A New, Better BET: Rescuing and Revising Basic Emotion Theory.
Basic Emotion Theory, or BET, has dominated the affective sciences for decades (Ekman, 1972, 1992, 1999; Ekman and Davidson, 1994; Griffiths, 2013; Scarantino and Griffiths, 2011). It has been highly influential, driving a number of empirical lines of research (e.g., in the context of facial expression detection, neuroimaging studies and evolutionary psychology). Nevertheless, BET has been criticized by philosophers, leading to calls for it to be jettisoned entirely (Colombetti, 2014; Hufendiek, 2016). This paper defuses those criticisms. In addition, it shows that we have good reason to retain BET. Finally, it reviews and puts to rest worries that BET's commitment to affect programs renders it outmoded. We propose that, with minor adjustments, BET can avoid such criticisms when conceived under a radically enactive account of emotions. Thus, rather than leaving BET behind, we show how its basic ideas can be revised, refashioned and preserved. Hence, we conclude, our new BET is still a good bet.


Fibrin structural and diffusional analysis suggests that fibers are permeable to solute transport.
Fibrin hydrogels are promising carrier materials in tissue engineering. They are biocompatible and easy to prepare, they can bind growth factors and they can be prepared from a patient's own blood. While fibrin structure and mechanics have been extensively studied, not much is known about the relation between structure and diffusivity of solutes within the network. This is particularly relevant for solutes with a size similar to that of growth factors. A novel methodological approach has been used in this study to retrieve quantitative structural characteristics of fibrin hydrogels, by combining two complementary techniques, namely confocal fluorescence microscopy with a fiber extraction algorithm and turbidity measurements. Bulk rheological measurements were conducted to determine the impact of fibrin hydrogel structure on mechanical properties. From these measurements it can be concluded that variations in the fibrin hydrogel structure have a large impact on the rheological response of the hydrogels (up to two orders of magnitude difference in storage modulus) but only a moderate influence on the diffusivity of dextran solutes (up to 25% difference). By analyzing the diffusivity measurements by means of the Ogston diffusion model we further provide evidence that individual fibrin fibers can be semi-permeable to solute transport, depending on the average distance between individual protofibrils. This can be important for reducing mass transport limitations, for modulating fibrinolysis and for growth factor binding, which are all relevant for tissue engineering. Fibrin is a natural biopolymer that has drawn much interest as a biomimetic carrier in tissue engineering applications. We hereby use a novel combined approach for the structural characterization of fibrin networks based on optical microscopy and light scattering methods that can also be applied to other fibrillar hydrogels, like collagen. Furthermore, our findings on the relation between solute transport and fibrin structural properties can lead to the optimized design of fibrin hydrogel constructs for controlled release applications. Finally, we provide new evidence for the fact that fibrin fibers may be permeable for solutes with a molecular weight comparable to that of growth factors. This finding may open new avenues for tailoring mass transport properties of fibrin carriers.


Apoptotic Effect of Melittin Purified from Iranian Honey Bee Venom on Human Cervical Cancer HeLa Cell Line.
Melittin, an amphipathic 26-residue peptide, is the main component of honey bee venom. Studies have been demonstrated that melittin has an inhibitory effect on proliferation of cancer cells. However, the precise mechanism of action is not completely understood. In the present study we have shown that purified melittin from Iranian honey bee venom shows anti-cancer effects on human cervical cancer cell line through induction of apoptosis. The venom was collected from Iranian honey bee (Apis mellifera meda) and melittin isolated using reversed phase HPLC. Biological activity of melittin was analyzed by hemolytic test on human red blood cells. In order to investigate whether melittin inhibits proliferation of cervical cancer cells, the viability of the melittin treated HeLa cell line was measured via MTT assay. Finally, cell death analysis was performed using Propidum iodide and Annexin V-FITC dual staining. The results showed that the half hemolytic concentration (HD50) induced by mellitin was 0.5 µg/ml in free FBS solution. IC50 obtained after 12 h at 1.8 µg/ml by MTT assay. According to flow cytometric analysis, melittin induced apoptosis at concentrations more than 1 µg/ml. These results suggest that melittin induces apoptotic cell death in cervical cancerous cells as observed by flow cytometric assay. It is concluded that melittin could be regarded as a potential candidate in future studies to discovery of new anticancer agents.


Sustained release of proteins from a modified vaginal ring device.
A new vaginal ring technology, the insert vaginal ring (InVR), is presented. The InVR overcomes the current shortfall of conventional vaginal rings (VRs) that are generally ineffectual for the delivery of hydrophilic and/or macromolecular actives, including peptides, proteins and antibodies, due to their poor permeation characteristics in the hydrophobic polymeric elastomers from which VRs are usually fabricated. Release of the model protein BSA from a variety of insert matrices for the InVR is demonstrated, including modified silicone rods, directly compressed tablets and lyophilised gels, which collectively provided controlled release profiles from several hours to beyond 4 weeks. Furthermore, the InVR was shown to deliver over 1 mg of the monoclonal antibody 2F5 from a single device, offering a potential means of protecting women against the transmission of HIV.


Concurrent administration of sexual assault prevention and risk reduction programming: outcomes for women.
The present study describes the 4- and 7-month postintervention outcomes of a sexual assault risk reduction program for women, which was part of an evaluation that included a prevention program for men. Relative to the control group, participants evidenced more relational sexual assertiveness and self-protective behavior, and were more likely to indicate that they utilized active verbal and physical self-defense strategies. Whether or not women experienced subsequent victimization did not differ between groups. Relative to control group women who were victimized, program participants who were victimized between the 4- and 7-month follow-up blamed the perpetrator more and evidenced less self-blame.


KRAS mutation status impacts diagnosis and treatment decision in a patient with two colon tumours: a case report.
KRAS mutation status predicts response to anti-EGFR therapy in colorectal cancer patients. Here we report an interesting case of discordant KRAS mutation status in a patient with two separate tumour foci. Tumour A in sigmoid colon invaded through muscularis propria into the subserosal fat with metastatic disease in regional lymph nodes (pT3N2b). Tumour B in ascending colon had a relatively lower stage and no metastasis (pT2N0). Both tumours showed similar morphology, immunohistochemical staining and microsatellite instability pattern. KRAS mutation, however, was detected only in tumour A. These findings indicate distinct clonal nature of these two tumours. The discordance of KRAS mutation status also suggests that a combination of anti-epidermal growth factor receptor and chemotherapy is likely the best treatment option for this patient. This case exemplifies a notion that comprehensive pathological work-up comprising molecular testing is critical to guide the diagnosis and treatment decisions for colorectal cancer patients with multiple tumours.


Ethical issues in the use of genetic information in the workplace: a review of recent developments.
In the wake of the Human Genome Project, the pace of genetic discovery has quickened. New genetic tests and other molecular technology have had immediate and wide relevance to American and European workers. These tests have the potential to provide improved workplace safety and protect workers' health, but they also carry the risk of genetic discrimination including loss of employment, promotion, insurance and health care. Ethical safeguards are necessary if the benefits are to outweigh the adverse consequences of genetics in the workplace. This review examines the major policy statements issued in Europe and the USA from 2000 to 2005 pertaining to genetic issues in occupational health. Recent findings stress that genetic testing can only be utilized with worker consent and that the workers should control access to genetic information. Such testing is only justified when the information is required to protect the safety of the worker or a third party. The progress of occupational genetic technology should not be permitted to shift the responsibility for a safe working environment from the employer to the employee. Genetic discrimination in all forms is neither supported scientifically nor warranted ethically. Increasingly, occupational physicians and clinicians treating workers will be faced with potentially stigmatizing genetic information and there is an urgent need for education and research to expand and implement the recommendations of major governmental and professional policy statements.


Overlooked Management and Risk Factors for Anemia in Patients with Intestinal Behçet's Disease in Actual Clinical Practice.
Anemia in patients with inflammatory bowel disease significantly affects the quality of life. The aim of this study was to investigate the frequency of and risk factors for anemia and to describe the management of anemia in patients with intestinal Behçet's disease (BD) in actual clinical practice. We included 64 patients with intestinal BD who visited the outpatient clinic of a tertiary referral center in June 2011 and had available laboratory data for the subsequent 6 months. Anemia was detected in 26 patients (40.6%). After 6 months, anemia was still present in 14 of these patients (53.8%). The cause of anemia was investigated in eight patients (30.8%), and oral iron supplementation was prescribed to four patients (15.4%). Of these four patients, two (50%) recovered completely within 6 months. Anemia was associated with a high Disease Activity Index for Intestinal Behçet's Disease (DAIBD, p=0.024), erythrocyte sedimentation rate (p=0.003), and C-reactive protein (p=0.049) in univariate analysis. In multivariate analysis, the factor predictive for anemia in patients with intestinal BD was a higher DAIBD (≥40; odds ratio, 4.08; 95% confidence interval, 1.21 to 13.71; p=0.023). Although anemia is common in intestinal BD patients, its clinical importance is overlooked in daily practice. Moderate to severe disease activity is predictive of anemia.


Innovative motor insurance schemes: A review of current practices and emerging challenges.
The objective of this paper is to provide a review of the most popular and often implemented methodologies related to Usage-based motor insurance (UBI). UBI schemes, such as Pay-as-you-drive (PAYD) and Pay-how-you-drive (PHYD), are a new innovative concept that has recently started to be commercialized around the world. The main idea is that instead of a fixed price, drivers have to pay a premium based on their travel and driving behaviour. Despite the fact that it has been implemented only for a few years, it appears to be a very promising practice with a significant potential impact on traffic safety as well as on traffic congestion mitigation and pollution emissions reduction. To this end, the existing literature on UBI schemes is reviewed and research gaps are identified Findings show that there is a multiplicity and diversity of several research studies accumulated in modern literature examining the correlation between PAYD (based on driver's travel behaviour and exposure) and PHYD (based on driving behaviour) schemes and crash risk in order to determine crash risk. Moreover, there is evidence that UBI implementation would eliminate the cross-subsidies phenomenon, which implies less insurance costs for less risky and exposed drivers. It would also provide a strong motivation for drivers to improve their driving behaviour, differentiate their travel behaviour and reduce their degree of exposure by receiving feedback and monitoring their driving preferences and performance, which would result in crash risk reduction both totally and individually. The paper finally discussed the current and emerging challenges on this research field.


