGR. A total of 242 monochorionic twin pregnancies were studied (102 TTTS, 36 sFGR and 104 controls). The median CRL discrepancy in the sFGR group (11.9%) was significantly higher (P < 0.001) than in the TTTS group (3.8%) and control group (3.5%). Median inter-twin NT discrepancies were not significantly different (P = 0.869) between sFGR and both TTTS and control groups (15.6%, 16.7% and 14.8%, respectively). Discrepancy in CRL performs well as a screening test for sFGR (area under ROC curve = 0.89), but not for TTTS (area under ROC curve = 0.58). First-trimester CRL discrepancy in monochorionic twins is a marker for subsequent development of sFGR rather than TTTS. Inter-twin NT discrepancy does not appear to be significantly different in these two groups from those with normal outcome.


Behavioral and physiological characterization of male mice under chronic psychosocial stress.
Social stress is a major factor in the etiology of several psychopathologies, with individuals greatly differing in vulnerability. The development of appropriate animal models of social stress is, thus, a major challenge of modern bio-medical research. Adult male mice were subjected to a new model of chronic psychosocial stress in which resident/intruder dyads live chronically in sensory contact and physically interact on a daily basis. Four behavioral categories were identified: Resident Dominants (RD), Resident Subordinates (RS), Intruder Dominants (InD), Intruder Subordinates (InS). Here we investigated: behavior during aggressive interactions; gross physiological components of mice metabolism; organ physiology; response to dexamethasone suppression test (DST). RD and InD mice showed persistently high levels of aggression. All four categories of mice showed robust lack of suppression of corticosterone level when challenged with the DST. Although food intake was not altered under chronic stress, body weight decreased in RD and InD mice while increased in InS and, even more so, in RS mice, suggesting an alteration of their metabolic functions. In conclusion, social status and territory ownership were factors determining individual vulnerability to stress exposure. Our model could, thus, be regarded as a valid model to investigate the biological basis of the individual differences in the response to stressful events.


Effects of a riboflavin source suitable for use in organic broiler diets on performance traits and health indicators.
Riboflavin (vitamin B2) is essential for monogastric animals. It is mainly produced by recombinant microorganisms (Candida famata, Bacillus subtilis and Ashbya gossypii). The availability of genetically modified organism (GMO)-free riboflavin, obligatory in European organic agriculture, is a major issue. Besides, requirements for organic livestock might differ from conventional production because other genotypes and feed formulations are used. The effects of a fermentation suspension with a high native content of riboflavin produced with unmodified A. gossypii by fermentation were investigated at graded dosages as an alternative to conventional (GMO-based) riboflavin in slow-growing broilers on performance traits and health and welfare indicators. In 2 runs with 800 animals each, Ranger Gold™ broilers were fed with 4 dietary treatments. For starter diets (day 1 to 18), treatments included a basal diet (1) without any riboflavin supplementation (negative control, N-C), (2) with conventional riboflavin supplementation (Cuxavit B2 80% riboflavin) at 9.6 mg/kg (positive control, P-C), (3) with riboflavin supplementation from the alternative source at 3.5 mg/kg (A-low) and (4) with riboflavin supplementation from the alternative source at 9.6 mg/kg (A-high). For the finisher diet (day 29 until slaughtering), P-C and A-high were supplemented with 8.0 mg/kg and A-low with 3.5 mg/kg. Diets were formulated according to organic regulations. Animals were kept in floor pens with 20 chickens per pen. Weekly, BW, feed and water consumption were recorded. Every second week, animal-based health and welfare indicators (feather score and footpad dermatitis) were scored. Slaughter traits were assessed for five males and females per pen at 62/63 days of age. Final body weight of A-high differed from N-C and A-low, but not from P-C. From week 2 until six years of age, A-high had a higher daily weight gain when compared to all other groups. With 74.4%, dressing percentage was higher in A-high compared with all other groups (73.3%). Breast percentage of A-low was lower than that of both control groups but did not differ from A-high. The highest frequency of liver scores indicating fatty liver syndrome was found in P-C, followed by N-C and A-low. Feather scores did not respond to treatment; the highest frequency of mild footpad dermatitis was observed in A-high, however at a low prevalence. In conclusion, the tested fermentation suspension with a high native content of riboflavin derived from fermentation of A. gossypii can be used at levels of commercial recommendations as alternative to riboflavin produced from GMO in broiler feeding. Further studies must verify whether riboflavin can be reduced without inducing riboflavin deficiency in slow-growing broilers.


