Modes and strategies for providing conventional mechanical ventilation in neonates.
Neonatal respiratory failure is a common and serious clinical problem which in a considerable proportion of infants requires invasive mechanical ventilation. The basic goal of mechanical ventilation is to restore lung function while limiting ventilator-induced lung injury, which is considered an important risk factor in the development of bronchopulmonary dysplasia (BPD). Over the last decades, new conventional mechanical ventilation (CMV) modalities have been introduced in clinical practice, aiming to assist clinicians in providing lung protective ventilation strategies. These modalities use more sophisticated techniques to improve patient-ventilator interaction and transfer control of ventilation from the operator to the patient. Knowledge on how these new modalities work and how they interact with lung physiology is essential for optimal and safe use. In this review, we will discuss some important basic lung physiological aspects for applying CMV, the basic principles of the old and new CMV modalities, and the evidence to support their use in daily clinical practice.


Reproducibility, accuracy, and predictors of accuracy for the detection of coronary atherosclerotic plaque composition by computed tomography: an ex vivo comparison to intravascular ultrasound.
To determine the reproducibility, accuracy, and predictors of accuracy of computed tomography (CT) angiography to detect and characterize coronary atherosclerotic plaque as compared with intravascular ultrasound. Ten ex vivo human coronary arteries were imaged in a moving phantom by dual-source CT (collimation: 0.6 mm, reconstructed slice thickness: 0.4 mm) and intravascular ultrasound (IVUS). Coregistered cross-sections were assessed at 0.4 mm intervals for the presence and composition of atherosclerotic plaque (noncalcified, mixed, and calcified) on CT and IVUS by independent readers to determine reader agreement and diagnostic accuracy. Quantitative measurements of lumen and plaque area, plaque eccentricity, and intimal thickness on IVUS were used to determine predictors for the detection of noncalcified plaque by CT. Within 1002 coregistered cross-sections, the interobserver agreement to detect plaque on CT was K = 0.48, K = 0.42, and K = 1.00 for noncalcified, mixed, and calcified plaque; respectively. The sensitivity and specificity of CT was 57% out of 84% for noncalcified, 32% of 92% for mixed, and 56% of 93% for calcified plaque when compared with IVUS; respectively. Misclassification occurred in 68% of mixed and 43% of noncalcified plaques. The odds of detecting noncalcified plaque in CT independently increased by 56% (95% CI: 47%-77%, P < 0.0001) with every 0.1 mm increase in maximum intimal thickness as measured by IVUS. Detection rate for noncalcified plaques was poor for plaques <1 mm (36%) but excellent for plaques >1 mm maximal intimal thickness (90%). Reader agreement and diagnostic accuracy for the detection of coronary atherosclerotic plaque vary with plaque composition. Intimal thickness independently predicts detection of noncalcified plaque by CT with excellent sensitivity for >1 mm thick plaques.


Cellular localization, expression and functional implications of the utero-placental endothelin system during maintenance and termination of canine gestation.
Utero-placental (Ut-Pl) angiogenesis and blood flow are fundamental for successful outcome of pregnancy. They are controlled by numerous vasodilator and vasoconstrictor systems such as endothelins (EDNs) and the renin angiotensin system. Dogs possess an invasive type of placentation, classified as endotheliochorial. Despite increasing knowledge regarding canine Ut-Pl function, little information exists on uterine and placental vascular activity during initiation, maintenance and termination of pregnancy in this species. The current study investigated expression of EDNs and their receptors (EDNRA and EDNRB) in the pre-implantation uterus and Ut-Pl compartments during gestation and at normal parturition, as well as in mid-pregnant dogs treated with the antigestagen aglepristone. The Ut-Pl mRNA expression of EDN1 and EDNRA was constant until mid-gestation and increased significantly during prepartum luteolysis. In contrast, EDN2 was highest pre-implantation and decreased following placentation, remaining low thereafter. Expression of the EDN-activating enzyme ECE1 and mRNA of EDNRB increased towards mid-gestation and was further elevated at prepartum luteolysis. Antigestagen treatment resulted in increased levels of EDN1 and EDNRA. At the cellular level, the uterine expression of EDN1, ECE1 and EDNRB was found predominantly in the endometrial surface and glandular epithelial cells; uterine signals for EDNRA were weak. In Ut-Pl all targets were mainly localized in the placenta fetalis, with syncytiotrophoblast staining stronger for ECE1 and EDNRB. In contrast, EDNRA stained strongly at the base of the placental labyrinth. Expression and localization of EDNs (EDN1, -2), EDN receptors and ECE1 in the placenta fetalis suggests their involvement in the trophoblast invasion and proliferation.


Plasma phospholipid very-long-chain SFAs in midlife and 20-year cognitive change in the Atherosclerosis Risk in Communities (ARIC): a cohort study.
Very-long-chain SFAs (VLSFAs) have recently gained considerable attention as having beneficial effects on health and aging. The objective of this study was to assess the associations of plasma phospholipid VLSFAs [arachidic acid (20:0), behenic acid (22:0), tricosanoic acid (23:0), and lignoceric acid (24:0)] with 20-y cognitive decline in the Atherosclerosis Risk in Communities (ARIC) participants. Furthermore, this study compared the associations of plasma phospholipid VLSFAs with 5 common groups of fatty acids [i.e., total SFAs, total MUFAs, total ω-3 (n-3) PUFAs, total marine-derived ω-3 PUFAs, total ω-6 PUFAs]. This study used a cohort study design of 3229 ARIC participants enrolled at the Minnesota field center. Fatty acids were measured at visit 1 (1987-1989); and cognition was assessed at visits 2 (1990-1992), 4 (1996-1998), and 5 (2011-2013) using 3 tests: the Delayed Word Recall Test (DWRT), the Digit-Symbol Substitution Test (DSST), and the Word Fluency Test (WFT). Higher proportions of plasma phospholipid total VLSFAs and each individual VLSFA were associated with less decline in WFT, a test of verbal fluency. For example, 1 SD higher in total VLSFAs at baseline was associated with 0.057 SD (95% CI: 0.018, 0.096, P = 0.004) less cognitive decline over 20 y as measured by WFT score. None of the 5 common fatty acid groups were associated with change in WFT, but a higher proportion of plasma phospholipid total MUFAs was associated with greater decline in DWRT; higher total ω-6 PUFAs with less decline in DWRT; and higher total ω-3 and total marine-derived ω-3 PUFAs with less decline in DSST. This study suggests that higher proportions of plasma phospholipid VLSFAs in midlife may be associated with less 20-y cognitive decline.


Commercial products to preserve specimens for tuberculosis diagnosis: a systematic review.
Eliminating tuberculosis in high-burden settings requires improved diagnostic capacity. Important tests such as Xpert® MTB/RIF and culture are often performed at centralised laboratories that are geographically distant from the point of specimen collection. Preserving specimen integrity during transportation, which could affect test performance, is challenging. To conduct a systematic review of commercial products for specimen preservation for a World Health Organization technical consultation. Databases were searched up to January 2018. Methodological quality was assessed using Quality Assessment of Technical Studies, a new technical study quality-appraisal tool, and Quality Assessment of Diagnostic Accuracy Studies-2. Studies were analysed descriptively in terms of the different products, study designs and diagnostic strategies used. Four products were identified from 16 studies: PrimeStore-Molecular-Transport-Medium (PS-MTM), FTA card, GENO•CARD (all for nucleic acid amplification tests [NAATs]) and OMNIgene•SPUTUM (OMS; culture, NAATs). PS-MTM, but not FTA card or GENO•CARD, rendered Mycobacterium tuberculosis non-culturable. OMS reduced Löwenstein-Jensen but not MGIT™ 960™ contamination, led to delayed MGIT time-to-positivity, resulted in Xpert performance similar to cold chain-transported untreated specimens, and obviated the need for N-acetyl-L-cysteine-sodium hydroxide decontamination. Data from paucibacillary specimens were limited. Evidence that a cold chain improves culture was mixed and absent for Xpert. The effect of the product alone could be discerned in only four studies. Limited evidence suggests that transport products result in test performance comparable to that seen in cold chain-transported specimens.


The Use of Recovery Strategies Among Participants of the BUPA Great North Run: A Cross-Sectional Survey.
To investigate half-marathon runners' frequency of use of recovery strategies, perceptions regarding the most beneficial recovery strategy, and reasons for using recovery strategies. Cross-sectional survey. 186 participants of the 13.1 mile BUPA Great North Run 2013. A questionnaire was developed which required participants to indicate how frequently they used 12 different recovery strategies, identify which recovery strategy they believed to be most beneficial, and rank 6 reasons for using recovery strategies in order of importance. Data were analyzed using a Friedman nonparametric ANOVA and additional nonparametric tests. All participants used recovery strategies. Stretching was the most commonly used recovery strategy (P < .001), whereas the use of nutritional supplements was the most commonly selected most beneficial recovery strategy. More than 50% of respondents indicated that they never used strategies such as kinesio tape (80%), hydrotherapy (78%), or ice baths (71%). A significant difference was observed between reasons for using recovery strategy (χ2 (5) = 292.29, P < .001). Reducing muscle tightness (rank 4.87) and reducing injury (rank 4.35) were the most frequently chosen most important reasons for using recovery strategies. Minor sex and age differences in the responses were identified. Recovery strategy usage appears to be widespread among half-marathon runners; however, disparities exist between the frequency of use and perceived effectiveness of different recovery strategies. Further research in this area is needed to facilitate the development of recovery strategy guidelines which are both evidence-based and practically relevant.


Autoantibody cancer biomarker: extracellular protein kinase A.
In cancer cells, cyclic AMP-dependent protein kinase (PKA) is secreted into the conditioned medium. This PKA, designated as extracellular protein kinase A (ECPKA), is markedly up-regulated in the sera of patients with cancer. The currently available tumor markers are based on the antigen determination method and lack specificity and sensitivity. Here, we present an ECPKA autoantibody detection method for a universal biomarker that detects cancer of various cell types. We tested sera from 295 patients with cancers of various cell types, 155 normal controls, and 55 patients without cancer. The specificity and sensitivity of this autoantibody enzyme immunoassay method were compared with the conventional antigen determination method by receiver-operating characteristic plots. In the sera, the presence of autoantibody directed against ECPKA was highly correlated with cancer. High anti-ECPKA autoantibody titers (frequency, 90%; mean titer, 3.0) were found in the sera of patients with various cancers, whereas low or negative titers (frequency, 12%; mean titer, 1.0) were found in the control group. The receiver-operating characteristic plot showed that autoantibody enzyme immunoassay exhibited 90% sensitivity and 88% specificity, whereas the enzymatic assay exhibited 83% sensitivity and 80% specificity. These results show that the autoantibody method distinguished between patients with cancer and controls better than the antigen method could. Our results show that autoantibody ECPKA is a universal serum biomarker for cancers of various cell types.