Initial bacterial attachment in slow flowing systems: effects of cell and substrate surface properties.
Bacterial biofilm can have significant effects on the behaviors and/or performance of natural and man-made systems. Understanding the factors governing initial bacterial attachment is critical to biofilm management. In this study, the initial attachment of three bacteria, Pseudomonas aeruginosa, Escherichia coli and Pseudomonas putida, on two substrates, glass and octadecyltrichlorosilane (OTS) modified glass, was examined in flow chambers. The flow chambers were designed and operated to mimic slow moving water bodies and minimize the gravitational settlement of cells. The hydrophobicity of bacterial surface was evaluated by partitioning of cells to the water-hexadecane interface and the liquid contact angles on cell layers collected on filter papers. On the more hydrophilic glass surface, the attachment trend was found to be E. coli>P. putida>P. aeruginosa, while the opposite trend was observed on the hydrophobic, OTS modified surface. The attachment trend on glass could be explained by the magnitude of the negative interaction energy at secondary minima, as predicted by the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory. The much higher attachments of P. aeruginosa and P. putida on the OTS-modified substrate, on the other hand, suggested that these cells could overcome the energy barrier between the primary and secondary minima of interaction energy to become attached to the primary minimum. The extent of primary-minimum attachment appeared to correlate with the scale of the energy barrier, with higher attachments in the bacteria-substrate combinations of lower energy barriers. The study generated important insights into the effects of cell and substrate surface properties on initial bacterial attachment.


Multiplanar transesophageal echocardiography for the evaluation and percutaneous management of ostium secundum atrial septal defects in the adult.
The purpose of this paper is to review the usefulness of multiplanar transesophageal echocardiography before, during and after percutaneous transcatheter closure of secundum atrial septal defects. Transesophageal echocardiography imaging techniques,including their role in patient selection, procedural guidance and immediate assessment of technical success and complications are described and discussed in this review. Percutaneous transcatheter closure is indicated for ostium secundum atrial septal defects of less than 40 mm in maximal diameter. The defect must have a favorable anatomy, with adequate rims of at least 5 mm to anchor the prosthesis. Transesophageal echocardiography plays a critical role before the procedure in identifying potential candidates for percutaneous closure and to exclude those with unfavorable anatomy or associated lesions, which could not be addressed percutaneously. Transesophageal echocardiography is also important during the procedure to guide the deployment of the device. After device deployment, the echocardiographer must assess the device (integrity, position and stability), residual shunt, atrio-ventricular valve regurgitation, obstruction to systemic or venous return and pericardial effusion, in order to determine procedural success and diagnose immediate complications.


Validating and expanding the Baveno VI criteria for esophageal varices in patients with advanced liver disease: a multicenter study.
According to the Baveno VI workshop, patients with compensated advanced liver disease, platelet count (PLT) >150,000/μL and liver stiffness measurement (LSM) <20 kPa can avoid screening endoscopy for high-risk varices (HRVs). The purpose of this study was to validate these criteria in a multicenter Greek cohort and consider other approaches that may further decrease the number of endoscopies. We prospectively enrolled patients with advanced liver disease (defined as LSM >12 kPa) and evaluated them according to the Baveno VI criteria. Exclusion criteria were splanchnic vein thrombosis, use of β-blockers, and esophageal varices. Screening endoscopy was conducted within 6 months of liver stiffness and laboratory measurements. One-hundred seven consecutive patients were enrolled in the study to undergo LSM and screening endoscopy. Of these, 13 met the Baveno VI criteria (12.1%); none of the latter had HRVs. Additional parameters were examined, among which the quotient PLT/log10LSM exhibited the largest area under the curve; concerning the latter, values ≤122,000 μL-1 x kPa-1 predicted high-risk varices with 100% sensitivity and negative predictive value (NPV), preventing 20.6% of patients from unneeded screening endoscopy (P=0.003). Moreover, values ≤92,000 μL-1 x kPa-1 exhibited 86% sensitivity and 94% NPV, preventing 44.9% of patients from unneeded screening endoscopy (P=0.001), while maintaining a tolerable percentage of overlooked patients with HRVs (6.3%). The Baveno VI criteria were successfully validated in our study. The quotient PLT/log10LSM can be used to further decrease the number of screening endoscopies in patients with advanced liver disease.


Luspatercept in the treatment of lower-risk myelodysplastic syndromes.
Transforming growth factor beta (TGF-β) signaling pathway is key to hematopoiesis regulation. Increased activation of this pathway contributes to ineffective terminal erythroid differentiation in myelodysplastic syndromes (MDS). Luspatercept is a novel fusion protein that traps TGF-β ligands preventing them from binding to Type II TGF-β receptors, thereby decreasing phosphorylated SMAD2/3 resulting in the downstream effect of promoting erythropoiesis. Seminal clinical trials using luspatercept, PACE-MD and MEDALIST, demonstrated impressive efficacy in the treatment of transfusion-dependent anemia in intermediate risk or lower MDS had led to the US FDA approval for this indication. This review summarizes luspatercept mechanisms of action, efficacy/safety data supporting its use and ongoing clinical trials in MDS.


CAM practitioners in the Australian health workforce: an underutilized resource.
CAM practitioners are a valuable but underutilizes resource in Australian health care. Despite increasing public support for complementary and alternative medicine (CAM) little is known about the CAM workforce. Apart from the registered professions of chiropractic, osteopathy and Chinese medicine, accurate information about the number of CAM practitioners in the workforce has been difficult to obtain. It appears that many non-registered CAM practitioners, although highly qualified, are not working to their full capacity. Increasing public endorsement of CAM stands in contrast to the negative attitude toward the CAM workforce by some members of the medical and other health professions and by government policy makers. The marginalisation of the CAM workforce is evident in prejudicial attitudes held by some members of the medical and other health professions and its exclusion from government policy making. Inconsistent educational standards has meant that non-registered CAM practitioners, including highly qualified and competent ones, are frequently overlooked. Legitimising their contribution to the health workforce could alleviate workforce shortages and provide opportunities for redesigned job roles and new multidisciplinary teams. Priorities for better utilisation of the CAM workforce include establishing a guaranteed minimum education standard for more CAM occupation groups through national registration, providing interprofessional education that includes CAM practitioners, developing courses to upgrade CAM practitioners' professional skills in areas of indentified need, and increasing support for CAM research. Marginalisation of the CAM workforce has disadvantaged those qualified and competent CAM practitioners who practise evidence-informed medicine on the basis of many years of university training. Legitimising and expanding the important contribution of CAM practitioners could alleviate projected health workforce shortages, particularly for the prevention and management of chronic health conditions and for health promotion.


Consequences of repeated blood-brain barrier disruption in football players.
The acknowledgement of risks for traumatic brain injury in American football players has prompted studies for sideline concussion diagnosis and testing for neurological deficits. While concussions are recognized etiological factors for a spectrum of neurological sequelae, the consequences of sub-concussive events are unclear. We tested the hypothesis that blood-brain barrier disruption (BBBD) and the accompanying surge of the astrocytic protein S100B in blood may cause an immune response associated with production of auto-antibodies. We also wished to determine whether these events result in disrupted white matter on diffusion tensor imaging (DT) scans. Players from three college football teams were enrolled (total of 67 volunteers). None of the players experienced a concussion. Blood samples were collected before and after games (n = 57); the number of head hits in all players was monitored by movie review and post-game interviews. S100B serum levels and auto-antibodies against S100B were measured and correlated by direct and reverse immunoassays (n = 15 players; 5 games). A subset of players underwent DTI scans pre- and post-season and after a 6-month interval (n = 10). Cognitive and functional assessments were also performed. After a game, transient BBB damage measured by serum S100B was detected only in players experiencing the greatest number of sub-concussive head hits. Elevated levels of auto-antibodies against S100B were elevated only after repeated sub-concussive events characterized by BBBD. Serum levels of S100B auto-antibodies also predicted persistence of MRI-DTI abnormalities which in turn correlated with cognitive changes. Even in the absence of concussion, football players may experience repeated BBBD and serum surges of the potential auto-antigen S100B. The correlation of serum S100B, auto-antibodies and DTI changes support a link between repeated BBBD and future risk for cognitive changes.


Pediatric Brain Tumors: Innovative Genomic Information Is Transforming the Diagnostic and Clinical Landscape.
Pediatric neuro-oncology has undergone an exciting and dramatic transformation during the past 5 years. This article summarizes data from collaborative group and institutional trials that have advanced the science of pediatric brain tumors and survival of patients with these tumors. Advanced genomic analysis of the entire spectrum of pediatric brain tumors has heralded an era in which stakeholders in the pediatric neuro-oncology community are being challenged to reconsider their current research and diagnostic and treatment strategies. The incorporation of this new information into the next-generation treatment protocols will unleash new challenges. This review succinctly summarizes the key advances in our understanding of the common pediatric brain tumors (ie, medulloblastoma, low- and high-grade gliomas, diffuse intrinsic pontine glioma, and ependymoma) and some selected rare tumors (ie, atypical teratoid/rhabdoid tumor and CNS primitive neuroectodermal tumor). The potential impact of this new information on future clinical protocols also is discussed. Cutting-edge genomics technologies and the information gained from such studies are facilitating the identification of molecularly defined subgroups within patients with particular pediatric brain tumors. The number of evaluable patients in each subgroup is small, particularly in the subgroups of rare diseases. Therefore, international collaboration will be crucial to draw meaningful conclusions about novel approaches to treating pediatric brain tumors.


