Comparison of calculated with measured oxygen consumption in children undergoing cardiac catheterization.
Our objective was to compare calculated (LaFarge) with measured oxygen consumption (VO(2)) using the AS/3 TM Compact Airway Module M-CAiOVX (Datex-Ohmeda, Helsinki, Finland; AS/3 TM) in children without cardiac shunts in a prospective, observational study. VO(2) was determined at the end of the routine diagnostic and/or interventional catheterization. VO(2 )was calculated according to the formula of LaFarge and Miettinen for each child and compared with the measured VO(2). Data were compared using simple regression and Bland Altman analysis. Fifty-two children aged from 0.5 to 16 years (median, 6.9 years) and weighing 3.4 to 59.4 kg (median, 22.9 kg) were investigated. Calculated VO(2 )values ranged from 59.0 to 230.8 ml/min, and measured VO(2) values from 62.7 to 282.2 ml/min. Comparison of calculated versus measured VO(2) values revealed a significant correlation (r = 0.90, p < 0.0001). Bias and precision were 8.9 and 48.3 ml/min, respectively (95% limits of agreement: -39.4 to 57.2 ml/min). Comparison of calculated VO(2) in children older than 3 years (n = 41), as restricted to the formula, with measured VO(2), revealed a slightly reduced correlation (r = 0.86, p < 0.0001). Bias and precision were 10.0 and 52.5 ml/min, respectively (95% limits of agreement: -42.4 to 62.5 ml/min). We conclude that calculation of VO(2) by the LaFarge formula does not provide reliable values compared to measured values. In clinical routine, measured rather than calculated VO(2) values should be used for the estimation of cardiac output and related variables.


Lumacaftor-Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del CFTR.
Cystic fibrosis is a life-limiting disease that is caused by defective or deficient cystic fibrosis transmembrane conductance regulator (CFTR) protein activity. Phe508del is the most common CFTR mutation. We conducted two phase 3, randomized, double-blind, placebo-controlled studies that were designed to assess the effects of lumacaftor (VX-809), a CFTR corrector, in combination with ivacaftor (VX-770), a CFTR potentiator, in patients 12 years of age or older who had cystic fibrosis and were homozygous for the Phe508del CFTR mutation. In both studies, patients were randomly assigned to receive either lumacaftor (600 mg once daily or 400 mg every 12 hours) in combination with ivacaftor (250 mg every 12 hours) or matched placebo for 24 weeks. The primary end point was the absolute change from baseline in the percentage of predicted forced expiratory volume in 1 second (FEV1) at week 24. A total of 1108 patients underwent randomization and received study drug. The mean baseline FEV1 was 61% of the predicted value. In both studies, there were significant improvements in the primary end point in both lumacaftor-ivacaftor dose groups; the difference between active treatment and placebo with respect to the mean absolute improvement in the percentage of predicted FEV1 ranged from 2.6 to 4.0 percentage points (P<0.001), which corresponded to a mean relative treatment difference of 4.3 to 6.7% (P<0.001). Pooled analyses showed that the rate of pulmonary exacerbations was 30 to 39% lower in the lumacaftor-ivacaftor groups than in the placebo group; the rate of events leading to hospitalization or the use of intravenous antibiotics was lower in the lumacaftor-ivacaftor groups as well. The incidence of adverse events was generally similar in the lumacaftor-ivacaftor and placebo groups. The rate of discontinuation due to an adverse event was 4.2% among patients who received lumacaftor-ivacaftor versus 1.6% among those who received placebo. These data show that lumacaftor in combination with ivacaftor provided a benefit for patients with cystic fibrosis homozygous for the Phe508del CFTR mutation. (Funded by Vertex Pharmaceuticals and others; TRAFFIC and TRANSPORT ClinicalTrials.gov numbers, NCT01807923 and NCT01807949.).