Moringa oleifera Lam prevents acetaminophen induced liver injury through restoration of glutathione level.
Initiation of acetaminophen (APAP) toxicities is believed to be promoted by oxidative stress during the event of overdosage. The aim of the present study was to evaluate the hepatoprotective action of Moringa oleifera Lam (MO), an Asian plant of high medicinal value, against a single high dose of APAP. Groups of five male Sprague-Dawley rats were pre-administered with MO (200 and 800 mg/kg) prior to a single dose of APAP (3g/kg body weight; p.o). Silymarin was used as an established hepatoprotective drug against APAP induced liver injury. The hepatoprotective activity of MO extract was observed following significant histopathological analysis and reduction of the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in groups pretreated with MO compared to those treated with APAP alone. Meanwhile, the level of glutathione (GSH) was found to be restored in MO-treated animals compared to the groups treated with APAP alone. These observations were comparable to the group pretreated with silymarin prior to APAP administration. Group that was treated with APAP alone exhibited high level of transaminases and ALP activities besides reduction in the GSH level. The histological hepatocellular deterioration was also evidenced. The results from the present study suggested that the leaves of MO can prevent hepatic injuries from APAP induced through preventing the decline of glutathione level.


How best to identify chromosomal interactions: a comparison of approaches.
Chromosome conformation capture (3C) methods are central to understanding the link between nuclear structure and function, and the physical interactions between distal regulatory elements and promoters. However, no one method is appropriate to address all biological questions, as each variant differs markedly in resolution, reproducibility, throughput and biases. A thorough appreciation of the strengths and weaknesses of each technique is critical when choosing the correct method for a specific application or for gauging how best to interpret different sources of data. In addition, the analysis method must be carefully considered, as this choice can profoundly affect the output. In this Review, we describe and compare the different available 3C-based approaches, with a focus on the analysis of mammalian genomes.


Prophylactic antidepressant treatment of interferon-induced depression in chronic hepatitis C: a systematic review and meta-analysis.
To assess the utility of prophylactic administration of antidepressants in preventing a major depressive episode during antiviral treatment for chronic hepatitis C. A computerized literature search was conducted in MEDLINE, PsycINFO, EMBASE, the Cochrane Library, and ClinicalTrials.gov to locate articles published in any language from the earliest available online year until October 2012, using the following phrase and Boolean logic algorithm: "hepatitis and c and (interferon-alpha OR peginterferon OR (pegylated and interferon)) and (depression OR mood) and (prevention OR prophylactic OR prophylaxis OR antidepressant)." Double-blind, randomized, placebo-controlled trials using antidepressants prophylactically before starting antiviral therapy for chronic hepatitis C were included. At baseline, none of the patients in the trials presented depression (DSM-IV-TR criteria). Using keywords and cross-referenced bibliographies, 144 studies were identified and examined in depth. 137 articles were rejected because inclusion criteria were not met. Finally, 7 studies were included. Data were extracted independently by 2 investigators. The primary outcome measure was the onset of a major depressive episode during the antiviral treatment. Depressive symptoms, other side effects, and sustained virologic response were also examined. A full review and meta-analysis were performed. Odds ratios (ORs), mean differences, and estimated numbers needed to treat (NNTs) with 95% confidence intervals (CIs) were calculated. 591 patients were randomly assigned to antiviral treatment and another intervention: escitalopram (n = 197), paroxetine (n = 42), citalopram (n = 53), or placebo (n = 299). Selective serotonin reuptake inhibitors (SSRIs), as a group, reduced the incidence of a major depressive episode during antiviral treatment (OR = 0.53; 95% CI, 0.33 to 0.84). The NNT was 12 (95% CI, 7.0 to 37.9). SSRIs reduced depressive symptoms at 24 weeks of treatment (mean difference -2.18; 95% CI, -4.25 to -0.10). With regard to side effects, only dizziness was associated with administration of antidepressants (OR = 2.65; 95% CI, 1.46 to 4.80). There were no differences in sustained virologic response (OR = 1.22; 95% CI, 0.58 to 2.57). Administration of SSRIs before starting antiviral treatment reduces the incidence of interferon-induced depression, with a relatively moderate prophylactic impact and good tolerability.