Outcomes after orthotopic liver transplantation in 15 HIV-infected patients.
Human immunodeficiency virus (HIV) infection has been associated with poor outcomes after orthotopic liver transplantation (OLT). Highly active antiretroviral therapy (HAART) has led to an increasing number of successful OLTs. The aim of this study was to examine survival and cause-specific mortality in HIV-infected patients after OLT at our institution. A retrospective analysis of all HIV patients that underwent OLT was compared to all non-HIV patients undergoing OLT during the same period. Cumulative patient and cause-specific survival were calculated using Kaplan-Meier methods; the log-rank test was used to compare the two cohorts. Fifteen HIV-infected patients and 857 non-HIV patients underwent OLT between June 1, 1999 and May 1, 2006. The actuarial 1-, 2-, and 3-year survival rates posttransplant (+/-standard error) were 73.3% (+/-11.4%) for the HIV group (unchanged from 1 to 3 years) versus 86.9% (+/-1.2%), 82.0% (+/-1.4%), and 79.4% (+/-1.5%) for the non-HIV group. Cumulative survival among HIV-infected recipients was not different from the non-HIV population (P=0.20). A difference was observed between the two groups in mortality rates due to infectious causes: the percentage of HIV patients dying from infection was 26.7% (4 of 15) vs. 8.2% (70 of 857) in the non-HIV group (P=0.006). PostOLT survival was comparable in HIV and non-HIV recipients; however, HIV patients had significantly higher mortality from infectious complications. This difference occurred despite adequate control of HIV postOLT. These findings suggest that OLT can be safely performed for HIV-infected patients; however, these patients are at higher risk of mortality from infectious complications.


[Institutional iatrogeny and maternal death: Semmelweis and puerperal fever].
Puerperal fever is a disease that becomes epidemic in the eighteenth century as a result of two factors: the urban working masses generated by the industrial revolution and the progressive hegemonization and medicalization of birth care in large public hospitals. Institutionalized maternal death reached figures above 30%, while in the case of birth care provided by midwives, it was than 2%. Semmelweis, an Hungarian physician, sustained that physicians contaminated women in labor due to insufficient hygiene after performing necropsies and established prophylactic measures in the Vienna Hospital that reduced mortality dramatically. However, his ideas were rejected because they affected the institutionalization process of medicine, based on altruism and honor, which would make it impossible to cause harm to patients. He was forced to leave Vienna Hospital and he continued his struggle in Budapest, but the rejection and disagreement of his peers with his doctrine affected his mental health. He died in an asylum, a few years before Pasteur and Koch proved the existence of the bacteria that caused diseases such as puerperal fever.


Functional Polarity of Microvascular Brain Endothelial Cells Supported by Neurovascular Unit Computational Model of Large Neutral Amino Acid Homeostasis.
The homeostatic regulation of large neutral amino acid (LNAA) concentration in the brain interstitial fluid (ISF) is essential for proper brain function. LNAA passage into the brain is primarily mediated by the complex and dynamic interactions between various solute carrier (SLC) transporters expressed in the neurovascular unit (NVU), among which SLC7A5/LAT1 is considered to be the major contributor in microvascular brain endothelial cells (MBEC). The LAT1-mediated trans-endothelial transport of LNAAs, however, could not be characterized precisely by available in vitro and in vivo standard methods so far. To circumvent these limitations, we have incorporated published in vivo data of rat brain into a robust computational model of NVU-LNAA homeostasis, allowing us to evaluate hypotheses concerning LAT1-mediated trans-endothelial transport of LNAAs across the blood brain barrier (BBB). We show that accounting for functional polarity of MBECs with either asymmetric LAT1 distribution between membranes and/or intrinsic LAT1 asymmetry with low intraendothelial binding affinity is required to reproduce the experimentally measured brain ISF response to intraperitoneal (IP) L-tyrosine and L-phenylalanine injection. On the basis of these findings, we have also investigated the effect of IP administrated L-tyrosine and L-phenylalanine on the dynamics of LNAAs in MBECs, astrocytes and neurons. Finally, the computational model was shown to explain the trans-stimulation of LNAA uptake across the BBB observed upon ISF perfusion with a competitive LAT1 inhibitor.


Growth variation in common marmoset monkeys (Callithrix jacchus) fed a purified diet: relation to care-giving and weaning behaviors.
Significant relations were observed between select infant-care and weaning behaviors and growth in body weight in common marmoset monkeys (Callithrix jacchus). The patterns of these relations suggest that earlier occurrence of developmental milestones, such as cessation of transport (being off carriers) and weaning to solid food, were associated with slower growth during the subsequent period. In contrast, more frequent nursing bouts during the period in which weaning was initiated were associated with higher growth rates. In the case of being off carriers, these effects did not carry over to older ages, suggesting that any deficits in growth were temporary. In the case of earlier, more frequent consumption of solid food, there was some suggestion that there were longer-term effects, followed by catch-up growth. The knee-to-heel length of subjects was not related to the measured behaviors. There was no relation between early weaning to solid food and leanness at day 75, suggesting that, although this behavior was affecting overall weight, it did not affect relative gains of fat versus lean mass. There were, however, significant correlations between cessation of transport or frequency of nursing bouts during the weaning period and leanness, with earlier cessation of transport and less frequent nursing associated with leaner infants, after weaning. Our results differed from those of a previous study that found a relation between linear growth and abuse in this species, with abuse defined as physical injury by other members of the group. We found no differences in growth between abused and nonabused infants. However, abused infants had lower birth weight.


Licensing delineates helper and effector NK cell subsets during viral infection.
Natural killer (NK) cells can be divided into phenotypic subsets based on expression of receptors that bind self-MHC-I molecules, a concept termed licensing or education. Here we show NK cell subsets with different migratory, effector, and immunoregulatory functions in dendritic cell and antigen (ag)-specific CD8+ T cell responses during influenza and murine cytomegalovirus infections. Shortly after infection, unlicensed NK cells localized in draining lymph nodes and produced GM-CSF, which correlated with the expansion and activation of dendritic cells, and resulted in greater and sustained ag-specific T cell responses. In contrast, licensed NK cells preferentially migrated to infected tissues and produced IFN-γ. Importantly, human NK cell subsets exhibited similar phenotypic characteristics. Collectively, our studies demonstrate a critical demarcation between the functions of licensed and unlicensed NK cell subsets, with the former functioning as the classical effector subset and the latter as the stimulator of adaptive immunity helping to prime immune responses.


Circulating CD10-/CD16low neutrophils provide a quantitative index of active bone marrow neutrophil release.
Circulating neutrophil phenotype and function are altered during neutrophilia associated with acute inflammatory states, however, the contribution of bone marrow neutrophil release to these changes has been difficult to quantify in humans. Accelerated release of neutrophils, with potentially distinct attributes, from the bone marrow and their dilution within the circulating pool may produce these apparent changes. Unfortunately selective analysis of these newly emergent neutrophils is difficult given their morphologic similarity to those already in the circulation and the coincident effect of soluble inflammatory mediators on circulating neutrophil phenotype and function. Using whole blood flow cytometry and cardiac surgery as an inflammatory stimulus, we demonstrate the emergence of a unique subpopulation of circulating neutrophils characterised as CD10(-)/CD16(low), indicative of active bone marrow neutrophil release peri-operatively. CD10(-)/CD16(low) neutrophils emerge at the same operative stages as band forms and a left shift, yet represent over 40% of circulating neutrophils postoperatively, and generate a greater stimulus-induced [Ca(2+)](i) flux than their CD10(+) counterparts. We conclude that CD10(-)/CD16(low) neutrophils represent a significant proportion of the circulating pool after cardiac surgery and that bone marrow release, a major contributor to neutrophilia, influences the phenotype and functional activity of circulating neutrophils following this acute inflammatory stimulus.


Phylogenetic position of mammoth and Steller's sea cow within Tethytheria demonstrated by mitochondrial DNA sequences.
Here we report DNA sequences from mitochondrial cytochrome b gene segments (1,005 base pairs per species) for the extinct woolly mammoth (Mammuthus primigenius) and Steller's sea cow (Hydrodamalis gigas) and the extant Asian elephant (Elephas maximus), the Western Indian manatee (Trichechus manatus), and the hyrax (Procavia capensis). These molecular data have allowed us to construct the phylogeny for the Tethytheria. Our molecular data resolve the trichotomy between the two species of living elephants and the mammoth and confirm that the mammoth was more closely related to the Asian elephant than to the African elephant. Our data also suggest that the sea cow-dugong divergence was likely as ancient as the dugong-manatee split, and it appears to have been much earlier (22 million years ago) than had been previously estimated (4-8 million years ago) by immunological comparison.


Estimation of Sex From Index and Ring Finger Lengths in An Indigenous Population of Eastern India.
Forensic anthropology involves the identification of human remains for medico-legal purposes. Estimation of sex is an essential element of medico-legal investigations when identification of unknown dismembered remains is involved. The present study was conducted with an aim to estimate sex from index and ring finger lengths of adult individuals belonging to an indigenous population of eastern India. A total of 500 unrelated adult individuals (18-60 years) from the Rajbanshi population (males: 250, females: 250) took part in the study. A total of 400 (males: 200, 200 female) participants were randomly used to develop sex estimation models using Binary Logistic Regression Analysis (BLR). A separate group of 200 adults (18-60 years) from the Karbi tribal population (males 100, females 100) were included to validate the results obtained on the Rajbanshi population. The univarate and bivariate models derived on the study group (n=400) were tested on hold-out sample of Rajbanshi participants (n=100) and the other test population of the Karbi (n=200) participants. The results indicate that Index Finger Length (IFL) and Ring Finger Length (RFL) of both hands were significantly longer in males as compared to females. The ring finger was longer than the index finger in both sexes. The study successfully highlights the existence of sex differences in IFL and RFL (p<0.05). No sex differences were however, observed for the index and ring finger ratio. The predictive accuracy of IFL and RFL in sex estimation ranged between 70%-75% (in the hold out sample from the Rajbanshi population) and 60-66% (in the test sample from the Karbi population). A Receiver Operating Curve (ROC) analysis was performed to test the predictive accuracy after predicting the probability of IFL and RFL in sex estimation. The predicted probabilities using ROC analysis were observed to be higher on the left side and in multivariate analysis. The study concludes that sex estimation from index and ring finger lengths could be of utility when more reliable means of sex estimation are not available during medico-legal investigations.


The Horn of Africa as a centre of barley diversification and a potential domestication site.
According to a widely accepted theory on barley domestication, wild barley (Hordeum vulgare ssp. spontaneum) from the Fertile Crescent is the progenitor of all cultivated barley (H. vulgare ssp. vulgare). To determine whether barley has undergone one or more domestication events, barley accessions from three continents have been studied (a) using 38 nuclear SSR (nuSSRs) markers, (b) using five chloroplast SSR (cpSSR) markers yielding 5 polymorphic loci and (c) by detecting the differences in a 468 bp fragment from the non-coding region of chloroplast DNA. A clear separation was found between Eritrean/Ethiopian barley and barley from West Asia and North Africa (WANA) as well as from Europe. The data from chloroplast DNA clearly indicate that the wild barley (H. vulgare ssp. spontaneum) as it is found today in the "Fertile Crescent" might not be the progenitor of the barley cultivated in Eritrea (and Ethiopia). Consequently, an independent domestication might have taken place at the Horn of Africa.


