Childhood stress, serotonin transporter gene and brain structures in major depression.
The underlying neurobiology of major depression (MD) is likely to represent an interaction between genetic susceptibility and environmental factors such as stress. We investigated, in a multimodal high-resolution magnetic resonance imaging (MRI) genetic study, whether reduced hippocampal volumes and other brain alterations are associated with the tri-allelic polymorphism of the serotonin transporter and childhood stress in patients with MD and healthy subjects. Patients with MD and healthy participants were investigated using high-resolution MRI and genotyping for serotonin transporter polymorphism in the promoter region of the serotonin transporter gene (SLC6A4, 5-HTTLPR). Region of interest analysis of the hippocampus, whole-brain voxel-based morphometry (VBM), and assessment of childhood stress were carried out. Patients carrying the risk S-allele developed smaller hippocampal volumes when they had a history of emotional neglect compared with patients who only had one risk factor (environmental or genetic). In patients, childhood stress also predicted further hippocampal white matter alterations independently from the genotype. Moreover, the left prefrontal cortex was smaller in patients, whereby childhood stress resulted in larger prefrontal volumes in those subjects carrying the non-risk L-allele, suggesting preventive effects. The findings indicate that subjects with both environmental and genetic risk factors are susceptible to stress-related hippocampal changes. Structural brain changes due to stress represent part of the mechanism by which the illness risk and outcome might be genetically mediated.


[Hepatitis A antibodies in children].
With the finding of HBV and HAV, it is possible at the present time to recognize according to its etiology three types of viral hepatitis: type A, Type B and type non-A-non-B. In this paper we have proved that a low socioeconomic status plays a very important role in the spreading of this disease. In a community with a low socioeconomic status, of forty children attending the fourth grade of a primary school, 97 percent were found to have anti-HA: while in a similar group of children with a higher socioeconomic background, we found only 40 per cent of positive cases. A positive sero convertion to anti-HA was found in 90 per cent of the 73 children with acute hepatitis; the remaining 10 per cent were non-A-non-B. A group of 61 children admitted to the L. Calvo Mackenna Children's Hospital with acute or chronic non hepatic disease were studied for anti-HA. We found 77% positive cases in the group of infants under 4 months old; in older infants, a low incidence of anti-HA was observed, but a notorious increase of positive cases was seen after the age of two years, reaching 100 percent of positive cases in children above four years of age. In ten patients studied with prolonged hepatitis, five of them could be possibly classified as having a non-A-non-B hepatitis.


Creating σ-holes through the formation of beryllium bonds.
Through the use of ab initio theoretical models based on MP2/aug-cc-pVDZ-optimized geometries and CCSD(T)/aug-cc-pVTZ and CCSD(T)/aug-c-pVDZ total energies, it has been shown that the significant electron density rearrangements that follow the formation of a beryllium bond may lead to the appearance of a σ-hole in systems that previously do not exhibit this feature, such as CH3 OF, NO2 F, NO3 F, and other fluorine-containing systems. The creation of the σ-hole is another manifestation of the bond activation-reinforcement (BAR) rule. The appearance of a σ-hole on the F atoms of CH3 OF is due to the enhancement of the electronegativity of the O atom that participates in the beryllium bond. This atom recovers part of the charge transferred to Be by polarizing the valence density of the F into the bonding region. An analysis of the electron density shows that indeed this bond becomes reinforced, but the F atom becomes more electron deficient with the appearance of the σ-hole. Importantly, similar effects are also observed even when the atom participating in the beryllium bond is not directly attached to the F atom, as in NO2 F, NO3 F, or NCF. Hence, whereas the isolated CH3 OF, NO2 F, and NO3 F are unable to yield F⋅⋅⋅Base halogen bonds, their complexes with BeX2 derivatives are able to yield such bonds. Significant cooperative effects between the new halogen bond and the beryllium bond reinforce the strength of both noncovalent interactions.


Phase I study of intravenous ribavirin treatment of respiratory syncytial virus pneumonia after marrow transplantation.
Respiratory syncytial virus (RSV) pneumonia in marrow transplant recipients is associated with significant mortality. Ribavirin is a nucleoside analog with activity against RSV and in its aerosolized formulation is the only drug approved for treatment of RSV pneumonia in the United States. The clinical use of aerosolized ribavirin has been limited by caregivers' concerns about drug exposure and potential teratogenic effects. Since there is lack of proven efficacy and safety of the aerosolized ribavirin in this setting, we performed a phase I study of intravenous ribavirin treatment. Between November 1993 and May 1994, 10 patients with clinically significant RSV pneumonia at the Fred Hutchinson Cancer Research Center were enrolled. Only 2 of the 10 survived (20%; 95% CI, 3-56). Two of the 10 patients developed acute hemolysis that necessitated discontinuation of the medication. In conclusion, treatment of marrow transplant recipients with RSV pneumonia with intravenous ribavirin did not improve mortality compared with historical controls treated with the aerosolized drug.


Estimating the value of novel interventions for Parkinson's disease: an early decision-making model with application to dopamine cell replacement.
A long-term cost-effectiveness model for early decision-making and estimation of outcomes of novel therapeutic procedures for Parkinson's disease (PD) was developed based on the Hoehn and Yahr (HY) stages of PD. Results provided support for model validity. Model application to a future dopamine cell replacement therapy indicated long-term cost offsets and gains in quality-adjusted life years (QALYs) in early onset PD (HY III-IV), as compared to standard drug therapy. The maximum price premium (i.e., profit or compensation for developmental costs) for the intervention to remain cost-effective was estimated to euro12000-64000 according to cost-per-QALY thresholds of euro38000-70000 and depending on whether all or only medical direct costs are considered. The study illustrates the value of early health economic modeling and the described model shows promise as a means to estimate outcomes and aid decision-making regarding novel interventions for PD.


Do terrestrial invertebrates experience floodplains as landscape mosaics? Immediate and longer-term effects of flooding on ant assemblages in a floodplain forest.
Considering the floodplain landscape as a mosaic of habitat patches at different successional stages is useful for understanding (1) the processes associated with individual floods and (2) the legacy of flood history. Here, we investigate the applicability of the mosaic model to opportunistic ant species inhabiting the floodplain. Ground-active ant assemblages in river red gum (Eucalyptus camaldulensis) floodplain forest in south-eastern Australia were sampled before and for two years after a major flood in 2000-2001 at 24 sites with different inundation histories. Despite the mobility and opportunistic life history traits of floodplain ants, flood history appeared to impose a persistent mosaic structure on ant assemblages. Increasing duration of inundation of the forest floor was associated with decreasing species richness. beta-diversity was low, with the ant species at the most inundation-prone sites being a subset of those at drier sites. Less extensive flooding occurred in 2002-2003, enabling the consistency of short-term responses to inundation to be assessed. Flooding acts as a resetting mechanism, creating a characteristic ant assemblage. After floodwaters receded, there was little evidence of convergence in the structure of ant assemblages through time between sites flooded for different durations. The persistence of dissimilarities in ant assemblages suggests that succession towards terrestrialization was either not occurring or that it was operating at a rate that was too slow to be detected.


Aberrant vimentin methylation is characteristic of upper gastrointestinal pathologies.
We have previously established aberrant DNA methylation of vimentin exon-1 (VIM methylation) as a common epigenetic event in colon cancer and as a biomarker for detecting colon neoplasia. We now examine vimentin methylation in neoplasia of the upper gastrointestinal tract. Using a quantitative real-time methylation-specific PCR assay, we tested for vimentin methylation in archival specimens of esophageal and gastric neoplasia. We find that acquisition of aberrant vimentin methylation is highly common in these neoplasms, but largely absent in controls. The highest frequency of vimentin methylation was detected in lesions of the distal esophagus, including 91% of Barrett's esophagus (n = 11), 100% of high-grade dysplasia (HGD, n = 5), and 81% of esophageal adenocarcinoma (EAC, n = 26) but absent in controls (n = 9). Vimentin methylation similarly was detected in 87% of signet ring (n = 15) and 53% of intestinal type gastric cancers (n = 17). Moreover, in tests of cytology brushings vimentin methylation proved detectable in 100% of Barrett's esophagus cases (n = 7), 100% of HGD cases (n = 4), and 83% of EAC cases (n = 18) but was absent in all controls (n = 5). These findings establish aberrant vimentin methylation as a highly common epigenetic alteration in neoplasia of the upper gastrointestinal tract and show that Barrett's esophagus, even without dysplasia, already contains epigenetic alterations characteristic of adenocarcinoma. These findings suggest vimentin methylation as a biomarker of upper gastrointestinal neoplasia with potential for development as molecular cytology in esophageal screening.


FGF23 gene regulation by 1,25-dihydroxyvitamin D: opposing effects in adipocytes and osteocytes.
In a closed endocrine loop, 1,25-dihydroxyvitamin D3 (1,25D) induces the expression of fibroblast growth factor 23 (FGF23) in bone, with the phosphaturic peptide in turn acting at kidney to feedback repress CYP27B1 and induce CYP24A1 to limit the levels of 1,25D. In 3T3-L1 differentiated adipocytes, 1,25D represses FGF23 and leptin expression and induces C/EBPβ, but does not affect leptin receptor transcription. Conversely, in UMR-106 osteoblast-like cells, FGF23 mRNA concentrations are upregulated by 1,25D, an effect that is blunted by lysophosphatidic acid, a cell-surface acting ligand. Progressive truncation of the mouse FGF23 proximal promoter linked in luciferase reporter constructs reveals a 1,25D-responsive region between -400 and -200 bp. A 0.6 kb fragment of the mouse FGF23 promoter, linked in a reporter construct, responds to 1,25D with a fourfold enhancement of transcription in transfected K562 cells. Mutation of either an ETS1 site at -346 bp, or an adjacent candidate vitamin D receptor (VDR)/Nurr1-element, in the 0.6 kb reporter construct reduces the transcriptional activity elicited by 1,25D to a level that is not significantly different from a minimal promoter. This composite ETS1-VDR/Nurr1 cis-element may function as a switch between induction (osteocytes) and repression (adipocytes) of FGF23, depending on the cellular setting of transcription factors. Moreover, experiments demonstrate that a 1 kb mouse FGF23 promoter-reporter construct, transfected into MC3T3-E1 osteoblast-like cells, responds to a high calcium challenge with a statistically significant 1.7- to 2.0-fold enhancement of transcription. Thus, the FGF23 proximal promoter harbors cis elements that drive responsiveness to 1,25D and calcium, agents that induce FGF23 to curtail the pathologic consequences of their excess.