Broadband multiple-cascaded integration of electroabsorption modulators and high impedance transmission lines by lowering standing-wave effect.
Standing wave effect of applied electrical field on optical modulation in multiple-cascaded integration (CI) electroabsorption modulator (EAM) and high-impedance transmission line (HITL) has been investigated in this paper. As modulation frequency is increased to the scale that electrical wavelength is in the order of optical modulator length, multiple electrical reflection and self-interference on impedance-mismatch boundaries becomes significant, leading to strong position-dependent field distribution and degrading modulation bandwidth. Sharp bandwidth roll of electrical-optical (EO) conversion by standing wave has been found experimentally in CI structure, consistent with simulation results. By comparing different segment number and length of CI- structure, larger section number of design can overcome such problem to get more flatten bandwidth response. Such simple CI for 300μm long EAM has been demonstrated with flat EO response of -3dB drop 45GHz and -10dB microwave reflection (up to 65GHz) in 6-segement device, suggesting this scheme design is quite useful for efficient broad band modulation.


Drosophila Tcf and Groucho interact to repress Wingless signalling activity.
Wingless/Wnt signalling directs cell-fate choices during embryonic development. Inappropriate reactivation of the pathway causes cancer. In Drosophila, signal transduction from Wingless stabilizes cytosolic Armadillo, which then forms a bipartite transcription factor with the HMG-box protein Drosophila Tcf (dTcf) and activates expression of Wingless-responsive genes. Here we report that in the absence of Armadillo, dTcf acts as a transcriptional repressor of Wingless-responsive genes, and we show that Groucho acts as a corepressor in this process. Reduction of dTcf activity partially suppresses wingless and armadillo mutant phenotypes, leading to derepression of Wingless-responsive genes. Furthermore, overexpression of wild-type dTcf enhances the phenotype of a weak wingless allele. Finally, mutations in the Drosophila groucho gene also suppress wingless and armadillo mutant phenotypes as Groucho physically interacts with dTcf and is required for its full repressor activity.


Distinct patterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas.
To compare lung adenocarcinoma (ADC) and lung squamous cell carcinoma (SqCC) and to identify new drivers of lung carcinogenesis, we examined the exome sequences and copy number profiles of 660 lung ADC and 484 lung SqCC tumor-normal pairs. Recurrent alterations in lung SqCCs were more similar to those of other squamous carcinomas than to alterations in lung ADCs. New significantly mutated genes included PPP3CA, DOT1L, and FTSJD1 in lung ADC, RASA1 in lung SqCC, and KLF5, EP300, and CREBBP in both tumor types. New amplification peaks encompassed MIR21 in lung ADC, MIR205 in lung SqCC, and MAPK1 in both. Lung ADCs lacking receptor tyrosine kinase-Ras-Raf pathway alterations had mutations in SOS1, VAV1, RASA1, and ARHGAP35. Regarding neoantigens, 47% of the lung ADC and 53% of the lung SqCC tumors had at least five predicted neoepitopes. Although targeted therapies for lung ADC and SqCC are largely distinct, immunotherapies may aid in treatment for both subtypes.