Molecular analysis of household transmission of Giardia lamblia in a region of high endemicity in Peru.
Giardia lamblia is ubiquitous in multiple communities of nonindustrialized nations. Genotypes A1, A2, and B (Nash groups 1, 2, and 3, respectively) are found in humans, whereas genotypes C and D are typically found in dogs. However, genotypes A and B have occasionally been identified in dogs. Fecal Giardia isolates from 22 families and their dogs, living in Pampas de San Juan, were collected over 7 weeks in 2002 and 6 weeks in 2003. Samples were genotyped, followed by sequencing and haplotyping of many of these isolates by using loci on chromosomes 3 and 5. Human infections were all caused by isolates of genotypes A2 and B. Human coinfections with genotypes A2 and B were common, and the reassortment pattern of different subtypes of A2 isolates supports prior observations that suggested recombination among genotype A2 isolates. All dogs had genotypes C and/or D, with one exception of a dog with a mixed B/D genotype infection. In a region of high endemicity where infected dogs and humans constantly commingle, different genotypes of Giardia are almost always found in dogs and humans, suggesting that zoonotic transmission is very uncommon.


Factors influencing self-reported anxiety or depression following stroke or TIA using linked registry and hospital data.
Approximately 30-50% of survivors experience problems with anxiety or depression post-stroke. It is important to understand the factors associated with post-stroke anxiety or depression to identify effective interventions. Patient-level data from the Australian Stroke Clinical Registry (years 2009-2013), from participating hospitals in Queensland (n = 23), were linked with Queensland Hospital Emergency and Admission datasets. Self-reported anxiety or depression was assessed using the EQ-5D-3L, obtained at 90-180 days post-stroke. Multivariable multilevel logistic regression, with manual stepwise elimination of variables, was used to investigate the association between self-reported anxiety or depression, patient factors and acute stroke processes of care. Comorbidities, including prior mental health problems (e.g. anxiety, depression and dementia) coded in previous hospital admissions or emergency presentations using ICD-10 diagnosis codes, were identified from 5 years prior to stroke event. 2853 patients were included (median age 74; 45% female; 72% stroke; 24% transient ischaemic attack). Nearly half (47%) reported some level of anxiety or depression post-stroke. The factors most strongly associated with anxiety or depression were a prior diagnosis of anxiety or depression [Adjusted Odds Ratio (aOR) 2.37, 95% confidence interval (95% CI) 1.66-3.39; p < 0.001], dementia (aOR 1.91, 95% CI 1.24-2.93; p = 0.003), being at home with support (aOR 1.41, 95% CI 1.12-1.69; p = < 0.001), and low socioeconomic advantage compared to high (aOR 1.59, 95% CI 1.21-2.10; p = 0.001). Acute stroke processes of care were not independently associated with anxiety or depression. Identification of those with prior mental health problems for early intervention and support may help reduce the prevalence of post-stroke anxiety or depression.


Apes in the Anthropocene: flexibility and survival.
We are in a new epoch, the Anthropocene, and research into our closest living relatives, the great apes, must keep pace with the rate that our species is driving change. While a goal of many studies is to understand how great apes behave in natural contexts, the impact of human activities must increasingly be taken into account. This is both a challenge and an opportunity, which can importantly inform research in three diverse fields: cognition, human evolution, and conservation. No long-term great ape research site is wholly unaffected by human influence, but research at those that are especially affected by human activity is particularly important for ensuring that our great ape kin survive the Anthropocene.


Towards the synthetic design of camelina oil enriched in tailored acetyl-triacylglycerols with medium-chain fatty acids.
The ability to manipulate expression of key biosynthetic enzymes has allowed the development of genetically modified plants that synthesise unusual lipids that are useful for biofuel and industrial applications. By taking advantage of the unique activities of enzymes from different species, tailored lipids with a targeted structure can be conceived. In this study we demonstrate the successful implementation of such an approach by metabolically engineering the oilseed crop Camelina sativa to produce 3-acetyl-1,2-diacyl-sn-glycerols (acetyl-TAGs) with medium-chain fatty acids (MCFAs). Different transgenic camelina lines that had been genetically modified to produce MCFAs through the expression of MCFA-specific thioesterases and acyltransferases were retransformed with the Euonymus alatus gene for diacylglycerol acetyltransferase (EaDAcT) that synthesises acetyl-TAGs. Concomitant RNAi suppression of acyl-CoA:diacylglycerol acyltransferase increased the levels of acetyl-TAG, with up to 77 mole percent in the best lines. However, the total oil content was reduced. Analysis of the composition of the acetyl-TAG molecular species using electrospray ionisation mass spectrometry demonstrated the successful synthesis of acetyl-TAG containing MCFAs. Field growth of high-yielding plants generated enough oil for quantification of viscosity. As part of an ongoing design-test-learn cycle, these results, which include not only the synthesis of 'designer' lipids but also their functional analysis, will lead to the future production of such molecules tailored for specific applications.


Long-term effects of dihydrotestosterone treatment on prostate growth in healthy, middle-aged men without prostate disease: a randomized, placebo-controlled trial.
Benign prostatic hypertrophy increases with age and can result in substantially decreased quality of life for older men. Surgery is often required to control symptoms. It has been hypothesized that long-term administration of a nonamplifiable pure androgen might decrease prostate growth, thereby decreasing or delaying the need for surgical intervention. To test the hypothesis that dihydrotestosterone (DHT), a nonamplifiable and nonaromatizable pure androgen, reduces late-life prostate growth in middle-aged men. Randomized, placebo-controlled, parallel-group trial. (Australian New Zealand Clinical Trials Registry number: ACTRN12605000358640) SETTING: Ambulatory care research center. Healthy men (n = 114) older than 50 years without known prostate disease. Transdermal DHT (70 mg) or placebo gel daily for 2 years. Prostate volume was measured by ultrasonography; bone mineral density (BMD) and body composition were measured by dual-energy x-ray absorptiometry; and blood samples and questionnaires were collected every 6 months, with data analyzed by mixed-model analysis for repeated measures. Over 24 months, there was an increase in total (29% [95% CI, 23% to 34%]) and central (75% [CI, 64% to 86%]; P < 0.01) prostate volume and serum prostate-specific antigen level (15% [CI, 6% to 24%]) with time on study, but DHT had no effect (P > 0.2). Dihydrotestosterone treatment decreased spinal BMD (1.4% [CI, 0.6% to 2.3%]; P < 0.001) at 24 months but not hip BMD (P > 0.2) and increased serum aminoterminal propeptide of type I procollagen in the second year of the study compared with placebo. Dihydrotestosterone increased serum DHT levels and its metabolites (5α-androstane-3α,17β-diol and 5α-androstane-3β,17β-diol) and suppressed serum testosterone, estradiol, luteinizing hormone, and follicle-stimulating hormone levels. Dihydrotestosterone increased hemoglobin levels (7% [CI, 5% to 9%]), serum creatinine levels (9% [CI, 5% to 11%]), and lean mass (2.4% [CI, 1.6% to 3.1%) but decreased fat mass (5.2% [CI, 2.6% to 7.7%]) (P <0.001 for all). Protocol-specific discontinuations due to DHT were asymptomatic increased hematocrit (n = 8), which resolved after stopping treatment, and increased prostate-specific antigen levels (n = 3; none with prostate cancer) in the DHT group. No serious adverse effects due to DHT occurred. Negative findings on prostate growth cannot exclude adverse effects on the natural history of prostate cancer. Dihydrotestosterone treatment for 24 months has no beneficial or adverse effect on prostate growth but causes a decrease in spinal but not hip BMD. These findings have important implications for the wider use of nonsteroidal pure androgens in older men. BHR Pharma.


Plasma phosphatidylcholine/free cholesterol ratio as an indicator for atherosclerosis.
Plasma lipid profiles were determined in two inbred strains of mice, C57BR/cdJ and CBA/J, fed either a normal chow or an atherogenic diet for a 15-week period, starting at 10 weeks of age. On the chow diet, the C57BR/cdJ had significantly higher mean free cholesterol, esterified cholesterol, and total lipid values, and a significantly lower mean phosphatidylcholine/free cholesterol ratio than the CBA/J mice. On the atherogenic diet, the C57BR/cdJ had significantly higher mean levels for all lipids classes, except triacylglycerols, than the CBA/J mice. The mean plasma free cholesterol and esterified cholesterol levels of the C57BR/cdJ were four times greater than those of the CBA/J strain on the atherogenic diet. The mean plasma phosphatidylcholine/free cholesterol ratio of the C57BR/cdJ mice on the high cholesterol diet was 0.87 compared to 1.91 for CBA/J mice. These plasma lipid changes were associated with a marked development of atheromatous deposits in the wall of the aortic sinus of the C57BR/cdJ compared to the CBA/J animals. The phosphatidylcholine/free cholesterol ratios of the liver lipids of both strains decreased from 2.5-2.7 on the chow diet to 1.0-1.1 on the high cholesterol diet. It is suggested that a plasma phosphatidylcholine/free cholesterol ratio less than 1 represents a supersaturation of the vascular system and the vessel wall with cholesterol, which leads to a destabilization of the plasma membranes of the endothelial and smooth muscle cells, and an infiltration of the vessel wall by the plasma lipids.


From Age 4 to 8, Children Become Increasingly Aware About Normative Situations for Adults to Consume Alcohol.
Limited cross-sectional studies have indicated that young children have some knowledge of the type of situations in which adults usually consume alcohol. However, it is unclear when and how this knowledge develops over time. This study tests the hypothesis that between the ages of 4 and 8, children become more knowledgeable about common drinking situations (e.g. 'partying') and uncommon situations (e.g. 'driving'). Data of two independent samples were used: a cross-sectional study (parents) and a three-wave longitudinal study (children). Parents and children were recruited via a convenience and random sampling strategy, respectively. To identify common, ambivalent, and uncommon drinking situations, parents (N = 158; 47% men) completed an online survey in which they indicated how common it is that any adult would drink alcohol in the 18 situations of the Dutch electronic appropriate beverage (eABT). Children (N = 329; 48.9% boys) completed the Dutch eABT to assess their knowledge of situations in which adults usually consume alcohol. General linear model repeated measures with post-hoc pairwise comparisons showed that parents' perceptions of common, ambivalent, and uncommon situations in which adults consume alcohol predicted the initial level and the change over time in children's knowledge of adults' alcohol use in these situations. Children aged 4-8 become increasingly knowledgeable about drinking norms in specific situations which implies that they know in what kind of situation alcohol consumption is a common human behavior. This knowledge may put them at risk for early alcohol initiation and frequent drinking later in life.