Translating genetic and preclinical findings into autism therapies.
Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by social deficits and repetitive/restrictive interests. ASD is associated with multiple comorbidities, including intellectual disability, anxiety, and epilepsy. Evidence that ASD is highly heritable has spurred major efforts to unravel its genetics, revealing possible contributions from hundreds of genes through rare and common variation and through copy-number changes. In this perspective, we provide an overview of the current state of ASD genetics and of how genetic research has spurred the development of in vivo and in vitro models using animals and patient cells to evaluate the impact of genetic mutations on cellular function leading to disease. Efforts to translate these findings into successful therapies have yet to bear fruit. We discuss how the valuable insight into the disorder provided by these new models can be used to better understand ASD and develop future clinical trials.


The nematode fauna of long-nosed mice Oxymycterus spp. from the Bolivian Yungas.
During a long-term survey of the parasites of mammals from all over Bolivia, 44 individuals of Oxymycterus inca and 6 of Oxymycterus paramensis were collected and examined for parasites from the foothills and Eastern Cordillera (Los Yungas) of the Andes of Bolivia. Three species of nematode were found including, from the cecum, a previously unknown genus and species of pinworm, Caroloxyuris boliviensis n. gen, n. sp., aspidoderids representing Nematomystes rodentophilus, and from the stomach, Protospirura numidica criceticola. These helminths occurred in prevalences of 25, 9, and 14%, respectively, in O. paramensis. Caroloxyuris boliviensis resembles species included in the genus Syphacia in the structure of the cephalic mask and copulatory organs, and the extension of lateral alae. However, males of this species possess only 2 mamelons on the ventral surface of the body. Caroloxyuris can be recognized as being distinct from both Helminthoxys and Rauschtineria in the ornamentation of the mamelons and the shape of cephalic mask. A redescription of N. rodentophilus is provided, and Nematomystes scapteromi is transferred from Ansiruptodera to Nematomystes.


Timing of Preharvest Infection of Pear Fruit by Botrytis cinerea and the Relationship to Postharvest Decay.
Botrytis cinerea causes significant levels of postharvest decay in the winter pear cultivar d'Anjou. The objectives of this study were to determine the timing of B. cinerea infection of pear stems and calyxes in the orchard during the growing season, to investigate the development of gray mold in storage, and to determine whether preharvest levels of B. cinerea in pear stems and calyxes can be used as predictors of gray mold levels observed in storage. Very low levels of B. cinerea were isolated from stem tissue prior to harvest. In a single year repeat experiment, stems sampled at harvest had higher levels of infection than those sampled earlier in the season. Little or no stem end gray mold was detected in fruit after 3 months in air-storage; however, incidence increased between 6 and 8 months. Calyx end gray mold was detected at low levels in fruit stored for up to 8 months. The mean incidence of stem end gray mold was 3.6 and 2.0%, and incidence of calyx end gray mold was 1.2 and 0.2%, in 1996 and 1997, respectively. Calyxes were susceptible to infection soon after full bloom; however, inoculation of calyxes in April or May did not result in higher levels of calyx end gray mold in storage. Therefore, preharvest level of calyx infection is a poor predictor of calyx end gray mold in storage. In addition, application of benomyl in the orchard reduced the level of B. cinerea in blossoms but had no effect on levels of calyx end gray mold of fruit in storage. Packing and shipping fruit within 3 to 6 months of harvest may mitigate economic losses due to gray mold.


DNA methylation profiles of donor nuclei cells and tissues of cloned bovine fetuses.
Methylation of DNA in CpG islands plays an important role during fetal development and differentiation because CpG islands are preferentially located in upstream regions of mammalian genomic DNA, including the transcription start site of housekeeping genes and are also associated with tissue-specific genes. Somatic nuclear transfer (NT) technology has been used to generate live clones in numerous mammalian species, but only a low percentage of nuclear transferred animals develop to term. Abnormal epigenetic changes in the CpG islands of donor nuclei after nuclear transfer could contribute to a high rate of abortion during early gestation and increase perinatal death. These changes have yet to be explored. Thus, we investigated the genome-wide DNA methylation profiles of CpG islands in nuclei donor cells and NT animals. Using Restriction Landmark Genomic Scanning (RLGS), we showed, for the first time, the epigenetic profile formation of tissues from NT bovine fetuses produced from cumulus cells. From approximately 2600 unmethylated NotI sites visualized on the RLGS profile, at least 35 NotI sites showed different methylation statuses. Moreover, we proved that fetal and placental tissues from artificially inseminated and cloned cattle have tissue-specific differences in the genome-wide methylation profiles of the CpG islands. We also found that possible abnormalities occurred in the fetal brain and placental tissues of cloned animals.


The Humoral Immune Response to HCV: Understanding is Key to Vaccine Development.
Hepatitis C virus (HCV) remains a global problem, despite advances in treatment. The low cost and high benefit of vaccines have made them the backbone of modern public health strategies, and the fight against HCV will not be won without an effective vaccine. Achievement of this goal will benefit from a robust understanding of virus-host interactions and protective immunity in HCV infection. In this review, we summarize recent findings on HCV-specific antibody responses associated with chronic and spontaneously resolving human infection. In addition, we discuss specific epitopes within HCV's envelope glycoproteins that are targeted by neutralizing antibodies. Understanding what prompts or prevents a successful immune response leading to viral clearance or persistence is essential to designing a successful vaccine.


Clonally expanded novel multipotent stem cells from human bone marrow regenerate myocardium after myocardial infarction.
We have identified a subpopulation of stem cells within adult human BM, isolated at the single-cell level, that self-renew without loss of multipotency for more than 140 population doublings and exhibit the capacity for differentiation into cells of all 3 germ layers. Based on surface marker expression, these clonally expanded human BM-derived multipotent stem cells (hBMSCs) do not appear to belong to any previously described BM-derived stem cell population. Intramyocardial transplantation of hBMSCs after myocardial infarction resulted in robust engraftment of transplanted cells, which exhibited colocalization with markers of cardiomyocyte (CMC), EC, and smooth muscle cell (SMC) identity, consistent with differentiation of hBMSCs into multiple lineages in vivo. Furthermore, upregulation of paracrine factors including angiogenic cytokines and antiapoptotic factors, and proliferation of host ECs and CMCs, were observed in the hBMSC-transplanted hearts. Coculture of hBMSCs with CMCs, ECs, or SMCs revealed that phenotypic changes of hBMSCs result from both differentiation and fusion. Collectively, the favorable effect of hBMSC transplantation after myocardial infarction appears to be due to augmentation of proliferation and preservation of host myocardial tissues as well as differentiation of hBMSCs for tissue regeneration and repair. To our knowledge, this is the first demonstration that a specific population of multipotent human BM-derived stem cells can induce both therapeutic neovascularization and endogenous and exogenous cardiomyogenesis.


Investigating sources of variability of monochromatic and transverse chromatic aberrations across eyes.
Schematic eye models have typically been used to explain the average monochromatic and chromatic imaging properties of the eye. Both monochromatic aberrations and transverse chromatic aberration are known to vary widely across subjects. However, to our knowledge, the ability of schematic eye models to predict these individual variations has not been tested experimentally. We used a spatially resolved refractometer to measure the monochromatic aberrations and the optical transverse chromatic aberration (oTCA) in a group of 15 eyes. By recording the 1st and 4th Purkinje images for five directions of gaze, we also estimated the tilt, misalignment of ocular surfaces (front surface of the cornea and back surface of the lens) and off-axis position of the fovea (angle alpha), as well as pupil centration. We conclude that, contrary to expectations none of those factors are major contributors to the variability in monochromatic aberrations and oTCA in this group of eyes. Simulations show that corneal curvature and corneal conicity are also unlikely to account for the observed relation between monochromatic aberrations and oTCA. Our results suggest an important contribution of corneal irregularities to those aberrations.


Implementing a complex rehabilitation intervention in a stroke trial: a qualitative process evaluation of AVERT.
The implementation of multidisciplinary stroke rehabilitation interventions is challenging, even when the intervention is evidence-based. Very little is known about the implementation of complex interventions in rehabilitation clinical trials. The aim of study was to better understand how the implementation of a rehabilitation intervention in a clinical trial within acute stroke units is experienced by the staff involved. This qualitative process evaluation was part of a large Phase III stroke rehabilitation trial (AVERT). A descriptive qualitative approach was used. We purposively sampled 53 allied health and nursing staff from 19 acute stroke units in Australia, New Zealand and Scotland. Semi-structured interviews were conducted by phone, voice-internet, or face to face. Digitally recorded interviews were transcribed and analysed by two researchers using rigorous thematic analysis. Our analysis uncovered ten important themes that provide insight into the challenges of implementing complex new rehabilitation practices within complex care settings, plus factors and strategies that assisted implementation. Themes were grouped into three main categories: staff experience of implementing the trial intervention, barriers to implementation, and overcoming the barriers. Participation in the trial was challenging but had personal rewards and improved teamwork at some sites. Over the years that the trial ran some staff perceived a change in usual care. Barriers to trial implementation at some sites included poor teamwork, inadequate staffing, various organisational barriers, staff attitudes and beliefs, and patient-related barriers. Participants described successful implementation strategies that were built on interdisciplinary teamwork, education and strong leadership to 'get staff on board', and developing different ways of working. The AVERT stroke rehabilitation trial required commitment to deliver an intervention that needed strong collaboration between nurses and physiotherapists and was different to current care models. This qualitative process evaluation contributes unique insights into factors that may be critical to successful trials teams, and as AVERT was a pragmatic trial, success factors to delivering complex intervention in clinical practice. AVERT registered with Australian New Zealand Clinical Trials Registry ACTRN12606000185561 .


Unraveling the intricacies of mammalian fertilization.
It is imperative to understand the molecular basis of various steps involved during fertilization. In the manuscript by Bianchi et al. a novel protein, Juno on egg membrane (oolemma) has been characterized that binds to sperm specific protein, Izumo-1. Monoclonal antibodies against Juno inhibited in vitro fertilization. Juno knock-out female mice failed to deliver litters on mating. It is rapidly shed from oolemma after fertilization, suggesting its role in preventing polyspermy. Taken together these studies will help in our understanding of sperm-egg recognition mechanisms and also facilitate development of new fertility treatment regimens and novel contraceptives.