Invariant natural killer T cells in adipose tissue: novel regulators of immune-mediated metabolic disease.
Adipose tissue (AT) represents a microenvironment where intersection takes place between immune processes and metabolic pathways. A variety of immune cells have been characterized in AT over the past decades, with the most recent addition of invariant natural killer T (iNKT) cells. As members of the T cell family, iNKT cells represent a subset that exhibits both innate and adaptive characteristics and directs ensuing immune responses. In disease conditions, iNKT cells have established roles that include disorders in the autoimmune spectrum in malignancies and infectious diseases. Recent work supports a role for iNKT cells in the maintenance of AT homeostasis through both immune and metabolic pathways. The deficiency of iNKT cells can result in AT metabolic disruptions and insulin resistance. In this review, we summarize recent work on iNKT cells in immune regulation, with an emphasis on AT-resident iNKT cells, and identify the potential mechanisms by which adipocytes can mediate iNKT cell activity.


Diagnosis and treatment of cytomegalovirus disease in transplant patients based on gastrointestinal tract manifestations.
Infection due to cytomegalovirus is a substantial cause of morbidity and mortality in immunocompromised patients. In particular, cytomegalovirus infection has been associated with a significant detrimental effect on patient and allograft survival after solid-organ transplantation. We are evaluating a new antiviral agent, ganciclovir 9-[1,3-dihydroxy-2-2 propoxymethyl] guanine (DHPG), used in solid-organ transplant recipients who developed life-threatening cytomegalovirus infections. Between March 1, 1987, and June 30, 1989, we treated 93 solid-organ transplant patients who developed tissue-invasive cytomegalovirus disease. From this group of patients we have identified 14 patients with primary gastrointestinal cytomegalovirus disease who received treatment with DHPG. Tissue diagnosis was made by endoscopy of the upper gastrointestinal tract (11 patients) or colonoscopy (three patients). Invasive cytomegalovirus disease was identified prior to severe complications of the gastrointestinal tract in all but one patient, who suffered colonic perforation prior to treatment with DHPG and subsequently died of bacterial sepsis. While 13 of the 14 patients improved after treatment with DHPG, four patients required additional treatments for recurrent cytomegalovirus disease and recovered. No DHPG toxicity was observed. We believe treatment with DHPG is indicated in this patient population, but that further studies are indicated to fully define the impact of this recommendation on both patient and allograft survival after solid-organ transplantation.


Removal of UroLume endoprosthesis: experience of the North American Study Group for detrusor-sphincter dyssynergia application.
We present the experience of the North American UroLume Multicenter Study Group with removal of the UroLume endoprosthesis. A total of 160 neurologically impaired patients were enrolled in the North American UroLume Multicenter Study Group for detrusor external sphincter dyssynergia application. Analysis was performed in 2 groups of patients in which the device was removed during insertion and after implantation, respectively. Device retrieval was required during insertion in 21 patients (13%) mainly due to misplacement or migration in 17. Extraction was done with minimal complications and in all but 2 cases subsequent UroLume implantation was successful. Of 158 men with the device in place 31 (19.6%) required removal. In 34 procedures 44 devices were removed, mainly due to migration. Time from implantation to removal ranged from 4 days to 66 months (mean 22 months). The UroLume was removed en bloc in 20 cases and in parts or wire by wire in 19. The majority of patients had no or minimal complications after extraction. Only 2 patients had serious temporary complications, including bleeding and urethral injury, with no lasting consequences. No malignancy developed as a result of UroLume insertion. While there is a potential for urethral injury and bleeding, UroLume endoprosthesis removal is largely a simple procedure with minimal complications and consequences.


The impact of having children on domain-specific life satisfaction: A quasi-experimental longitudinal investigation using the Socio-Economic Panel (SOEP) data.
Longitudinal studies have documented improvements in parents' life satisfaction due to childbearing, followed by postpartum adaptation back to baseline. However, the details underlying this process remain largely unexplored. Based on past literature, set-point theory, and results from an exploratory sample, we investigated empirically how first childbirth affected satisfaction with specific domains of life. In a preregistered study, we compared parents with matched childless respondents in their trajectories of life satisfaction, and also satisfaction with family life, health, sleep, work, housework, leisure, dwelling, household income, and personal income. First-time parents and childless respondents were matched in a procedure combining exact and propensity score matching. Using the population-representative German SOEP data (N = 3,370), longitudinal multilevel models revealed heterogeneous effects of childbirth on different domains of satisfaction: Both mothers' and fathers' satisfaction with family life increased temporarily in a similar fashion to life satisfaction before going back to baseline within five years after childbirth. However, only mothers experienced drastic losses to satisfaction with sleep and satisfaction with personal income. For the remaining domains, parents' satisfaction largely resembled that of the matched childless respondents. These divergent domain trajectories underscore the need for multivariate analyses in life satisfaction research. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Impact of genetically modified crops and their management on soil microbially mediated plant nutrient transformations.
One of the potential environmental effects of the recent rapid increase in the global agricultural area cultivated with transgenic crops is a change in soil microbially mediated processes and functions. Among the many essential functions of soil biota are soil organic matter decomposition, nutrient mineralization and immobilization, oxidation-reduction reactions, biological N fixation, and solubilization. However, relatively little research has examined the direct and indirect effects of transgenic crops and their management on microbially mediated nutrient transformations in soils. The objectives of this paper are to review the available literature related to the environmental effects of transgenic crops and their management on soil microbially mediated nutrient transformations, and to consider soil properties and climatic factors that may affect the impact of transgenic crops on these processes. Targeted genetic traits for improved plant nutrition include greater plant tolerance to low Fe availability in alkaline soils, enhanced acquisition of soil inorganic and organic P, and increased assimilation of soil N. Among the potential direct effects of transgenic crops and their management are changes in soil microbial activity due to differences in the amount and composition of root exudates, changes in microbial functions resulting from gene transfer from the transgenic crop, and alteration in microbial populations because of the effects of management practices for transgenic crops, such as pesticide applications, tillage, and application of inorganic and organic fertilizer sources. Possible indirect effects of transgenic crops, including changes in the fate of transgenic crop residues and alterations in land use and rates of soil erosion, deserve further study. Despite widespread public concern, no conclusive evidence has yet been presented that currently released transgenic crops, including both herbicide and pest resistant crops, are causing significant direct effects on stimulating or suppressing soil nutrient transformations in field environments. Further consideration of the effects of a wide range of soil properties, including the amount of clay and its mineralogy, pH, soil structure, and soil organic matter, and variations in climatic conditions, under which transgenic crops may be grown, is needed in evaluating the impact of transgenic crops on soil nutrient transformations. Future environmental evaluation of the impact of the diverse transgenic crops under development could lead to an improved understanding of soil biological functions and processes.


Protein kinases as targets for anti-parasitic chemotherapy.
Parasitic protozoa infecting humans have a staggering impact on public health, especially in the developing world. Furthermore, several protozoan species are major pathogens of domestic animals and have a considerable impact on food production. In many instances, the parasites have developed resistance against available chemotherapeutic agents, making the search for alternative drugs a priority. In line with the current interest in protein kinases inhibitors as potential drugs against a variety of diseases, the possibility that protein kinases may represent targets for novel anti-parasitic agents is being explored. Research into parasite protein kinases has benefited greatly from genome and EST sequencing projects, with the genomes of a few species fully sequenced (notably that of the human malaria parasite Plasmodium falciparum) and several more under way. The overall picture that emerged from research in this area shows that the phylogenetic isolation of parasitic protozoa is reflected by atypical structural and functional properties of many of their protein kinase homologues. Likewise, evidence is emerging, which suggests that the organisation of some otherwise well-conserved signal transduction pathways is divergent in some parasitic species. The differences between protein kinases of a parasite and their homologues in its host cell suggest that specific inhibition of the former can be achieved. The development of anti-parasitic drugs based on protein kinase inhibition is being pursued following two avenues: one consists of screening chemical libraries on recombinant enzymes; several protein kinases from parasitic protozoa are now available for this approach. The second approach relies on the identification of the molecular targets of kinase inhibitors which display anti-parasitic properties. This has led to promising developments in a few instances, in particular regarding PKG as a drug target against Eimeria and Toxoplasma, and purvalanol B, a purine-based CDK inhibitor which appears to affect unexpected targets in several protozoan parasites. The recent resolution of the structure of a Plasmodium protein kinase complexed with small inhibitory molecules opens the way to a rational approach towards the design of anti-parasitic drugs based on kinase inhibition.


Spatially integrated assessment reveals widespread changes in penguin populations on the Antarctic Peninsula.
As important marine mesopredators and sensitive indicators of Antarctic ecosystem change, penguins have been a major focus of long-term biological research in the Antarctic. However, the vast majority of such studies have been constrained by logistics and relate mostly to the temporal dynamics of individual breeding populations from which regional trends have been inferred, often without regard for the complex spatial heterogeneity of population processes and the underlying environmental conditions. Integrating diverse census data from 70 breeding sites across 31 years in a robust, hierarchical analysis, we find that trends from intensely studied populations may poorly reflect regional dynamics and confuse interpretation of environmental drivers. Results from integrated analyses confirm that Pygoscelis adeliae (Adélie Penguins) are decreasing at almost all locations on the Antarctic Peninsula. Results also resolve previously contradictory studies and unambiguously establish that P. antarctica (Chinstrap Penguins), thought to benefit from decreasing sea ice, are instead declining regionally. In contrast, another open-water species, P. papua (Gentoo Penguin), is increasing in abundance and expanding southward. These disparate population trends accord with recent mechanistic hypotheses of biological change in the Southern Ocean and highlight limitations of the influential but oversimplified "sea ice" hypothesis. Aggregating population data at the regional scale also allows us to quantify rates of regional population change in a way not previously possible.