Irritable bowel syndrome: methods, mechanisms, and pathophysiology. The confluence of increased permeability, inflammation, and pain in irritable bowel syndrome.
Irritable bowel syndrome (IBS) is one of the most common gastrointestinal ailments among those seeking health care for gastrointestinal disorders. Despite its prevalence, IBS pathophysiology is still not completely understood. Continued elucidation of IBS etiological mechanisms will lead to a greater appreciation of possible therapeutic targets. In the past decade, there has been increasing focus on the possible connection between increased intestinal mucosal permeability, inflammation, and visceral hypersensitivity. Increased permeability in subsets of IBS patients has been observed and the possible mechanisms underlying this defect are just beginning to be understood. The objectives of this review are to summarize the role of the healthy intestinal epithelium as a barrier between the lumen and the rest of the body with a focus on tight junctions; to examine the lines of evidence that suggest that different triggers lead to increased intestinal mucosal permeability and disruption of tight junctions in IBS patients; and to explore how this increased permeability may elicit immune responses that affect afferent nerves, resulting in the pain associated with IBS.


Late passage human fibroblasts induced to pluripotency are capable of directed neuronal differentiation.
It is possible to generate induced pluripotent stem (iPS) cells from mouse and human somatic cells by ectopic expression of defined sets of transcription factors. However, the recommendation that somatic cells should be utilized at early passages for induced reprogramming limits their therapeutic application. Here we report successful reprogramming of human fibroblasts after more than 20 passages in vitro, to a pluripotent state with four transcription factors: Oct4, Sox2, Klf4, and c-Myc. The late passage-derived human iPS cells resemble human embryonic stem cells in morphology, cell surface antigens, pluripotent gene expression profiles, and epigenetic states. Moreover, these iPS cells differentiate into cell types representative of the three germ layers in teratomas in vivo, and directed neuronal differentiation in vitro.


Shape discrimination in age-related macular degeneration.
Recent studies suggest that a global shape-discrimination task is sensitive to neural undersampling and/or irregular sampling, but is not affected by normal aging. In this study, the ability of patients with age-related macular degeneration (AMD) to perform the shape-discrimination task was examined. Twenty patients with AMD (age range, 66-81 years) were selected on the basis of Snellen visual acuity of 20/50 or better in at least one eye and prior clinical documentation. A control group consisted of 10 older subjects (age range, 61-93 years) with normal findings in a fundus examination. Radial frequency (RF) patterns were used as stimuli. A spatial paradigm and a temporal two-alternative, forced-choice (2AFC) staircase paradigm were used. In each trial, two RF patterns (one deformed and one undeformed) were presented, and patients were asked to identify the deformed pattern. The peak spatial frequency of RF patterns was 5 cyc/deg; the radial modulation frequency was 8 cyc/360 degrees; mean radii were 0.5 degrees, 1 degrees, 2.0 degrees, or 2.5 degrees; and stimulus contrast was 80%. Thresholds for detecting the deformation were estimated by a maximum-likelihood fitting procedure. Thirty-five of 40 eyes with AMD had 20/50 or better acuity. Among them, 29 eyes had early AMD (drusen, hyperpigmentation, hypopigmentation), 5 had extrafoveal geographic atrophy, and 1 had exudative AMD. With the spatial 2AFC, 91% (32/35) of eyes with AMD showed significant elevation of the threshold for detecting radial deformation of RF patterns when compared with normal control eyes. With the temporal 2AFC, 97% (31/32) of eyes with AMD showed significant threshold elevations, and the degree of the deficit in the shape discrimination did not correlate significantly with visual acuity loss (r = 0.3, P = 0.094). Comparison of the severity of AMD with shape-discrimination performance revealed that the average detection threshold of the eyes with extrafoveal geographic atrophy was significantly higher than that of the eyes with drusen only (P < 0.01), even though average acuity showed no significant difference. Patients with AMD had significant deficits in performing the global shape-discrimination task. The dissociation of shape discrimination with visual acuity suggests that the shape-discrimination task may provide distinguishable information about the integrity of the photoreceptor mosaic in AMD.


Specific inhibition of hypoxia inducible factor 1 exaggerates cell injury induced by in vitro ischemia through deteriorating cellular redox environment.
Hypoxia inducible factor 1 (HIF-1) has been suggested to play a critical role in the fate of cells exposed to hypoxic stress. However, the mechanism of HIF-1-regulated cell survival is still not fully understood in ischemic conditions. Redox status is critical for decisions of cell survival, death and differentiation. We investigated the effects of inhibiting HIF-1 on cellular redox status in SH-SY5Y cells exposed to hypoxia or oxygen and glucose deprivation (OGD), coupled with cell death analyses. Our results demonstrated that inhibiting HIF-1alpha expression by HIF-1alpha specific small interfering RNA (siRNA) transfection increased reactive oxygen species generation, and transformed the cells to more oxidizing environments (low GSH/GSSG ratio, low NADPH level) under either hypoxic or OGD exposure. Cell death increased dramatically in the siRNA transfected cells, compared to non-transfected cells after hypoxic/OGD exposures. In contrast, increasing HIF-1alpha expression by desferrioxamine, a metal chelator and hydroxylase inhibitor, induced a more reducing environment (high GSH/GSSG ratio, high NADPH level) and reduced cell death. Further studies showed that HIF-1 regulated not only glucose transporter-1 expression, but also the key enzymes of the pentose phosphate pathway such as glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. These enzymes are important in maintaining cellular redox homeostasis by generating NADPH, the primary reducing agent in cells. Moreover, catalase significantly decreased cell death in the siRNA-transfected cells induced by hypoxia and OGD. These results suggest that maintenance of cellular redox status by HIF-1 protects cells from hypoxia and ischemia mediated injuries.


Improving the assessment of medication adherence: Challenges and considerations with a focus on low-resource settings.
Improving patient survival and quality of life in chronic diseases requires prolonged and often lifelong medication intake. Less than half of patients with chronic diseases globally are adherent to their prescribed medications which preclude the full benefit of treatment, worsens therapeutic outcomes, accelerates disease progression, and causes enormous economic losses. The accurate assessment of medication adherence is pivotal for both researchers and clinicians. Medication adherence can be assessed through both direct and indirect measures. Indirect measures include both subjective (self-report measures such as questionnaire and interview) and objective (pill count and secondary database analysis) measures and constitute the mainstay of assessing medication adherence. However, the lack of an inexpensive, ubiquitous, universal gold standard for assessment of medication adherence emphasizes the need to utilize a combination of measures to differentiate adherent and nonadherent patients. The global heterogeneity in health systems precludes the development of a universal guideline for evaluating medication adherence. Methods based on the secondary database analysis are mostly ineffectual in low-resource settings lacking electronic pharmacy and insurance databases and allowing refills without updated, valid prescriptions from private pharmacies. This significantly restricts the choices for assessing adherence until digitization of medical data takes root in much of the developing world. Nevertheless, there is ample scope for improving self-report measures of adherence. Effective interview techniques, especially accounting for suboptimal patient health literacy, validation of adherence questionnaires, and avoiding conceptual fallacies in reporting adherence can improve the assessment of medication adherence and promote understanding of its causal factors.


Proposed minimal standards for the genus Mycobacterium and for description of new slowly growing Mycobacterium species.
In accordance with Recommendation 30b of the International Code of Nomenclature of Bacteria, which calls for the development of recommended minimal standards for describing new species, we propose minimal standards for describing the genus Mycobacterium and new slowly growing species of this genus. The minimal standards for assignment of a strain to the genus Mycobacterium include acid-alcohol fastness, a DNA G+C content in the range from 61 to 71 mol%, and mycolic acid detection with characterization of C22 to C26 pyrolysis esters. The recommended minimal standards for describing a new slowly growing Mycobacterium species are based on the results of phenotypic and genomic studies and include the results of the following conventional tests: growth at 25, 30, 33, 37, 42, and 45 degrees C; pigmentation; resistance to isoniazid, thiophene-2-carboxylic acid hydrazide, hydroxylamine, p-nitrobenzoic acid, sodium chloride, thiacetazone, picrate, and oleate; catalase activity; Tween hydrolysis; urease activity; niacin detection; and nitrate reductase, acid phosphatase, arylsulfatase, pyrazinamidase, and alpha-esterase activities. In addition, a mycolic acid profile should be determined, and DNA-DNA hybridization experiments in which the difference between the denaturation temperature of the homologous reaction and the denaturation temperature of the heterologous reaction is determined should be performed. This proposal has been endorsed by the members of the Subcommittee for Taxonomy of the Mycobacteria of the International Committee on Systematic Bacteriology.


Aerodynamics of hovering flight in the long-eared bat Plecotus auritus.
Steady-state aerodynamic and momentum theories were used for calculations of the lift and drag coefficients of Plecotus auritus in hovering flight. The lift coefficient obtained varies between 3-1 and 6-4, and the drag coefficient between --5-0 and 10-5, for the possible assumptions regarding the effective angles of attack during the upstroke. This demonstrates that hovering flight in Plecotus auritus can not be explained by quasi-steady-state aerodynamics. Thus, non-steady-state aerodynamics must prevail.


An estimate of the turnover rate of bone-derived plasma alkaline phosphatase in Paget's disease.
A through-knee amputation for suspected osteogenic sarcoma was performed on a patient with Paget's disease of the tibia. Plasma alkaline phosphatase (AP) activity fell to normal values, apparently as a single exponential function of time, with a half-life of 1.7 days. The daily turnover of bone-derived plasma AP was estimated to be approximately 200 I.U. from the Pagetic tibia and 20 I.U. from the rest of the skeleton.


Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples.
Campylobacter spp., especially Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli), are recognized as the leading human foodborne pathogens in developed countries. Livestock animals carrying Campylobacter pose an important risk for human contamination. Pigs are known to be frequently colonized with Campylobacter, especially C. coli, and to excrete high numbers of this pathogen in their faeces. Molecular tools, notably real-time PCR, provide an effective, rapid, and sensitive alternative to culture-based methods for the detection of C. coli and C. jejuni in various substrates. In order to serve as a diagnostic tool supporting Campylobacter epidemiology, we developed a quantitative real-time PCR method for species-specific detection and quantification of C. coli and C. jejuni directly in faecal, feed, and environmental samples. With a sensitivity of 10 genome copies and a linear range of seven to eight orders of magnitude, the C. coli and C. jejuni real-time PCR assays allowed a precise quantification of purified DNA from C. coli and C. jejuni. The assays were highly specific and showed a 6-log-linear dynamic range of quantification with a quantitative detection limit of approximately 2.5 × 10² CFU/g of faeces, 1.3 × 10² CFU/g of feed, and 1.0 × 10³ CFU/m² for the environmental samples. Compared to the results obtained by culture, both C. coli and C. jejuni real-time PCR assays exhibited a specificity of 96.2% with a kappa of 0.94 and 0.89 respectively. For faecal samples of experimentally infected pigs, the coefficients of correlation between the C. coli or C. jejuni real-time PCR assay and culture enumeration were R² = 0.90 and R² = 0.93 respectively. The C. coli and C. jejuni real-time quantitative PCR assays developed in this study provide a method capable of directly detecting and quantifying C. coli and C. jejuni in faeces, feed, and environmental samples. These assays represent a new diagnostic tool for studying the epidemiology of Campylobacter by, for instance, investigating the carriage and excretion of C. coli and C. jejuni by pigs from conventional herds.


Subcellular diversion of cholesterol by gain- and loss-of-function mutations in PMP22.
Abnormalities of the peripheral myelin protein 22 (PMP22) gene, including duplication, deletion and point mutations are a major culprit in Type 1 Charcot-Marie-Tooth (CMT) diseases. The complete absence of PMP22 alters cholesterol metabolism in Schwann cells, which likely contributes to myelination deficits. Here, we examined the subcellular trafficking of cholesterol in distinct models of PMP22-linked neuropathies. In Schwann cells from homozygous Trembler J (TrJ) mice carrying a Leu16Pro mutation, cholesterol was retained with TrJ-PMP22 in the Golgi, alongside a corresponding reduction in its plasma membrane level. PMP22 overexpression, which models CMT1A caused by gene duplication, triggered cholesterol sequestration to lysosomes, and reduced ATP-binding cassette transporter-dependent cholesterol efflux. Conversely, lysosomal targeting of cholesterol by U18666A treatment increased wild type (WT)-PMP22 levels in lysosomes. Mutagenesis of a cholesterol recognition motif, or CRAC domain, in human PMP22 lead to increased levels of PMP22 in the ER and Golgi compartments, along with higher cytosolic, and lower membrane-associated cholesterol. Importantly, cholesterol trafficking defects observed in PMP22-deficient Schwann cells were rescued by WT but not CRAC-mutant-PMP22. We also observed that myelination deficits in dorsal root ganglia explants from heterozygous PMP22-deficient mice were improved by cholesterol supplementation. Collectively, these findings indicate that PMP22 is critical in cholesterol metabolism, and this mechanism is likely a contributing factor in PMP22-linked hereditary neuropathies. Our results provide a basis for understanding how altered expression of PMP22 impacts cholesterol metabolism.


Mining significant substructure pairs for interpreting polypharmacology in drug-target network.
A current key feature in drug-target network is that drugs often bind to multiple targets, known as polypharmacology or drug promiscuity. Recent literature has indicated that relatively small fragments in both drugs and targets are crucial in forming polypharmacology. We hypothesize that principles behind polypharmacology are embedded in paired fragments in molecular graphs and amino acid sequences of drug-target interactions. We developed a fast, scalable algorithm for mining significantly co-occurring subgraph-subsequence pairs from drug-target interactions. A noteworthy feature of our approach is to capture significant paired patterns of subgraph-subsequence, while patterns of either drugs or targets only have been considered in the literature so far. Significant substructure pairs allow the grouping of drug-target interactions into clusters, covering approximately 75% of interactions containing approved drugs. These clusters were highly exclusive to each other, being statistically significant and logically implying that each cluster corresponds to a distinguished type of polypharmacology. These exclusive clusters cannot be easily obtained by using either drug or target information only but are naturally found by highlighting significant substructure pairs in drug-target interactions. These results confirm the effectiveness of our method for interpreting polypharmacology in drug-target network.


Environmental DNA as an innovative technique to identify the origins of falsified antimalarial tablets-a pilot study of the pharmabiome.
Falsified medicines are a major threat to global health. Antimalarial drugs have been particularly targeted by criminals. As DNA analysis has revolutionized forensic criminology, we hypothesized that these techniques could also be used to investigate the origins of falsified medicines. Medicines may contain diverse adventitious biological contamination, and the sealed nature of blister-packages may capture and preserve genetic signals from the manufacturing processes allowing identification of production source(s). We conducted a blinded pilot study to determine if such environmental DNA (eDNA) could be detected in eleven samples of falsified and genuine artesunate antimalarial tablets, collected in SE Asia, which could be indicative of origin. Massively Parallel Sequencing (MPS) was used to characterize microbial and eukaryote diversity. Two mitochondrial DNA analysis approaches were explored to detect the presence of human DNA. Trace eDNA from these low biomass samples demonstrated sample specific signals using two target markers. Significant differences in bacterial and eukaryote DNA community structures were observed between genuine and falsified tablets and between different packaging types of falsified artesunate. Human DNA, which was indicative of likely east Asian ancestry, was found in falsified tablets. This pilot study of the 'pharmabiome' shows the potential of environmental DNA as a powerful forensic tool to assist with the identification of the environments, and hence location and timing, of the source and manufacture of falsified medicines, establish links between seizures and complement existing tools to build a more complete picture of criminal trade routes. The finding of human DNA in tablets raises important ethical issues that need to be addressed.


Ultra-low microcurrent in the management of diabetes mellitus, hypertension and chronic wounds: report of twelve cases and discussion of mechanism of action.
Oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus and cardiovascular diseases including hypertension. The low levels of antioxidants accompanied by raised levels of markers of free radical damage play a major role in delaying wound healing. Ultra-low microcurrent presumably has an antioxidant effect, and it was shown to accelerate wound healing. The purpose of the study is to investigate the efficacy of ultra-low microcurrent delivered by the Electro Pressure Regeneration Therapy (EPRT) device (EPRT Technologies-USA, Simi Valley, CA) in the management of diabetes, hypertension and chronic wounds. The EPRT device is an electrical device that sends a pulsating stream of electrons in a relatively low concentration throughout the body. The device is noninvasive and delivers electrical currents that mimic the endogenous electric energy of the human body. It is a rechargeable battery-operated device that delivers a direct current (maximum of 3 milliAmperes) of one polarity for 11.5 minutes, which then switched to the opposite polarity for another 11.5 minutes. The resulting cycle time is approximately 23min or 0.000732 Hz and delivers a square wave bipolar current with a voltage ranging from 5V up to a maximum of 40 V. The device produces a current range of 3 mA down to 100 nA. Twelve patients with long standing diabetes, hypertension and unhealed wounds were treated with EPRT. The patients were treated approximately for 3.5 h/day/5 days a week. Assessment of ulcer was based on scale used by National Pressure Ulcer Advisory Panel Consensus Development Conference. Patients were followed-up with daily measurement of blood pressure and blood glucose level, and their requirement for medications was recorded. Treatment continued from 2-4 months according to their response. Results showed that diabetes mellitus and hypertension were well controlled after using this device, and their wounds were markedly healed (30-100%). The patients either reduced their medication or completely stopped after the course of treatment. No side effects were reported. The mechanism of action was discussed.


Dynamics of galectin-3 in the nucleus and cytoplasm.
This review summarizes selected studies on galectin-3 (Gal3) as an example of the dynamic behavior of a carbohydrate-binding protein in the cytoplasm and nucleus of cells. Within the 15-member galectin family of proteins, Gal3 (M(r) approximately 30,000) is the sole representative of the chimera subclass in which a proline- and glycine-rich NH(2)-terminal domain is fused onto a COOH-terminal carbohydrate recognition domain responsible for binding galactose-containing glycoconjugates. The protein shuttles between the cytoplasm and nucleus on the basis of targeting signals that are recognized by importin(s) for nuclear localization and exportin-1 (CRM1) for nuclear export. Depending on the cell type, specific experimental conditions in vitro, or tissue location, Gal3 has been reported to be exclusively cytoplasmic, predominantly nuclear, or distributed between the two compartments. The nuclear versus cytoplasmic distribution of the protein must reflect, then, some balance between nuclear import and export, as well as mechanisms of cytoplasmic anchorage or binding to a nuclear component. Indeed, a number of ligands have been reported for Gal3 in the cytoplasm and in the nucleus. Most of the ligands appear to bind Gal3, however, through protein-protein interactions rather than through protein-carbohydrate recognition. In the cytoplasm, for example, Gal3 interacts with the apoptosis repressor Bcl-2 and this interaction may be involved in Gal3's anti-apoptotic activity. In the nucleus, Gal3 is a required pre-mRNA splicing factor; the protein is incorporated into spliceosomes via its association with the U1 small nuclear ribonucleoprotein (snRNP) complex. Although the majority of these interactions occur via the carbohydrate recognition domain of Gal3 and saccharide ligands such as lactose can perturb some of these interactions, the significance of the protein's carbohydrate-binding activity, per se, remains a challenge for future investigations.