Experience of a faculty development workshop in mentoring at an Indian medical college.
Our medical college is running a mentoring programme for undergraduate medical students since 2009. The academic leadership of the college identified the need to change the focus of the programme from mere problem- solving to professional and personal development of mentees. A core group of faculty designed and implemented a workshop on mentoring for 28 mentors. The workshop included reflections on the participants' previous experiences about mentoring, discussion on perceptions of mentees about the existing mentoring programme, self-analysis of mentoring skills, overview of the Surrendering, Accepting, Gifting and Extending (SAGE) model and demonstration of effective mentoring skills using role plays and a film. We collected written anonymous feedback from participants at the end of the workshop to elicit their responses regarding various aspects of the programme, change in their views about mentoring and suggestions for future workshops. A majority of the participants (17, 60.7%) said that role plays and reflection on role plays were the most valuable part of workshop as they provided clarity on the concepts about mentoring. The most frequently identified take-home messages were: building trust with the mentee (7, 25%), balance in life and approach towards the mentee (6, 21.4%), and understanding that mentoring is a process geared towards personal and professional development of the mentee (6, 21.4%). The participants' reaction to the workshop was positive. The responses of participants suggested that the workshop was successful in changing their views regarding the purpose of the mentoring programme.


Selective perceptual impairments after perirhinal cortex ablation.
It has been suggested that the primate perirhinal cortex contributes exclusively to memory. However, recent studies in macaque monkeys have implied that the perirhinal cortex may also contribute to object perception. To investigate whether the perirhinal cortex does contribute to perception, we devised several perceptual oddity tasks in which monkeys had to choose which stimulus of several presented concurrently on a touch screen was different. Macaques with bilateral perirhinal cortex ablations were selectively impaired relative to controls at perceptually discriminating the odd stimulus when the odd stimulus was a different object and when the discrimination could not be done on the basis of simple differences in features between the stimuli. They remained unimpaired relative to controls on discriminating the odd stimulus when the odd stimulus was a different color, a different shape, or a different size even when these discriminations were extremely difficult. They were also impaired on human and monkey face oddity tasks and oddity tasks with scenes containing objects. Therefore, we reject the notion that the macaque perirhinal cortex has a role exclusive to memory and conclude that the macaque perirhinal cortex does contribute to perception. We argue that the perirhinal cortex is neither specialized for perception nor memory processes alone, but rather, is specialized for processing stimuli that require processing at a more abstract level such as at the level of an object and that the perirhinal cortex contributes to both memory and perception of such stimuli.


Psychotic experiences co-occur with sleep problems, negative affect and mental disorders in preadolescence.
Knowledge on the significance of childhood psychotic symptoms and experiences (PE) is still limited. This study aimed to investigate the prevalence and clinical significance of PE in preadolescent children from the general population by use of in-depth psychopathological interviews and comprehensive diagnostic assessments. We investigated 1,632 children from the general population-based Copenhagen Child Cohort 2000. PE were measured by semistructured interviews using the K-SADS-PL-items on psychotic and affective symptoms, each symptom scored as not present versus likely or definitely present. The Development and Well-Being Assessment (DAWBA) was used independently to diagnose DSM-IV-mental disorders. Puberty development and sleep disturbance were self-reported. The associations between PE (any lifetime hallucination and/or delusion) and various mental problems and disorders were examined by multivariable binomial regression analyses, adjusting for gender and onset of puberty. The weighted life time prevalence of PE at age 11-12 years was 10.9% (CI 9.1-12.7). The majority of children with PE (n = 172) either had a diagnosable DSM-IV-mental disorder (31.4%) or self-reported mental health difficulties in absence of a diagnosis (31.4%). The risk of delusions increased with onset of puberty. The risk of PE increased with emotional and neurodevelopmental disorders, subthreshold depressive symptoms, sleep problems and lack of sleep, regardless of whether PE were expressed as hallucinations and/or delusions. The highest correlations were seen for emotional and multiple disorders. Psychotic experiences are particularly prevalent in the context of affective dysregulation and sleep disturbance, increase with onset of puberty and represent a trans-diagnostic marker of psychopathology.


Closed-cell crystalline foams: self-assembling, resonant metamaterials.
Internal degrees of freedom and periodic structure are critical requirements in the design of acoustic/elastic metamaterials since they can give rise to extraordinary properties like negative effective mass and stiffness. However, they are challenging to realize in three dimensions. Closed-cell, crystalline foams are a particularly advantageous basis to develop metamaterials as they intrinsically have a complex microstructure, exhibiting internal resonances. Recently self-assembly techniques have been implemented to produce such foams: a Kelvin (body centered cubic) foam, a face centered cubic foam, and a Weaire-Phelan structure. Numerical models are employed to demonstrate that such foams are superanisotropic, selectively behaving as a fluid or a solid, pentamode solids as a result of fluid-structure interaction, in addition to having regimes characterized by film resonances and high density of states. Microstructural deformations obtained from numerical models allow the derivation of equivalent mechanical models.


Deletion of microsomal cytochrome b5 profoundly affects hepatic and extrahepatic drug metabolism.
We demonstrated recently that cytochrome b(5) plays an important in vivo role in hepatic cytochrome P450 (P450) function [J Biol Chem 283:31385-31393, 2008]. We have now generated a model in which cytochrome b(5) has been deleted in all tissues [cytochrome b(5) complete null (BCN)], which surprisingly results in a viable mouse despite the putative in vivo roles of this protein in lipid and steroid hormone metabolism and the reduction of methemoglobin. In contrast to the liver-specific deletion, complete deletion of cytochrome b(5) leads to a neonatal increase in the expression of many hepatic P450s at both the protein and mRNA level. In extrahepatic tissues, some changes in P450 expression were also observed that were isoform-dependent. In vitro cytochrome P450 activities in liver, kidney, lung, and small intestine of BCN mice were determined for a range of model substrates and probe drugs; a profound reduction in the metabolism of some substrates, particularly in lung, kidney, and small intestine, was observed. In vivo, the metabolism of metoprolol was significantly altered in BCN mice, in contrast to the previous finding in the liver-specific cytochrome b(5) deletion, suggesting that extrahepatic cytochrome b(5) plays a significant role in its disposition. Testicular Cyp17 hydroxylase and lyase activities were also significantly reduced by cytochrome b(5) deletion, leading to significantly lower levels of testicular testosterone. The BCN mouse provides an additional model system with which to further investigate the functions of cytochrome b(5), particularly in extrahepatic tissues.


Adipose levels of polybrominated diphenyl ethers and risk of breast cancer.
We conducted a case-control study to evaluate the risk of breast cancer associated with adipose concentrations of polybrominated diphenyl ethers (PBDEs) among women undergoing surgical breast biopsies in the San Francisco Bay Area of California (n=78 cases; 56 controls). Adipose tissue was analyzed for the five major congeners of PBDEs. Unconditional logistic regression was used to estimate age- and race-adjusted exposure-specific odds ratios (ORs) and 95% confidence intervals (95% CI). Adipose levels of PBDEs were among the highest ever reported. Adjusted ORs for the highest compared with lowest levels of exposures were as follows: 0.56 (95% CI 0.19-1.68) for BDE-47; 1.19 (95% CI 0.35-4.10) for BDE-99; 0.91 (95% CI 0.33-2.53) for BDE-100; 0.52 (95% CI 0.19-1.39) for BDE-153; 1.67 (95% CI 0.44-6.29) for BDE-154; 2.04 (95% CI 0.45-9.20) for total BDEs. These results provide no evidence of an association between PBDE adipose concentrations measured at or near the time of diagnosis and breast cancer risk. Our study was limited by a small sample size. Given the high levels of PBDEs found in this population of California women, future studies are warranted. Such studies would benefit from a larger sample size, a more representative control series, and/or a prospective design.


Towards elimination of lymphatic filariasis in India.
The global initiatives to eliminate lymphatic filariasis as a public health problem by the year 2020 have generated a great deal of debate in India, the largest endemic country. This has led to a shift in the focus from control to elimination of the disease. Although the campaign to eliminate filariasis has begun, much more needs to be done. Several recent research studies have provided an insight into various operational issues and prospects of elimination of lymphatic filariasis. In this article, the current scenario, recent research results, logistics and the prospects of eliminating lymphatic filariasis in India will be discussed.


Lathosterolosis: an inborn error of human and murine cholesterol synthesis due to lathosterol 5-desaturase deficiency.
Lathosterol 5-desaturase catalyzes the conversion of lathosterol to 7-dehydrocholesterol in the next to last step of cholesterol synthesis. Inborn errors of cholesterol synthesis underlie a group of human malformation syndromes including Smith-Lemli-Opitz syndrome, desmosterolosis, CHILD syndrome, CDPX2 and lathosterolosis. We disrupted the lathosterol 5-desaturase gene (Sc5d ) in order to further our understanding of the pathophysiological processes underlying these disorders and to gain insight into the corresponding human disorder. Sc5d (-/-) pups were stillborn, had elevated lathosterol and decreased cholesterol levels, had craniofacial defects including cleft palate and micrognathia, and limb patterning defects. Many of the malformations found in Sc5d (-/-) mice are consistent with impaired hedgehog signaling, and appear to be a result of decreased cholesterol rather than increased lathosterol. A patient initially described as atypical SLOS with mucolipidosis was shown to have lathosterolosis by biochemical and molecular analysis. We identified a homozygous mutation of SC5D (137A>C, Y46S) in this patient. An unique aspect of the lathosterolosis phenotype is the combination of a malformation syndrome with an intracellular storage defect.