Outcome of intracranial flow diversion according to the antiplatelet regimen used: a systematic review and meta-analysis.
Thromboembolic complications are not uncommon in patients undergoing neurointerventional procedures. The use of flow diverting stents is associated with higher risks of these complications despite current dual antiplatelet regimens. To explore contemporary evidence on the safety of emerging dual antiplatelet regimens in flow diverting stenting procedures. We performed a systematic review and meta-analysis to identify relevant articles in electronic databases, and relevant references. Studies reporting the complications and mortality of flow diverting stenting procedures using acetyl salicylic acid (ASA) + ticagrelor or ASA + prasugrel compared with ASA + clopidogrel were included. Of 452 potentially relevant studies, we identified 49 studies (2526 patients) which reported the safety of ticagrelor or prasugrel for pooled analysis, and five studies (1005 patients) for meta-analysis. The pooled overall mortality in all studies was 2.14%, ischemic complications 6.89%, and hemorrhagic complications 3.68%. The use of ticagrelor or prasugrel was associated with a lower risk of mortality compared with clopidogrel (RR=4.57, 95% CI 1.23 to 16.99; p=0.02). Considering ischemic events, ASA + clopidogrel was as safe as ASA + prasugrel (RR=0.55, 95% CI 0.11 to 2.74; p=0.47) and ASA + ticagrelor (RR=0.74, 95% CI 0.32 to 1.74; p=0.49). ASA +ticagrelor was not associated with a higher risk of hemorrhagic complications (RR=0.92, 95% CI 0.27 to 3.16; p=0.89). Evidence suggests that dual antiplatelet regimens including ticagrelor or prasugrel are safe for patients undergoing flow diversion procedures. Regimens using ticagrelor were associated with better survival than those using clopidogrel in the included studies.


Complementary activities of DOT1L and Menin inhibitors in MLL-rearranged leukemia.
Chromosomal rearrangements of the mixed lineage leukemia (MLL/KMT2A) gene leading to oncogenic MLL-fusion proteins occur in ~10% of acute leukemias and are associated with poor clinical outcomes, emphasizing the need for new treatment modalities. Inhibition of the DOT1-like histone H3K79 methyltransferase (DOT1L) is a specific therapeutic approach for such leukemias that is currently being tested in clinical trials. However, in most MLL-rearranged leukemia models responses to DOT1L inhibitors are limited. Here, we performed deep-coverage short hairpin RNA sensitizer screens in DOT1L inhibitor-treated MLL-rearranged leukemia cell lines and discovered that targeting additional nodes of MLL complexes concomitantly with DOT1L inhibition bears great potential for superior therapeutic results. Most notably, combination of a DOT1L inhibitor with an inhibitor of the MLL-Menin interaction markedly enhanced induction of differentiation and cell killing in various MLL disease models including primary leukemia cells, while sparing normal hematopoiesis and leukemias without MLL rearrangements. Gene expression analysis on human and murine leukemic cells revealed that target genes of MLL-fusion proteins and MYC were suppressed more profoundly upon combination treatment. Our findings provide a strong rationale for a novel targeted combination therapy that is expected to improve therapeutic outcomes in patients with MLL-rearranged leukemia.


Safety and tolerability of chikungunya virus-like particle vaccine in healthy adults: a phase 1 dose-escalation trial.
Chikungunya virus--a mosquito-borne alphavirus--is endemic in Africa and south and southeast Asia and has recently emerged in the Caribbean. No drugs or vaccines are available for treatment or prevention. We aimed to assess the safety, tolerability, and immunogenicity of a new candidate vaccine. VRC 311 was a phase 1, dose-escalation, open-label clinical trial of a virus-like particle (VLP) chikungunya virus vaccine, VRC-CHKVLP059-00-VP, in healthy adults aged 18-50 years who were enrolled at the National Institutes of Health Clinical Center (Bethesda, MD, USA). Participants were assigned to sequential dose level groups to receive vaccinations at 10 μg, 20 μg, or 40 μg on weeks 0, 4, and 20, with follow-up for 44 weeks after enrolment. The primary endpoints were safety and tolerability of the vaccine. Secondary endpoints were chikungunya virus-specific immune responses assessed by ELISA and neutralising antibody assays. This trial is registered with ClinicalTrials.gov, NCT01489358. 25 participants were enrolled from Dec 12, 2011, to March 22, 2012, into the three dosage groups: 10 μg (n=5), 20 μg (n=10), and 40 μg (n=10). The protocol was completed by all five participants at the 10 μg dose, all ten participants at the 20 μg dose, and eight of ten participants at the 40 μg dose; non-completions were for personal circumstances unrelated to adverse events. 73 vaccinations were administered. All injections were well tolerated, with no serious adverse events reported. Neutralising antibodies were detected in all dose groups after the second vaccination (geometric mean titres of the half maximum inhibitory concentration: 2688 in the 10 μg group, 1775 in the 20 μg group, and 7246 in the 40 μg group), and a significant boost occurred after the third vaccination in all dose groups (10 μg group p=0·0197, 20 μg group p<0·0001, and 40 μg group p<0·0001). 4 weeks after the third vaccination, the geometric mean titres of the half maximum inhibitory concentration were 8745 for the 10 μg group, 4525 for the 20 μg group, and 5390 for the 40 μg group. The chikungunya VLP vaccine was immunogenic, safe, and well tolerated. This study represents an important step in vaccine development to combat this rapidly emerging pathogen. Further studies should be done in a larger number of participants and in more diverse populations. Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, and National Institutes of Health.


The association between Toxoplasma gondii infection and hypertensive disorders in T2DM patients: a case-control study in the Han Chinese population.
Type 2 diabetes mellitus (T2DM) is a major global health problem. The rate of infection with Toxoplasma gondii (T. gondii) is more than one-third of the total world population. The effects of T. gondii infection on the risk of diabetic complications and comorbidities are unclear. This study aims to determine the relationship between T. gondii infection and complications of T2DM in the Han Chinese population. We collected 1580 blood samples from T2DM patients and measured the levels of specific IgG antibodies against T. gondii in the sera of these patients using an ELISA assay. A logistic regression analysis was performed to estimate the effect of T. gondii infection on the complications of T2DM, while adjusting for age, gender, and triglyceride level (TG). We applied the multifactor dimensionality reduction (MDR) method to detect the interactions between T. gondii infections, demographic indexes and biochemical indicators among the different complications. Gender (the odds ratio (OR) = 0.63, 95%CI =0.45-0.89, P = 0.008) and TG level (OR = 0.64, 95%CI =0.45-0.89, P = 0.009) were influencing factors in T. gondii infections. T2DM patients who were infected with T. gondii had a 2.34 times risk of developing hypertension than those patients without T. gondii infection (OR = 2.34, 95%CI = 1.12-4.88, P = 0.024). The multiplicative interaction analysis and the additive interaction analysis did not reveal any evidence of interactive effects on diabetic complications and comorbidities. T. gondii might be a factor associated with hypertension in T2DM patients.


Ontogeny of regulatory mechanisms for beta-adrenoceptor control of rat cardiac adenylyl cyclase: targeting of G-proteins and the cyclase catalytic subunit.
Fetal and neonatal tissues are resistant to catecholamine-induced desensitization of essential physiological responses. We examined the mechanisms underlying the ontogeny of desensitization in neonatal rat heart for the beta-adrenergic receptor/adenylyl cyclase signaling cascade. Animals of different ages received isoproterenol daily or 4 days and cardiac membrane preparations were evaluated on the 5th day (6, 15, 25 days old and adults). Measurements were made of basal activity, activity stimulated by two agonists (isoproterenol or glucagon) that operate at different receptors but that share Gs as the transduction intermediate, or by forskolin-Mn' to assess total catalytic capacity of the cyclase subunit; we also assessed inhibition of activity by carbachol which acts via muscarinic cholinergic receptors and G. Adult rats exhibited robust desensitization of the adenylyl cyclase response but the effect was heterologous in that equivalent loss of activity was seen for basal, isoproterenol- and glucagon-stimulated activity forskolin-Mn(2+)-stimulated activity was also decreased. Two factors contributed to desensitization; generalized reduction in membrane protein concentrations caused by cell enlargement (reduced surface-to-volume ratio), and specific interference with the G-protein component that couples receptors to the cyclase. Thus, after adjustment for changes in membrane protein, the desensitization of the forskolin-Mn2, response was no longer evident, but the effects on the other measures were still present. In addition, isoproterenol treatment produced crosstalk with the carbachol/Gi signaling pathway, with significant reductions in the ability of carbachol to inhibit adenylyl cyclase activity. Heterologous desensitization by isoproterenol was also present in 15 and 25 day old rats, but involved only selective components of the effects seen in adults. At 25 days, uncoupling of signals operating through Gs and Gi was obtained without a reduction in forskolin-Mn(2+)-stimulated activity. At 15 days, only the effect on Gs coupling was seen. At 6 days, agonist-induced desensitization was not detectable and instead, heterologous sensitization was found. In these youngest animals, isoproterenol treatment produced a parallel increase in basal, isoproterenol-, glucagon- and forskolin-Mn(2+)-stimulated activity, unaccompanied by changes in membrane protein concentrations, indicating an increase in adenylyl cyclase catalytic activity. These results indicate that the ability to elicit desensitization is not an inherent property of cardiac cells but rather is acquired in distinct stages during development. Sensitization by agonists early in development may be important in preserving physiological responsiveness during ontogenetic surges of adrenergic activity.


Time trends in coroners' use of different verdicts for possible suicides and their impact on officially reported incidence of suicide in England: 1990-2005.
Official suicide statistics for England are based on deaths given suicide verdicts and most cases given an open verdict following a coroner's inquest. Previous research indicates that some deaths given accidental verdicts are considered to be suicides by clinicians. Changes in coroners' use of different verdicts may bias suicide trend estimates. We investigated whether suicide trends may be over- or underestimated when they are based on deaths given suicide and open verdicts. Method Possible suicides assessed by 12 English coroners in 1990/91, 1998 and 2005 and assigned open, accident/misadventure or narrative verdicts were rated by three experienced suicide researchers according to the likelihood that they were suicides. Details of all suicide verdicts given by these coroners were also recorded. In 1990/91, 72.0% of researcher-defined suicides received a suicide verdict from the coroner, this decreased to 65.4% in 2005 (p trend < 0.01); equivalent figures for combined suicide and open verdicts were 95.4% (1990/91) and 86.7% (2005). Researcher-defined suicides with a verdict of accident/misadventure doubled over that period, from 4.6% to 9.1% (p < 0.01). Narrative verdict cases rose from zero in 1990/91 to 25 in 2005 (4.2% of researcher-defined suicides that year). In 1998 and 2005, 50.0% of the medicine poisoning deaths given accidental/misadventure verdicts were rated as suicide by the researchers. Between 1990/91 and 2005, the proportion of researcher-defined suicides given a suicide verdict by coroners decreased, largely due to an increased use of accident/misadventure verdicts, particularly for deaths involving poisoning. Consideration should be given to the inclusion of 'accidental' deaths by poisoning with medicines in the statistics available for monitoring suicides rates.