Utilizing social media data for pharmacovigilance: A review.
Automatic monitoring of Adverse Drug Reactions (ADRs), defined as adverse patient outcomes caused by medications, is a challenging research problem that is currently receiving significant attention from the medical informatics community. In recent years, user-posted data on social media, primarily due to its sheer volume, has become a useful resource for ADR monitoring. Research using social media data has progressed using various data sources and techniques, making it difficult to compare distinct systems and their performances. In this paper, we perform a methodical review to characterize the different approaches to ADR detection/extraction from social media, and their applicability to pharmacovigilance. In addition, we present a potential systematic pathway to ADR monitoring from social media. We identified studies describing approaches for ADR detection from social media from the Medline, Embase, Scopus and Web of Science databases, and the Google Scholar search engine. Studies that met our inclusion criteria were those that attempted to extract ADR information posted by users on any publicly available social media platform. We categorized the studies according to different characteristics such as primary ADR detection approach, size of corpus, data source(s), availability, and evaluation criteria. Twenty-two studies met our inclusion criteria, with fifteen (68%) published within the last two years. However, publicly available annotated data is still scarce, and we found only six studies that made the annotations used publicly available, making system performance comparisons difficult. In terms of algorithms, supervised classification techniques to detect posts containing ADR mentions, and lexicon-based approaches for extraction of ADR mentions from texts have been the most popular. Our review suggests that interest in the utilization of the vast amounts of available social media data for ADR monitoring is increasing. In terms of sources, both health-related and general social media data have been used for ADR detection-while health-related sources tend to contain higher proportions of relevant data, the volume of data from general social media websites is significantly higher. There is still very limited amount of annotated data publicly available , and, as indicated by the promising results obtained by recent supervised learning approaches, there is a strong need to make such data available to the research community.


Majorization-minimization algorithms for wavelet-based image restoration.
Standard formulations of image/signal deconvolution under wavelet-based priors/regularizers lead to very high-dimensional optimization problems involving the following difficulties: the non-Gaussian (heavy-tailed) wavelet priors lead to objective functions which are nonquadratic, usually nondifferentiable, and sometimes even nonconvex; the presence of the convolution operator destroys the separability which underlies the simplicity of wavelet-based denoising. This paper presents a unified view of several recently proposed algorithms for handling this class of optimization problems, placing them in a common majorization-minimization (MM) framework. One of the classes of algorithms considered (when using quadratic bounds on nondifferentiable log-priors) shares the infamous "singularity issue" (SI) of "iteratively reweighted least squares" (IRLS) algorithms: the possibility of having to handle infinite weights, which may cause both numerical and convergence issues. In this paper, we prove several new results which strongly support the claim that the SI does not compromise the usefulness of this class of algorithms. Exploiting the unified MM perspective, we introduce a new algorithm, resulting from using l1 bounds for nonconvex regularizers; the experiments confirm the superior performance of this method, when compared to the one based on quadratic majorization. Finally, an experimental comparison of the several algorithms, reveals their relative merits for different standard types of scenarios.


Signals from the nucleus: activation of NF-kappaB by cytosolic ATM in the DNA damage response.
In response to genotoxic stress induced by DNA double-stranded breaks (DSBs), the inhibitor of κB kinase (IKK) to nuclear factor κB (NF-κB) pathway is activated, which can promote cancer progression and increase the resistance of cancer cells to ionizing radiation or chemotherapeutic drugs. The kinase ataxia telangiectasia mutated (ATM) has a critical role in the activation of NF-κB in response to genotoxic stress. Two reports reveal key cytoplasmic functions of ATM in triggering IKK activation upon DNA damage. After induction of DSBs, ATM is exported from the nucleus and stimulates the ubiquitin ligase activity of tumor necrosis factor receptor-associated factor 6 (TRAF6) or X-linked inhibitor of apoptosis protein, which catalyze the auto-polyubiquitylation of TRAF6 and the polyubiquitylation of the IKK adaptor ELKS, respectively. Ubiquitylation promotes the assembly of signalosomes containing the kinase TAK1 (transforming growth factor b-activated kinase 1). These signalosomes are the site of activation of the cytosolic IKK complex, which stimulates NF-κB-dependent induction of a proliferative and antiapoptotic gene program. These studies show that ATM executes essential functions outside the nucleus in response to DSBs.