Variant Ciz1 is a circulating biomarker for early-stage lung cancer.
There is an unmet need for circulating biomarkers that can detect early-stage lung cancer. Here we show that a variant form of the nuclear matrix-associated DNA replication factor Ciz1 is present in 34/35 lung tumors but not in adjacent tissue, giving rise to stable protein quantifiable by Western blot in less than a microliter of plasma from lung cancer patients. In two independent sets, with 170 and 160 samples, respectively, variant Ciz1 correctly identified patients who had stage 1 lung cancer with clinically useful accuracy. For set 1, mean variant Ciz1 level in individuals without diagnosed tumors established a threshold that correctly classified 98% of small cell lung cancers (SCLC) and non-SCLC patients [receiver operator characteristic area under the curve (AUC) 0.958]. Within set 2, comparison of patients with stage 1 non-SCLC with asymptomatic age-matched smokers or individuals with benign lung nodules correctly classified 95% of patients (AUCs 0.913 and 0.905), with overall specificity of 76% and 71%, respectively. Moreover, using the mean of controls in set 1, we achieved 95% sensitivity among patients with stage 1 non-SCLC patients in set 2 with 74% specificity, demonstrating the robustness of the classification. RNAi-mediated selective depletion of variant Ciz1 is sufficient to restrain the growth of tumor cells that express it, identifying variant Ciz1 as a functionally relevant driver of cell proliferation in vitro and in vivo. The data show that variant Ciz1 is a strong candidate for a cancer-specific single marker capable of identifying early-stage lung cancer within at-risk groups without resort to invasive procedures.


"Moving forward: a cross sectional baseline study of staff and student attitudes towards a totally smoke free university campus".
Baseline data were collected to inform the adoption, implementation and institutionalisation phases of a completely smoke free campus policy at a large Western Australian university with a diverse student and staff community. An online survey was randomly emailed to staff and students to measure the attitudes towards and the acceptability and enforcement of the policy prior to implementation. In total, 969 respondents completed the survey. General attitudes towards smoking were negative. While smokers, ex-smokers and non-smokers were supportive of smoke free policy on campus, 65.7% of respondents felt the campus should be completely smoke free. Respondents indicated a smoke free policy should be stringently enforced. The majority of respondents reported that they had been exposed to second-hand smoke on campus (n = 768; 79.5%). Theory of Organisational Change provides a useful framework to support the implementation of the completely smoke free policy in the University setting. The implementation process needs to consider the broad range of issues associated with implementing a completely smoke free policy and address issues such as safety of smokers, ensuring smokers are not marginalised and ensuring a comprehensive program is implemented. These baseline findings can be used to advocate for the implementation of a comprehensive range of strategies that recognise the addictive nature of tobacco smoking and address attitude and behaviour change, environmental adaptations and effective implementation of the policy. Administration should consider smokers and non-smokers when policy is implemented.


Validation of the OSCAR 2 oscillometric 24-hour ambulatory blood pressure monitor according to the International Protocol for the validation of blood pressure measuring devices.
The accuracy of blood pressure measuring devices is of prime importance and should be validated before devices are used clinically. We carried out an independent evaluation of the OSCAR 2 oscillometric ambulatory blood pressure monitor (SunTech Medical model 222) according to the European Society of Hypertension International Protocol. Adult subjects were recruited from patients and staff at The James Cook University Hospital, Middlesbrough, UK. Nine sequential same-arm blood pressure measurements were taken alternating between simultaneous readings by two independent, trained observers using mercury sphygmomanometers and the device operated by a third observer. In phase one, 15 subjects participated (five in each of the low, medium and high blood pressure ranges) with 18 subjects participating in phase two. Data from 33 subjects (11 in each of the three blood pressure ranges) were analysed for systolic (19 male, 14 female, mean age 56.0 years) and for diastolic (17 male, 16 female, mean age 51.1 years) blood pressure. The OSCAR 2 passed the first phase of the validation process. In phase 2.1, the OSCAR 2 monitor had 71 readings within 5 mmHg, 86 within 10 mmHg and 94 within 15 mmHg for systolic blood pressure (SBP) and 72 readings within 5 mmHg, 88 within 10 mmHg and 96 within 15 mmHg for diastolic blood pressure (DBP). Mean (+/-SD) differences between observers and device were 0.9+/-2.3 mmHg for SBP and -0.5+/-2.3 mmHg for DBP. In phase 2, 24 subjects had at least two of the differences within 5 mmHg and three subjects had no differences within 5 mmHg for SBP while for DBP 25 subjects had at least two of the differences within 5 mmHg and two subjects had no differences within 5 mmHg. The OSCAR 2 passes all requirements for validation by the International Protocol and can be recommended for clinical use in an adult population.


Spectroscopical and mechanical characterization of normal and thalassemic red blood cells by Raman Tweezers.
In this work, the effects of thalassemia, a blood disease quite diffuse in the Mediterranean sea region, have been investigated at single cell level using a Raman Tweezers system. By resonant excitation of hemoglobin Raman bands, we have examined the oxygenation capability of beta-thalassemic erythrocytes. A reduction of this fundamental erythrocyte function has been found. The measurements have been performed on a significant number of red blood cells; the relative statistical analysis is presented. Moreover, the response to photo-induced oxidative stress of diseased cells with respect to the normal ones has been analyzed. Finally, the deformability of thalassemic erythrocytes has been quantified by measuring the membrane shear modulus by using a double-trap system: the measurements have revealed an increase in membrane rigidity of more than 40%, giving evidence that the genetic defect associated to thalassemia, which manly relies on hemoglobin structure, also strongly affects the erythrocyte mechanical properties. Our results demonstrate that the developed set-up may have potential for the monitoring of blood diseases and their response to drug therapies.


Isolation of three novel rat and mouse papillomaviruses and their genomic characterization.
Despite a growing knowledge about the biological diversity of papillomaviruses (PV), only little is known about non-human PV in general and about PV mice models in particular. We cloned and sequenced the complete genomes of two novel PV types from the Norway rat (Rattus norvegicus; RnPV2) and the wood mouse (Apodemus sylvaticus; AsPV1) as well as a novel variant of the recently described MmuPV1 (originally designated as MusPV) from a house mouse (Mus musculus; MmuPV1 variant). In addition, we conducted phylogenetic analyses using a systematically representative set of 79 PV types, including the novel sequences. As inferred from concatenated amino acid sequences of six proteins, MmuPV1 variant and AsPV1 nested within the Beta+Xi-PV super taxon as members of the Pi-PV. RnPV2 is a member of the Iota-PV that has a distant phylogenetic position from Pi-PV. The phylogenetic results support a complex scenario of PV diversification driven by different evolutionary forces including co-divergence with hosts and adaptive radiations to new environments. PV types particularly isolated from mice and rats are the basis for new animal models, which are valuable to study PV induced tumors and new treatment options.


Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor Aegilops tauschii.
Development of a high quality reference sequence is a daunting task in crops like wheat with large (~17Gb), highly repetitive (>80%) and polyploid genome. To achieve complete sequence assembly of such genomes, development of a high quality physical map is a necessary first step. However, due to the lack of recombination in certain regions of the chromosomes, genetic mapping, which uses recombination frequency to map marker loci, alone is not sufficient to develop high quality marker scaffolds for a sequence ready physical map. Radiation hybrid (RH) mapping, which uses radiation induced chromosomal breaks, has proven to be a successful approach for developing marker scaffolds for sequence assembly in animal systems. Here, the development and characterization of a RH panel for the mapping of D-genome of wheat progenitor Aegilops tauschii is reported. Radiation dosages of 350 and 450 Gy were optimized for seed irradiation of a synthetic hexaploid (AABBDD) wheat with the D-genome of Ae. tauschii accession AL8/78. The surviving plants after irradiation were crossed to durum wheat (AABB), to produce pentaploid RH1s (AABBD), which allows the simultaneous mapping of the whole D-genome. A panel of 1,510 RH1 plants was obtained, of which 592 plants were generated from the mature RH1 seeds, and 918 plants were rescued through embryo culture due to poor germination (<3%) of mature RH1 seeds. This panel showed a homogenous marker loss (2.1%) after screening with SSR markers uniformly covering all the D-genome chromosomes. Different marker systems mostly detected different lines with deletions. Using markers covering known distances, the mapping resolution of this RH panel was estimated to be <140kb. Analysis of only 16 RH lines carrying deletions on chromosome 2D resulted in a physical map with cM/cR ratio of 1:5.2 and 15 distinct bins. Additionally, with this small set of lines, almost all the tested ESTs could be mapped. A set of 399 most informative RH lines with an average deletion frequency of ~10% were identified for developing high density marker scaffolds of the D-genome. The RH panel reported here is the first developed for any wild ancestor of a major cultivated plant species. The results provided insight into various aspects of RH mapping in plants, including the genetically effective cell number for wheat (for the first time) and the potential implementation of this technique in other plant species. This RH panel will be an invaluable resource for mapping gene based markers, developing a complete marker scaffold for the whole genome sequence assembly, fine mapping of markers and functional characterization of genes and gene networks present on the D-genome.


Conformational consequences of cooperative binding of a coiled-coil peptide motif to poly(N-(2-hydroxypropyl) methacrylamide) HPMA copolymers.
Small-angle neutron scattering and pulsed-gradient spin-echo NMR have been used to examine the solution conformation of a series of water soluble poly(N-(2-hydroxypropyl) methacrylamide) P(HPMA) co-polymer drug delivery vehicles incorporating a coiled-coil peptide motif as a novel pH sensitive non-covalent linker. The conformation of the HPMA homopolymer is well-described by a Gaussian coil model and changing pH from pH 7 to pH 5 has little effect on the solution conformation, as quantified via the radius of gyration. Copolymerisation with 5-10mol% of the K3 peptide bearing methacrylate monomer (K3-MA), gave a series of copolymers that exhibited an increase in radius of gyration at both pH's, despite being typically 30% lower in molecular weight, indicating that the K3-MA causes a perturbation (expansion) of the copolymer conformation. Subsequent addition of an equimolar amount of the complementary peptide E3 makes little further difference to the conformation, indicative of the intimate binding (coiled-coil motif) between the two peptides. Again, the effects of pH are small. Only the addition of a large aromatic structure such as methotrexate causes a further perturbation of the structure - the hydrophobic interaction between the MTX units causes a significant collapse of the polymer coil. These findings further elaborate the understanding of those factors that determine the solution conformation of novel polymer therapeutics.


