Transparent and accurate reporting increases reliability, utility, and impact of your research: reporting guidelines and the EQUATOR Network.
Although current electronic methods of scientific publishing offer increased opportunities for publishing all research studies and describing them in sufficient detail, health research literature still suffers from many shortcomings. These shortcomings seriously undermine the value and utility of the literature and waste scarce resources invested in the research. In recent years there have been several positive steps aimed at improving this situation, such as a strengthening of journals' policies on research publication and the wide requirement to register clinical trials.The EQUATOR (Enhancing the QUAlity and Transparency Of health Research) Network is an international initiative set up to advance high quality reporting of health research studies; it promotes good reporting practices including the wider implementation of reporting guidelines. EQUATOR provides free online resources http://www.equator-network.org supported by education and training activities and assists in the development of robust reporting guidelines. This paper outlines EQUATOR's goals and activities and offers suggestions for organizations and individuals involved in health research on how to strengthen research reporting.


Evaluation of the benefits and risks of introducing Ebola community care centers, Sierra Leone.
In some parts of western Africa, Ebola treatment centers (ETCs) have reached capacity. Unless capacity is rapidly scaled up, the chance to avoid a generalized Ebola epidemic will soon diminish. The World Health Organization and partners are considering additional Ebola patient care options, including community care centers (CCCs), small, lightly staffed units that could be used to isolate patients outside the home and get them into care sooner than otherwise possible. Using a transmission model, we evaluated the benefits and risks of introducing CCCs into Sierra Leone's Western Area, where most ETCs are at capacity. We found that use of CCCs could lead to a decline in cases, even if virus transmission occurs between CCC patients and the community. However, to prevent CCC amplification of the epidemic, the risk of Ebola virus-negative persons being exposed to virus within CCCs would have to be offset by a reduction in community transmission resulting from CCC use.


Seasat scanning multichannel microwave radiometer: results of the gulf of alaska workshop.
The scanning multichannel microwave radiometer results for the Gulf of Alaska Seasat Experiment Workshop are quite encouraging, especially in view of the immaturity of the data-processing algorithms. For open ocean, rain-free cells of highest-quality surface truth wind determinations exhibit standard deviations of 3 meters per second about a bias of 1.5 meters per second. The sea-surface temperature shows a standard deviation of approximately 1.5 degrees C about a bias of 3 degrees to 5 degrees C under a variety of changing meteorological conditions.


Molecular genetics of carbapenem antibiotic biosynthesis.
Carbapenems are potent beta-lactam antibiotics with a broad spectrum of activity against both Gram positive and Gram negative bacteria. As naturally produced metabolites, they have been isolated from species of Streptomyces, Erwinia and Serratia. The latter two members of the Enterobacteriaceae have proved to be genetically amenable and a growing body of research on these organisms now exists concerning the genes responsible for carbapenem biosynthesis and the regulatory mechanisms controlling their expression. A cluster of nine carbapenem (car) genes has been identified on the chromosome of Erwinia carotovora. These genes encode the enzymes required for construction of carbapenem and the proteins responsible for a novel beta-lactam resistance mechanism, conferring carbapenem immunity in the producing host. Although sharing no homology with the well known enzymes of penicillin biosynthesis, two of the encoded proteins are apparently similar to enzymes of the clavulanic acid biosynthetic pathway implying a common mechanism for construction of the beta-lactam ring. In addition, a transcriptional activator is encoded as the first gene of the carbapenem cluster and this allows positive expression of the remaining downstream genes in response to a quorum sensing, N-acyl homoserine lactone, signalling molecule.


Carcinogenic tobacco-specific N-nitrosamines in US cigarettes: three decades of remarkable neglect by the tobacco industry.
Modification of tobacco curing methods and other changes in cigarette manufacturing techniques could substantially reduce the levels of tobacco-specific nitrosamines (TSNA), a group of potent carcinogens, in cigarette smoke. In 1999, two major US cigarette manufacturers stated their intent to move towards using tobaccos low in TSNA. There is no information available on current TSNA levels in tobacco of various cigarettes available in the US, particularly in the newer varieties introduced over the past decade. Seventeen brands of cigarettes were purchased in April of 2010 from retail stores in Minnesota. TSNA levels were measured in the tobacco filler and smoke of these cigarettes. In all brands, the sum of two potent carcinogenic TSNA--4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-nitrosonornicotine--in cigarette filler averaged 2.54 (± 1.05) μg/g tobacco. This value is virtually identical to the sum of these two carcinogens reported for the tobacco of a US filtered cigarette in 1979. TSNA levels in smoke positively correlated with those in tobacco filler of the same cigarettes. We found no indication that any meaningful attempt was made to reduce or at least control TSNA levels in the new varieties of the popular brands Marlboro and Camel introduced over the last decade. In light of the recently granted regulatory authority to the FDA over tobacco products, regulation of TSNA levels in cigarette tobacco should be strongly considered to reduce the levels of these potent carcinogens in cigarette smoke.


Micronuclei-based model system reveals functional consequences of chromothripsis in human cells.
Cancer cells often harbor chromosomes in abnormal numbers and with aberrant structure. The consequences of these chromosomal aberrations are difficult to study in cancer, and therefore several model systems have been developed in recent years. We show that human cells with extra chromosome engineered via microcell-mediated chromosome transfer often gain massive chromosomal rearrangements. The rearrangements arose by chromosome shattering and rejoining as well as by replication-dependent mechanisms. We show that the isolated micronuclei lack functional lamin B1 and become prone to envelope rupture, which leads to DNA damage and aberrant replication. The presence of functional lamin B1 partly correlates with micronuclei size, suggesting that the proper assembly of nuclear envelope might be sensitive to membrane curvature. The chromosomal rearrangements in trisomic cells provide growth advantage compared to cells without rearrangements. Our model system enables to study mechanisms of massive chromosomal rearrangements of any chromosome and their consequences in human cells.


Mortality after Inpatient Treatment for Severe Pneumonia in Children: a Cohort Study.
Although pneumonia is a leading cause of inpatient mortality, deaths may also occur after discharge from hospital. However, prior studies have been small, in selected groups or did not fully evaluate risk factors, particularly malnutrition and HIV. We determined 1-year post-discharge mortality and risk factors among children diagnosed with severe pneumonia. A cohort study of children aged 1-59 months admitted to Kilifi County Hospital with severe pneumonia (2007-12). The primary outcome was death <1 year after discharge, determined through Kilifi Health and Demographic Surveillance System (KHDSS) quarterly census rounds. Of 4184 children (median age 9 months) admitted with severe pneumonia, 1041 (25%) had severe acute malnutrition (SAM), 267 (6.4%) had a positive HIV antibody test, and 364 (8.7%) died in hospital. After discharge, 2279 KHDSS-resident children were followed up; 70 (3.1%) died during 2163 child-years: 32 (95% confidence interval (CI) 26, 41) deaths per 1000 child years. Post-discharge mortality was greater after admission for severe pneumonia than for other diagnoses, hazard ratio 2.5 (95% CI 1.2, 5.3). Malnutrition, HIV status, age and prolonged hospitalisation, but not signs of pneumonia severity, were associated with post-discharge mortality. Fifty-two per cent (95% CI 37%, 63%) of post-discharge deaths were attributable to low mid-upper arm circumference and 11% (95% CI 3.3%, 18%) to a positive HIV test. Admission with severe pneumonia is an important marker of vulnerability. Risk stratification and better understanding of the mechanisms underlying post-discharge mortality, especially for undernourished children, are needed to reduce mortality after treatment for pneumonia.


Climate-suitable planting as a strategy for maintaining forest productivity and functional diversity.
Within the time frame of the longevity of tree species, climate change will change faster than the ability of natural tree migration. Migration lags may result in reduced productivity and reduced diversity in forests under current management and climate change. We evaluated the efficacy of planting climate-suitable tree species (CSP), those tree species with current or historic distributions immediately south of a focal landscape, to maintain or increase aboveground biomass productivity, and species and functional diversity. We modeled forest change with the LANDIS-II forest simulation model for 100 years (2000-2100) at a 2-ha cell resolution and five-year time steps within two landscapes in the Great Lakes region (northeastern Minnesota and northern lower Michigan, USA). We compared current climate to low- and high-emission futures. We simulated a low-emission climate future with the Intergovernmental Panel on Climate Change (IPCC) 2007 B1 emission scenario and the Parallel Climate Model Global Circulation Model (GCM). We simulated a high-emission climate future with the IPCC A1FI emission scenario and the Geophysical Fluid Dynamics Laboratory (GFDL) GCM. We compared current forest management practices (business-as-usual) to CSP management. In the CSP scenario, we simulated a target planting of 5.28% and 4.97% of forested area per five-year time step in the Minnesota and Michigan landscapes, respectively. We found that simulated CSP species successfully established in both landscapes under all climate scenarios. The presence of CSP species generally increased simulated aboveground biomass. Species diversity increased due to CSP; however, the effect on functional diversity was variable. Because the planted species were functionally similar to many native species, CSP did not result in a consistent increase nor decrease in functional diversity. These results provide an assessment of the potential efficacy and limitations of CSP management. These results have management implications for sites where diversity and productivity are expected to decline. Future efforts to restore a specific species or forest type may not be possible, but CSP may sustain a more general ecosystem service (e.g., aboveground biomass).


Membrane scaling and flux decline during fertiliser-drawn forward osmosis desalination of brackish groundwater.
Fertiliser-drawn forward osmosis (FDFO) desalination has been recently studied as one feasible application of forward osmosis (FO) for irrigation. In this study, the potential of membrane scaling in the FDFO process has been investigated during the desalination of brackish groundwater (BGW). While most fertilisers containing monovalent ions did not result in any scaling when used as an FO draw solution (DS), diammonium phosphate (DAP or (NH4)2HPO4) resulted in significant scaling, which contributed to severe flux decline. Membrane autopsy using scanning electron microscopy (SEM), energy-dispersive x-ray spectroscopy (EDS), and x-ray diffraction (XRD) analysis indicated that the reverse diffusion of DAP from the DS to the feed solution was primarily responsible for scale formation during the FDFO process. Physical cleaning of the membrane with deionised water at varying crossflow velocities was employed to evaluate the reversibility of membrane scaling and the extent of flux recovery. For the membrane scaled using DAP as DS, 80-90% of the original flux was recovered when the crossflow velocity for physical cleaning was the same as the crossflow velocity during FDFO desalination. However, when a higher crossflow velocity or Reynolds number was used, the flux was recovered almost completely, irrespective of the DS concentration used. This study underscores the importance of selecting a suitable fertiliser for FDFO desalination of brackish groundwater to avoid membrane scaling and severe flux decline.