Effect of the "Recruitment" Maneuver on Respiratory Mechanics in Laparoscopic Sleeve Gastrectomy Surgery.
LSG surgery is used for surgical treatment of morbid obesity. Obesity, anesthesia, and pneumoperitoneum cause reduced pulmoner functions and a tendency for atelectasis. The alveolar "recruitment" maneuver (RM) keeps airway pressure high, opening alveoli, and increasing arterial oxygenation. The aim of our study is to research the effect on respiratory mechanics and arterial blood gases of performing the RM in LSG surgery. Sixty patients undergoing LSG surgery were divided into two groups (n = 30) Patients in group R had the RM performed 5 min after desufflation with 100% oxygen, 40 cmH2O pressure for 40 s. Group C had standard mechanical ventilation. Assessments of respiratory mechanics and arterial blood gases were made in the 10th min after induction (T1), 10th min after insufflation (T2), 5th min after desufflation (T3), and 15th min after desufflation (T4). Arterial blood gases were assessed in the 30th min (T5) in the postoperative recovery unit. In group R, values at T5, PaO2 were significantly high, while PaCO2 were significantly low compared with group C. Compliance in both groups reduced with pneumoperitoneum. At T4, the compliance in the recruitment group was higher. In both groups, there was an increase in PIP with pneumoperitoneum and after desufflation this was identified to reduce to levels before pneumoperitoneum. Adding the RM to PEEP administration for morbidly obese patients undergoing LSG surgery is considered to be effective in improving respiratory mechanics and arterial blood gas values and can be used safely.


Controlled growth mechanism of poly (3-hexylthiophene) nanowires.
Synthesis of 1D-polymer nanowires by a self-assembly method using marginal solvents is an attractive technique. While the formation mechanism is poorly understood, this method is essential in order to control the growth of nanowires. Here we visualized the time-dependent assembly of poly (3-hexyl-thiophene-2,5-diyl) (P3HT) nanowires by atomic force microscopy and scanning tunneling microscopy. The assembly of P3HT nanowires was carried out at room temperature by mixing cyclohexanone (CHN), as a poor solvent, with polymer solution in 1,2-dichlorobenzene (DCB). Both π-π stacking and planarization, obtained at the mix volume ratio of P3HT (in DCB):CHN (10:7), were considered during the investigation. We find that the length of nanowires was determined by the ordering of polymers in the polymer repetition direction. Additionally, our density functional theory calculations revealed that the presence of DCB and CHN molecules that stabilize the structural distortions due to tail group of polymers was essential for the core-wire formation.


[Multimodality imaging in transcatheter aortic valve implantation procedures: the pivotal role of echocardiography].
Alternative percutaneous approaches to the management of symptomatic aortic stenosis in elderly and high-risk patients have become more attractive and raised profound interest in recent years. The superiority of transcatheter aortic valve implantation (TAVI) compared with medical therapy for patients deemed unsuitable for surgery has recently been established by the PARTNER trial, and preliminary randomized data in high-risk patients have confirmed that TAVI is non-inferior to surgical valve replacement in terms of safety and effectiveness. This finding will probably lead to an exponential increase in TAVI procedures over the next decade. In the clinical setting of the percutaneous approach, the choice of a specific valve is crucial to optimize the success rate and minimize the procedure-related complications and requires accurate preprocedural diagnostic assessment. According to the high comorbidities of candidates currently undergoing a TAVI procedure, a tailored diagnostic framework in the elderly population is highly recommended. The present review discusses the role of echocardiography in the setting of multimodality imaging before the TAVI procedure and throughout the entire process, from patient selection to procedural guidance and post-procedural follow-up, suggesting a practical algorithm necessary for the best clinical outcome.


Zebrafish with mutations in mismatch repair genes develop neurofibromas and other tumors.
Defective mismatch repair (MMR) in humans causes hereditary nonpolyposis colorectal cancer. This genetic predisposition to colon cancer is linked to heterozygous familial mutations, and loss-of-heterozygosity is necessary for tumor development. In contrast, the rare cases with biallelic MMR mutations are juvenile patients with brain tumors, skin neurofibromas, and café-au-lait spots, resembling the neurofibromatosis syndrome. Many of them also display lymphomas and leukemias, which phenotypically resembles the frequent lymphoma development in mouse MMR knockouts. Here, we describe the identification and characterization of novel knockout mutants of the three major MMR genes, mlh1, msh2, and msh6, in zebrafish and show that they develop tumors at low frequencies. Predominantly, neurofibromas/malignant peripheral nerve sheath tumors were observed; however, a range of other tumor types was also observed. Our findings indicate that zebrafish mimic distinct features of the human disease and are complementary to mouse models.