Creation and validation of the Picture-Set of Young Children's Affective Facial Expressions (PSYCAFE).
The immediate detection and correct processing of affective facial expressions are one of the most important competences in social interaction and thus a main subject in emotion and affect research. Generally, studies in these research domains, use pictures of adults who display affective facial expressions as experimental stimuli. However, for studies investigating developmental psychology and attachment behaviour it is necessary to use age-matched stimuli, where it is children that display affective expressions. PSYCAFE represents a newly developed picture-set of children's faces. It includes reference portraits of girls and boys aged 4 to 6 years averaged digitally from different individual pictures, that were categorized to six basic affects (fear, disgust, happiness, sadness, anger and surprise) plus a neutral facial expression by cluster analysis. This procedure led to deindividualized and affect prototypical portraits. Individual affect expressive portraits of adults from an already validated picture-set (KDEF) were used in a similar way to create affect prototypical images also of adults. The stimulus set has been validated on human observers and entail emotion recognition accuracy rates and scores for intensity, authenticity and likeability ratings of the specific affect displayed. Moreover, the stimuli have also been characterized by the iMotions Facial Expression Analysis Module, providing additional data on probability values representing the likelihood that the stimuli depict the expected affect. Finally, the validation data from human observers and iMotions are compared to data on facial mimicry of healthy adults in response to these portraits, measured by facial EMG (m. zygomaticus major and m. corrugator supercilii).


Butyrophilin 3A (BTN3A, CD277)-specific antibody 20.1 differentially activates Vγ9Vδ2 TCR clonotypes and interferes with phosphoantigen activation.
Phosphoantigens (PAgs)-like HMBPP ((E)-4-hydroxy-3-methyl-but-2-enyl diphosphate) and butyrophilin 3 (BTN3A, CD277)-specific monoclonal antibody 20.1 induce TCR-mediated activation of Vγ9Vδ2 T cells. Here, we compared murine reporter cells transduced with Vγ9Vδ2 TCRs G115, D1C55, and MOP for the activation in culture with human RAJI cells and PAgs or mAb 20.1 and its single-chain (sc) derivative. All transductants responded readily to PAg but only TCR MOP γ-chain-expressing cells responded to mAb/sc 20.1. Furthermore, both antagonist and agonist mAb and sc of the agonist mAb inhibited the PAg response of TCR-transduced murine reporter cells. These findings suggest that, in contrast to stimulation by physiological stimulators (PAg), the responsiveness to mAb 20.1 depends strongly on CDR3 sequences of the TCR, and that mAb 20.1 can interfere with the PAg-response. Mouse or human origin of reporter cells might affect the mAb 20.1 response since all three TCR-mediated mAb 20.1-induced activation of TCR-transduced Jurkat cells. The pronounced differences between PAg and mAb 20.1-induced activation observed here help to understand the often contradictory published data. This study provides novel perspectives on the physiological mechanism of Vγ9Vδ2 T-cell activation, and highlights the complex mode of action of BTN3A-specific antibodies as agents in cancer immunotherapy.


Analysis of actin dynamics at the leading edge of crawling cells: implications for the shape of keratocyte lamellipodia.
Leading edge protrusion is one of the critical events in the cell motility cycle and it is believed to be driven by the assembly of the actin network. The concept of dendritic nucleation of actin filaments provides a basis for understanding the organization and dynamics of the actin network at the molecular level. At a larger scale, the dynamic geometry of the cell edge has been described in terms of the graded radial extension model, but this level of description has not yet been linked to the molecular dynamics. Here, we measure the graded distribution of actin filament density along the leading edge of fish epidermal keratocytes. We develop a mathematical model relating dendritic nucleation to the long-range actin distribution and the shape of the leading edge. In this model, a steady-state graded actin distribution evolves as a result of branching, growth and capping of actin filaments in a finite area of the leading edge. We model the shape of the leading edge as a product of the extension of the actin network, which depends on actin filament density. The feedback between the actin density and edge shape in the model results in a cell shape and an actin distribution similar to those experimentally observed. Thus, we explain the stability of the keratocyte shape in terms of the self-organization of the branching actin network.


An organized approach to improvement in guideline adherence for acute myocardial infarction: results with the Get With The Guidelines quality improvement program.
Evidence-based guidelines from the American Heart Association are voluntary, and adherence is highly variable across the country. Get With The Guidelines (GWTG) is a national quality improvement program sponsored and developed by the American Heart Association. The objective of this study was to evaluate whether participation in GWTG is associated with greater adherence to guidelines for coronary artery disease (CAD). Data on adherence to guidelines were obtained from Hospital Compare, grouping hospitals according to participation in the GWTG-CAD program on January 1, 2004: GWTG-CAD hospitals, n = 223; non-GWTG-CAD hospitals, n = 3407. The GWTG program uses a patient management tool, education, and benchmarked quality reports to improve guideline adherence. Adherence to 8 national measures, including the use of aspirin and beta-blockers early and at discharge and timeline reperfusion, was analyzed. A composite score was also calculated. Multivariable logistic regression was performed for comparing composite adherence rates between groups. Adherence to the overall Hospital Compare composite measure was higher in GWTG-CAD hospitals than in non-GWTG-CAD hospitals (mean [SD], 89.7% [10.0%] vs 85.0 [15.0%]; absolute increase, 4.7%; P < .001). Adherence to the GWTG-CAD performance measures (PM) composite was also higher (89.5% [11.0%] vs 83.0% [18.0%]; P < .001). In multivariate analysis, GWTG-CAD participation was associated with a modest absolute increase in adherence to the PM composite by 2.52% (95% confidence interval [CI], 0.19%-4.85%). Larger acute myocardial infarction volume by quartile (absolute increase, 14.2%; 95% CI, 12.2%-16.3%), geographic location in the Northeast, and teaching hospital status (absolute increase, 2.87%; 95% CI, 0.43-5.32) were also associated with improved adherence to the PM composite. As a control, evaluation of unrelated quality measures for pneumonia, showed lower adherence among GWTG-CAD participating hospitals (74.8% [7.3%] vs 76.1% [9.7%]; P = .005). Participation in GWTG-CAD was independently associated with improvements in guideline adherence beyond that associated with public reporting.


High-Speed Videography Reveals How Honeybees Can Turn a Spatial Concept Learning Task Into a Simple Discrimination Task by Stereotyped Flight Movements and Sequential Inspection of Pattern Elements.
Honey bees display remarkable visual learning abilities, providing insights regarding visual information processing in a miniature brain. It was discovered that bees can solve a task that is generally viewed as spatial concept learning in primates, specifically the concept of "above" and "below." In these works, two pairs of visual stimuli were shown in the two arms of a Y-maze. Each arm displayed a "referent" shape (e.g., a cross, or a horizontal line) and a second geometric shape that appeared either above or below the referent. Bees learning the "concept of aboveness" had to choose the arm of the Y-maze in which a shape-any shape-occurred above the referent, while those learning the "concept of belowness" had to pick the arm in which there was an arbitrary item beneath the referent. Here, we explore the sequential decision-making process that allows bees to solve this task by analyzing their flight trajectories inside the Y-maze. Over 368 h of high-speed video footage of the bees' choice strategies were analyzed in detail. In our experiments, many bees failed the task, and, with the possible exception of a single forager, bees as a group failed to reach significance in picking the correct arm from the decision chamber of the maze. Of those bees that succeeded in choosing correctly, most required a close-up inspection of the targets. These bees typically employed a close-up scan of only the bottom part of the pattern before taking the decision of landing on a feeder. When rejecting incorrect feeders, they repeatedly scanned the pattern features, but were still, on average, faster at completing the task than the non-leaners. This shows that solving a concept learning task could actually be mediated by turning it into a more manageable discrimination task by some animals, although one individual in this study appeared to have gained the ability (by the end of the training) to solve the task in a manner predicted by concept learning.


Surgical interventions for treating intracapsular hip fractures in older adults: a network meta-analysis.
Hip fractures are a major healthcare problem, presenting a considerable challenge and burden to individuals and healthcare systems. The number of hip fractures globally is rising rapidly. The majority of intracapsular hip fractures are treated surgically. To assess the relative effects (benefits and harms) of all surgical treatments used in the management of intracapsular hip fractures in older adults, using a network meta-analysis of randomised trials, and to generate a hierarchy of interventions according to their outcomes. We searched CENTRAL, MEDLINE, Embase, Web of Science, and five other databases in July 2020. We also searched clinical trials databases, conference proceedings, reference lists of retrieved articles and conducted backward-citation searches. We included randomised controlled trials (RCTs) and quasi-RCTs comparing different treatments for fragility intracapsular hip fractures in older adults. We included total hip arthroplasties (THAs), hemiarthroplasties (HAs), internal fixation, and non-operative treatments. We excluded studies of people with hip fracture with specific pathologies other than osteoporosis or resulting from high-energy trauma. Two review authors independently assessed studies for inclusion. One review author completed data extraction which was checked by a second review author. We collected data for three outcomes at different time points: mortality and health-related quality of life (HRQoL) - both reported within 4 months, at 12 months, and after 24 months of surgery, and unplanned return to theatre (at end of study follow-up). We performed a network meta-analysis (NMA) with Stata software, using frequentist methods, and calculated the differences between treatments using risk ratios (RRs) and standardised mean differences (SMDs) and their corresponding 95% confidence intervals (CIs). We also performed direct comparisons using the same codes. We included 119 studies (102 RCTS, 17 quasi-RCTs) with 17,653 participants with 17,669 intracapsular fractures in the review; 83% of fractures were displaced. The mean participant age ranged from 60 to 87 years and 73% were women. After discussion with clinical experts, we selected 12 nodes that represented the best balance between clinical plausibility and efficiency of the networks: cemented modern unipolar HA, dynamic fixed angle plate, uncemented first-generation bipolar HA, uncemented modern bipolar HA, cemented modern bipolar HA, uncemented first-generation unipolar HA, uncemented modern unipolar HA, THA with single articulation, dual-mobility THA, pins, screws, and non-operative treatment. Seventy-five studies (with 11,855 participants) with data for at least two of these treatments contributed to the NMA. We selected cemented modern unipolar HA as a reference treatment against which other treatments were compared. This was a common treatment in the networks, providing a clinically appropriate comparison. In order to provide a concise summary of the results, we report only network estimates when there was evidence of difference between treatments. We downgraded the certainty of the evidence for serious and very serious risks of bias and when estimates included possible transitivity, particularly for internal fixation which included more undisplaced fractures. We also downgraded for incoherence, or inconsistency in indirect estimates, although this affected few estimates. Most estimates included the possibility of benefits and harms, and we downgraded the evidence for these treatments for imprecision. We found that cemented modern unipolar HA, dynamic fixed angle plate and pins seemed to have the greatest likelihood of reducing mortality at 12 months. Overall, 23.5% of participants who received the reference treatment died within 12 months of surgery. Uncemented modern bipolar HA had higher mortality than the reference treatment (RR 1.37, 95% CI 1.02 to 1.85; derived only from indirect evidence; low-certainty evidence), and THA with single articulation also had higher mortality (network estimate RR 1.62, 95% CI 1.13 to 2.32; derived from direct evidence from 2 studies with 225 participants, and indirect evidence; very low-certainty evidence). In the remaining treatments, the certainty of the evidence ranged from low to very low, and we noted no evidence of any differences in mortality at 12 months. We found that THA (single articulation), cemented modern bipolar HA and uncemented modern bipolar HA seemed to have the greatest likelihood of improving HRQoL at 12 months. This network was comparatively sparse compared to other outcomes and the certainty of the evidence of differences between treatments was very low. We noted no evidence of any differences in HRQoL at 12 months, although estimates were imprecise. We found that arthroplasty treatments seemed to have a greater likelihood of reducing unplanned return to theatre than internal fixation and non-operative treatment. We estimated that 4.3% of participants who received the reference treatment returned to theatre during the study follow-up. Compared to this treatment, we found low-certainty evidence that more participants returned to theatre if they were treated with a dynamic fixed angle plate (network estimate RR 4.63, 95% CI 2.94 to 7.30; from direct evidence from 1 study with 190 participants, and indirect evidence). We found very low-certainty evidence that more participants returned to theatre when treated with pins (RR 4.16, 95% CI 2.53 to 6.84; only from indirect evidence), screws (network estimate RR 5.04, 95% CI 3.25 to 7.82; from direct evidence from 2 studies with 278 participants, and indirect evidence), and non-operative treatment (RR 5.41, 95% CI 1.80 to 16.26; only from indirect evidence). There was very low-certainty evidence of a tendency for an increased risk of unplanned return to theatre for all of the arthroplasty treatments, and in particular for THA, compared with cemented modern unipolar HA, with little evidence to suggest the size of this difference varied strongly between the arthroplasty treatments. There was considerable variability in the ranking of each treatment such that there was no one outstanding, or subset of outstanding, superior treatments. However, cemented modern arthroplasties tended to more often yield better outcomes than alternative treatments and may be a more successful approach than internal fixation. There is no evidence of a difference between THA (single articulation) and cemented modern unipolar HA in the outcomes measured in this review. THA may be an appropriate treatment for a subset of people with intracapsular fracture but we have not explored this further.