Microbiology of controlled rotting of egusi (Colocynthis citrullus L.) fruits for the harvesting of the seeds.
Microorganisms isolated from naturally rotting egusi fruits which are used as a source of dietary seeds, were mainly bacteria: Bacillus subtilis, B. licheniformis, B. polymyxa, B. megaterium, B. pumilus, Lactobacillus plantarum, La. brevis, Leuconostoc mesenteroides, Enterobacter aerogenes, E. cloacae, Klebsiella aerogenes, K. pneumoniae, Staphylococcus epidermidis, S. aureus, Micrococcus and Candida spp. The rotting period of egusi fruits could be decreased from 120 to 72 h by using a mixture of B. subtilis and B. licheniformis.


Promotion of couples' voluntary counselling and testing for HIV through influential networks in two African capital cities.
Most new HIV infections in Africa are acquired from cohabiting heterosexual partners. Couples' Voluntary Counselling and Testing (CVCT) is an effective prevention strategy for this group. We present our experience with a community-based program for the promotion of CVCT in Kigali, Rwanda and Lusaka, Zambia. Influence Network Agents (INAs) from the health, religious, non-governmental, and private sectors were trained to invite couples for CVCT. Predictors of successful promotion were identified using a multi-level hierarchical analysis. In 4 months, 9,900 invitations were distributed by 61 INAs, with 1,411 (14.3%) couples requesting CVCT. INAs in Rwanda distributed fewer invitations (2,680 vs. 7,220) and had higher response rates (26.9% vs. 9.6%), than INAs in Zambia. Context of the invitation event, including a discreet location such as the INA's home (OR 3.3-3.4), delivery of the invitation to both partners in the couple (OR 1.6-1.7) or to someone known to the INA (OR 1.7-1.8), and use of public endorsement (OR 1.7-1.8) were stronger predictors of success than INA or couple-level characteristics. Predictors of successful CVCT promotion included strategies that can be easily implemented in Africa. As new resources become available for Africans with HIV, CVCT should be broadly implemented as a point of entry for prevention, care and support.


Neuroprotection and repair.
Neuroprotection can be applied to treatments that have a primary role in protecting nerves and axons from damage. Alternatively it can refer to treatments which act up-stream on the primary pathology and which have a secondary neuroprotective effect. Although there are many potential neuroprotective agents, clinical evidence to prove their hypothetical action is lacking. Studying natural brain repair processes is important in understanding how repair and recovery a) is limited in disease and can b) contribute to the development of treatments to enhance this process. Different mechanisms underlie repair and recovery and they vary in the time taken to produce improvement and the degree of translation into a clinical effect. Additionally individuals may vary in their ability to repair and this may contribute to the heterogeneity of diseases such as MS.


Comparison of dogs and humans in visual scanning of social interaction.
Previous studies have demonstrated similarities in gazing behaviour of dogs and humans, but comparisons under similar conditions are rare, and little is known about dogs' visual attention to social scenes. Here, we recorded the eye gaze of dogs while they viewed images containing two humans or dogs either interacting socially or facing away: the results were compared with equivalent data measured from humans. Furthermore, we compared the gazing behaviour of two dog and two human populations with different social experiences: family and kennel dogs; dog experts and non-experts. Dogs' gazing behaviour was similar to humans: both species gazed longer at the actors in social interaction than in non-social images. However, humans gazed longer at the actors in dog than human social interaction images, whereas dogs gazed longer at the actors in human than dog social interaction images. Both species also made more saccades between actors in images representing non-conspecifics, which could indicate that processing social interaction of non-conspecifics may be more demanding. Dog experts and non-experts viewed the images very similarly. Kennel dogs viewed images less than family dogs, but otherwise their gazing behaviour did not differ, indicating that the basic processing of social stimuli remains similar regardless of social experiences.


Illuminating the developing brain: the past, present and future of functional near infrared spectroscopy.
A decade has passed since near infrared spectroscopy (NIRS) was first applied to functional brain imaging in infants. As part of the team that published the first functional near infrared spectroscopy (fNIRS) infant study in 1998, we have continued to develop and refine both the technology and methods associated with these measurements. The increasing international interest that this technology is generating among neurodevelopmental researchers and the recent technical developments in biomedical optics have prompted us to compile this review of the challenges that have been overcome in this field, and the practicalities of performing fNIRS in infants. We highlight the increasingly diverse and ambitious studies that have been undertaken and review the technological and methodological advances that have been made in the study design, optical probe development, and interpretation and analyses of the haemodynamic response. A strong emphasis is placed on the potential of the technology and future prospects of fNIRS in the field of developmental neuroscience.


Transplantation of Human Embryonic Stem Cells in Patients with Multiple Sclerosis and Lyme Disease.
BACKGROUND Multiple sclerosis (MS) is an inflammatory and neurodegenerative disease in which the myelin sheath of nerve cells is damaged. It can cause delayed neurologic symptoms similar to those seen in Lyme disease (LD) patients. Thymus derived T-cells (myelin reactive) migrate to the blood brain barrier and stimulate an inflammatory cascade in the central nervous system. Cell based therapies play an important role in treating neurological diseases such as MS and LD. CASE REPORT Human embryonic stem cell (hESC) therapy was used to treat two patients with both MS and LD. The hESCs were administered via different routes including intramuscular, intravenous, and supplemental routes (e.g., deep spinal, caudal, intercostal through eye drops) to regenerate the injured cells. Both the patients showed remarkable improvement in their functional skills, overall stamina, cognitive abilities, and muscle strength. Furthermore, the improvement in the patients' conditions were assessed by magnetic resonance tractography and single photon emission computed tomography (SPECT). CONCLUSIONS Therapy with hESCs might emerge as an effective and safe treatment for patients with both MS and LD. Well-designed clinical trials and follow-up studies are needed to prove the long-term efficacy and safety of hESC therapy in the treatment of patients with MS and LD.


Quantifying antimicrobial resistance at veal calf farms.
This study was performed to determine a sampling strategy to quantify the prevalence of antimicrobial resistance on veal calf farms, based on the variation in antimicrobial resistance within and between calves on five farms. Faecal samples from 50 healthy calves (10 calves/farm) were collected. From each individual sample and one pooled faecal sample per farm, 90 selected Escherichia coli isolates were tested for their resistance against 25 mg/L amoxicillin, 25 mg/L tetracycline, 0.5 mg/L cefotaxime, 0.125 mg/L ciprofloxacin and 8/152 mg/L trimethoprim/sulfamethoxazole (tmp/s) by replica plating. From each faecal sample another 10 selected E. coli isolates were tested for their resistance by broth microdilution as a reference. Logistic regression analysis was performed to compare the odds of testing an isolate resistant between both test methods (replica plating vs. broth microdilution) and to evaluate the effect of pooling faecal samples. Bootstrap analysis was used to investigate the precision of the estimated prevalence of resistance to each antimicrobial obtained by several simulated sampling strategies. Replica plating showed similar odds of E. coli isolates tested resistant compared to broth microdilution, except for ciprofloxacin (OR 0.29, p ≤ 0.05). Pooled samples showed in general lower odds of an isolate being resistant compared to individual samples, although these differences were not significant. Bootstrap analysis showed that within each antimicrobial the various compositions of a pooled sample provided consistent estimates for the mean proportion of resistant isolates. Sampling strategies should be based on the variation in resistance among isolates within faecal samples and between faecal samples, which may vary by antimicrobial. In our study, the optimal sampling strategy from the perspective of precision of the estimated levels of resistance and practicality consists of a pooled faecal sample from 20 individual animals, of which 90 isolates are tested for their susceptibility by replica plating.


Sugar additives for MALDI matrices improve signal allowing the smallest nucleotide change (A:T) in a DNA sequence to be resolved.
Sample preparation for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) of DNA is critical for obtaining high quality mass spectra. Sample impurity, solvent content, substrate surface and environmental conditions (temperature and humidity) all affect the rate of matrix-analyte co-crystallization. As a result, laser fluence threshold for desorption/ionization varies from spot to spot. When using 3-hydroxypicolinic acid (3-HPA) as the matrix, laser fluence higher than the threshold value reduces mass resolution in time-of-flight (TOF) MS as the excess energy transferred to DNA causes metastable decay. This can be overcome by either searching for 'hot' spots or adjusting the laser fluence. However, both solutions may require a significant amount of operator manipulation and are not ideal for automatic measurements. We have added various sugars for crystallization with the matrix to minimize the transfer of excess laser energy to DNA molecules. Fructose and fucose were found to be the most effective matrix additives. Using these additives, mass resolution for DNA molecules does not show noticeable deterioration as laser energy increases. Improved sample preparation is important for the detection of single nucleotide polymorphisms (SNPs) using primer extension with a single nucleotide. During automatic data acquisition it is difficult to routinely detect heterozygous A/T mutations, which requires resolving a mass difference of 9 Da, unless a sugar is added during crystallization.


Assessment and Management of Delirium in Pediatric Patients.
The Psychiatric Consultation Service at Massachusetts General Hospital sees medical and surgical inpatients with comorbid psychiatric symptoms and conditions. During their twice-weekly rounds, Dr Stern and other members of the Consultation Service discuss diagnosis and management of hospitalized patients with complex medical or surgical problems who also demonstrate psychiatric symptoms or conditions. These discussions have given rise to rounds reports that will prove useful for clinicians practicing at the interface of medicine and psychiatry.


Spectral coarse graining and synchronization in oscillator networks.
Coarse graining techniques offer a promising alternative to large-scale simulations of complex dynamical systems, as long as the coarse-grained system is truly representative of the initial one. Here, we investigate how the dynamical properties of oscillator networks are affected when some nodes are merged together to form a coarse-grained network. Moreover, we show that there exists a way of grouping nodes preserving as much as possible some crucial aspects of the network dynamics. This coarse graining approach provides a useful method to simplify complex oscillator networks, and more generally, networks whose dynamics involves a Laplacian matrix.