Multichromophore light harvesting in hybrid solar cells.
A new technologically relevant method for multichromophore sensitizing of hybrid blend solar cells is presented. Two dyes having complementary absorption in the UV-visible regions are individually adsorbed on nanocrystalline TiO(2) powder. These dyed TiO(2) nanoparticles are blended with an organic hole-conductor (HC) Spiro-OMeTAD in desired compositions and applied on a conducting substrate by doctor-blading at room temperature to fabricate multichromophore-sensitized hybrid blend solar cells. The external quantum efficiency (EQE) of the single hybrid layer system fabricated with two dyes, that absorb mainly UV (TPD dye) and visible regions (Ru-TPA-NCS dye), exhibited a clear panchromatic response with the sum of the EQE characteristics of each single dye cell. The first results of a multichromophore-sensitized solid-state solar cell showed J(sc) of 2.1 mA cm(-2), V(oc) of 645 mV, FF of 47% and efficiency of 0.65% at AM 1.5 G, 100 mW cm(-2) illumination intensity. The J(sc) of the multichromophore cell is the sum of the individually dyed solar cells. The process described here is technically very innovative and very simple in procedure. It has potentials to be adopted for panchromatic sensitization using more than two dyes in a single hybrid layer or layer-wise fabrication of a tandem structure at room temperature.


Group based diabetes self-management education compared to routine treatment for people with type 2 diabetes mellitus. A systematic review with meta-analysis.
Diabetes self-management education (DSME) can be delivered in many forms. Group based DSME is widespread due to being a cheaper method and the added advantages of having patient meet and discuss with each other. assess effects of group-based DSME compared to routine treatment on clinical, lifestyle and psychosocial outcomes in type-2 diabetes patients. A systematic review with meta-analysis. Computerised bibliographic database were searched up to January 2008 for randomised controlled trials evaluating group-based DSME for adult type-2 diabetics versus routine treatment where the intervention had at least one session and =/>6 months follow-up. At least two reviewers independently extracted data and assessed study quality. In total 21 studies (26 publications, 2833 participants) were included. Of all the participants 4 out of 10 were male, baseline age was 60 years, BMI 31.6, HbA1c 8.23%, diabetes duration 8 years and 82% used medication. For the main clinical outcomes, HbA1c was significantly reduced at 6 months (0.44% points; P=0.0006, 13 studies, 1883 participants), 12 months (0.46% points; P=0.001, 11 studies, 1503 participants) and 2 years (0.87% points; P<0.00001, 3 studies, 397 participants) and fasting blood glucose levels were also significantly reduced at 12 months (1.26 mmol/l; P<0.00001, 5 studies, 690 participants) but not at 6 months. For the main lifestyle outcomes, diabetes knowledge was improved significantly at 6 months (SMD 0.83; P=0.00001, 6 studies, 768 participants), 12 months (SMD 0.85; P<0.00001, 5 studies, 955 participants) and 2 years (SMD 1.59; P=0.03, 2 studies, 355 participants) and self-management skills also improved significantly at 6 months (SMD 0.55; P=0.01, 4 studies, 534 participants). For the main psychosocial outcomes, there were significant improvement for empowerment/self-efficacy (SMD 0.28, P=0.01, 2 studies, 326 participants) after 6 months. For quality of life no conclusion could be drawn due to high heterogeneity. For the secondary outcomes there were significant improvements in patient satisfaction and body weight at 12 months for the intervention group. There were no differences between the groups in mortality rate, body mass index, blood pressure and lipid profile. Group-based DSME in people with type 2 diabetes results in improvements in clinical, lifestyle and psychosocial outcomes.


Cytochromes P450, oxygen, and evolution.
The role of the cytochrome P450 superfamily of heme-thiolate enzymes in oxidative metabolism is discussed in the context of evolutionary development. Concordances between the rise in atmospheric oxygen content, elaboration of the P450 phylogenetic tree and the accepted timescale for the emergence of animal phyla are described. The unique ability of the P450 monooxygenase system to activate molecular oxygen via the consecutive input of two reducing equivalents is explored, such that the possibility of oxygen radical generation and its toxic consequences can be explained in mechanistic terms, together with an appreciation of the ways in which this oxygen activating ability has been utilized by evolving biological systems in their adaptation to an increasing atmospheric oxygen concentration over the past two billon years.