Thinking and decision making, bacterial style: Bacterial Neural Networks, Obernai, France, 7th-12th June 2002.
Bacteria exhibit a bewildering range of behavioural responses and permutations of metabolic pathways for maximum exploitation of their environment. These are based on sensory perception of external and internal signals through batteries of surface and cytoplasmic receptors, evaluation of complex information flows and rapid decision making. Appreciation of the diversity of bacterial behaviour and adaptation capacities requires the study of a broad range of organisms and at this meeting we sampled more than 30 species with new findings which included the nature of gaseous receptors, advances in chemotaxis, subversion of host defences by pathogens, adaptation to high salt, community life and its obvious benefits, cell to cell communications and even the nature of bacterial circadian rhythms. With around 80 bacterial genomes now completed, and many more almost there, it was appropriate to complete the meeting with an introduction to Systems Biology and prospects for simulating the virtual cell. The versatility and seemingly 'intelligent' behaviour of bacteria will continue to fascinate, and this meeting on Bacterial Neural Networks fully reflected the excitement of this field.


Comprehensive treatment of late-onset tibia vara.
Late-onset tibia vara (Blount disease) can be difficult to treat because of frequent morbid obesity and associated deformities, including distal femoral varus, proximal tibial procurvatum, and distal tibial valgus, that contribute to lower extremity malalignment. We present a comprehensive approach that addresses all components of the deformity and allows restoration of the anatomic and mechanical axes. Fifteen consecutive patients (nineteen lower extremities) with late-onset tibia vara were managed with this comprehensive approach. The mean age of the patients at the time of surgery was 14.9 years, and the mean weight was 113 kg. Standing anteroposterior and lateral radiographs were made preoperatively and at the time of the final follow-up. Preoperatively, the mean mechanical axis deviation was 108 mm, the mean lateral distal femoral angle was 95 degrees , and the mean mechanical medial proximal tibial angle was 71 degrees . In all nineteen extremities, the proximal tibial varus deformity was corrected by means of a valgus osteotomy and application of an Ilizarov ring external fixator. Distal femoral varus was corrected by means of either hemiepiphyseal stapling or valgus osteotomy with blade-plate fixation in thirteen of the nineteen extremities. Distal tibial valgus was treated either with hemiepiphyseal stapling or with varus osteotomy and gradual correction with use of the Ilizarov external fixator in eleven of the nineteen extremities. After a mean duration of follow-up of 5.0 years, the mean mechanical axis deviation had improved to 1 mm (range, 20 to -30 mm), the lateral distal femoral angle had improved to 87 degrees (range, 83 degrees to 98 degrees), and the mechanical medial proximal tibial angle had improved to 88 degrees (range, 83 degrees to 98 degrees ). The mean time required for correction of the proximal tibial varus deformity was thirty-one days, and the external fixator was removed at a mean of 4.5 months postoperatively. All patients had development of one or more superficial pin-track infections (mean, 1.9 pin-site infections per patient). No wound infections, nonunions, or neurovascular complications occurred. Eighteen of the nineteen extremities were pain-free at the time of the final follow-up. This comprehensive approach allowed restoration of the mechanical and anatomic axes of the lower extremity in patients with late-onset tibia vara, resulting in a resolution of symptoms as a result of normalization of the weight-bearing forces across the knee and ankle. We believe that this approach will decrease the risk of early degenerative arthritis of the knee.


Patterns of HIV disclosure and condom use among HIV-infected young racial/ethnic minority men who have sex with men.
Recent findings highlight the continued rise in cases of HIV infection among racial/ethnic minority young men who have sex with men (YMSM). In adults, disclosure of HIV status has been associated with decreased sexual risk behaviors but this has not been explored among YMSM. In this study of 362 HIV-infected racial/ethnic minority YMSM, rates of disclosure were high, with almost all disclosing their status to at least one person at baseline. The majority had disclosed to a family member, with higher disclosure rates to female relatives compared with males. After adjustment for site, disclosure to sex partners and boyfriends was associated with an increase in condom use during both oral and anal sex. Future studies should consider skills training to assist youth in the disclosure process, facilitate how to determine who in their family and friend social network can be safely disclosed to and support family-based interventions.


Pointing towards colonoscopy: sharp foreign body removal via colonoscopy.
Removal of sharp foreign bodies via upper endoscopy is common; however, management in the setting of distal migration is not well-documented. We report two cases of objects beyond the ligament of Treitz, including successful extraction of a 4.4 cm sewing pin from the cecum using hot biopsy forceps with a protector hood to shield colonic mucosa, and in a separate case, a 3.4 cm glass shard from the ascending colon using a Roth Net retriever. We demonstrate that monitoring with serial radiographs and examination may allow for supervised passage of sharp objects into the colon, where removal can be performed safely via colonoscopy.


Space-Time Geostatistical Assessment of Hypoxia in the Northern Gulf of Mexico.
Nearly every summer, a large hypoxic zone forms in the northern Gulf of Mexico. Research on the causes and consequences of hypoxia requires reliable estimates of hypoxic extent, which can vary at submonthly time scales due to hydro-meteorological variability. Here, we use an innovative space-time geostatistical model and data collected by multiple research organizations to estimate bottom-water dissolved oxygen (BWDO) concentrations and hypoxic area across summers from 1985 to 2016. We find that 27% of variability in BWDO is explained by deterministic trends with location, depth, and date, while correlated stochasticity accounts for 62% of observational variance within a range of 185 km and 28 days. Space-time modeling reduces uncertainty in estimated hypoxic area by 30% when compared to a spatial-only model, and results provide new insights into the temporal variability of hypoxia. For years with shelf-wide cruises in multiple months, hypoxia is most severe in July in 59% of years, 29% in August, and 12% in June. Also, midsummer cruise estimates of hypoxic area are only modestly correlated with summer-wide (June-August) average estimates ( r2 = 0.5), suggesting midsummer cruises are not necessarily reflective of seasonal hypoxic severity. Furthermore, summer-wide estimates are more strongly correlated with nutrient loading than midsummer estimates.


Hierarchically structured hydrogels utilizing multifunctional assembling peptides for 3D cell culture.
Approaches for the creation of soft materials, particularly hydrogels, with hierarchical structure are of interest in a variety of applications owing to their unique properties. In the context of tissue mimics, hydrogels with multiscale structures more accurately capture the complexities of tissues within the body (e.g., fibrous collagen-rich microenvironments). However, cytocompatible fabrication of such materials with hierarchical structures and independent control of mechanical and biochemical properties remains challenging and is needed for probing and directing cell-microenvironment interactions for three-dimensional (3D) cell encapsulation and culture applications. To address this, we have designed innovative multifunctional assembling peptides: these unique peptides contain a core block that mimics the structure of collagen for achieving relevant melting temperatures; 'sticky' ends to promote assembly of long fibrils; and a biocompatible reactive handle that is orthogonal to assembly to allow the formation of desired multiscale structures and their subsequent rapid, light-triggered integration within covalently crosslinked synthetic hydrogels. Nano- to micro-fibrils were observed to form in physiologically-relevant aqueous solutions, where both underlying peptide chemical structure and assembly conditions were observed to impact the resulting fibril sizes. These assembled structures were 'locked' into place and integrated as linkers within cell-degradable, bioactive hydrogels formed with photoinitiated thiol-ene 'click' chemistry. Hydrogel compositions were identified for achieving robust mechanical properties like those of soft tissues while also retaining higher ordered structures after photopolymerization. The utility of these innovative materials for 3D cell culture was demonstrated with human mesenchymal stem cells, where cell morphologies reminiscent of responses to assembled native collagen were observed now with a fully synthetic material. Using a bottom-up approach, a new materials platform has been established that combines the advantageous properties of covalent and assembling chemistries for the creation of synthetic hydrogels with controllable nanostructure, mechanical properties, and biochemical content.


Kinetic stabilization against the oxidation reaction induced by a silaalkane cage in a thiophene-bridged molecular gyroscope.
Macrocage molecules with a bridged rotor have been synthesized as molecular gyroscopes. The kinetics of the oxidation reaction of the thiophene-bridged molecular gyroscope, whose thiophene ring was bridged inside a silaalkane cage, was investigated. A remarkable kinetic stabilization against the oxidation of the thiophene moiety induced by the molecular cage framework was observed.