Antigenotoxic and antioxidant potentials of newly derivatized compound naringenin-oxime relative to naringenin on human mononuclear cells.
We investigated antigenotoxic and antioxidative effects of newly derivatized compound naringenin-oxime (NG-Ox) compared to its mother compound naringenin (NG) against oxidative damage induced by hydrogen peroxide (HP) in human peripheral blood mononuclear cells (PBMC). Antigenotoxic activity was assessed using alkaline single cell gel electrophoresis assay (comet assay). Oxidative status was evaluated by measurement of total antioxidant status, total oxidant status and lipid hydroperoxide levels in the cells. Oxidative stress index was also calculated. Both NG and NG-Ox show a protective effect against HP-induced oxidative damage on PBMC and are able to reduce oxidative stress. The percentage of antigenotoxic and antioxidant potential progressively increased in a dose-dependent manner. However, these activities were found to be more significant in NG-Ox-treated cells than in NG-treated cells. Taken together, these observations provide evidences indicating that both NG and NG-Ox are able to protect cells against oxidative damage and apparently NG-Ox is more effective than NG.


Orthostatic Tremor: An Update on a Rare Entity.
Orthostatic tremor (OT) remains among the most intriguing and poorly understood of movement disorders. Compared to Parkinson's disease or even essential tremor, there are very few articles addressing more basic science issues. In this review, we will discuss the findings of main case series on OT, including data on etiology, pathophysiology, diagnostic approach, treatment strategies, and outcome. Data for this review were identified by searching PUBMED (January 1966 to August 2016) for the terms "orthostatic tremor" or "shaky leg syndrome," which yielded 219 entries. We did not exclude papers on the basis of language, country, or publication date. The electronic database searches were supplemented by articles in the authors' files that pertained to this topic. Owing to its rarity, the current understanding of OT is limited and is mostly based on small case series or case reports. Despite this, a growing body of evidence indicates that OT might be a progressive condition that is clinically heterogeneous (primary vs. secondary cases) with a broader spectrum of clinical features, mainly cerebellar signs, and possible cognitive impairment and personality disturbances. Along with this, advanced neuroimaging techniques are now demonstrating distinct anatomical and functional changes, some of which are consistent with neuronal loss. OT might be a family of diseases, unified by the presence of leg tremor, but further characterized by etiological and clinical heterogeneity. More work is needed to understand the pathogenesis of this condition.


Corticosteroid Tapering Regimens in Rheumatic Disease: A Systematic Review.
Corticosteroids have long been used to effectively treat rheumatic disorders, but adverse effects associated with extended-duration regimens generate disagreement among clinicians regarding optimal tapering strategies. The objective of this systematic review was to assess clinical outcomes of differing tapering regimens after corticosteroid monotherapy in adults with rheumatic disorders. A systematic review of Medline/PubMed, Embase, Cochrane, International Pharmaceutical Abstracts, Web of Science, Scopus, Global Index Medicus, American College of Rheumatology, gray literature, and reference lists up to June 27, 2018, was conducted by 2 authors. Randomized controlled trials, case-control studies, and prospective observational studies comparing at least 2 tapering strategies of medium- to high-dose (>7.5 mg but ≤100 mg oral prednisone equivalent daily), extended-duration (≥10 days) corticosteroids were included if they reported at least 1 efficacy and 1 adverse effect parameter. Two studies met criteria for the review, which included 62 patients. One study examined a prednisolone versus a modified release prednisone taper for giant cell arteritis and suggested 80% (n = 4) and 85.7% (n = 6) remission rates, respectively, at 26 weeks. The other study examined a methylprednisolone versus a prednisone taper for polymyalgia rheumatica and reported 100% and 89% remission rates, respectively, at 26 weeks. Adverse effects reported between the 2 studies included sleep, hyperglycemia, infection, and fractures. However, the studies were not powered to detect differences in these outcomes. There is no high-level evidence to guide tapering until discontinuation after extended courses of medium- to high-dose treatment regimens, as current guidelines rely heavily on expert opinion and small case series with a trial-and-error approach. This review supports the need for additional research to shift tapering recommendations to a more evidence-based practice.


Sympathetic innervation of inguinal white adipose tissue in the mouse.
Adipose tissue plays an important role in metabolic homeostasis and its prominent role as endocrine organ is now well recognized. Adipose tissue is controlled via the sympathetic nervous system (SNS). New viral, molecular-genetic tools will soon allow a more detailed study of adipose tissue innervation in metabolic function, yet, the precise anatomical extent of preganglionic and postganglionic inputs to the inguinal white adipose tissue (iWAT) is limited. Furthermore, several viral, molecular-genetic tools will require the use of cre/loxP mouse models, while the available studies on sympathetic iWAT innervation were established in larger species. In this study, we generated a detailed map for the sympathetic innervation of iWAT in male and female mice. We adapted iDISCO tissue clearing to process large, whole-body specimens for an unprecedented view of the natural abdominal SNS. Combined with pseudorabies virus retrograde tracing from the iWAT, we defined the preganglionic and postganglionic sympathetic input to iWAT. We used fluorescence-guided anatomical dissections of sympathetic nerves in reporter mice to further clarify that postganglionic axons connect to iWAT via lateral cutaneous rami (dorsolumbar iWAT portion) and the lumbar plexus (inguinal iWAT portion). Importantly, these rami carry axons that branch to iWAT, as well as axons that travel further to innervate the skin and vasculature, and their functional impact will require consideration in denervation studies. Our study may serve as a comprehensive map for future experiments that employ virally driven neuromodulation techniques to predict anatomy-based viral labeling.


Diffusion MRI and tensor tractography in ischemic optic neuropathy.
Ischemic optic neuropathies (IONs) are among the most prevalent diseases causing visual impairment in middle-aged and elderly people. While arteritic ION is an ocular emergency and requires early diagnosis and immediate treatment with systemic high-dose corticosteroids to prevent further visual loss, treatment options for non-arteritic ION remain limited. We describe the case of a woman with unilateral right-sided non-arteritic posterior ischemic optic neuropathy. The diagnosis was made on clinical and radiographic grounds. Diffusion-weighted sequences and apparent diffusion coefficient maps revealed markedly restricted diffusion in the right optic nerve. It was very helpful to precise the posterior topography of the optic nerve lesion. Furthermore, we reported the diffusion tensor tractography study which appears to be an objective tool to assess the incomplete visual recovery. These MRI techniques including tensor tractography remain to be evaluated in large cohort of ION patients' particularly in future therapeutic trials.


Central nervous system metastatic epithelial ovarian cancer. Clinical parameters and prognostic factors: a multicenter study of Anatolian Society of Medical Oncology.
Central nervous system (CNS) metastasis is a rare event in the course of late stage epithelial ovarian cancer (EOC); however its incidence is increasing in parallel with prolonged survival of patients. The authors assessed the clinical parameters and potential prognostic features in patients with CNS metastatic disease. Clinical data of the 33 patients from the participating centers were retrospectively collected and analyzed. Median age at the time of CNS metastasis was 57 years. Median time from the diagnosis of primary EOC until CNS metastatic disease was 22 months. Nearly half (45.5%) of the patients had single CNS metastatic lesions and all patients in the study group except two received radiotherapy as palliative treatment. Median overall survival (OS) from the time of CNS metastasis was 15 months (0-66). At univariate analysis only number of brain metastatic lesions (p = 0.001) and presence of extracranial disease (p = 0.004) were strongly associated with OS whereas multimodal treatment, size of metastatic lesions, platinum sensitivity, age, grade, and disease stage at presentation were not. Development of CNS metastasis carries a poor prognosis, however patients with single metastatic lesions and only intracranial metastatic disease can have prolonged survival after appropriate palliative management of their disease.


Association of metabolic syndrome and its components with suicidal ideation and depression in adults: A nationally representative sample of the Korean population.
Few studies have evaluated the association of depressive symptoms and suicidal ideation with metabolic syndrome (MetS) and its components. This study examined the associations of MetS and its components with suicidal ideation and depression and determined the mediation effect of self-rated health (SRH) on these associations. This study was conducted using cross-sectional data on 9867 people (aged ≥ 19 years old) collected in 2014 and 2016 from a nationally representative sample of the Korean National Health and Nutrition Examination Survey. The associations of MetS with suicidal ideation and depression were estimated after adjusting for potential confounding factors using logistic regression analyses, and a mediation analysis by Hayes and Preacher was used to estimate the mediation pathway between such associations through SRH. After adjusting for confounding factors, suicidal ideation was associated with MetS. Depressive symptoms and suicidal ideation were associated with two components of MetS: high triglyceride level and high fasting plasma glucose level. In the mediation analysis, we found a significant mediating effect of SRH on the association between high triglyceride level and suicidal ideation. We cannot elucidate causal relationships because of the cross-sectional design. Our study provides the implication that a high triglyceride level and elevated fasting plasma glucose level may contribute suicidal risk and depressive symptoms in adults and that SRH may contribute to the development of suicidal ideation in adults via the apparent interaction with one of the components of MetS-high triglyceride level.


Cathepsin B, cathepsin L, and cystatin C in the porcine uterus and placenta: potential roles in endometrial/placental remodeling and in fluid-phase transport of proteins secreted by uterine epithelia across placental areolae.
Cathepsins (CTSB and CTSL1) and their inhibitor, cystatin C (CST3), remodel uterine endometrium and placenta for transport of gases, micronutrients, and macromolecules essential for development and growth of the conceptus (embryo/fetus and placental membranes). We examined the temporal/spatial control of expression for CTSB, CTSL1, and CST3 mRNAs in endometria and placentae of pigs using three developmental models: 1) pigs were hysterectomized during the estrous cycle or pregnancy; 2) cyclic pigs were injected with estrogen to induce pseudopregnancy and were hysterectomized; and 3) pigs were ovariectomized, injected with progesterone, and hysterectomized. The abundance of CTSB, CTSL1, and CST3 mRNAs increased in endometrial epithelia during pregnancy and in response to exogenous progesterone but not estrogen. CST3 was also expressed in cells scattered within the stratum compactum stroma. Progesterone decreased epithelial but increased stromal compartment expression of CST3. CTSB increased in all chorionic epithelia, but CTSL1 was limited to chorionic epithelia that form areolae to absorb secretions from uterine glands. Based on the placental and endometrial distribution of CTSL1, we examined expression in the neonatal enterocytes known to transport immunoglobulins from colostrum. CTSL1 was also expressed in enterocytes of intestine from neonatal piglets. Therefore, CTSL1 is expressed by endometrial epithelia, placental areolae, and neonatal intestine, and it may function in the transport of macromolecules across these epithelia. Our results support the idea that reciprocal interactions between CSTL1, CTSB, and CST3 may be required to remodel endometrial and placental tissues for close apposition between maternal and fetal vasculatures and to facilitate transplacental transport of gases, micronutrients (amino acids, glucose), and macromolecules (proteins). Cysteine proteases and their inhibitors may also specifically modify proteins for successful utilization and fluid-phase transport across uterine, placental, and neonatal gut epithelia.