Posterior paresis and osteolysis in guinea pigs (Cavia porcellus) secondary to Freund's adjuvant immunization.
Bilateral hindlimb paresis occurred in 3 guinea pigs after immunization with an adjuvant-antigen mixture containing complete Freund's adjuvant. Doses were injected into unanaesthetized animals, divided among 3 or 4 sites, and given slightly off midline in the subcutaneous tissues of the back. Neurologic examination of affected animals revealed intact flexor and panniculus responses and limited voluntary movement of the hindlimbs. Histopathologic interpretation of 2 affected animals showed fibrogranulomatous material effacing the skeletal muscle and vertebral bone, with marked bone lysis and infiltration into the marrow space and spinal canal. In addition, multiple granulomas in the pulmonary parenchyma were noted. A postmortem radiograph of the excised thoracolumbar spine of 1 animal revealed a soft tissue swelling and "moth-eaten" and geographic osteolysis of 2 spinous processes. Hindlimb paresis and osteolysis likely resulted from accidental injection of the adjuvant-antigen mixture into the epaxial musculature and subsequent extension of injection site granulomas into the spinal canal.


Multiscale modeling of thermal conductivity of polycrystalline graphene sheets.
We developed a multiscale approach to explore the effective thermal conductivity of polycrystalline graphene sheets. By performing equilibrium molecular dynamics (EMD) simulations, the grain size effect on the thermal conductivity of ultra-fine grained polycrystalline graphene sheets is investigated. Our results reveal that the ultra-fine grained graphene structures have thermal conductivity one order of magnitude smaller than that of pristine graphene. Based on the information provided by the EMD simulations, we constructed finite element models of polycrystalline graphene sheets to probe the thermal conductivity of samples with larger grain sizes. Using the developed multiscale approach, we also investigated the effects of grain size distribution and thermal conductivity of grains on the effective thermal conductivity of polycrystalline graphene. The proposed multiscale approach on the basis of molecular dynamics and finite element methods could be used to evaluate the effective thermal conductivity of polycrystalline graphene and other 2D structures.


The N-terminus of the intrinsically disordered protein α-synuclein triggers membrane binding and helix folding.
Alpha-synuclein (αS) is a 140-amino-acid protein that is involved in a number of neurodegenerative diseases. In Parkinson's disease, the protein is typically encountered in intracellular, high-molecular-weight aggregates. Although αS is abundant in the presynaptic terminals of the central nervous system, its physiological function is still unknown. There is strong evidence for the membrane affinity of the protein. One hypothesis is that lipid-induced binding and helix folding may modulate the fusion of synaptic vesicles with the presynaptic membrane and the ensuing transmitter release. Here we show that membrane recognition of the N-terminus is essential for the cooperative formation of helical domains in the protein. We used circular dichroism spectroscopy and isothermal titration calorimetry to investigate synthetic peptide fragments from different domains of the full-length αS protein. Site-specific truncation and partial cleavage of the full-length protein were employed to further characterize the structural motifs responsible for helix formation and lipid-protein interaction. Unilamellar vesicles of varying net charge and lipid compositions undergoing lateral phase separation or chain melting phase transitions in the vicinity of physiological temperatures served as model membranes. The results suggest that the membrane-induced helical folding of the first 25 residues may be driven simultaneously by electrostatic attraction and by a change in lipid ordering. Our findings highlight the significance of the αS N-terminus for folding nucleation, and provide a framework for elucidating the role of lipid-induced conformational transitions of the protein within its intracellular milieu.