Evaluating the Psychometric Properties of the Eating Assessment Tool (EAT-10) Using Rasch Analysis.
Early and reliable screening for oropharyngeal dysphagia (OD) symptoms in at-risk populations is important and a crucial first stage in effective OD management. The Eating Assessment Tool (EAT-10) is a commonly utilized screening and outcome measure. To date, studies using classic test theory methodologies report good psychometric properties, but the EAT-10 has not been evaluated using item response theory (e.g., Rasch analysis). The aim of this multisite study was to evaluate the internal consistency and structural validity and conduct a preliminary investigation of the cross-cultural validity of the EAT-10; floor and ceiling effects were also checked. Participants involved 636 patients deemed at risk of OD, from outpatient clinics in Spain, Turkey, Sweden, and Italy. The EAT-10 and videofluoroscopic and/or fiberoptic endoscopic evaluation of swallowing were used to confirm OD diagnosis. Patients with esophageal dysphagia were excluded to ensure a homogenous sample. Rasch analysis was used to investigate person and item fit statistics, response scale, dimensionality of the scale, differential item functioning (DIF), and floor and ceiling effect. The results indicate that the EAT-10 has significant weaknesses in structural validity and internal consistency. There are both item redundancy and lack of easy and difficult items. The thresholds of the rating scale categories were disordered and gender, confirmed OD, and language, and comorbid diagnosis showed DIF on a number of items. DIF analysis of language showed preliminary evidence of problems with cross-cultural validation, and the measure showed a clear floor effect. The authors recommend redevelopment of the EAT-10 using Rasch analysis.


Nitrogen addition pulse has minimal effect in big sagebrush (Artemisia tridentata) communities on the Pinedale Anticline, Wyoming (USA).
Nitrogen additions are known to elicit variable responses in semi-arid ecosystems, with responses increasing with precipitation. The response of semi-arid ecosystems to nitrogen are important to understand due to their large spatial extent worldwide and the global trend of increasingly available nitrogen. In this study, we evaluated the impact of a single nitrogen addition pulse on a semi-arid big sagebrush (Artemisia tridentata) ecosystem in western Wyoming. This is important given that sagebrush ecosystems are poorly understood, despite their prevalence in the western US. In addition, large-scale nitrogen additions have begun on sagebrush landscapes in Wyoming in order to mitigate population declines in mule deer (Odocoileus hemionus). The study objectives were (1) to evaluate the effectiveness of a nitrogen fertilization pulse in increasing sagebrush biomass and forage quality, and (2) to assess effects of nitrogen addition on soil biogeochemistry and vegetation community structure. We fertilized 15 plots across 5 locations in western Wyoming using a single pulse of urea (5.5g N m-2). In addition, we immobilized available nitrogen through surface hay treatments (250g hay/m2). Nitrogen additions failed to increase growth of sagebrush, alter nitrogen content of sagebrush leaders, or alter greenhouse gas efflux from soils. The plant community also remained unchanged; total cover, species richness, and community composition were all unaffected by our treatment application. Over the two years of this study, we did not find indications of nitrogen limitation of ecosystem processes, despite a wet growing season in 2014. Thus, we have found a general lack of response to nitrogen in sagebrush ecosystems and no treatment effect of a single pulse of N to sagebrush biomass or forage quality.


Type 1 Diabetes and the HLA Region: Genetic Association Besides Classical HLA Class II Genes.
Type 1 diabetes is an autoimmune disease with rising incidence in high-income countries. Genetic and environmental predisposing factors contribute to the etiology of the disease, although their interaction is not sufficiently understood to allow for preventive action. Strongest known associations with genetic variation map to classical HLA class II genes. Because of its genetic complexity, the HLA region has been under-represented in genome-wide association studies, having potentially hindered the identification of relevant associations underlying the etiology of the disease. Here, we performed a comprehensive HLA-wide genetic association analysis of type 1 diabetes including multi-allelic and rare variants. We used high-density whole-exome sequencing data of the HLA region in the large UK Biobank dataset to apply gene-based association tests with a carefully defined type 1 diabetes phenotype (97 cases and 48,700 controls). Exon-based and single-variant association tests were used to complement the analysis. We replicated the known association of type 1 diabetes with the classical HLA-DQ gene. Tailoring the analysis toward rare variants, we additionally identified the lysine methyl transferase EHMT2 as associated. Deeper insight into genetic variation associated with disease as presented and discussed in detail here can help unraveling mechanistic details of the etiology of type 1 diabetes. More specifically, we hypothesize that genetic variation in EHMT2 could impact autoimmunity in type 1 diabetes development.


Nailfold Capillaroscopy and Retinal Findings in Patients with Systemic Sclerosis: Is There An Association?
Systemic sclerosis (SSc) is characterized by fibrosis and intimal proliferation of cutaneous and visceral small vessels. These architectural abnormalities can be visualized with nailfold capillaroscopy (NFC); the changes being quite characteristic. At the same time, morphological alterations in retinal vascular bed are expected but sparsely described. We aimed to characterize the frequency and type of retinal microvascular changes in patients with SSc and to analyze any association with NFC changes. With institutional ethical committee approval, we recruited 45 consecutive patients with SSc (diagnosed based on American College of Rheumatology and European League against Rheumatism [ACR/EULAR-2013] criteria). NFC was done for all of them with a Universal Serial Bus (USB) dermatoscope; additionally, fundoscopy, fundus photography, and optical coherence tomography (OCT) were analyzed. Disease characteristics in patients with and without retinal disease were compared. Among the 45 SSc patients, 12 (26.67%) had limited cutaneous SSc (lSSc) while 33 (73.33%) had diffuse cutaneous disease (dSSc). Retinal microvascular changes seen as mild arteriolar alteration and arteriovenous crossing changes were recorded in 13 patients (28.89%); mostly in those with dSSc (12/13). The NFC architectural changes were more severe in patients with retinal disease, though the difference was not statistically significant. Patients with SSc can often have retinal microvascular abnormalities commensurate with the vascular changes characteristic of SSc. The severity of retinal changes correlates with changes in NFC. NFC, which is now an essential tool for the management of SSc, could be a surrogate marker for retinal involvement in these patients.


Twenty-year trajectories of alcohol consumption during midlife and atherosclerotic thickening in early old age: findings from two British population cohort studies.
Epidemiological evidence indicates a protective effect of light-moderate drinking on cardiovascular disease and an increased risk for heavier drinking. Nevertheless, the effect of alcohol on atherosclerotic changes in vessel walls is disputed. Most previous studies have only looked at the cross-sectional relationship between alcohol and carotid intima media thickness (cIMT) - a surrogate marker of atherosclerosis. Single measurements of alcohol assume that alcohol exposure is stable and ignore the possible cumulative effects of harm, leading to possibly incorrect inferences. Data were retrieved from two UK population based cohort studies: the Whitehall II cohort of civil servants and the MRC National Survey of Health and Development (combined sample size of 5403 men and women). Twenty year-drinking trajectories during midlife were linked to measures of cIMT when participants were in early old age, and adjusted for age, sex, socioeconomic position, ethnicity and smoking. Those who consistently drank heavily had an increased cIMT compared to stable moderate drinkers (pooled difference in cIMT 0.021 mm; 95 % CI 0.002 to 0.039), after adjustment for covariates. This was not detected in cross-sectional analyses. Former drinkers also had an increased cIMT compared to moderate drinkers (pooled difference in cIMT 0.021; 95 % CI 0.005 to 0.037). There were no appreciable differences in cIMT between non-drinkers and consistent moderate drinkers. The drinking habits among adults during midlife affect the atherosclerotic process and sustained heavy drinking is associated with an increased cIMT compared to stable moderate drinkers. This finding was not seen when only using cross-sectional analyses, thus highlighting the importance of taking a life course approach. There was no evidence of a favourable atherosclerotic profile from stable moderate drinking compared to stable non-drinking.


Identification of disulfide reductases in Campylobacterales: a bioinformatics investigation.
Disulfide reductases of host-colonising bacteria are involved in the expression of virulence factors, resistance to drugs, and elimination of toxic compounds. Large-scale genome analyses of 281 prokaryotes identified CXXC and CXXC-derived motifs in each microorganism. The total number of these motifs showed correlations with genome size and oxygen tolerance of the prokaryotes. Specific bioinformatic analyses served to identify putative disulfide reductases in the Campylobacterales Campylobacter jejuni, Helicobacter pylori, Wolinella succinogenes and Arcobacter butzleri which colonise the gastrointestinal tract of higher animals. Three filters applied to the genomes of these species yielded 35, 25, 28 and 34 genes, respectively, encoding proteins with the characteristics of disulfide reductases. Ten proteins were common to the four species, including four belonging to the thioredoxin system. The presence of thioredoxin reductase activities was detected in the four bacterial species by observing dithiobis-2-nitrobenzoic acid reduction with beta-nicotinamide adenine dinucleotide phosphate as cofactor. Phylogenetic analyses of the thioredoxin reductases TrxB(1) and TrxB(2) of the four Campylobacterales were performed. Their TrxB(1) proteins were more closely related to those of Firmicutes than to the corresponding proteins of other Proteobacteria. The Campylobacterales TrxB(2) proteins were closer to glutathione reductases of other organisms than to their respective TrxB(1) proteins. The phylogenetic features of the Campylobacterales thioredoxin reductases suggested a special role for these enzymes in the physiology of these bacteria.