An event-related fMRI Study of exogenous facilitation and inhibition of return in the auditory modality.
The orienting of attention to different locations in space is fundamental to most organisms and occurs in all sensory modalities. Orienting has been extensively studied in vision, but to date, few studies have investigated neuronal networks underlying automatic orienting of attention and inhibition of return to auditory signals. In the current experiment, functional magnetic resonance imaging and behavioral data were collected while healthy volunteers performed an auditory orienting task in which a monaurally presented tone pip (cue) correctly or incorrectly cued the location of a target tone pip. The stimulus onset asynchrony (SOA) between the cue and target was 100 or 800 msec. Behavioral results were consistent with previous studies showing that valid auditory cues produced facilitation at the short SOA and inhibition of return at the long SOA. Functional results indicated that the reorienting of attention (100 msec SOA) and inhibition of return (800 msec SOA) were mediated by both common and distinct neuronal structures. Both attention mechanisms commonly activated a network consisting of fronto-oculomotor areas, the left postcentral gyrus, right premotor area, and bilateral tonsil of the cerebellum. Several distinct areas of frontal and parietal activation were identified for the reorienting condition, whereas the right inferior parietal lobule was the only structure uniquely associated with inhibition of return.


Recommended Cancer Screening in Accountable Care Organizations: Trends in Colonoscopy and Mammography in the Medicare Shared Savings Program.
Accountable care organizations (ACOs) are a delivery and payment model designed to encourage integrated, high-value care. We designed a study to test the association between ACOs and two recommended cancer screening tests, colonoscopy for colorectal cancer and mammography for breast cancer. Using the random 20% sample of Medicare claims, beneficiaries were attributed to ACO or non-ACO cohorts on the basis of providers' enrollment in the Medicare Shared Savings Program. An inverse probability of treatment weighting was used to balance patient characteristics between ACO and non-ACO cohorts. A propensity score-weighted, difference-in-differences analysis was then performed using the same provider groups in 2010-pre-ACO-as a baseline. A secondary analysis for older-nonrecommended-age ranges was performed. Prevalence of colonoscopy in recommended age ranges in ACOs from 2010 to 2014 increased from 15.3% (95% CI, 14.9% to 15.6%) to 17.9% (95% CI, 17.3% to 18.5%). This differed significantly from the change in non-ACOs (difference in differences, 1.2%; P < .001). Among women in ACOs, mammography prevalence rose from 53.7% (95% CI, 53.0% to 54.4%) to 54.9% (95% CI, 54.2% to 55.7%). In contrast to colonoscopy, the difference in mammography prevalence was not significantly different in ACO versus non-ACOs (difference in differences, 0.49%; P < .13). A similar pattern was also observed in older-nonrecommended-age ranges with significant difference in differences (ACO v non-ACO) in colonoscopy, but not mammography. The impact of ACOs on cancer screening varies between screening tests. Our results are consistent with a greater effect of ACOs on high-cost, high-complexity screening services, which may be more sensitive to integrated care delivery models.


Valinomycin biosynthetic gene cluster in Streptomyces: conservation, ecology and evolution.
Many Streptomyces strains are known to produce valinomycin (VLM) antibiotic and the VLM biosynthetic gene cluster (vlm) has been characterized in two independent isolates. Here we report the phylogenetic relationships of these strains using both parsimony and likelihood methods, and discuss whether the vlm gene cluster shows evidence of horizontal transmission common in natural product biosynthetic genes. Eight Streptomyces strains from around the world were obtained and sequenced for three regions of the two large nonribosomal peptide synthetase genes (vlm1 and vlm2) involved in VLM biosynthesis. The DNA sequences representing the vlm gene cluster are highly conserved among all eight environmental strains. The geographic distribution pattern of these strains and the strict congruence between the trees of the two vlm genes and the housekeeping genes, 16S rDNA and trpB, suggest vertical transmission of the vlm gene cluster in Streptomyces with no evidence of horizontal gene transfer. We also explored the relationship of the sequence of vlm genes to that of the cereulide biosynthetic genes (ces) found in Bacillus cereus and found them highly divergent from each other at DNA level (genetic distance values >or= 95.6%). It is possible that the vlm gene cluster and the ces gene cluster may share a relatively distant common ancestor but these two gene clusters have since evolved independently.


Cytokine expression in CD3+ cells in an infant with food protein-induced enterocolitis syndrome (FPIES): case report.
Food protein-induced enterocolitis syndrome (FPIES) is a non-IgE-mediated food allergy characterized by severe vomiting, diarrhea, and often failure to thrive in infants. Symptoms typically resolve after the triggering food-derived protein is removed from the diet and recur within few hours after the re-exposure to the causal protein. The diagnosis is based on clinical symptoms and a positive food challenge. In this study, we report a case of FPIES to rice in an 8-month-old boy. We performed a double-blind placebo-controlled food challenge (DBPCFC) to rice and we measured the intracellular T cell expression of interleukin-4 (IL-4); IL-10, and interferon gamma (IFN-gamma) pre-and post-challenge during an acute FPIES reaction and when tolerance to rice had been achieved. For the first time we describe an increase in T cell IL-4 and decrease in IFN-gamma expression after a positive challenge with rice (i.e. rice triggered a FPIES attack) and an increase in T cell IL-10 expression after rice challenge 6 months later after a negative challenge (i.e., the child had acquired tolerance to rice) in an 8 month old with documented FPIES to rice. A Th2 activation associated with high IL-4 levels may contribute to the pathophysiology of the disease. On the other hand, T cell-derived IL-10 may play a role in the acquisition of immunotolerance by regulating the Th1 and Th2 responses.


Death with graft function in elderly patients after cadaveric renal transplantation: effect of waiting time.
Survival after kidney transplantation is better than on the waiting list, even in the elderly. However, the effects of a prolonged waiting time for an organ on death with graft function have not been critically examined in this patient group. We conducted a single-center retrospective analysis of our cadaveric renal transplant experience in patients older than 60 years who received a kidney between January 1, 1990 and December 31, 2003. Besides waiting time, the effects of recipient age, gender, and diabetes were also examined. Cox proportional hazards analysis using patient death as a time-dependent outcome was used to estimate the hazard ratio of death posttransplantation. Using Kaplan-Meier survival methodology, patients with waiting times < or =5 years had significantly better survival times posttransplantation compared with those with waiting times >5 years (6.2 vs 2.8 years; P <.001). Each year of waiting was associated with hazard ratio 1.16 (95% confidence interval [CI], 1.06-1.27) for death. Prolonged waiting time on dialysis is deleterious to patient survival in recipients older than 60 years at transplantation. Early transplantation thus should be strongly encouraged in this group of patients.


Order within a mosaic distribution of mitochondrial c-type cytochrome biogenesis systems?
Mitochondrial cytochromes c and c(1) are present in all eukaryotes that use oxygen as the terminal electron acceptor in the respiratory chain. Maturation of c-type cytochromes requires covalent attachment of the heme cofactor to the protein, and there are at least five distinct biogenesis systems that catalyze this post-translational modification in different organisms and organelles. In this study, we use biochemical data, comparative genomic and structural bioinformatics investigations to provide a holistic view of mitochondrial c-type cytochrome biogenesis and its evolution. There are three pathways for mitochondrial c-type cytochrome maturation, only one of which is present in prokaryotes. We analyze the evolutionary distribution of these biogenesis systems, which include the Ccm system (System I) and the enzyme heme lyase (System III). We conclude that heme lyase evolved once and, in many lineages, replaced the multicomponent Ccm system (present in the proto-mitochondrial endosymbiont), probably as a consequence of lateral gene transfer. We find no evidence of a System III precursor in prokaryotes, and argue that System III is incompatible with multi-heme cytochromes common to bacteria, but absent from eukaryotes. The evolution of the eukaryotic-specific protein heme lyase is strikingly unusual, given that this protein provides a function (thioether bond formation) that is also ubiquitous in prokaryotes. The absence of any known c-type cytochrome biogenesis system from the sequenced genomes of various trypanosome species indicates the presence of a third distinct mitochondrial pathway. Interestingly, this system attaches heme to mitochondrial cytochromes c that contain only one cysteine residue, rather than the usual two, within the heme-binding motif. The isolation of single-cysteine-containing mitochondrial cytochromes c from free-living kinetoplastids, Euglena and the marine flagellate Diplonema papillatum suggests that this unique form of heme attachment is restricted to, but conserved throughout, the protist phylum Euglenozoa.


[Analysis of the clinical diagnosis of asbestosis complicated with malignant mesothelioma].
To discuss the clinical characteristics, diagnosis and treatment of the clinical characteristics and prognosis of asbestosis complicated with malignant mesothelioma patients. In 3 cases of asbestosis complicated with malignant mesothelioma were retrospectively analyzed. In the 3 patients, 2 cases of pleural mesothelioma, with chest tightness, chest and back pain as initial symptom; 1 case of peritoneal mesothelioma, with abdominal distention, abdominal pain, dysuria as initial symptom. One case of the pleural mesothelioma misdiagnosed as tuberculous pleurisy. 3 patients were in CT or B ultrasound guided biopsy pathology confirmed to be malignant mesothelioma. 2 patients received systemic chemotherapy, another received symptomatic and supportive treatment. Up till now, 3 patients have died. The disease is a high degree of malignant, the early clinical manifestations are not specific, easily missed diagnosis and misdiagnosis. The treatment effect is not ideal, the prognosis is poor. Biopsy is a reliable method for diagnosis of MM.