Interleukin-6 primarily produced by non-hematopoietic cells mediates the lipopolysaccharide-induced febrile response.
Interleukin-6 (IL-6) is critical for the lipopolysaccharide (LPS)-induced febrile response. However, the exact source(s) of IL-6 involved in regulating the LPS-elicited fever is still to be identified. One known source of IL-6 is hematopoietic cells, such as monocytes. To clarify the contribution of hematopoietically derived IL-6 to fever, we created chimeric mice expressing IL-6 selectively either in cells of hematopoietic or, conversely, in cells of non-hematopoietic origin. This was performed by extinguishing hematopoietic cells in wild-type (WT) or IL-6 knockout (IL-6 KO) mice by whole-body irradiation and transplanting them with new stem cells. Mice on a WT background but lacking IL-6 in hematopoietic cells displayed normal fever to LPS and were found to have similar levels of IL-6 protein in the cerebrospinal fluid (CSF) and in plasma and of IL-6 mRNA in the brain as WT mice. In contrast, mice on an IL-6 KO background, but with intact IL-6 production in cells of hematopoietic origin, only showed a minor elevation of the body temperature after peripheral LPS injection. While they displayed significantly elevated levels of IL-6 both in plasma and CSF compared with control mice, the increase was modest compared with that seen in LPS injected mice on a WT background, the latter being approximately 20 times larger in magnitude. These results suggest that IL-6 of non-hematopoietic origin is the main source of IL-6 in LPS-induced fever, and that IL-6 produced by hematopoietic cells only plays a minor role.


High-throughput automated organoid culture via stem-cell aggregation in microcavity arrays.
Stem-cell-derived epithelial organoids are routinely used for the biological and biomedical modelling of tissues. However, the complexity, lack of standardization and quality control of stem cell culture in solid extracellular matrices hampers the routine use of the organoids at the industrial scale. Here, we report the fabrication of microengineered cell culture devices and scalable and automated methods for suspension culture and real-time analysis of thousands of individual gastrointestinal organoids trapped in microcavity arrays within a polymer-hydrogel substrate. The absence of a solid matrix substantially reduces organoid heterogeneity, which we show for mouse and human gastrointestinal organoids. We use the devices to screen for anticancer drug candidates with patient-derived colorectal cancer organoids, and apply high-content image-based phenotypic analyses to reveal insights into mechanisms of drug action. The scalable organoid-culture technology should facilitate the use of organoids in drug development and diagnostics.


Cross-reactivity of antibodies against leptospiral recurrent uveitis-associated proteins A and B (LruA and LruB) with eye proteins.
Infection by Leptospira interrogans has been causally associated with human and equine uveitis. Studies in our laboratories have demonstrated that leptospiral lipoprotein LruA and LruB are expressed in the eyes of uveitic horses, and that antibodies directed against LruA and LruB react with equine lenticular and retinal extracts, respectively. These reactivities were investigated further by performing immunofluorescent assays on lenticular and retinal tissue sections. Incubation of lens tissue sections with LruA-antiserum and retinal sections with LruB-antiserum resulted in positive fluorescence. By employing two-dimensional gel analyses followed by immunoblotting and mass spectrometry, lens proteins cross-reacting with LruA antiserum were identified to be alpha-crystallin B and vimentin. Similarly, mass spectrometric analyses identified beta-crystallin B2 as the retinal protein cross-reacting with LruB-antiserum. Purified recombinant human alpha-crystallin B and vimentin were recognized by LruA-directed antiserum, but not by control pre-immune serum. Recombinant beta-crystallin B2 was likewise recognized by LruB-directed antiserum, but not by pre-immune serum. Moreover, uveitic eye fluids contained significantly higher levels of antiibodies that recognized alpha-crystallin B, beta-crystallin B2 and vimentin than did normal eye fluids. Our results indicate that LruA and LruB share immuno-relevant epitopes with eye proteins, suggesting that cross-reactive antibody interactions with eye antigens may contribute to immunopathogenesis of Leptospira-associated recurrent uveitis.


A comparison of opioids and benzodiazepines dispensing in Australia.
Inappropriate utilization of prescription opioids and benzodiazepines is a public health problem. This study examined and compared user-types and trends in dispensing of these medicines, and identified associated factors related to the duration of dispensing in Australia. A random 10% sample of unit-record data of opioids and benzodiazepines dispensed nationally during 2013-2016 was analyzed. Users were categorized into four types: single-quarter (i.e., three months), medium-episodic (dispensed 2-6 quarters), long-episodic (dispensed 7-11 quarters), chronic (dispensed 12-16 quarters). Dispensing quantity was computed in defined daily dose (DDD). Generalized multilevel ordinal models were developed to examine the factors associated with the duration of dispensing. There were similarities in terms of trends of dispensing of opioids and benzodiazepines in Australia. Overall, more people were dispensed opioids than benzodiazepines. Around 52% of opioids users and 46% of benzodiazepines users were dispensed these medicines for a single quarter. However, chronic users were dispensed 60% of opioids and 50% of benzodiazepines in DDD/1000 people/day, respectively. On average, 16.6 DDD/1000 people/day of opioids and 14.2 DDD/1000 people/day of benzodiazepines were dispensed in Australia during the study period. Tasmania was dispensed the highest quantity (in DDD/1000 people/day) of these medicines, followed by South Australia and Queensland. Women compared to men, and clients of age-group 20-44, 45-64 and 65+ compared to age-group 0-19, were significantly more likely to have dispensed opioids/benzodiazepine for a relatively long duration. Clients with a history of dispensing of one of these two medicines were significantly more likely to have dispensed the other for a relatively long period. There were similarities in patterns of dispensing of opioids and benzodiazepines in terms of user characteristics and structural variables. Consistent use of real-time drug monitoring program and tailored intervention are recommended.


Compliance to antihypertensive therapy.
To determine compliance, factors affecting compliance to antihypertensive therapy and to compare compliant and non-compliant groups, in a tertiary care setting. Analytical (cross-sectional) study. The outpatient clinics at the Aga Khan University from May 2004 to February 2005. Two hundred patients presenting to the outpatients clinic were included. All patients 18 years and above, who had stage 1 and 2 hypertension, had one clinic visit to a medicine clinic, 6 months prior to presentation and started on antihypertensive medicines, were included. Sixty six percent were males and 33.5 % were females. Mean age was 58.1(+/-12) years and mean duration of hypertension was 7.2 (+/- 6.7) years. Fifty-seven percent were compliant and 43 % were noncompliant. In the noncompliant group, 53.4 % had mild noncompliance, 24.4 % had severe non-compliance, while 22% had moderate noncompliance. Factors of noncompliance were 56.8% missed doses due to forgetfulness, 12.7% deliberately missed their doses, 11.6% could not take the medicine due to side effects, 10.4% did not take the dose due to increased number of tablets, 4.6% were not properly counseled by the physician and 3.48% did not take medicines due to cost issues. The mean systolic blood pressure was 126 +/- 19.2 mmHg in the compliant group while it was 133 +/-16.5 mmHg in the noncompliant group p-value 0.004). The mean diastolic blood pressure in the compliant group was 76 +/- 11.9 mmHg, while in the noncompliant group it was 81.9 +/-10.9 mmHg (p-value 0.001). Compliance to antihypertensive therapy in a tertiary care center is significantly good. Forgetfulness was the major reason for noncompliance. The mean blood pressure control was better in the compliant group.


Caenorhabditis elegans mutants defective in the functioning of the motor neurons responsible for egg laying.
We have isolated and characterized 45 Caenorhabditis elegans mutants presumed to be defective in the functioning of the hermaphrodite-specific neurons (HSNs). Like hermaphrodites that lack the HSN motor neurons, these mutants are egg-laying defective and do not lay eggs in response to exogenous imipramine but do lay eggs in response to exogenous serotonin. Twenty of the 45 mutations define 10 new egl genes; the other 25 mutations are alleles of five previously defined genes, four of which are known to affect the HSNs. Seven mutations in three genes cause the HSNs to die in hermaphrodites, as they normally do in males. These genes appear to be involved in the determination of the sexual phenotype of the HSNs, and one of them (egl-41) is a newly identified gene that may function generally in sex determination. Five of the 15 genes are defined only by mutations that have dominant effects on egg laying. One gene egl(n1108), is defined by a temperature-sensitive allele that has a temperature-sensitive period after HSN development is complete, suggesting that egl(n1108) may be involved in HSN synaptic transmission. Four of the genes are defined by single alleles, which suggests that other such genes remain to be discovered. Mutations in no more than 4 of the 15 genes specifically affect the HSNs, indicating that there are few genes with functions needed only in this single type of nerve cell.


Granulosa cell tumors express erbB4 and are sensitive to the cytotoxic action of heregulin-beta2/PE40.
The molecular genetic events involved in the etiology of human granulosa cell (GC) tumors, which represent approximately 7% of all malignant ovarian neoplasms, are unknown. Amplification and/or overexpression of the ERBB genes are a feature of many cancer types, and overexpression of erbB2 correlates with poor prognosis in epithelial ovarian cancer. In the present study, we used immunohistochemistry to determine the level and frequency of expression of different erbB receptors in GC tumors. Ten of 12 tumors expressed erbB4 at moderate to high levels in >50% of cancer cells, whereas erbB2 (6 of 12) and erbB3 (2 of 12) were expressed less frequently. Western blot experiments showed that the only available GC tumor cell line, COV434, also expressed erbB receptors. Heregulin (HRG)-beta2, a ligand for erbB3 and erbB4 receptors, stimulated tyrosine phosphorylation of the erbB receptors, which was accompanied by activation of Erk1 and Erk2, two mitogen-activated protein kinases with a functional role in mitogenesis. Importantly, HRG increased cell proliferation in COV434 cells, and treatment with HRG/PE40, a ligand toxin shown previously to be cytotoxic against human breast cancer cells overexpressing erbB receptors, led to a dramatic and irreversible decrease in cell number. These results indicate that erbB receptor signaling pathways may be critical in the control of GC tumor cell proliferation and that HRG/PE40 is a potential therapeutic agent for the treatment of GC tumors.