Short RNA half-lives in the slow-growing marine cyanobacterium Prochlorococcus.
RNA turnover plays an important role in the gene regulation of microorganisms and influences their speed of acclimation to environmental changes. We investigated whole-genome RNA stability of Prochlorococcus, a relatively slow-growing marine cyanobacterium doubling approximately once a day, which is extremely abundant in the oceans. Using a combination of microarrays, quantitative RT-PCR and a new fitting method for determining RNA decay rates, we found a median half-life of 2.4 minutes and a median decay rate of 2.6 minutes for expressed genes - twofold faster than that reported for any organism. The shortest transcript half-life (33 seconds) was for a gene of unknown function, while some of the longest (approximately 18 minutes) were for genes with high transcript levels. Genes organized in operons displayed intriguing mRNA decay patterns, such as increased stability, and delayed onset of decay with greater distance from the transcriptional start site. The same phenomenon was observed on a single probe resolution for genes greater than 2 kb. We hypothesize that the fast turnover relative to the slow generation time in Prochlorococcus may enable a swift response to environmental changes through rapid recycling of nucleotides, which could be advantageous in nutrient poor oceans. Our growing understanding of RNA half-lives will help us interpret the growing bank of metatranscriptomic studies of wild populations of Prochlorococcus. The surprisingly complex decay patterns of large transcripts reported here, and the method developed to describe them, will open new avenues for the investigation and understanding of RNA decay for all organisms.


Impact of Caregiver Type for 3-Year-Old Children on Subsequent Between-Meal Eating Habits and Being Overweight From Childhood to Adulthood: A 20-Year Follow-up of the Ibaraki Children's Cohort (IBACHIL) Study.
Because of the increasing number of mothers who continue to work after childbirth, participation in childcare has diversified. However, the impact of the main caregiver on children's habits has not been determined. We sought to examine the effect of caregiver differences on childhood habituation of between-meal eating and body mass index (BMI). The Ibaraki Children's Cohort Study involved 4592 Japanese children whose parents answered health questionnaires at age 3. Follow-up questionnaires were distributed to parents when children were 6 and 12 years old and to study subjects directly when they were 22 years old. We compared prevalence of between-meal eating and overweight as well as mean BMI at ages 6, 12, and 22 years, by their main daytime caregiver at age 3. Compared to children cared for by mothers, those cared for by grandparents had a higher prevalence of between-meal eating before dinner for boys and girls at ages 6 and 12 years. At age 22 years, boys cared for by grandparents had a higher prevalence of overweight than those cared for by mothers (18.5% versus 11.2%, P = 0.037), but no such difference was noted in girls. However, both boys and girls cared for by grandparents had higher mean BMI over time than those cared for by mothers (coefficient = 0.47 kg/m(2) for boys and coefficient = 0.35 kg/m(2) for girls). Being cared for by grandparents at age 3 was associated with subsequent between-meal eating habits, being overweight, and increased mean BMI from childhood to adulthood.


Adolescent sibling relationships in Mexican American families: exploring the role of familism.
To address a significant gap in the literature on normative processes in minority families, the authors studied adolescents' sibling relationships in two-parent Mexican American families and explored connections between sibling relationship characteristics and familism. Participants were 246 adolescent Mexican American sibling pairs who participated in (a) home interviews during which adolescents described their sibling relationships and familism values and (b) a series of 7 nightly phone calls during which adolescents reported their daily activities, including time spent with siblings and family members. Siblings described their relationships as both intimate and conflictual, and daily activity data revealed that they spent an average of 17.2 hr per 7 days in shared activities. Sibling relationship qualities were linked to familism values and practices, and stronger patterns of association emerged for sisters than brothers. Discussion highlights the significance of studying the processes that underlie within-group variations among families of different cultural backgrounds.


Crowding, intermolecular interactions, and shear flow effects in the diffusion model of chemical reactions.
In the diffusion model of chemical reactions, the encounter (reaction) rate between reactant particles is governed by the Smoluchowski equation, which is a diffusion equation in a field of forces. We consider crowded environments where the particles diffuse through a "liquid" of like particles. Assuming that the liquid-like short-range structure around the reactant gives rise to an effective (osmotic) barrier leads us to map a complicated many-body problem to a one-dimensional problem. This allows us to describe theoretically such complex systems that are encountered in many applications where crowding, intermolecular interactions, and flow are simultaneously present. A particularly important effect is discovered which is due to the interplay between shear and crowding. This effect is responsible for unexpected peaks in the reactivity at low flow intensity which may explain, among other things, the bizarre colloidal stability behavior of concentrated protein suspensions.


Antitumor effects of Osthol from Cnidium monnieri: an in vitro and in vivo study.
Cnidium monnieri (L.) Cusson is a Chinese medicine which is used widely by traditional medicine doctors. Osthol is a major bio-activity compound of the herb. In this study, osthol was isolated from C. monnieri and its in vitro and in vivo antitumor effects studied. The results of the in vitro study showed: that osthol inhibited the growth of HeLa, in a time- and concentration-dependent manner, with IC(50) values of 77.96 and 64.94 microm for 24 and 48 h, respectively; that osthol had lower cytotoxic effects in primary cultured normal cervical fibroblasts; and that increased DNA fragmentation and activated PARP in HeLa after treatment with osthol which could induce apoptosis. The results of the in vivo model showed that the survival days of the P-388 D1 tumor-bearing CDF(1) mice were prolonged (ILS% = 37) after osthol (30 mg/kg) was given once a day for 9 days. Based on these results, it is suggested that osthol could inhibit P-388 D1 cells in vivo and induce apoptosis in HeLa cells in vitro, and that osthol is good lead compound for developing antitumor drugs. However, C. formosanum Yabe of Taiwan's endemic plants contained little osthol, with no imperatorin, and its major components were different from that of C. monnieri. Therefore, it is suggested that C. formosanum also may possess economic worth.


A Machine Learning Approach for Drug-target Interaction Prediction using Wrapper Feature Selection and Class Balancing.
Drug-Target interaction (DTI) plays a crucial role in drug discovery, drug repositioning and understanding the drug side effects which helps to identify new therapeutic profiles for various diseases. However, the exponential growth in the genomic and drugs data makes it difficult to identify the new associations between drugs and targets. Therefore, we use computational methods as it helps in accelerating the DTI identification process. Usually, available data driven sources consisting of known DTI is used to train the classifier to predict the new DTIs. Such datasets often face the problem of class imbalance. Therefore, in this study we address two challenges faced by such datasets, i. e., class imbalance and high dimensionality to develop a predictive model for DTI prediction. The study is carried out on four protein classes namely Enzyme, Ion Channel, G Protein-Coupled Receptor (GPCR) and Nuclear Receptor. We encoded the target protein sequence using the dipeptide composition and drug with a molecular descriptor. A machine learning approach is employed to predict the DTI using wrapper feature selection and synthetic minority oversampling technique (SMOTE). The ensemble approach achieved at the best an accuracy of 95.9 %, 93.4 %, 90.8 % and 90.6 % and 96.3 %, 92.8 %, 90.1 %, and 90.2 % of precision on Enzyme, Ion Channel, GPCR and Nuclear Receptor datasets, respectively, when evaluated excluding SMOTE samples with 10-fold cross validation. Furthermore, our method could predict new drug-target interactions not contained in training dataset. Selected features using wrapper feature selection may be important to understand the DTI for the protein categories under this study. Based on our evaluation, the proposed method can be used for understanding and identifying new drug-target interactions. We provide the readers with a standalone package available at https://github.com/shwetagithub1/predDTI which will be able to provide the DTI predictions to user for new query DTI pairs.


Three Patterns of Cutaneous Involvement in Granulomatosis with Polyangiitis.
Skin involvement in granulomatosis with polyangiitis (GPA) is common and can appear as an initial presentation of the disease or more commonly through its course. We report a case of a 24-year-old male patient, previously diagnosed as having GPA, admitted with fever, hemoptysis, generalized hemorrhagic blisters associated with arthralgia, fatigue, myalgia, nasal crusting, and vertigo. Three weeks prior to admission, he developed erythematous papules on both elbows, and purpuric papules on both lower limbs. Histopathological examination revealed: interstitial granulomatous dermatitis (elbows) and foci of dermal hemorrhage, foci of interstitial histiocytes and zones of altered necrobiotic collagen (lower limbs) consistent with cutaneous lesions of GPA. Two weeks later, his rash progressed to widespread purpura associated with hemorrhagic blisters. Another biopsy revealed leukocytoclastic vasculitis with fibrinoid necrosis of the vessel walls associated with perivascular infiltrate of neutrophils, nuclear dust and extravasated erythrocytes without an associated granulomatous inflammation or necrobiosis. The constellation of the results of the three biopsies together with clinical correlation pointed to a flare of GPA. Skin involvement in GPA is quite common, and it can manifest in different forms in the same patient. Our patient developed three different skin pathologies within a short period of time.


BCL::EMAS--enantioselective molecular asymmetry descriptor for 3D-QSAR.
Stereochemistry is an important determinant of a molecule's biological activity. Stereoisomers can have different degrees of efficacy or even opposing effects when interacting with a target protein. Stereochemistry is a molecular property difficult to represent in 2D-QSAR as it is an inherently three-dimensional phenomenon. A major drawback of most proposed descriptors for 3D-QSAR that encode stereochemistry is that they require a heuristic for defining all stereocenters and rank-ordering its substituents. Here we propose a novel 3D-QSAR descriptor termed Enantioselective Molecular ASymmetry (EMAS) that is capable of distinguishing between enantiomers in the absence of such heuristics. The descriptor aims to measure the deviation from an overall symmetric shape of the molecule. A radial-distribution function (RDF) determines a signed volume of tetrahedrons of all triplets of atoms and the molecule center. The descriptor can be enriched with atom-centric properties such as partial charge. This descriptor showed good predictability when tested with a dataset of thirty-one steroids commonly used to benchmark stereochemistry descriptors (r² = 0.89, q² = 0.78). Additionally, EMAS improved enrichment of 4.38 versus 3.94 without EMAS in a simulated virtual high-throughput screening (vHTS) for inhibitors and substrates of cytochrome P450 (PUBCHEM AID891).