Keratin expression profiling of transitional epithelium in the painful bladder syndrome/interstitial cystitis.
Painful bladder syndrome/interstitial cystitis (PBS/IC) is a severely debilitating condition. Its cause is poorly understood; therapy is symptomatic and often unsuccessful. To study urothelial involvement, we characterized the keratin phenotype of bladder urothelium in 18 patients with PBS/IC using a panel of 11 keratin antibodies recognizing simple keratins found in columnar epithelia (keratins 7, 8, 18, and 20) and keratins associated with basal cell compartments of squamous epithelia (keratins 5, 13, 14, and 17). We also tested 2 antibodies recognizing more than 1 keratin also directed against basal cell compartments of squamous epithelia (D5/16 B4 and 34betaE12). Bladder urothelium in PBS/IC showed distinct differences in the profiles of keratins 7, 8, 14, 17, 18, and 20 compared with literature reports for normal bladder urothelium. These were characterized by a shift from the normal bladder urothelial keratin phenotype to a more squamous keratin profile, despite the lack of morphologic evidence of squamous epithelial differentiation and a loss of compartmentalization of keratin expression. The severity of these changes varied between biopsy specimens. Whether these changes are primary or secondary to another underlying condition remains to be determined.


Musical beauty and information compression: Complex to the ear but simple to the mind?
The biological origin of music, its universal appeal across human cultures and the cause of its beauty remain mysteries. For example, why is Ludwig Van Beethoven considered a musical genius but Kylie Minogue is not? Possible answers to these questions will be framed in the context of Information Theory. The entire life-long sensory data stream of a human is enormous. The adaptive solution to this problem of scale is information compression, thought to have evolved to better handle, interpret and store sensory data. In modern humans highly sophisticated information compression is clearly manifest in philosophical, mathematical and scientific insights. For example, the Laws of Physics explain apparently complex observations with simple rules. Deep cognitive insights are reported as intrinsically satisfying, implying that at some point in evolution, the practice of successful information compression became linked to the physiological reward system. I hypothesise that the establishment of this "compression and pleasure" connection paved the way for musical appreciation, which subsequently became free (perhaps even inevitable) to emerge once audio compression had become intrinsically pleasurable in its own right. For a range of compositions, empirically determine the relationship between the listener's pleasure and "lossless" audio compression. I hypothesise that enduring musical masterpieces will possess an interesting objective property: despite apparent complexity, they will also exhibit high compressibility. Artistic masterpieces and deep Scientific insights share the common process of data compression. Musical appreciation is a parasite on a much deeper information processing capacity. The coalescence of mathematical and musical talent in exceptional individuals has a parsimonious explanation. Musical geniuses are skilled in composing music that appears highly complex to the ear yet transpires to be highly simple to the mind. The listener's pleasure is influenced by the extent to which the auditory data can be resolved in the simplest terms possible.


Competence for Genetic Transformation in Streptococcus pneumoniae: Mutations in σA Bypass the ComW Requirement for Late Gene Expression.
Streptococcus pneumoniae is able to integrate exogenous DNA into its genome by natural genetic transformation. Transient accumulation of high levels of the only S. pneumoniae alternative σ factor is insufficient for development of full competence without expression of a second competence-specific protein, ComW. The ΔcomW mutant is 10(4)-fold deficient in the yield of recombinants, 10-fold deficient in the amount of σ(X) activity, and 10-fold deficient in the amount of σ(X) protein. The critical role of ComW during transformation can be partially obviated by σ(A) mutations clustered on surfaces controlling affinity for core RNA polymerase (RNAP). While strains harboring σ(A) mutations in the comW mutant background were transforming at higher rates, the mechanism of transformation restoration was not clear. To investigate the mechanism of transformation restoration, we measured late gene expression in σ(A)* suppressor strains. Restoration of late gene expression was observed in ΔcomW σ(A)* mutants, indicating that a consequence of the σ(A)* mutations is, at least, to restore σ(X) activity. Competence kinetics were normal in ΔcomW σ(A)* strains, indicating that strains with restored competence exhibit the same pattern of transience as wild-type (WT) strains. We also identified a direct interaction between ComW and σ(X) using the yeast two-hybrid (Y2H) assay. Taken together, these data are consistent with the idea that ComW increases σ(X) access to core RNAP, pointing to a direct role of ComW in σ factor exchange during genetic transformation. However, the lack of late gene shutoff in ΔcomW mutants also points to a potential new role for ComW in competence shutoff. The sole alternative sigma factor of the streptococci, SigX, regulates development of competence for genetic transformation, a widespread mechanism of adaptation by horizontal gene transfer in this genus. The transient appearance of this sigma factor is strictly controlled at the levels of transcription and stability. This report shows that it is also controlled at the point of its substitution for SigA by a second transient competence-specific protein, ComW.


Activation of mesangial cell MAPK in response to homocysteine.
Alteration in mesangial cell function is central to the progression of glomerular disease in numerous models of chronic renal failure (CRF). Animal models of chronic glomerular disease are characterized by mesangial cell proliferation and elaboration of extracellular matrix protein (ECM), resulting in glomerulosclerosis. Elevated plasma levels of homocysteine (Hcy) are seen in both animal models and humans with CRF, and have been proposed to contribute to the high prevalence of vascular disease in this group. Some of the pathogenetic effects of Hcy are thought to be mediated via the induction of endoplasmic reticulum stress. Thus, Hcy effects on mesangial cells could contribute to the progression of CRF. Previous work has shown Hcy- mediated induction of Erk mitogen-activated protein kinase (MAPK) in vascular smooth muscle cells (VSMCs). Erk induces increases in activator protein-1 (AP-1) transcription factor activity which may augment mesangial cell proliferation and ECM protein production. Consequently, we studied the effect of Hcy on mesangial cell Erk signaling. Mesangial cells were exposed to Hcy after 24 hours of serum starvation and Erk activity assessed. Nuclear translocation of phospho-Erk was visualized by confocal microscopy. AP-1 nuclear protein binding was measured in response to Hcy by mobility shift assay. Hcy-induced mesangial cell calcium flux was measured in Fura-2 loaded cells. Mesangial cell DNA synthesis in response to Hcy was assessed by [3H]-thymidine incorporation and proliferation by Western blotting for proliferating cell nuclear antigen (PCNA). Expression of endoplasmic reticulum stress response genes were determined by Northern and Western analysis. Hcy led to an increase in Erk activity that was maximal at 50 micromol/L and 20 minutes of treatment. Subsequent experiments used this concentration and time point. Erk activity in response to Hcy was insensitive to n-acetylcysteine and catalase, indicating oxidative stress did not play a role. However, Hcy50 micromol/L induced a brief increase in intracellular mesangial cell calcium within 5 minutes, and the calcium ionophores A23187 and ionomycin increased Erk activity while chelation of intracellular calcium with BAPTA-AM abrogated the Erk response to Hcy. Confocal microscopy of activated Erk nuclear translocation mirrored these results as did mesangial cell nuclear protein binding to AP-1 consensus sequences. Hcy- induced increases in thymidine incorporation and PCNA expression at 24 hours were Erk dependent. The expression of endoplasmic reticulum stress response genes was significantly elevated by Hcy in an Erk-dependent manner. Hcy increases Erk activity in mesangial cells via a calcium-dependent mechanism, resulting in increased AP-1 nuclear protein binding, cell DNA synthesis and proliferation and induction of endoplasmic reticulum stress. These observations suggest potential mechanisms by which Hcy may contribute to progressive glomerular injury.


Physical activity interventions in African American women: A systematic review.
Background: African American women are at high risk of acquiring chronic diseases due to sedentary lifestyles. This objective of this article was to perform a narrative systematic review of physical activity interventions among African American women published between 2009 and 2015. Methods: A review of literature in following databases: Academic Search Premier, CINAHL (Cumulative Index to Nursing & Allied Health), ERIC (Education Resources Information Center), MEDLINE, PsychInfo, and SPORTDiscus was performed to locate interventions promoting physical activity among African American women. Results: The search yielded 13 interventions. All the studies were conducted within the United States. It was found that walking coupled with healthy food choices were salient strategies in the interventions. Studies using social support along with healthy diet were found to be more efficacious in fostering physical activity among African American women. Conclusion: Walking, social support and a healthy diet were found to be significant strategies promoting physical activity in African American women. Physical activity for African American women must build on the constructs of healthier food choices and social support.


Glucosylceramide acyl chain length is sensed by the glycolipid transfer protein.
The glycolipid transfer protein, GLTP, can be found in the cytoplasm, and it has a FFAT-like motif (two phenylalanines in an acidic tract) that targets it to the endoplasmic reticulum (ER). We have previously shown that GLTP can bind to a transmembrane ER protein, vesicle-associated membrane protein-associated protein A (VAP-A), which is involved in a wide range of ER functions. We have addressed the mechanisms that might regulate the association between GLTP and the VAP proteins by studying the capacity of GLTP to recognize different N-linked acyl chain species of glucosylceramide. We used surface plasmon resonance and a lipid transfer competition assay to show that GLTP prefers shorter N-linked fully saturated acyl chain glucosylceramides, such as C8, C12, and C16, whereas long C18, C20, and C24-glucosylceramides are all bound more weakly and transported more slowly than their shorter counterparts. Changes in the intrinsic GLTP tryptophan fluorescence blueshifts, also indicate a break-point between C16- and C18-glucosylceramide in the GLTP sensing ability. It has long been postulated that GLTP would be a sensor in the sphingolipid synthesis machinery, but how this mechanistically occurs has not been addressed before. It is unclear what proteins the GLTP VAP association would influence. Here we found that if GLTP has a bound GlcCer the association with VAP-A is weaker. We have also used a formula for identifying putative FFAT-domains, and we identified several potential VAP-interactors within the ceramide and sphingolipid synthesis pathways that could be candidates for regulation by GLTP.


Peripheral nervous system manifestations in systemic autoimmune diseases.
The peripheral nervous system refers to parts of the nervous system outside the brain and spinal cord. Systemic autoimmune diseases can affect both the central and peripheral nervous systems in a myriad of ways and through a heterogeneous number of mechanisms leading to many different clinical manifestations. As a result, neurological complications of these disorders can result in significant morbidity and mortality. The most common complication of peripheral nervous system (PNS) involvement is peripheral neuropathy, with symptoms of numbness, sensory paresthesias, weakness, or gait imbalance. The neuropathy may be multifocal and asymmetric or, less frequently, distal and symmetric.