Phenotype microarrays for high-throughput phenotypic testing and assay of gene function.
The bacterium Escherichia coli is used as a model cellular system to test and validate a new technology called Phenotype MicroArrays (PMs). PM technology is a high-throughput technology for simultaneous testing of a large number of cellular phenotypes. It consists of preconfigured well arrays in which each well tests a different cellular phenotype and an automated instrument that continuously monitors and records the response of the cells in all wells of the arrays. For example, nearly 700 phenotypes of E. coli can be assayed by merely pipetting a cell suspension into seven microplate arrays. PMs can be used to directly assay the effects of genetic changes on cells, especially gene knock-outs. Here, we provide data on phenotypic analysis of six strains and show that we can detect expected phenotypes as well as, in some cases, unexpected phenotypes.


The impact of systemic and copper pesticide applications on the phyllosphere microflora of tomatoes.
Contamination of tomatoes by Salmonella can occur in agricultural settings. Little is currently understood about how agricultural inputs such as pesticide applications may impact epiphytic crop microflora and potentially play a role in contamination events. We examined the impact of two materials commonly used in Virginia tomato agriculture: acibenzolar-S-methyl (crop protectant) and copper oxychloride (pesticide) to identify the effects these materials may exert on baseline tomato microflora and on the incidence of three specific genera; Salmonella, Xanthomonas and Paenibacillus. Approximately 186 441 16S rRNA gene and 39 381 18S rRNA gene sequences per independent replicate were used to analyze the impact of the pesticide applications on tomato microflora. An average of 3 346 677 (634 892 974 bases) shotgun sequences per replicate were used for metagenomic analyses. A significant decrease in the presence of Gammaproteobacteria was observed between controls and copper-treated plants, suggesting that copper is effective at suppressing growth of certain taxa in this class. A higher mean abundance of Salmonella and Paenibacillus in control samples compared to treatments may suggest that both systemic and copper applications diminish the presence of these genera in the phyllosphere; however, owing to the lack of statistical significance, this could also be due to other factors. The most distinctive separation of shared membership was observed in shotgun data between the two different sampling time-points (not between treatments), potentially supporting the hypothesis that environmental pressures may exert more selective pressures on epiphytic microflora than do certain agricultural management practices.


Gene expression changes and signaling events associated with the direct antimelanoma effect of IFN-gamma.
IFN-gamma plays a role in the response to melanoma indirectly through its effect on the immune system and directly through its antiproliferative and proapoptotic effects on melanoma cells. To understand the molecular basis for the direct antimelanoma effect of IFN-gamma, we studied IFN-induced changes in gene expression and signaling among three human melanoma cell lines (DM6, DM93, and 501mel). These were resistant to the antimelanoma effect of IFN-alpha, and only DM6 cells exhibited growth inhibition and apoptosis with IFN-gamma. Through DNA microarray analysis, we found that the antimelanoma effect of IFN-gamma in DM6 was associated with the down-regulation of multiple genes involved in G-protein signaling and phospholipase C activation (including Rap2B and calpain 3) as well as the down-regulation of genes involved in melanocyte/melanoma survival (MITF and SLUG), apoptosis inhibition (Bcl2A1 and galectin-3), and cell cycling (CDK2). The antimelanoma effect of IFN-gamma was also associated with the up-regulation of the proapoptotic dependence receptor UNC5H2 and the Wnt inhibitor Dkk-1. Whereas both IFNs were able to activate Stat1 in all cell lines, the delayed activation of the extracellular signal-regulated kinase, p38, and c-Jun NH2-terminal kinase mitogen-activated protein kinases occurred only in DM6 with IFN-gamma, and the effect of IFN-gamma on cell growth and survival as well as gene expression in DM6 was dependent on the coordinate activation of MEK1 and p38. These findings provide new insights into the signaling events and gene expression changes associated with growth inhibition and apoptosis in melanoma and may thereby assist in identifying new targets for the treatment of melanoma.


Symmetry breaking and structural polymorphism in a bacterial microcompartment shell protein for choline utilization.
Bacterial microcompartments are protein-based organelles that carry out specialized metabolic functions in diverse bacteria. Their outer shells are built from several thousand protein subunits. Some of the architectural principles of bacterial microcompartments have been articulated, with lateral packing of flat hexameric BMC proteins providing the basic foundation for assembly. Nonetheless, a complete understanding has been elusive, partly owing to polymorphic mechanisms of assembly exhibited by most microcompartment types. An earlier study of one homologous BMC shell protein subfamily, EutS/PduU, revealed a profoundly bent, rather than flat, hexameric structure. The possibility of a specialized architectural role was hypothesized, but artifactual effects of crystallization could not be ruled out. Here we report a series of crystal structures of an orthologous protein, CutR, from a glycyl-radical type choline-utilizing microcompartment from the bacterium Streptococcus intermedius. Depending on crystal form, expression construct, and minor mutations, a range of novel quaternary architectures was observed, including two spiral hexagonal assemblies. A new graphical approach helps illuminate the variations in BMC hexameric structure, with results substantiating the idea that the EutS/PduU/CutR subfamily of BMC proteins may endow microcompartment shells with flexible modes of assembly.


Evaluation of standardized doctor's orders as an educational tool for undergraduate medical students: a prospective cohort study.
Standardized doctor's orders are replacing traditional order writing in teaching hospitals. The impact of this shift in practice on medical education is unknown. It is possible that preprinted orders interfere with knowledge acquisition and retention by not requiring active decision-making. The objective of the study was to evaluate the impact of standardized admission orders on disease-specific knowledge among undergraduate medical trainees. This prospective cohort study enrolled Year 3 (n = 121) and Year 4 (n = 54) medical students at two academic hospitals in Toronto (Ontario, Canada) during their general internal medicine rotation. We used standardized orders for patient admissions for alcohol withdrawal (AW) and for acute exacerbations of chronic obstructive pulmonary disease (AECOPD) as the intervention and manual order writing as the control. Educational outcomes were assessed through end-of-rotation questionnaires assessing disease-specific knowledge of AW and AECOPD. Of 175 students, 105 had exposure to patients with alcohol withdrawal during their rotation, and 68 students wrote admission orders. Among these 68 students, 48 used standardized orders (intervention, n = 48) and 20 used manual order writing (control, n = 20). Only 3 students used standardized orders for AECOPD, precluding analysis. There was no significant difference found in mean total score of questionnaires between those who used AW standardized orders and those who did not (11.8 vs. 11.0, p = 0.4). Students who had direct clinical experience had significantly higher mean total scores (11.6 vs. 9.0, p < 0.0001 for AW; 13.8 vs. 12.6, p = 0.02 for AECOPD) compared to students who did not. When corrected for overall knowledge, this difference only persisted for AW. No significant differences were found in total scores between students who used standardized admission orders and traditional manual order writing. Clinical exposure was associated with increase in disease-specific knowledge.


Retinoic acid-related orphan receptor alpha reprograms glucose metabolism in glutamine-deficient hepatoma cells.
The metabolism of glutamine and glucose is recognized as a promising therapeutic target for the treatment of cancer; however, targeted molecules that mediate glutamine and glucose metabolism in cancer cells have not been addressed. Here, we show that restricting the supply of glutamine in hepatoma cells, including HepG2 and Hep3B cells, markedly increased the expression of retinoic acid-related orphan receptor alpha (RORα). Up-regulation of RORα in glutamine-deficient hepatoma cells resulted from an increase in the level of cellular reactive oxygen species and in the nicotinamide adenine dinucleotide phosphate/nicotinamide adenine dinucleotide phosphate reduced (NADP+ /NADPH) ratio, which was consistent with a reduction in the glutathione/glutathione disulfide (GSH/GSSG) ratio. Adenovirus (Ad)-mediated overexpression of RORα (Ad-RORα) or treatment with the RORα activator, SR1078, reduced aerobic glycolysis and down-regulated biosynthetic pathways in hepatoma cells. Ad-RORα and SR1078 reduced the expression of pyruvate dehydrogenase kinase 2 (PDK2) and inhibited the phosphorylation of pyruvate dehydrogenase and subsequently shifted pyruvate to complete oxidation. The RORα-mediated decrease in PDK2 levels was caused by up-regulation of p21, rather than p53. Furthermore, RORα inhibited hepatoma growth both in vitro and in a xenograft model in vivo. We also found that suppression of PDK2 inhibited hepatoma growth in a xenograft model. These findings mimic the altered glucose utilization and hepatoma growth caused by glutamine deprivation. Finally, tumor tissue from 187 hepatocellular carcinoma patients expressed lower levels of RORα than adjacent nontumor tissue, supporting a potential beneficial effect of RORα activation in the treatment of liver cancer. RORα mediates reprogramming of glucose metabolism in hepatoma cells in response to glutamine deficiency. The relationships established here between glutamine metabolism, RORα expression and signaling, and aerobic glycolysis have implications for therapeutic targeting of liver cancer metabolism.


Whole-genome patterns of common DNA variation in three human populations.
Individual differences in DNA sequence are the genetic basis of human variability. We have characterized whole-genome patterns of common human DNA variation by genotyping 1,586,383 single-nucleotide polymorphisms (SNPs) in 71 Americans of European, African, and Asian ancestry. Our results indicate that these SNPs capture most common genetic variation as a result of linkage disequilibrium, the correlation among common SNP alleles. We observe a strong correlation between extended regions of linkage disequilibrium and functional genomic elements. Our data provide a tool for exploring many questions that remain regarding the causal role of common human DNA variation in complex human traits and for investigating the nature of genetic variation within and between human populations.


Changes in Physiology before, during, and after Yawning.
The ultimate function of yawning continues to be debated. Here, we examine physiological measurements taken before, during, and after yawns in humans, in an attempt to identify key proximate mechanisms associated with this behavior. In two separate studies we measured changes in heart rate, lung volume, eye closure, skin conductance, ear pulse, respiratory sinus arrhythmia, and respiratory rate. Data were depicted from 75 s before and after yawns, and analyzed at baseline, during, and immediately following yawns. Increases in heart rate, lung volume, and eye muscle tension were observed during or immediately following yawning. Patterns of physiological changes during yawning were then compared to data from non-yawning deep inhalations. In one study, respiration period increased following the execution of a yawn. Much of the variance in physiology surrounding yawning was specific to the yawning event. This was not the case for deep inhalation. We consider our findings in light of various hypotheses about the function of yawning and conclude that they are most consistent with the brain cooling hypothesis.