Medication continuation and compliance: a comparison of patients treated with clozapine and haloperidol.
This study compares medication continuation and regimen compliance with the atypical antipsychotic medication clozapine versus the conventional antipsychotic haloperidol. Data from a 15-site double-blind, randomized clinical trial (N = 423) were used to compare patients with DSM-III-R schizophrenia assigned to clozapine or haloperidol in terms of duration of participation while taking the randomly assigned study drug (continuation) and the proportion of prescribed pills that were taken (compliance). Multiple regression analysis was used to determine the relationship of baseline characteristics and measures of clinical change to continuation for the entire sample and for patients assigned to each medication. Patients assigned to clozapine continued taking the study drug for a mean of 35.5 weeks as compared with only 27.2 among patients assigned to haloperidol (F = 4.45, df = 1,422; p = .0001). No differences were found between the groups in the proportion of prescribed pills that were returned at any timepoint. Among patients assigned to haloperidol, poorer continuation was associated with being older and greater continuation with receiving public support. Among patients on clozapine treatment, continuation was poorer among African American patients and greater among patients who showed reduced clinical symptoms and akathisia. Continuation with clozapine was greater even after adjusting for these factors. Continuation with medication is greater with clozapine than haloperidol and is partly explained by greater symptom improvement and reduced side effects. No differences were found in regimen compliance.


Staphylococcus hyicus virulence in relation to exudative epidermitis in pigs.
Staphylococcus hyicus strains with different phage types, plasmid profiles, and antibiotic resistance patterns were isolated from piglets with exudative epidermitis. The strains could be divided into virulent strains, producing exudative epidermitis, and avirulent strains, producing no dermal changes when injected in experimental piglets. The results showed that both virulent and avirulent strains were present simultaneously on diseased piglets. This constitutes a diagnostic problem. Concentrated culture supernatants from nine virulent strains injected in the skin of healthy piglets produced a crusting reaction in all piglets. Acanthosis was observed in the histopathological examination of the crustaceous skin. Concentrated culture supernatants from nine avirulent strains produced no macroscopic or microscopic skin changes. Protein profiles from all virulent strains and seven out of nine avirulent strains showed a high degree of protein band homology. An approximately 30 kDa protein present in all concentrated culture supernatants capable of producing skin changes, could not be detected in samples that did not produce skin changes. No other protein showed a similar association. It is concluded that crusting reaction of piglet skin is a suitable indicator of virulence in S. hyicus in relation to exudative epidermitis, and that virulent strains produce a 30 kDa protein, absent in concentrated culture supernatants from avirulent strains. This 30 kDa protein might be an exfoliative toxin.


Vision and perception of community on the use of recycled water for household laundry: a case study in Australia.
This study investigates the community perception of household laundry as a new end use of recycled water in three different locations of Australia through a face to face questionnaire survey (n=478). The study areas were selected based on three categories of (1) non-user, (2) perspective user and (3) current user of recycled water. The survey results indicate that significantly higher number (70%) of the respondents supported the use of recycled water for washing machines (χ(2)=527.40, df=3; p=0.000). Significant positive correlation between the overall support for the new end use and the willingness of the respondents to use recycled water for washing machine was observed among all users groups (r=0.43, p=0.000). However, they had major concerns regarding the effects of recycled water on the aesthetic appearance of cloth, cloth durability, machine durability, odour of the recycled water and cost along with the health issues. The perspective user group had comparatively more reservations and concerns about the effects of recycled water on washing machines than the non-users and the current users (χ(2)=52.73, df=6; p=0.000). Overall, community from all three study areas are willing to welcome this new end use as long as all their major concerns are addressed and safety is assured.


An updated protocol to detect invalid entries in an online survey of men who have sex with men (MSM): how do valid and invalid submissions compare?
Researchers use protocols to screen for suspicious survey submissions in online studies. We evaluated how well a de-duplication and cross-validation process detected invalid entries. Data were from the Sexually Explicit Media Study, an Internet-based HIV prevention survey of men who have sex with men. Using our protocol, 146 (11.6 %) of 1254 entries were identified as invalid. Most indicated changes to the screening questionnaire to gain entry (n = 109, 74.7 %), matched other submissions' payment profiles (n = 56, 41.8 %), or featured an IP address that was recorded previously (n = 43, 29.5 %). We found few demographic or behavioral differences between valid and invalid samples, however. Invalid submissions had lower odds of reporting HIV testing in the past year (OR 0.63), and higher odds of requesting no payment compared to check payments (OR 2.75). Thus, rates of HIV testing would have been underestimated if invalid submissions had not been removed, and payment may not be the only incentive for invalid participation.


Genetic Factors Interact With Tobacco Smoke to Modify Risk for Inflammatory Bowel Disease in Humans and Mice.
The role of tobacco smoke in the etiology of inflammatory bowel disease (IBD) is unclear. We investigated interactions between genes and smoking (gene-smoking interactions) that affect risk for Crohn's disease (CD) and ulcerative colitis (UC) in a case-only study of patients and in mouse models of IBD. We used 55 Immunochip-wide datasets that included 19,735 IBD cases (10,856 CD cases and 8879 UC cases) of known smoking status. We performed 3 meta-analyses each for CD, UC, and IBD (CD and UC combined), comparing data for never vs ever smokers, never vs current smokers, and never vs former smokers. We studied the effects of exposure to cigarette smoke in Il10-/- and Nod2-/- mice, as well as in Balb/c mice without disruption of these genes (wild-type mice). Mice were exposed to the smoke of 5 cigarettes per day, 5 days a week, for 8 weeks, in a ventilated smoking chamber, or ambient air (controls). Intestines were collected and analyzed histologically and by reverse transcription polymerase chain reaction. We identified 64 single nucleotide polymorphisms (SNPs) for which the association between the SNP and IBD were modified by smoking behavior (meta-analysis Wald test P < 5.0 × 10-5; heterogeneity Cochrane Q test P > .05). Twenty of these variants were located within the HLA region at 6p21. Analysis of classical HLA alleles (imputed from SNP genotypes) revealed an interaction with smoking. We replicated the interaction of a variant in NOD2 with current smoking in relation to the risk for CD (frameshift variant fs1007insC; rs5743293). We identified 2 variants in the same genomic region (rs2270368 and rs17221417) that interact with smoking in relation to CD risk. Approximately 45% of the SNPs that interact with smoking were in close vicinity (≤1 Mb) to SNPs previously associated with IBD; many were located near or within genes that regulate mucosal barrier function and immune tolerance. Smoking modified the disease risk of some variants in opposite directions for CD vs UC. Exposure of Interleukin 10 (il10)-deficient mice to cigarette smoke accelerated development of colitis and increased expression of interferon gamma in the small intestine compared to wild-type mice exposed to smoke. NOD2-deficient mice exposed to cigarette smoke developed ileitis, characterized by increased expression of interferon gamma, compared to wild-type mice exposed to smoke. In an analysis of 55 Immunochip-wide datasets, we identified 64 SNPs whose association with risk for IBD is modified by tobacco smoking. Gene-smoking interactions were confirmed in mice with disruption of Il10 and Nod2-variants of these genes have been associated with risk for IBD. Our findings from mice and humans revealed that the effects of smoking on risk for IBD depend on genetic variants.


Alzheimer's disease brain-derived amyloid-β-mediated inhibition of LTP in vivo is prevented by immunotargeting cellular prion protein.
Synthetic amyloid-β protein (Aβ) oligomers bind with high affinity to cellular prion protein (PrP(C)), but the role of this interaction in mediating the disruption of synaptic plasticity by such soluble Aβ in vitro is controversial. Here we report that intracerebroventricular injection of Aβ-containing aqueous extracts of Alzheimer's disease (AD) brain robustly inhibits long-term potentiation (LTP) without significantly affecting baseline excitatory synaptic transmission in the rat hippocampus in vivo. Moreover, the disruption of LTP was abrogated by immunodepletion of Aβ. Importantly, intracerebroventricular administration of antigen-binding antibody fragment D13, directed to a putative Aβ-binding site on PrP(C), prevented the inhibition of LTP by AD brain-derived Aβ. In contrast, R1, a Fab directed to the C terminus of PrP(C), a region not implicated in binding of Aβ, did not significantly affect the Aβ-mediated inhibition of LTP. These data support the pathophysiological significance of SDS-stable Aβ dimer and the role of PrP(C) in mediating synaptic plasticity disruption by soluble Aβ.


A functional micro-electrode mapping of ventral thalamus in essential tremor.
Deep brain stimulation enables the delivery of therapeutic interventions to otherwise inaccessible areas of the brain while, at the same time, offering the unique opportunity to record from these same regions in awake patients. The posterior ventrolateral thalamus has become a reliable deep brain stimulation target for medically-refractory patients suffering from essential tremor. However, the contribution of the thalamus in essential tremor, and even whether posterior ventrolateral thalamus is the optimal target, remains a matter of ongoing debate. There are several lines of evidence supporting clusters of activity within the posterior ventrolateral thalamus that are important for tremor emergence. In this study we sought to map the functional properties of these clusters through microelectrode recordings during deep brain stimulation surgery. Data were obtained from 10 severely affected patients with essential tremor (12 hemispheres) undergoing deep brain stimulation surgery. Our results demonstrate power and coherence maxima located in the inferior posterior ventrolateral thalamus and immediate ventral region. Moreover, we identified distinct yet overlapping clusters of predominantly efferent (driving) and afferent (feedback) activity, with a preference for more efferent contributors, consistent with a net role in the driving of tremor output. Finally, we demonstrate that resolvable thalamic spiking activity directly relates to background activity and that the strength of tremor may be dictated by phase relationships between efferent and afferent pockets in the posterior ventrolateral thalamus. Taken together, these results provide important evidence for the role of the inferior posterior ventrolateral thalamus and its border region in essential tremor pathophysiology. Such results progress our mechanistic understanding and promote the adoption of next-generation therapies such as high resolution segregated deep brain stimulation electrodes.


Emiliania huxleyi coccolith calcite mass modulation by morphological changes and ecology in the Mediterranean Sea.
To understand the response of marine calcifying organisms under high CO2 scenarios, it is critical to study their calcification patterns in the natural environment. This paper focuses on a major calcifying phytoplankton group, the coccolithophores, through the analysis of water samples collected along a W-E Mediterranean transect during two research cruises, in April 2011 (Meteor cruise M84/3) and May 2013 (MedSeA cruise 2013). The Mediterranean Sea is a marginal sea characterized by large biogeochemical gradients. Currently, it is undergoing both warming and ocean acidification, processes which are rapidly modifying species distribution and calcification. The species Emiliania huxleyi largely dominates the total coccolithophore production in present day oceans and marine basins, including the Mediterranean Sea. A series of morphometric measurements were performed on the coccoliths of this species to estimate their mass, length and calculate a calcification index (proxy for the size-normalized calcification degree). The most abundant morphotype of E. huxleyi in the Mediterranean Sea is Type A. Coccoliths of this morphotype were additionally analyzed based on scanning electron microscopy images: four calcification varieties were quantified, according to the relationship between slit length-tube width, and the state of the central area (open or closed). The average E. huxleyi coccolith mass along the Mediterranean oceanographic transect depended strongly on both the average coccolith length and calcification index. The variability in average coccolith length and calcification index across samples reflected oscillations in the relative abundance of the calcification varieties. We also demonstrated that the distribution of the calcification varieties followed the main environmental gradients (carbonate chemistry, salinity, temperature, nutrient concentrations). Hence, shifts in the distribution of the calcification varieties and of the average E. huxleyi coccolith mass are to be expected in the Mediterranean Sea under climate change. These physiological and ecological responses will modulate the net coccolithophore calcification and, ultimately, the regional carbonate export to the seafloor.