Increase in Psychoactive Drug Prescriptions in the Years Following Autism Spectrum Diagnosis: A Population-Based Cohort Study.
Psychoactive medications are commonly prescribed to autistic individuals, but little is known about how their use changes after diagnosis. This study describes the use of psychoactive drugs in children and young adults newly diagnosed with autism spectrum, between the year before and up to 5 years after diagnosis. Multivariable logistic regression was used to examine the relationship between the use psychoactive drugs before the first diagnosis of autism spectrum condition (from 1998 to 2010), and the clinical and demographic characteristics, identified from public health care databases in Quebec. The types of drugs prescribed and psychoactive polypharmacy were evaluated over 5 years of follow-up. Generalized estimating equations (GEE) were used to examine the association of age and time with the use of psychoactive drugs. In our cohort of 2,989 individuals, diagnosis of another psychiatric disorder before autism spectrum strongly predicted psychoactive drug use. We observed that the proportion of users of psychoactive drugs increased from 35.6% the year before, to 53.2% 5 years after the autism spectrum diagnosis. Psychoactive polypharmacy (≥2 psychoactive drug classes) also increased from 9% to 22% in that time. Age and time since diagnosis strongly associated with the types and combinations of psychoactive drugs prescribed. Psychoactive drug use and polypharmacy increases substantially over time after autism spectrum disorder diagnosis in children.


Hsp70 regulates the interaction between the peroxisome targeting signal type 1 (PTS1)-receptor Pex5p and PTS1.
The peroxisome targeting signal type 1 (PTS1) receptor, Pex5p, of the tetratricopeptide repeat (TPR) motif family is located mostly in the cytosol and mediates the translocation of PTS1 proteins to peroxisomes. As a step towards understanding the mechanisms of protein import into peroxisomes, we investigated the molecular mechanisms involved in PTS1 recognition by Pex5p with regard to requirement of energy and cytosolic factors, using cell-free synthesized acyl-CoA oxidase (AOx) as a PTS1 cargo protein, together with Pex5p and heat-shock protein (Hsp)70 from rat liver. Pex5p was partly associated with peroxisomes of rat liver, was resistant to washing with a high concentration of salt and to alkaline extraction and was inaccessible to protease added externally. Pex5p bound to AOx in an ATP-dependent manner. AOx synthesized in a cell-free translating system from rabbit reticulocyte lysate was imported into peroxisomes without being supplemented with Pex5p and Hsp70, implying that peroxisome-associated Pex5p was released from the membranes and functional in this in vitro import assay. Antibodies against Pex5p and Hsp70 inhibited AOx import. In contrast, AOx synthesized in a wheat-germ lysate required the external addition of Pex5p for import, in which Hsp70 augmented the AOx import. The TPR domain of Pex5p was revealed to bind to the N-terminal part in an Hsp70-independent manner, whereas mutual interaction of the TPR region was noted in the presence of Hsp70. Hsp70 interacted with the TPR domain of Pex5p. Moreover, Hsp70 and ATP synergistically enhanced the binding of Pex5p to the C-terminal PTS1-containing part of AOx, implying that Pex5p recognizes its cargo PTS1 protein by chaperone-assisted as well as energy-dependent mechanisms in vivo.


Delayed globin synthesis leads to excess heme and the macrocytic anemia of Diamond Blackfan anemia and del(5q) myelodysplastic syndrome.
Diamond Blackfan anemia (DBA) and myelodysplastic syndrome (MDS) with isolated del(5q) are severe macrocytic anemias; although both are associated with impaired ribosome assembly, why the anemia occurs is not known. We cultured marrow cells from DBA (n = 3) and del(5q) MDS (n = 6) patients and determined how heme (a toxic chemical) and globin (a protein) are coordinated. We show that globin translation initiates slowly, whereas heme synthesis proceeds normally. This results in insufficient globin protein, excess heme and excess reactive oxygen species in early erythroid precursors, and CFU-E (colony-forming unit-erythroid)/proerythroblast cell death. The cells that can more rapidly and effectively export heme or can slow heme synthesis preferentially survive and appropriately mature. Consistent with these observations, treatment with 10 μM succinylacetone, a specific inhibitor of heme synthesis, improved the erythroid cell output of DBA and del(5q) MDS marrow cultures by 68 to 95% (P = 0.03 to 0.05), whereas the erythroid cell output of concurrent control marrow cultures decreased by 4 to 13%. Our studies demonstrate that erythropoiesis fails when heme exceeds globin. Our data further suggest that therapies that decrease heme synthesis (or facilitate heme export) could improve the red blood cell production of persons with DBA, del(5q) MDS, and perhaps other macrocytic anemias.


PDGFRα Is a Key Regulator of T1 and T3's Differential Effect on SMA Expression in Human Corneal Fibroblasts.
The goal of this study was to examine the mechanism behind the unique differential action of transforming growth factor β3 (TGF-β3) and TGF-β1 on SMA expression. It was our hypothesis that platelet-derived growth factor receptor α (PDGFRα) played a key role in determining TGF-β3's response to wounding. A stable cell line, human corneal fibroblast (HCF)-P, was created from HCFs by knocking down PDGFRα expression using a lentivirus-delivered shRNA sequence. A three-dimensional (3D) in vitro model was constructed by culturing HCF or HCF-P on poly-transwell membranes for 4 weeks in the presence and absence of 0.1 ng/mL TGF-β1 or -β3. At the end of 4 weeks, the constructs were processed for immunofluorescence and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, HCF and HCF-P cell migration was evaluated. In HCF, TGF-β3 treatment resulted in significantly lower α-smooth muscle actin (SMA) mRNA expression and immunolocalization when compared to TGF-β1, while in HCF-P, both TGF-β1 and -β3 treatment increased the SMA mRNA expression and immunolocalization compared to both the untreated HCF-P control and TGF-β3-treated HCF. Human corneal fibroblast-P also had a lower migration rate and construct thickness when compared to HCF. These results show that TGF-β3 decreases SMA in HCF, while remarkably increasing SMA in HCF-P, thus indicating that the presence or absence of PDGFRα elicits contrasting responses to the same TGF-β3 treatment. Understanding the role of PDGFRα in TGF-β3's ability to stimulate SMA may potentially help in understanding the differential functions of TGF-β1 and TGF-β3 in corneal wound healing.


T-maze forced alternation and left-right discrimination tasks for assessing working and reference memory in mice.
Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of α-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that α-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that α-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.


Biological effects of prostate specific antigen as an insulin-like growth factor binding protein-3 protease.
Prostate specific antigen (PSA) is an insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) protease found in seminal plasma and produced by prostatic epithelial cells (PC-E) in vivo. We examined the effects of PSA-proteolysis of IGFBP-3 on the affinity of IGFBP-3 fragments for IGFs and on the mitogenic action of IGFs on PC-E. Recombinant human IGFBP-3 was cleaved by PSA, then incubated with 125I-IGF-I or -II in the presence of varying concentrations of unlabelled peptides, and then cross-linking electrophoresis and densitometric analysis were performed. While the affinity of IGF-II for the PSA-generated IGFBP-3 fragments fell slightly compared to intact IGFBP-3, the affinity of the PSA-generated IGFBP-3 fragments for IGF-I fell by ten fold. The addition of IGF-I or -II to PC-E in serum-free culture conditions resulted in a two-fold stimulation of cell number compared to control. The presence of IGFBP-3 in the media blocked the IGF-induced stimulation, but had no independent effect in the absence of IGFs. When PSA was added to PC-E cultures to which both IGF-I or -II and IGFBP-3 were added, the inhibitory effects of IGFBP-3 on IGF mitogenesis were reversed. We conclude that PSA decreases the affinity of IGFBP-3 for IGF and can potentiate IGF action in the presence of inhibitory IGFBP-3. This phenomenon may contribute to normal and malignant prostate growth.


How First Shell-Second Shell Interactions and Metal Substitution Modulate Protein Function.
Hydrogen bonds to metal-ligands in proteins play a vital role in biological function. They help to stabilize/protect the metal complex and enhance metal-binding affinity/specificity, enzyme-substrate recognition, and enzyme activation. Yet, knowledge of the preferred hydrogen-bonding partners of metal ligands in different metalloproteins is lacking. Using well-calibrated methods, we have determined the preferred hydrogen-bonding partners of Cys- bound to native Zn2+ or xenobiotic Cd2+ in Zn-fingers of varying net charge and solvent accessibility as well as the key factors underlying the observed preference. We show how secondary hydrogen-bonding interactions with metal-bound thiolates might exert a significant impact on Zn2+→Cd2+ substitution and thus protein function. Knowing which Zn-fingers may be vulnerable to structural deformation by Cd2+ is important since this would lead to their inactivation, which might impair cell growth, differentiation, cell-cycle control, and DNA repair.


Factors affecting breastfeeding cessation after discontinuation of antiretroviral therapy to prevent mother-to-child transmission of HIV.
In the Kisumu Breastfeeding Study (KiBS), prevention of mother-to-child HIV transmission study, highly active antiretroviral therapy (HAART) is provided from 34 weeks gestation, through delivery to six months postpartum. The study recommends that women practice exclusive breastfeeding for six months, then wean abruptly. We sought to explore factors such as, education, family support, cultural norms, and sources of information about perinatal HIV transmission, which may influence a mother's decision to comply or not comply with the study's recommendation to stop breastfeeding when HAART is discontinued. We used semi-structured interviews of a purposive sample of 18 mothers participating in the KiBS. By interviewing 10 mothers who stopped breastfeeding and eight mothers who continued, it was possible to examine how different factors may have affected the groups of participants. All participants stated that it was not traditional to stop breastfeeding at six months. Participants who stopped breastfeeding reported more family support, were more educated, and were more likely to disclose their HIV status. Participants who continued breastfeeding more often expressed concern about stigma. Participants learned about mother-to-child transmission from clinics, churches, community groups, and other HIV-positive mothers. This substudy suggests that family support, education, and cultural norms are important factors that may influence a mother's decision regarding breastfeeding cessation. Thus, counseling and family support may play integral roles in the promotion of early breastfeeding cessation.