Genomic Landscape of Esophageal Squamous Cell Carcinoma in a Japanese Population.
Esophageal squamous cell carcinoma (ESCC) is the predominant form of esophageal cancer in Japan. Smoking and drinking alcohol are environmental risk factors for ESCC, whereas single nucleotide polymorphisms in ADH1B and ALDH2, which increase harmful intermediates produced by drinking alcohol, are genetic risk factors. We conducted a large-scale genomic analysis of ESCCs from patients in Japan to determine the mutational landscape of this cancer. We performed whole-exome sequence analysis of tumor and nontumor esophageal tissues collected from 144 patients with ESCC who underwent surgery at 5 hospitals in Japan. We also performed single-nucleotide polymorphism array-based copy number profile and germline genotype analyses of polymorphisms in ADH1B and ALDH2. Polymorphisms in CYP2A6, which increase harmful effects of smoking, were analyzed. Functions of TET2 mutants were evaluated in KYSE410 and HEK293FT cells. A high proportion of mutations in the 144 tumor samples were C to T substitution in CpG dinucleotides (called the CpG signature) and C to G/T substitutions with a flanking 5' thymine (called the APOBEC signature). Based on mutational signatures, patients were assigned to 3 groups, which associated with environmental (drinking and smoking) and genetic (polymorphisms in ALDH2 and CYP2A6) factors. Many tumors contained mutations in genes that regulate the cell cycle (TP53, CCND1, CDKN2A, FBXW7); epigenetic processes (MLL2, EP300, CREBBP, TET2); and the NOTCH (NOTCH1, NOTCH3), WNT (FAT1, YAP1, AJUBA) and receptor-tyrosine kinase-phosphoinositide 3-kinase signaling pathways (PIK3CA, EGFR, ERBB2). Mutations in EP300 and TET2 correlated with shorter survival times, and mutations in ZNF750 associated with an increased number of mutations of the APOBEC signature. Expression of mutant forms of TET2 did not increase cellular levels of 5-hydroxymethylcytosine in HEK293FT cells, whereas knockdown of TET2 increased the invasive activity of KYSE410 ESCC cells. Computational analyses associated the mutations in NFE2L2 we identified with transcriptional activation of its target genes. We associated environmental and genetic factors with base substitution patterns of somatic mutations and provide a registry of genes and pathways that are disrupted in ESCCs. These findings might be used to design specific treatments for patients with esophageal squamous cancers.


The 64-kilodalton capsid protein homolog of Beet yellows virus is required for assembly of virion tails.
The filamentous virion of the closterovirus Beet yellows virus (BYV) consists of a long body formed by the major capsid protein (CP) and a short tail composed of the minor capsid protein (CPm) and the virus-encoded Hsp70 homolog. By using nano-liquid chromatography-tandem mass spectrometry and biochemical analyses, we show here that the BYV 64-kDa protein (p64) is the fourth integral component of BYV virions. The N-terminal domain of p64 is exposed at the virion surface and is accessible to antibodies and mild trypsin digestion. In contrast, the C-terminal domain is embedded in the virion and is inaccessible to antibodies or trypsin. The C-terminal domain of p64 is shown to be homologous to CP and CPm. Mutation of the signature motifs of capsid proteins of filamentous RNA viruses in p64 results in the formation of tailless virions, which are unable to move from cell to cell. These results reveal the dual function of p64 in tail assembly and BYV motility and support the concept of the virion tail as a specialized device for BYV cell-to-cell movement.


Not-patient and not-visitor: a metasynthesis fathers' encounters with pregnancy, birth and maternity care.
the active engagement of fathers in maternity care is associated with long-term health and social benefits for the mother, baby and family. The maternity care expectations and experiences of expectant and new fathers have received little attention to date. to identify and synthesise good quality qualitative research that explores the views and experiences of fathers who have encountered maternity care in high resource settings. based on a pre-determined search strategy, relevant databases were searched for papers published between January 1999 and January 2010. Backchaining of the reference lists in included papers was undertaken. good quality qualitative research studies exploring fathers' involvement in maternity care through pregnancy, birth, and up to 6 months postnatally, that were undertaken in high resource countries. No language restrictions were imposed. ANALYTIC STRATEGY: the analysis was based on the metaethnographic techniques of Noblit and Hare (1988) as amended by Downe et al. (2007). from 856 hits 23 papers were included. The emerging themes were as follows: risk and uncertainty, exclusion, fear and frustration, the ideal and the reality, issues of support and experiencing transition. fathers feel themselves to be 'partner and parent' but their experience of maternity care services is as 'not-patient and not-visitor'. This situates them in an interstitial and undefined space (both emotionally and physically) with the consequence that many feel excluded and fearful. fathers cannot support their partner effectively in achieving the ideal of transition to a successful pregnancy, joyful birth and positive parenthood experience unless they are themselves supported, included, and prepared for the reality of risk and uncertainty in pregnancy, labour and parenthood and for their role in this context.