Implementing an SMS-based intervention for persons living with human immunodeficiency virus.
This article describes a process evaluation conducted as part of a proof-of-concept study to develop, implement, and test a text messaging program to promote medication and appointment adherence, sexual and substance use risk reduction, general health and well-being, social support, and patient involvement. The text-messaging program was implemented in Chicago, Illinois, at an outpatient medical clinic that promotes the well-being of gay, lesbian, bisexual, and transgender persons. We collected and analyzed qualitative data from patients, providers, and research staff to answer the following questions: (1) What factors of the organizational context were important for implementation? (2) How are implementation policies and practices, organizational climate, and perceptions of implementation effectiveness described by intervention stakeholders? (3) What types of issues related to fidelity occurred during implementation? (4) What recommendations for improvement do stakeholders suggest? The study coordinator, providers, and the patients themselves confirmed that patients liked the messages and program overall. The program was implemented with high fidelity. The primary recommendations for improvements were to enhance confidentiality and implement strategies to lessen message fatigue. The findings from this process evaluation demonstrate the feasibility and acceptability of the intervention from the perspectives of patients, providers, and research staff. A larger-scale intervention study that incorporates these stakeholders' suggestions for improvement is warranted.


[Survey on bioethics in a pediatric hospital. Basis for new strategies].
Although medical ethics is part of medicine since its beginning, it is only in the last decades that it is recognized as a necessary discipline. A survey of pediatricians of Hospital de Niños Ricardo Gutiérrez was carried out to determine a) the knowledge of basic concepts of bioethics b) the place and function of the committee. A total of 150 surveys were given out, 90 responses were obtained (60%). Basic concepts of bioethics (definition of bioethics, end of life, autonomy, justice, beneficence, and informed consent) were correctly answered by 75.2%; 97.8% knew of the existence of the committee in the hospital, it had been consulted by 61.1%. From the group that had consulted the recommendations suggested by the committee to solve the ethical dilemma was considered: useful by 23.6%, partially useful by 45.5% and not useful by 27.3%; 37.8% had never consulted the committee. Of these, 64.7% had never had a dilemma, 11.7% considered not useful any possible answer of the committee, and 11.7% did not give a reason. The majority of the responders had appropriate information of the basic bioethical concepts. Both the index of consultation to the committee and the assessment of its judgements as totally or partially useful (69.1%) guarantee its presence. It is peculiar that the principal argument for not consulting the committee was the absence of dilemmas. Both the committee in the hospital and the universities should intensify the teaching of medical ethics (mainly practical ethics), in order to facilitate the detection of ethical dilemmas by the health team.


Observation of bosonic condensation in a hybrid monolayer MoSe2-GaAs microcavity.
Bosonic condensation belongs to the most intriguing phenomena in physics, and was mostly reserved for experiments with ultra-cold quantum gases. More recently, it became accessible in exciton-based solid-state systems at elevated temperatures. Here, we demonstrate bosonic condensation driven by excitons hosted in an atomically thin layer of MoSe2, strongly coupled to light in a solid-state resonator. The structure is operated in the regime of collective strong coupling between a Tamm-plasmon resonance, GaAs quantum well excitons, and two-dimensional excitons confined in the monolayer crystal. Polariton condensation in a monolayer crystal manifests by a superlinear increase of emission intensity from the hybrid polariton mode, its density-dependent blueshift, and a dramatic collapse of the emission linewidth, a hallmark of temporal coherence. Importantly, we observe a significant spin-polarization in the injected polariton condensate, a fingerprint for spin-valley locking in monolayer excitons. Our results pave the way towards highly nonlinear, coherent valleytronic devices and light sources.


[Oral feeding of long-term ventilated patients with a tracheotomy tube. Underestimated danger of dysphagia].
In long-term mechanically ventilated patients, dysphagia is a common and potentially life-threatening complication, which can lead to aspiration and pneumonia. Nevertheless, many intensive care unit (ICU) patients are fed by mouth without evaluation of their deglutition capability. The goal of this work was to evaluate the prevalence of aspiration due to swallowing disorders in long-term ventilated patients who were fed orally in the ICU while having a blocked tracheotomy tube. In all, 43 patients participated-each underwent a fiberoptic investigation of deglutition on the day of admission to the rehabilitation clinic. A total of 65 % of the patients aspirated, 71 % of these silently. There were no associations between aspiration and any of the following: gender, indication for mechanical ventilation (underlying disease) or the duration of intubation and ventilation by tracheotomy tube. However, the association between aspiration and age was statistically significant (p = 0.041). Aspirating patients were older (arithmetic mean = 70 years, median = 74 years) than patients who did not aspirate (arithmetic mean = 66 years, median = 67 years). Intubation and add-on tracheotomies can lead to potentially life-threatening swallowing disorders that cause aspiration, independent of the underlying disease. Before feeding long-term mechanically ventilated patients by mouth, their ability to swallow needs to be investigated using fiberoptic endoscopic evaluation of swallowing (FEES) or a saliva dye test, if a cuff deflation and the use of a speaking valve are possible during spontaneous respiration.


Carcharhinus humani sp. nov., a new whaler shark (Carcharhiniformes:Carcharhinidae) from the western Indian Ocean.
A new species of whaler shark, Carcharhinus humani sp. nov., is described based on five type specimens from the western Indian Ocean near the Socotra Islands, off Kuwait, Mozambique, and South Africa. The new species represents the fifth species of the C. dussumieri/sealei group and the third species of the C. sealei subgroup. The new species is the only species of the C. sealei subgroup known from the western Indian Ocean. Within the C. sealei subgroup, C. humani differs from C. sealei in having a sharply demarcated black apical marking on the second dorsal fin which does not extend onto body surface (vs. black marking diffuse-edged and usually extending onto upper sides of trunk), a longer horizontal prenarial length (4.1-4.7 vs. 3.4-4.2% TL), and a longer preoral length (6.8-7.6 vs. 5.7-6.5% TL); C. humani differs from C. coatesi in having a taller second dorsal fin (its height 4.0-4.5 vs. 2.9-3.6% TL), a shorter first dorsal fin (its length 13.4-14.6 vs. 14.8-17.3% TL), and more vertebrae (total centra 152-167 vs. 134-147).


Concise review: mind the gap: challenges in characterizing and quantifying cell- and tissue-based therapies for clinical translation.
There are many challenges associated with characterizing and quantifying cells for use in cell- and tissue-based therapies. From a regulatory perspective, these advanced treatments must not only be safe and effective but also be made by high-quality manufacturing processes that allow for on-time delivery of viable products. Although sterility assays can be adapted from conventional bioprocessing, cell- and tissue-based therapies require more stringent safety assessments, especially in relation to use of animal products, immune reaction, and potential instability due to extended culture times. Furthermore, cell manufacturers who plan to use human embryonic stem cells in their therapies need to be particularly stringent in their final purification steps, due to the unrestricted growth potential of these cells. This review summarizes the current issues in characterization and quantification for cell- and tissue-based therapies, dividing these challenges into the regulatory themes of safety, potency, and manufacturing quality. It outlines current assays in use, as well as highlights the limits of many of these product release tests. Mode of action is discussed, with particular reference to in vitro surrogate assays that can be used to provide information to correlate with proposed in vivo patient efficacy. Importantly, this review highlights the requirement for basic research to improve current knowledge on the in vivo fate of these treatments; as well as an improved stakeholder negotiation process to identify the measurement requirements that will ensure the manufacture of the best possible cell- and tissue-based therapies within the shortest timeframe for the most patient benefit.


Multi-institutional Oncogenic Driver Mutation Analysis in Lung Adenocarcinoma: The Lung Cancer Mutation Consortium Experience.
Molecular genetic analyses of lung adenocarcinoma have recently become standard of care for treatment selection. The Lung Cancer Mutation Consortium was formed to enable collaborative multi-institutional analyses of 10 potential oncogenic driver mutations. Technical aspects of testing and clinicopathologic correlations are presented. Mutation testing in at least one of the eight genes (epidermal growth factor receptor [EGFR], KRAS, ERBB2, AKT1, BRAF, MEK1, NRAS, and PIK3CA) using SNaPshot, mass spectrometry, Sanger sequencing+/- peptide nucleic acid and/or sizing assays, along with anaplastic lymphoma kinase (ALK) and/or MET fluorescence in situ hybridization, were performed in six labs on 1007 patients from 14 institutions. In all, 1007 specimens had mutation analysis performed, and 733 specimens had all 10 genes analyzed. Mutation identification rates did not vary by analytic method. Biopsy and cytology specimens were inadequate for testing in 26% and 35% of cases compared with 5% of surgical specimens. Among the 1007 cases with mutation analysis performed, EGFR, KRAS, ALK, and ERBB2 alterations were detected in 22%, 25%, 8.5%, and 2.4% of cases, respectively. EGFR mutations were highly associated with female sex, Asian race, and never-smoking status; and less strongly associated with stage IV disease, presence of bone metastases, and absence of adrenal metastases. ALK rearrangements were strongly associated with never-smoking status and more weakly associated with presence of liver metastases. ERBB2 mutations were strongly associated with Asian race and never-smoking status. Two mutations were seen in 2.7% of samples, all but one of which involved one or more of PIK3CA, ALK, or MET. Multi-institutional molecular analysis across multiple platforms, sample types, and institutions can yield consistent results and novel clinicopathological observations.