Neural crest-derived neurons invade the ovary but not the testis during mouse gonad development.
Testes and ovaries undergo sex-specific morphogenetic changes and adopt strikingly different morphologies, despite the fact that both arise from a common precursor, the bipotential gonad. Previous studies showed that recruitment of vasculature is critical for testis patterning. However, vasculature is not recruited into the early ovary. Peripheral innervation is involved in patterning development of many organs but has been given little attention in gonad development. In this study, we show that while innervation in the male reproductive complex is restricted to the epididymis and vas deferens and never invades the interior of the testis, neural crest-derived innervation invades the interior of the ovary around E16.5. Individual neural crest cells colonize the ovary, differentiate into neurons and glia, and form a dense neural network within the ovarian medulla. Using a sex-reversing mutant mouse line, we show that innervation is specific to ovary development, is not dependent on the genetic sex of gonadal or neural crest cells, and may be blocked by repressive guidance signals elevated in the male pathway. This study reveals another aspect of sexually dimorphic gonad development, establishes a precise timeline and structure of ovarian innervation, and raises many questions for future research.


Mental illness and the provision of mental health services in prisons.
Around 11 million people are held in prisons internationally, and criminal justice systems are overburdened with a high prevalence of multiple psychiatric disorders. In England and Wales over 200 000 people enter prisons each year, and in many cases, this facilitates their first contact with mental health services. Research, evaluations, government reports and independent reviews. Screening, Triage, Assessment, Intervention and Re-integration (STAIR) are necessary components of prison mental health provision, offering an opportunity to improve the wellbeing of a complex population. There are serious problems with service provision across many parts of the world, with human rights abuses occurring in some States. Screening and service delivery models still require substantial development. In England and Wales, self-harm, self-inflicted deaths and violence are increasing. Introducing comprehensive mental health models throughout prisons would offer a massive public health initiative, providing new services for the socially disadvantaged. A rights-based framework would be useful in ensuring systemic improvements, especially in low and middle-income countries. Mechanisms for screening and triage, specific interventions across a broad range of conditions, and practical re-integration models, should be submitted to research across international sites.


Dimethyl adipimidate: a new antisickling agent.
A new approach to the prevention of sickling in vitro by use of the bifunctional crosslinking reagent, dimethyl adipimidate, is described. Prior treatment of sickle erythrocytes with dimethyl adipimidate will inhibit sickling in completely deoxygenated erythrocytes. Treated erythrocytes do not demonstrate the potassium loss and viscosity increase that usually accompany sickling. The oxygen affinity of hemoglobin in these cells is increased independently from changes in the concentration of 2,3-diphosphoglycerate. The hemoglobin obtained from treated erythrocytes contains a high-molecular-weight component as well as additional positively charged components. The relative degree to which chemical modification and/or crosslinking is an essential part of the antisickling properties of the material is not known.


Impact of COVID-19 on the quantile connectedness between energy, metals and agriculture commodities.
With many studies highlighting the heterogeneous impact of the COVID-19 pandemic on different commodity markets, this study provides evidence of quantile connectedness between energy, metals, and agriculture commodity markets before and during the COVID-19 outbreak. Since mean-based measures of connectedness are not necessarily suitable to measure connectedness in the crisis period, especially in the tails of the return distribution, thus in this study, we use the newly developed approach of quantile-based connectedness. The full-sample analysis results show that return shocks only propagate within the energy commodity group. The findings manifest that transmission of return spillovers is stronger in the left and right tails of the conditional return distribution. In addition, the results unveil that degree of tail-dependence between energy, metals, and agriculture commodities are time-varying. Meanwhile, our sub-sample analysis clearly shows that the commodity market return connectedness demonstrates a significant shift over time due to COVID-19 shocks. There is evidence of strong transmission of return shocks between energy, metals, and agriculture commodities during the COVID-19 fiasco. Finally, the results also illustrate that softs and livestock commodities hold significant diversification benefits for energy market investors.


Oligomeric Architecture of Mouse Activating Nkrp1 Receptors on Living Cells.
Mouse activating Nkrp1 proteins are commonly described as type II transmembrane receptors with disulfide-linked homodimeric structure. Their function and the manner in which Nkrp1 proteins of mouse strain (C57BL/6) oligomerize are still poorly understood. To assess the oligomerization state of Nkrp1 proteins, mouse activating EGFP-Nkrp1s were expressed in mammalian lymphoid cells and their oligomerization evaluated by Förster resonance energy transfer (FRET). Alternatively, Nkrp1s oligomers were detected by Western blotting to specify the ratio between monomeric and dimeric forms. We also performed structural characterization of recombinant ectodomains of activating Nkrp1 receptors. Nkrp1 isoforms c1, c2 and f were expressed prevalently as homodimers, whereas the Nkrp1a displays larger proportion of monomers on the cell surface. Cysteine-to-serine mutants revealed the importance of all stalk cysteines for protein dimerization in living cells with a major influence of cysteine at position 74 in two Nkrp1 protein isoforms. Our results represent a new insight into the oligomerization of Nkrp1 receptors on lymphoid cells, which will help to determine their function.


Fast and accurate determination of the relative binding affinities of small compounds to HIV-1 protease using non-equilibrium work.
The fast pulling ligand (FPL) out of binding cavity using non-equilibrium molecular dynamics (MD) simulations was demonstrated to be a rapid, accurate and low CPU demand method for the determination of the relative binding affinities of a large number of HIV-1 protease (PR) inhibitors. In this approach, the ligand is pulled out of the binding cavity of the protein using external harmonic forces, and the work of pulling force corresponds to the relative binding affinity of HIV-1 PR inhibitor. The correlation coefficient between the pulling work and the experimental binding free energy of R=-0.95 shows that FPL results are in good agreement with experiment. It is thus easier to rank the binding affinities of HIV-1 PR inhibitors, that have similar binding affinities because the mean error bar of pulling work amounts to δW=7%. The nature of binding is discovered using the FPL approach. © 2016 Wiley Periodicals, Inc.


TLC-based detection of methylated cytosine: application to aging epigenetics.
5-Methylcytosine (m(5)C) has a plethora of functions and roles in various biological processes including human diseases and aging. A TLC-based fast and simple method for quantitative determination of total genomic levels of m(5)C in DNA is described, which can be applicable to aging research with respect to rapid and high throughput screening and comparison. Using this method, an example of the analysis of global alternations of m(5)C in serially passaged human skin fibroblasts is provided, which shows age-related global hypomethylation during cellular aging in vitro. This method can be useful for screening potential modulators of aging at the level of epigenetic alterations.


Distribution of Serotonin-Immunoreactive Neurons in the Brain and Gnathal Ganglion of Caterpillar Helicoverpa armigera.
Serotonin (5-hydroxytryptamine, 5-HT) is an important biogenic amine that acts as a neural circuit modulator. It is widespread in the central nervous system of insects. However, little is known about the distribution of serotonin in the nervous system of the cotton bollworm Helicoverpa armigera. In the present study, we performed immunohistochemical experiments with anti-serotonin serum to examine the distribution of serotonin in the central nervous system of H. armigera larvae. We found about 40 serotonin-immunoreactive neurons in the brain and about 20 in the gnathal ganglion. Most of these neurons are wide-field neurons giving rise to processes throughout the neuropils of the brain and the gnathal ganglion. In the central brain, serotonin-immunoreactive processes are present bilaterally in the tritocerebrum, the deutocerebrum, and major regions of the protocerebrum, including the central body (CB), lateral accessory lobes (LALs), clamps, crepine, superior protocerebrum, and lateral protocerebrum. The CB, anterior ventrolateral protocerebrum (AVLP), and posterior optic tubercle (POTU) contain extensive serotonin-immunoreactive process terminals. However, the regions of mushroom bodies, the lateral horn, and protocerebral bridges (PBs) are devoid of serotonin-immunoreactivity. In the gnathal ganglion, the serotonin-immunoreactive processes are also widespread throughout the neuropil, and some process projections extend to the tritocerebrum. Our results provide the first comprehensive description of the serotonergic neuronal network in H. armigera larvae, and they reveal the neural architecture and the distribution of neural substances, allowing us to explore the neural mechanisms of behaviors by using electrophysiological and pharmacological approaches on the target regions.


Is visual memory predictive of below-average academic achievement in second through fourth graders?
Controversy exists regarding the relation between visual memory and academic achievement. A masked investigation of the relation between visual memory and academics was performed in 155 second-through fourth-grade children (mean age = 8.83 years). Visual memory ability was assessed with the Test of Visual Perceptual Skills visual memory subtest. The school administered the Otis-Lennon School Ability Test and Stanford Achievement Test. Age and verbal ability were controlled in all regression analyses. Visual memory score was significantly predictive of below-average word decoding (p = 0.027), total math score (p = 0.031), and Stanford complete battery score (p = 0.018). Visual memory score showed a positive trend in predicting reading comprehension (p = 0.093). Poor visual memory ability (as measured by the Test of Visual Perceptual Skills) is significantly related to below-average reading decoding, math, and overall academic achievement (as measured by the Stanford Achievement Test) in second- through fourth-grade children, while controlling for age and verbal ability.


Impact of Previous Somatostatin Analogue Use on the Activity of Everolimus in Patients with Advanced Neuroendocrine Tumors: Analysis from the Phase III RADIANT-2 Trial.
The phase III placebo-controlled RADIANT-2 trial investigated the efficacy of everolimus plus octreotide long-acting repeatable (LAR) in patients with advanced neuroendocrine tumors (NET) associated with carcinoid syndrome. Here we report a secondary analysis based on the previous somatostatin analogue (SSA) exposure status of patients enrolled in RADIANT-2. Patients were randomly assigned to receive oral everolimus 10 mg/day plus octreotide LAR 30 mg intramuscularly (i.m.) or to receive matching placebo plus octreotide LAR 30 mg i.m. every 28 days. SSA treatment before study enrollment was permitted. Patient characteristics and progression-free survival (PFS) were analyzed by treatment arm and previous SSA exposure status. Of the 429 patients enrolled in RADIANT-2, 339 were previously exposed to SSA (95% received octreotide); 173 of 339 patients were in the everolimus plus octreotide LAR arm. All patients had a protocol-specified history of secretory symptoms, but analysis by type showed that more patients who previously received SSA therapy had a history of flushing symptoms (77%), diarrhea (86%), or both (63%) compared with SSA-naive patients (62, 62, and 24%, respectively). Patients who received everolimus plus octreotide LAR had longer median PFS regardless of previous SSA exposure (with: PFS 14.3 months, 95% confidence interval, CI, 12.0-20.1; without: 25.2 months, 95% CI, 12.0-not reached) compared with patients who received placebo plus octreotide LAR (with: 11.1 months, 95% CI, 8.4-14.6; without: 13.6 months, 95% CI, 8.2-22.7). Everolimus in combination with octreotide improves PFS in patients with advanced NET associated with carcinoid syndrome, regardless of previous SSA exposure.


Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea.
One attractive strategy to prevent or control allograft rejection is to genetically modify the donor tissue before transplantation. In this study, we have examined the feasibility of gene transfer to human corneal endothelium, using a number of recombinant adenovirus constructs. Ex vivo infection of human corneas with adenoviral vectors containing lacZ, under transcriptional control of either cytomegalovirus (CMV) or Rous sarcoma virus (RSV) promoters, provided high-level gene expression, which was largely restricted to endothelium. Expression of the reporter gene persisted at relatively high levels for up to 7 days, followed by a decline to indetectable levels by 28 days. RT-PCR analysis of lacZ transcription showed a similar picture with a short period (3-7 days) of RNA transcription after infection. In contrast, adenoviral DNA persisted for at least 56 days. Subsequently, we examined the expression of a potential therapeutic gene, CTLA-4 Ig fusion protein. Following infection of human corneas with adenoviral vectors encoding CTLA-4 Ig protein, high levels of the fusion protein were detected in corneal culture supernatants for up to 28 days. This protein was functionally active, as determined by binding to B7.1 (CD80)-expressing transfectants. This study suggests that genetic alteration of donor cornea before transplantation is a feasible approach for preventing or controlling allograft rejection. Similar gene-based strategies might also be feasible to prevent rejection of other transplanted tissues or organs.


First report of peritonitis caused by the vancomycin-resistant coccus Pediococcus pentosaceus in a patient on continuous ambulatory peritoneal dialysis.
Worldwide, about one-tenth of end-stage renal disease (ESRD) patients are on peritoneal dialysis (PD). Peritonitis is a major cause of PD failure and change of therapy to haemodialysis. An update on peritoneal dialysis-related infections has recommended the use of a first generation cephalosporin or vancomycin as an empirical therapy for Gram-positive organisms. Pediococcus spp. is a Gram-positive environmental cocci that have been increasingly reported from various nosocomial infections but very rarely from peritoneal dialysis infections. It is intrinsically resistant to Vancomycin but sensitive to ampicillin. So, diagnosis of this bacteria is important if isolated from PD infections. An elderly female patient of ESRD on continuous ambulatory peritoneal dialysis (CAPD) was admitted with complaints of high fever and cloudy PD effluent for 2 days. She was started with vancomycin and imipenem empirically but did not improve even after 4 days. Pus cells were seen when PD fluid was examined microscopically. BACTEC culture of PD fluid isolated growth of Gram-positive cocci, which was confirmed as Pediococcus pentosaceus . It was resistant to vancomycin. The antibiotic of the patient was changed to ciprofloxacin IV. The patient responded in 2 days and was discharged after 7 days. This is the first case report of Pediococcus pentosaceus peritonitis in an ESRD patient on CAPD. Accurate diagnosis and antibiotic sensitivity test of the bacteria is important especially if isolated in critical patients as it is intrinsically resistant to vancomycin.


KARAOKE: Krill oil versus placebo in the treatment of knee osteoarthritis: protocol for a randomised controlled trial.
Knee osteoarthritis (OA) is a common and important cause of pain and disability, but interventions aimed at modifying structures visible on imaging have been disappointing. While OA affects the whole joint, synovitis and effusion have been recognised as having a role in the pathogenesis of OA. Krill oil reduces knee pain and systemic inflammation and could be used for targeting inflammatory mechanisms of OA. We will recruit 260 patients with clinical knee OA, significant knee pain and effusion-synovitis present on MRI in five Australian cities (Hobart, Melbourne, Sydney, Adelaide and Perth). These patients will be randomly allocated to the two arms of the study, receiving 2 g/day krill oil or inert placebo daily for 6 months. MRI of the study knee will be performed at screening and after 6 months. Knee symptoms, function and MRI structural abnormalities will be assessed using validated methods. Safety data will be recorded. Primary outcomes are absolute change in knee pain (assessed by visual analog score) and change in size of knee effusion-synovitis over 24 weeks. Secondary outcomes include improvement in knee pain over 4, 8, 12, 16 and 20 weeks. The primary analyses will be intention-to-treat analyses of primary and secondary outcomes. Per protocol analyses adjusting for missing data and for treatment compliance will be performed as the secondary analyses. This study will provide high-quality evidence to assess whether krill oil 2 g/day reduces pain and effusion-synovitis size in older adults with clinical knee OA and knee effusion-synovitis. If krill oil is effective and confirmed to be safe, we will provide compelling evidence that krill oil improves pain and function, changes disease trajectory and slows disease progression in OA. Given the lack of approved therapies for slowing disease progression in OA, and moderate cost of krill oil, these findings will be readily translated into clinical practice. Australian New Zealand Clinical Trials Registry, ACTRN12616000726459. Registered on 02 June 2016. Universal Trial Number (UTN) U1111-1181-7087.


Insights into gait disorders: walking variability using phase plot analysis, Huntington's disease.
Huntington's disease (HD) is a progressive inherited neurodegenerative disorder. Identifying sensitive methodologies to quantitatively measure early motor changes have been difficult to develop. This exploratory observational study investigated gait variability and symmetry in HD using phase plot analysis. We measured the walking of 22 controls and 35 HD gene carriers (7 premanifest (PreHD)), 16 early/mid (HD1) and 12 late stage (HD2) in Oxford and Cardiff, UK. The unified Huntington's disease rating scale-total motor scores (UHDRS-TMS) and disease burden scores (DBS) were used to quantify disease severity. Data was collected during a clinical walk test (8.8 or 10 m) using an inertial measurement unit attached to the trunk. The 6 middle strides were used to calculate gait variability determined by spatiotemporal parameters (co-efficient of variation (CoV)) and phase plot analysis. Phase plots considered the variability in consecutive wave forms from vertical movement and were quantified by SDA (spatiotemporal variability), SDB (temporal variability), ratio ∀ (ratio SDA:SDB) and Δangleβ (symmetry). Step time CoV was greater in manifest HD (p<0.01, both manifest groups) than controls, as was stride length CoV for HD2 (p<0.01). No differences were found in spatiotemporal variability between PreHD and controls (p>0.05). Phase plot analysis identified differences between manifest HD and controls for SDB, Ratio ∀ and Δangle (all p<0.01, both manifest groups). Furthermore Ratio ∀ was smaller in PreHD compared with controls (p<0.01). Ratio ∀ also produced the strongest correlation with UHDRS-TMS (r=-0.61, p<0.01) and was correlated with DBS (r=-0.42, p=0.02). Phase plot analysis may be a sensitive method of detecting gait changes in HD and can be performed quickly during clinical walking tests.


Zinc induces expression of the BH3-only protein PUMA through p53 and ERK pathways in SH-SY5Y neuroblastoma cells.
Accumulating evidence suggests that zinc (Zn2+) contributes to neuronal death in pathologic states such as ischemia. p53-upregulated modulator of apoptosis (PUMA), which is a BH3-only protein, is known to promote apoptosis through a tumor suppressor p53-dependent and -independent mechanism. In this study, we examined the effect of Zn2+ on the induction of the PUMA gene in human neuroblastoma SH-SY5Y cells. The expression of PUMA was induced by Zn2+ in a dose- and time-dependent manner. A reporter assay revealed that Zn2+ activated the PUMA promoter. In addition, the mutation of the p53 binding site in the PUMA promoter region reduced promoter activation by Zn2+. These findings suggest that p53 participates in Zn2+-induced PUMA expression. Furthermore, we also demonstrated here that Zn2+ stimulates the phosphorylation of ERK and that the MEK-ERK pathway inhibitor, U0126, suppressed Zn2+-induced PUMA expression. Taken together, these results indicate that Zn2+ regulates the induction of PUMA through p53 and ERK pathways.


Contrast response in a comprehensive network model of macaque V1.
The response to contrast is one of the most important functions of the macaque primary visual cortex, V1, but up to now there has not been an adequate theory for it. To fill this gap in our understanding of cortical function, we built and analyzed a new large-scale, biologically constrained model of the input layer, 4Cα, of macaque V1. We called the new model CSY2. We challenged CSY2 with a three-parameter family of visual stimuli that varied in contrast, orientation, and spatial frequency. CSY2 accurately simulated experimental data and made many new predictions. It accounted for 1) the shapes of firing-rate-versus-contrast functions, 2) orientation and spatial frequency tuning versus contrast, and 3) the approximate contrast-invariance of cortical activity maps. Post-analysis revealed that the mechanisms that were needed to produce the successful simulations of contrast response included strong recurrent excitation and inhibition that find dynamic equilibria across the cortical surface, dynamic feedback between L6 and L4, and synaptic dynamics like inhibitory synaptic depression.


Recreational participation and the development of social competence in preschool aged children with disabilities: a cross-sectional study.
To explore the association between participation and social competence for preschool aged children with and without disabilities. The sample was drawn from the Longitudinal Study of Australian Children (n = 4983) which included children aged 4-5 years reported to have hearing problems (n = 164), vision problems (n = 146) and other physical disabilities (n = 114). Chi-square, correlation and logistic regression analyses were used to describe the participation and social competence of children with and without these conditions, as well as examine the association between these constructs. Children with disabilities had similar levels of participation but lower social competence than children without these disabilities. Further analyses revealed a small negative correlation between the two variables (ranging from -0.120 to -0.300 for the three groups) and that children who have low participation are more likely to have abnormal levels of social competence than children with higher participation. The association between participation and social competence may not be as strong for this age group as anticipated in the literature, additional factors may be influential. Examination of the social competence scores identified two factors which may assist in explaining the variance in scores: (1) the experience of disability and (2) the quality of interactions. Children with disabilities who participate in similar activities to their typically developing peers may not necessarily develop commensurate levels of social competence. As well as focusing on increasing the participation of children in activities other factors that may have a stronger influence on social competence should be considered, such as supporting (1) the social experience of disability and (2) the quality of interactions that children with disabilities experience.