[Duration of anticoagulant therapy in venous thromboembolic complications].
Adequate anticoagulant therapy is a general approach to treatment of deep vein thrombosis. However, the duration of anticoagulant therapy is not strictly specified in everyday clinical practice. The present article deals with various approaches to selecting the duration of therapy with anticoagulants based on the findings of studies, national and foreign clinical guidelines. The minimal duration of therapy for deep vein thrombosis and pulmonary thromboembolism amounts to 3 months in accordance with the national and American recommendations. For some cohorts of patients, continuation of therapy above 3 months is considered: patients with idiopathic thrombosis (the recommended duration of therapy of not less than 6 months), patients having persisting risk factor for relapse of thrombosis on termination of the main therapeutic course, oncological patients (6 month therapy followed by assessing the risk and benefit of continuing therapy with anticoagulants). Prolonged therapy of venous thromboembolism using unfractionated heparin or low-molecular-weight heparin followed by changing over to vitamin K antagonists is associated with decreased risk for thrombosis relapse approximately by 90%, however increasing the risk of haemorrhage. Currently, as an alternative, it is possible to consider administration of novel oral anticoagulants (rivaroxaban, dabigatran, apixaban) which beside high efficacy are associated with less risk of bleeding. The route of administration, no necessity to control the INR, and the minimal number of drug and food interactions make administration of new oral anticoagulants an attractive alternative to therapy with heparins and vitamin K antagonists.


Structure and Energetic Contributions of a Designed Modular Peptide-Binding Protein with Picomolar Affinity.
Natural armadillo repeat proteins (nArmRP) like importin-α or β-catenin bind their target peptides such that each repeat interacts with a dipeptide unit within the stretched target peptide. However, this modularity is imperfect and also restricted to short peptide stretches of usually four to six consecutive amino acids. Here we report the development and characterization of a regularized and truly modular peptide-specific binding protein, based on designed armadillo repeat proteins (dArmRP), binding to peptides of alternating lysine and arginine residues (KR)n. dArmRP were obtained from nArmRP through cycles of extensive protein engineering, which rendered them more uniform. This regularity is reflected in the consistent binding of dArmRP to (KR)-peptides, where affinities depend on the lengths of target peptides and the number of internal repeats in a very systematic manner, thus confirming the modularity of the interaction. This exponential dependency between affinity and recognition length suggests that each module adds a constant increment of binding energy to sequence-specific recognition. This relationship was confirmed by comprehensive mutagenesis studies that also reveal the importance of individual peptide side chains. The 1.83 Å resolution crystal structure of a dArmRP with five identical internal repeats in complex with the cognate (KR)5 peptide proves a modular binding mode, where each dipeptide is recognized by one internal repeat. The confirmation of this true modularity over longer peptide stretches lays the ground for the design of binders with different specificities and tailored affinities by the assembly of dipeptide-specific modules based on armadillo repeats.


Risk and protective factors for comorbid internalizing and externalizing problems among economically disadvantaged African American youth.
Comorbidity of internalizing and externalizing problems and its risk and protective factors have not been well incorporated into developmental research, especially among racial minority youth from high-poverty neighborhoods. The present study identified a latent comorbid factor as well as specific factors underlying internalizing and externalizing problems among 592 African American adolescents living in economically disadvantaged neighborhoods (291 male; M age = 15.9 years, SD = 1.43 years). Stressful life events and racial discrimination were associated with higher comorbid problems, whereas stressful life events and exposure to violence were associated with higher specific externalizing problems. Collective efficacy was associated with both lower specific externalizing problems and lower comorbid problems. Moreover, high collective efficacy buffered the risk effects of stressful life events and racial discrimination on comorbid problems. Our results demonstrated the advantages of latent variable modeling to understanding comorbidity by articulating impacts of risk factors on comorbid and specific components underlying internalizing and externalizing problems. They also highlighted the protective effect of collective efficacy in mitigating risks for these problems. These findings broadly call for more studies on comorbidities in developmental psychopathology among youth from diverse sociocultural backgrounds.


Relationship of restriction fragment length polymorphisms (RFLP) at the bovine calpastatin locus to calpastatin activity and meat tenderness.
Restriction fragment length polymorphisms (RFLP) have been identified at the bovine calpastatin locus. The objective of the present study was to determine whether these polymorphisms are related to variations in calpastatin activity or beef tenderness in unrelated animals of mixed breeding. A sample of 83 crossbred steers from sires representing eight different breeds was examined to determine this relationship. A 2.2-kb cDNA coding for domains 2 through 4 plus a 3' untranslated region of bovine skeletal muscle calpastatin was used as a probe for calpastatin RFLP. Polymorphisms were found using the restriction enzymes BamHI and EcoRI. Polymorphic restriction fragments for BamHI were 9.0 and 5.0 kb and for EcoRI were 6.0 and 4.0 kb. Allelic frequencies for BamHI restriction fragments were .53 for the 9.0-kb allele and .47 for the 5.0-kb allele. Allelic frequencies for EcoRI restriction fragments were .43 for the 6.0-kb allele and >57 for the 4.0-kb allele. No polymorphisms were identified using the restriction enzymes BglII, DraI, or PstI. No associations between EcoRI and BamHI RFLP and 24-h calpastatin activity of Warner-Bratzler shear force at 14 d postmortem were detected. Therefore, the polymorphic EcoRI and BamHI restriction sites within the bovine calpastatin locus do not detect DNA sequence differences responsible for variation in calpastatin activity or tenderness of aged beef. Therefore, these polymorphisms cannot be used to predict tenderness of aged beef from unrelated animals of mixed breeding. These results do not exclude the possibility that other DNA sequences in or near the bovine calpastatin gene are responsible for variation in calpastatin activity or meat tenderness. The lack of a relationship between these calpastatin RFLP and meat tenderness must be distinguished from the well-documented relationship between calpastatin activity and meat tenderness. Therefore, further development of calpastatin-based methods for predicting beef tenderness in unrelated animals of mixed breeding should focus on basic factors influencing calpastatin activity at the molecular and cellular level.


Nuts and bolts of running a pulmonary embolism response team: results from an organizational survey of the National PERT™ Consortium members.
Pulmonary embolism response teams (PERT) are developing rapidly to operationalize multi-disciplinary care for acute pulmonary embolism patients. Our objective is to describe the core components of PERT necessary for newly developing programs. An online organizational survey of active National PERT™ Consortium members was performed between April and June 2016. Analysis, including descriptive statistics and Kruskal-Wallis tests, was performed on centers self-reporting a fully operational PERT program. The survey response rate was 80%. Of the 31 institutions that responded (71% academic), 19 had fully functioning PERT programs. These programs were run by steering committees (17/19, 89%) more often than individual physicians (2/19, 11%). Most PERT programs involved 3-5 different specialties (14/19, 74%), which did not vary based on hospital size or academic affiliation. Of programs using multidisciplinary discussions, these occurred via phone or conference call (12/18, 67%), with a minority of these utilizing 'virtual meeting' software (2/12, 17%). Guidelines for appropriate activations were provided at 16/19 (84%) hospitals. Most PERT programs offered around-the-clock catheter-based or surgical care (17/19, 89%). Outpatient follow up usually occurred in personal physician clinics (15/19, 79%) or dedicated PERT clinics (9/19, 47%), which were only available at academic institutions. PERT programs can be implemented, with similar structures, at small and large, community and academic medical centers. While all PERT programs incorporate team-based multi-disciplinary care into their core structure, several different models exist with varying personnel and resource utilization. Understanding how different PERT programs impact clinical care remains to be investigated.


Tapping the translation potential of cAMP signalling: molecular basis for selectivity in cAMP agonism and antagonism as revealed by NMR.
Eukaryotic CBDs (cAMP-binding domains) control multiple cellular functions (e.g. phosphorylation, guanine exchange and ion channel gating). Hence the manipulation of cAMP-dependent signalling pathways has a high translational potential. However, the ubiquity of eukaryotic CBDs also poses a challenge in terms of selectivity. Before the full translational potential of cAMP signalling can be tapped, it is critical to understand the structural basis for selective cAMP agonism and antagonism. Recent NMR investigations have shown that structurally homologous CBDs respond differently to several CBD ligands and that these unexpected differences arise at the level of either binding (i.e. affinity) or allostery (i.e. modulation of the autoinhibitory equilibria). In the present article, we specifically address how the highly conserved CBD fold binds cAMP with markedly different affinities in PKA (protein kinase A) relative to other eukaryotic cAMP receptors, such as Epac (exchange protein directly activated by cAMP) and HCN (hyperpolarization-activated cyclic-nucleotide-modulated channel). A major emerging determinant of cAMP affinity is hypothesized to be the position of the autoinhibitory equilibrium of the apo-CBD, which appears to vary significantly across different CBDs. These analyses may assist the development of selective CBD effectors that serve as potential drug leads for the treatment of cardiovascular diseases.


Current and Emerging Technologies for Probing Molecular Signatures of Traumatic Brain Injury.
Traumatic brain injury (TBI) is understood as an interplay between the initial injury, subsequent secondary injuries, and a complex host response all of which are highly heterogeneous. An understanding of the underlying biology suggests a number of windows where mechanistically inspired interventions could be targeted. Unfortunately, biologically plausible therapies have to-date failed to translate into clinical practice. While a number of stereotypical pathways are now understood to be involved, current clinical characterization is too crude for it to be possible to characterize the biological phenotype in a truly mechanistically meaningful way. In this review, we examine current and emerging technologies for fuller biochemical characterization by the simultaneous measurement of multiple, diverse biomarkers. We describe how clinically available techniques such as cerebral microdialysis can be leveraged to give mechanistic insights into TBI pathobiology and how multiplex proteomic and metabolomic techniques can give a more complete description of the underlying biology. We also describe spatially resolved label-free multiplex techniques capable of probing structural differences in chemical signatures. Finally, we touch on the bioinformatics challenges that result from the acquisition of such large amounts of chemical data in the search for a more mechanistically complete description of the TBI phenotype.