Molecular absorption spectrometry in flames and furnaces: a review.
Molecular absorption spectrometry (MAS), originally developed in the 1970s, is a technique to determine non-metals in flames and graphite furnaces by monitoring the absorbance of diatomic molecules. Early studies employed low resolution instruments designed for line source atomic absorption, which provided a limited choice of analytical wavelengths, insufficient spectral resolution, and spectral interferences. However, the development of high-resolution continuum source atomic absorption spectrometry (HR-CS AAS) instrumentation has allowed the analysis of challenging samples for non-metals as well as some difficult elements to determine by AAS, such as aluminum and phosphorus. In this review, theory and analytical considerations for MAS are discussed. The principles and limitations of low resolution MAS are described, along with its applications. HR-CS AAS instrumentation is reviewed, emphasizing performance characteristics most relevant for MAS. Applications of flame and HR-CS GFMAS are reviewed, highlighting the most significant work to date. The paper concludes with an evaluation of the enhanced analytical capabilities provided by HR-CS MAS.


Acetyl-L-carnitine in neuropathic pain: experimental data.
Acetyl-L-carnitine (ALC) has gained clinical interest for its analgesic effect in different forms of neuropathies associated with chronic pain, such as diabetic and HIV-related peripheral neuropathies. The antinociceptive effect of ALC has been confirmed in several experimental models of neuropathic pain, including streptozotocin- and chemotherapy-induced neuropathy, and the sciatic nerve chronic constriction injury model. In these models, prophylactic administration of ALC has proven to be effective in preventing the development of neuropathic pain. In addition, ALC is known to produce a strong antinociceptive effect when given after neuropathic pain has been established. ALC can also improve the function of peripheral nerves by increasing nerve conduction velocity, reducing sensory neuronal loss, and promoting nerve regeneration. Analgesia requires repeated administrations of ALC, suggesting that the drug regulates neuroplasticity across the pain neuraxis. Recent evidence indicates that ALC regulates processes that go beyond its classical role in energy metabolism. These processes involve the activation of muscarinic cholinergic receptors in the forebrain, and an increased expression of type-2 metabotropic glutamate (mGlu2) receptors in dorsal root ganglia neurons. Induction of mGlu2 receptors is mediated by acetylation mechanisms that involve transcription factors of the nuclear factor (NF)-kappaB family.


EGF activates TTP expression by activation of ELK-1 and EGR-1 transcription factors.
Tristetraprolin (TTP) is a key mediator of processes such as inflammation resolution, the inhibition of autoimmunity and in cancer. It carries out this role by the binding and degradation of mRNA transcripts, thereby decreasing their half-life. Transcripts modulated by TTP encode proteins such as cytokines, pro-inflammatory agents and immediate-early response proteins. TTP can also modulate neoplastic phenotypes in many cancers. TTP is induced and functionally regulated by a spectrum of both pro- and anti-inflammatory cytokines, mitogens and drugs in a MAPK-dependent manner. So far the contribution of p38 MAPK to the regulation of TTP expression and function has been best described. Our results demonstrate the induction of the gene coding TTP (ZFP36) by EGF through the ERK1/2-dependent pathway and implicates the transcription factor ELK-1 in this process. We show that ELK-1 regulates ZFP36 expression by two mechanisms: by binding the ZFP36 promoter directly through ETS-binding site (+ 883 to +905 bp) and by inducing expression of EGR-1, which in turn increases ZFP36 expression through sequences located between -111 and -103 bp. EGF activates TTP expression via ELK-1 and EGR-1 transcription factors.


Molecular genetic relationships of the extinct dusky seaside sparrow.
Mitochondrial DNA from the extinct dusky seaside sparrow (Ammodramus maritimus nigrescens) was compared in terms of nucleotide sequence divergence to mitochondrial DNAs from extant populations of seaside sparrows. Analyses of restriction sites revealed a close phylogenetic affinity of A. m. nigrescens to other sparrow populations along the Atlantic coast of the United States but considerable genetic distance from Gulf coast birds. Concerns and applied management strategies for the seaside sparrow have been based on a morphological taxonomy that does not adequately reflect evolutionary relationships within the complex.


Postnatal development of GABAergic signalling in the rat lateral geniculate nucleus: presynaptic dendritic mechanisms.
Diverse forms of GABAergic inhibition are found in the mature brain. To understand how this diversity develops, we studied the changes in morphology of inhibitory interneurons and changes in interneuron-mediated synaptic transmission in the rat dorsal lateral geniculate nucleus (dLGN). We found a steady expansion of the dendritic tree of interneurons over the first three postnatal weeks. During this period, the area around a thalamocortical cell from which GABA(A) inhibition could be elicited also expanded. Dendritic branching and burst firing in interneurons evolved more slowly. The distal dendrites of interneurons began to branch extensively after the third week, and at the same time burst firing appeared. The appearance of burst firing and an elaborated dendritic tree were accompanied by a pronounced GABA(B) inhibition of thalamocortical cells. Thus, GABA inhibition of thalamocortical cells developed from one type of GABA(A) inhibition (spatially restricted) in the young animal into two distinct types of GABA(A) inhibition (short- and long-range) and GABA(B) inhibition in the adult animal. The close temporal relationships between the development of the diverse forms of inhibition and the postnatal changes in morphology of local GABAergic interneurons in the dLGN suggest that postnatal dendritic maturation is an important presynaptic factor for the developmental time course of the various types of feedforward inhibition in thalamus.


Expression of R345W-Fibulin-3 Induces Epithelial-Mesenchymal Transition in Retinal Pigment Epithelial Cells.
To investigate the role of protein misfolding in retinal pigment epithelial (RPE) cell dysfunction, the effects of R345W-Fibulin-3 expression on RPE cell phenotype were studied. Primary RPE cells were cultured to confluence on Transwells and infected with lentivirus constructs to express wild-type (WT)- or R345W-Fibulin-3. Barrier function was assessed by evaluating zonula occludens-1 (ZO-1) distribution and trans-epithelial electrical resistance (TER). Polarized secretion of vascular endothelial growth factor (VEGF), was measured by Enzyme-linked immunosorbent assay (ELISA). Differentiation status was assessed by qPCR of genes known to be preferentially expressed in terminally differentiated RPE cells, and conversion to an epithelial-mesenchymal transition (EMT) phenotype was assessed by a migration assay. Compared to RPE cells expressing WT-Fibulin-3, ZO-1 distribution was disrupted and TER values were significantly lower in RPE cells expressing R345W-Fibulin-3. In cells expressing mutant Fibulin-3, VEGF secretion was attenuated basally but not in the apical direction, whereas Fibulin-3 secretion was reduced in both the apical and basal directions. Retinal pigment epithelial signature genes were downregulated and multiple genes associated with EMT were upregulated in the mutant group. Migration assays revealed a faster recovery rate in ARPE-19 cells overexpressing R345W-Fibulin-3 compared to WT. The results suggest that expression of R345W-Fibulin-3 promotes EMT in RPE cells.


[Treatment of refractory ascites using a transjugular intrahepatic portosystemic shunt (TIPS). Clinical case].
Approximately, 10% of cirrhotic patients with ascites become refractory to medical treatment, a condition that reduces overall survival. TIPS is a therapeutic alternative for the management of refractory ascites. It corrects sinusoidal portal hypertension, becoming a rational and relatively safe therapeutic approach. The most frequent complications of TIPS are the development of hepatic encephalopathy and a high incidence of shunt stenosis or occlusion after one year of follow-up. We report a 43 years old alcoholic cirrhotic male with ascites refractory to medical treatment, that required multiple large volume paracentesis, whose clinical condition deteriorated progressively. A TIPS was successfully placed with a rapid and complete resolution of ascites and marked improvement of his clinical condition. During the ensuing six months the patient remained in excellent conditions, but was lost from follow-up and resumed excessive alcohol intake. Eight months after TIPS placement he had an upper gastrointestinal bleeding and died.


Downstream of tyrosine kinase 1 and 2 play opposing roles in CD200 receptor signaling.
The CD200 receptor (CD200R) negatively regulates myeloid cells by interacting with its widely expressed ligand CD200. CD200R signals through a unique inhibitory pathway involving a direct interaction with the adaptor protein downstream of tyrosine kinase 2 (Dok2) and the subsequent recruitment and activation of Ras GTPase-activating protein (RasGAP). Ligand engagement of CD200R also results in tyrosine phosphorylation of Dok1, but this protein is not essential for inhibitory CD200R signaling in human myeloid cells. In this paper, we show that CD200R-induced phosphorylation of Dok2 precedes phosphorylation of Dok1, and that Dok2 and Dok1 recruit different downstream proteins. Compared with Dok2, Dok1 recruits substantially less RasGAP. In addition to binding RasGAP, Dok2 recruits the adaptor molecule Nck in response to ligand engagement of CD200R. CD200R-induced phosphorylation of Dok1 results in the recruitment of CT10 sarcoma oncogene cellular homologue-like (CrkL), whereas the closely related CT10 sarcoma oncogene cellular homologue interacts constitutively with Dok1. Knockdown of Dok1 or CrkL expression in U937 cells resulted in increased Dok2 phosphorylation and RasGAP recruitment to Dok2. These data are consistent with a model in which Dok1 negatively regulates Dok2-mediated CD200R signaling through the recruitment of CrkL.


K-bZIP of Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) binds KSHV/HHV-8 Rta and represses Rta-mediated transactivation.
The regulatory circuit for Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) gene expression bears resemblance to that of Epstein-Barr virus (EBV), but with interesting differences. Based on protein sequence similarities and synteny to their EBV counterparts, two KSHV/HHV-8 viral regulatory factors, HHV-8 Rta and K-bZIP, encoded by open reading frame (ORF) 50 and ORF K8, respectively, have been identified. Rta is an immediate early transcriptional activator that activates lytic viral replication and mediates viral reactivation from latency, while ORF K8 is an early gene activated by Rta. Extensive splicing of ORF K8 mRNA leads to the production of K-bZIP, a protein of the basic domain-leucine zipper (bZIP) family. The role of K-bZIP in viral replication, however, remains unresolved. Here, we report that K-bZIP is a nuclear protein that binds Rta directly both in vivo and in vitro and represses Rta-mediated transactivation of the K-bZIP promoter. We further demonstrate that the leucine zipper domain of K-bZIP is required for Rta binding and a K-bZIP mutant lacking the leucine zipper does not repress Rta activity. Finally, the K-bZIP-mediated repression of Rta transactivation cannot be restored by overexpression of the transcriptional coactivator p300 or the p300-CBP-associated factor, P/CAF. Our results suggest that K-bZIP is involved in a feedback circuit to turn off its own expression and possibly the expression of other early genes activated by Rta.