Remodeling of the mammary microenvironment after lactation promotes breast tumor cell metastasis.
The mammary gland microenvironment during postlactational involution shares similarities with inflammation, including high matrix metalloproteinase activity, fibrillar collagen deposition, and release of bioactive fragments of fibronectin and laminin. Because inflammation can promote tumorigenesis, we evaluated whether the tissue microenvironment of the involuting gland is also promotional. Extracellular matrix was isolated from mammary glands of nulliparous rats or rats with mammary glands undergoing weaning-induced involution. Using these matrices as substratum, nulliparous matrix was found to promote ductal organization of normal mammary epithelial MCF-12A cells in three-dimensional culture and to suppress invasion of mammary tumor MDA-MB-231 cells in transwell filter assays. Conversely, involution matrix failed to support ductal development in normal cells and promoted invasiveness in tumor cells. To evaluate the effects of these matrices on metastasis in vivo, MDA-MB-231 cells, premixed with Matrigel, nulliparous matrix, or involution matrix, were injected into mammary fat pads of nude mice. Metastases to lung, liver, and kidney were increased in the involution matrix group, and correlated with a twofold increase in tumor vascular endothelial growth factor expression and increased angiogenesis. These data suggest that the mammary gland microenvironment becomes promotional for tumor cell dissemination during involution, thus providing a plausible mechanism to explain the high rate of metastases that occur with pregnancy-associated breast cancer.


Possible association between a genetic polymorphism at 8q24 and risk of upper gastrointestinal cancer.
Over recent years, genome-wide association studies have contributed to our understanding of genetic susceptibility to sporadic cancer. In this study, we assessed the association between upper gastrointestinal cancer risk and four genome-wide association studies-identified single nucleotide polymorphisms (SNPs), implicated earlier in prostate and colorectal cancer susceptibility. Genotyping for each SNP was performed in two independent Caucasian population-based case-control studies. The first study comprised 290 gastric cancer cases and 374 controls. The second study included 185 noncardia gastric cancers, 123 cardia cancers, 158 oesophageal cancers and 209 controls. Odds ratios (ORs) were computed from logistic models and adjusted for potential confounding variables. An inverse association was observed between the SNP rs1447295, located at 8q24, and gastric cancer risk in the first study population (OR=0.63; 95% confidence interval: 0.41-0.97). A positive association was observed for the same SNP and oesophageal squamous cell carcinoma in the second study population (OR=7.43; 95% confidence interval: 1.37-49.98). No significant associations were detected in either study for the three remaining SNPs (rs6983297, rs10505477 and rs719725). Our data represent novel findings on heritable susceptibility to gastric and oesophageal cancer and warrant validation in additional populations.


A Case of Refractory Hypoxemic Respiratory Failure due to Antineutrophil Cytoplasmic Antibodies-associated Diffuse Alveolar Hemorrhage Rescued by Extracorporeal Membrane Oxygenation.
Diffuse alveolar hemorrhage (DAH) is a rare but life-threatening disease. Mortality is very high in those patients who require mechanical ventilation. Traditionally, active bleeding has been considered a contraindication for extracorporeal membrane oxygenation (ECMO) support. There is limited evidence for ECMO in DAH as rescue therapy. Herein, we describe a case of antineutrophil cytoplasmic antibodies-associated DAH with intractable hypoxemic respiratory failure. An appropriate ventilator strategy failed to improve her hypoxemia leading to imminent risk to her life. The patient was rescued with veno-venous ECMO targeting lower than usual range of anticoagulation. ECMO proved to be lifesaving in our patient who was initiated on prompt immunosuppressive therapy and plasmapheresis along with continuous veno-venous hemodiafiltration and hemodynamic support. We feel that ECMO could be considered as adjunctive therapy in severe hypoxemic respiratory failure associated with DAH after careful consideration of the risk of bleeding and a restrictive anticoagulation strategy. How to cite this article: Goel MK, Chauhan M, Kumar A, Wadwa P, Maitra G, Talegaonkkar M, et al. A Case of Refractory Hypoxemic Respiratory Failure due to Antineutrophil Cytoplasmic Antibodies-associated Diffuse Alveolar Hemorrhage Rescued by Extracorporeal Membrane Oxygenation. Indian J Crit Care Med 2020;24(9):879-881.


The British Journal of Clinical Pharmacology: The first 50 years.
The British Journal of Clinical Pharmacology celebrates its 50th anniversary of publication in 2023. Here four previous Editors-in-Chief and the current Editor reflect on the Journal's history and the changes that have occurred during that time.


Gum Arabic coated magnetic nanoparticles with affinity ligands specific for antibodies.
A novel magnetic support based on gum Arabic (GA) coated iron oxide magnetic nanoparticles (MNP) has been endowed with affinity properties towards immunoglobulin G (IgG) molecules. The success of the in situ triazine ligand synthesis was confirmed by fluorescence assays. Two synthetic ligands previously developed for binding to IgG, named as ligand 22/8 (artificial Protein A) and ligand 8/7 (artificial Protein L) were immobilized on to MNPs coated with GA (MNP_GA). The dimension of the particles core was not affected by the surface functionalization with GA and triazine ligands. The hydrodynamic diameters of the magnetic supports indicate that the coupling of GA leads to the formation of larger agglomerates of particles with about 1 microm, but the introduction of the triazine ligands leads to a decrease on MNPs size. The non-functionalized MNP_GA bound 28 mg IgG/g, two times less than bare MNP (60 mg IgG/g). MNP_GA modified with ligand 22/8 bound 133 mg IgG/g support, twice higher than the value obtained for ligand 8/7 magnetic adsorbents (65 mg/g). Supports modified with ligand 22/8 were selected to study the adsorption and the elution of IgG. The adsorption of human IgG on this support followed a Langmuir behavior with a Q(máx) of 344 mg IgG/g support and K(a) of 1.5 x 10(5) M. The studies on different elution conditions indicated that although the 0.05 M citrate buffer (pH 3) presented good recovery yields (elution 64% of bound protein), there was occurrence of iron leaching at this acidic pH. Therefore, a potential alternative would be to elute bound protein with a 0.05 M glycine-NaOH (pH 11) buffer.


Acute effect of a cod protein hydrolysate on postprandial acylated ghrelin concentration and sensations associated with appetite in healthy subjects: a double-blind crossover trial.
Fish protein hydrolysates are suggested to contain bioactive sequences capable of affecting metabolic pathways involved in the regulation of glucose metabolism and body weight when consumed in low doses. Modulation of the appetite-regulating hormone ghrelin may explain suppression of insulin secretion and weight loss observed in previous studies with fish protein hydrolysates. This study aimed to assess the effect of a single, low dose of cod protein hydrolysate (CPH) before a breakfast meal on postprandial acylated ghrelin concentration and sensations associated with appetite in healthy subjects. In this explorative trial with a crossover design, 41 healthy individuals (15 males and 26 females, age 51 ± 6 years) completed 2 study days separated by 4-7 days of washout. On both study days, a test drink containing 20 mg CPH or casein (control) per kg body weight was given immediately before a standardized breakfast meal. Acylated ghrelin concentrations were measured before test drink/breakfast (baseline) and at time 0, 20, 40, 80, and 180 min postprandially. Sensations associated with appetite were measured by a Visual Analog Scale (100 mm) at baseline and 0, 20, 40, and 180 min postprandially. Statistically, no difference was observed between CPH and control for postprandial acylated ghrelin concentrations (mean difference geometric mean: 1.05 pg/mL, 95% confidence interval [CI]: 0.97-1.13, P = 0.266), or between the total area under the curve (tAUC) for acylated ghrelin after CPH (tAUC = 17518 pg/mL × min, 95% CI: 0-47941) and control (tAUC = 17272 pg/mL × min, 95% CI: 0-48048, P = 0.991). No differences were found between CPH and control for sensation of appetite, according to tAUC of postprandial scores for satiety (P = 0.794) and the feeling of fullness (P = 0.996). We did not find an effect of a single dose of CPH on postprandial concentrations of acylated ghrelin or sensations related to feeling of hunger, compared to control. Further studies should aim to evaluate the effect of a supplement with CPH given daily over a period of time.


Chemotactic response of periodontal ligament cells decreases with donor age: association with reduced expression of c-fos.
To understand the effects of aging on cellular motility of human periodontal ligament (PDL) cells, and to determine the possible link between cell proliferation and migration in relation to cellular aging. The chemotactic response of PDL cells from three juvenile and four older donors were compared. Then, migrated or unmigrated cells were examined for their cell cycle by morphological and immunocytochemical procedures. Finally, migrated or unmigrated cells were examined for the expression of c-fos and c-myc by in situhybridization. PDL cells from older donors showed lower chemotaxis compared with the cells from juvenile donors (P < 0.05). Cells undergoing migration were found not to be in the S- or M-phase of the cell cycle. However, all migrated cells were found to express c-fos, while many of the cells which did not migrate were found not to express c-fos. Cellular motility of PDL cells decreases with donor age as well as cell proliferation. Since the cells reaching senescence fail to express c-fos, the mechanisms linked to cellular senescence may be a possible underlying mechanism for low migration seen in the older cells.


Identification and Herc5-mediated ISGylation of novel target proteins.
ISG15, a protein containing two ubiquitin-like domains, is an interferon-stimulated gene product that functions in antiviral response and is conjugated to various cellular proteins (ISGylation) upon interferon stimulation. ISGylation occurs via a pathway similar to the pathway for ubiquitination that requires the sequential action of E1/E2/E3: the E1 (UBE1L), E2 (UbcH8), and E3 (Efp/Herc5) enzymes for ISGylation have been hitherto identified. In this study, we identified six novel candidate target proteins for ISGylation by a proteomic approach. Four candidate target proteins were demonstrated to be ISGylated in UBE1L- and UbcH8-dependent manners, and ISGylation of the respective target proteins was stimulated by Herc5. In addition, Herc5 was capable of binding with the respective target proteins. Thus, these results suggest that Herc5 functions as a general E3 ligase for protein ISGylation.