Antibody Testing in Estimating Past Exposure to Chlamydia trachomatis in the Netherlands Chlamydia Cohort Study.
The asymptomatic course of Chlamydia trachomatis (CT) infections can result in underestimated CT lifetime prevalence. Antibody testing might improve this estimate. We assessed CT antibody positivity and predictive factors thereof in the Netherlands Chlamydia Cohort Study. Women who had ≥1 CT Nucleic Acid Amplification Test (NAAT) in the study (2008-2011) and who provided self-reported information on NAATs were tested for CT major outer membrane protein specific IgG in serum (2016). CT antibody positivity was assessed and predictive factors were identified using multivariable logistic regressions, separately for CT-positive women (≥1 positive NAAT or ≥1 self-reported positive CT test) and CT-negative women (negative by study NAAT and self-report). Of the 3,613 women studied, 833 (23.1%) were CT -positive. Among the CT-negative women, 208 (7.5%, 95%CI 6.5-8.5) tested positive for CT antibodies. This increased CT lifetime prevalence with 5.8% (95%CI 5.0-6.5). Among women with a CT-positive history, 338 (40.6%, 95%CI 38.5-44.1) tested positive. Predictive factors for antibody positivity related to lower social economic status, sexual risk behavior, multiple infections, higher body mass index, and non-smoking. CT antibody testing significantly increased the lifetime prevalence. Combining NAAT outcomes, self-reported positive tests, and antibody testing reduced misclassification in CT prevalence estimates.


Shoulder arthroplasty in osteoarthritis: current concepts in biomechanics and surgical technique.
Shoulder arthroplasty is a technically demanding procedure to restore shoulder function in patients with severe osteoarthritis of the glenohumeral joint. The modern prosthetic system exploit the benefits of modularity and the availibility of additional sizes of the prosthetic components. In this paper we describe the biomechanics of shoulder arthroplasty and the technique for shoulder replacement including total shoulder arthroplasty (TSA) with all-polyethylene and metal-backed glenoid component, humeral head resurfacing and stemless humeral replacement.


Recent advances in the stereoselective synthesis of acyclic all-carbon tetrasubstituted alkenes.
Alkenes bearing four carbon-based groups are ubiquitous motifs in chemical sciences due to their various applications from medicinal to materials chemistry, and as chemical platforms for the synthesis of complex, chiral molecules. As such, tremendous research efforts are currently ongoing in order to develop general procedures for the challenging stereoselective synthesis of all-carbon tetrasubstituted alkenes, especially for acyclic structures. Since classical approaches to carbon-carbon double bonds are not suitable for the high steric demand around tetrasubstituted alkenes, a variety of unique approaches to access these privileged functional groups have been developed in recent years. This review article highlights the most significant developments in the field from 2007 to 2020, with an emphasis on the mechanisms and remaining limitations of these contemporary methods. Specifically, recent advances in internal alkyne carbofunctionalizations, in multicomponent couplings or other cross-couplings from nucleophilic or electrophilic alkenyl partners, and in the development of miscellaneous methods, are discussed.


The mechanism of islet amyloid polypeptide toxicity is membrane disruption by intermediate-sized toxic amyloid particles.
NIDDM is characterized by islet amyloid deposits and decreased beta-cell mass. Islet amyloid is derived from the locally expressed protein islet amyloid polypeptide (IAPP). While it is now widely accepted that abnormal aggregation of IAPP has a role in beta-cell death in NIDDM, the mechanism remains unknown. We hypothesized that small IAPP aggregates, rather than mature large amyloid deposits, are cytotoxic. Consistent with this hypothesis, freshly dissolved human (h)-IAPP was cytotoxic when added to dispersed mouse and human islet cells, provoking the formation of abnormal vesicle-like membrane structures in association with vacuolization and cell death. Human islet cell death occurred by both apoptosis and necrosis, predominantly between 24 and 48 h after exposure to h-IAPP. In contrast, the addition to dispersed islet cells of matured h-IAPP containing large amyloid deposits of organized fibrils was seldom associated with vesicle-like structures or features of cell death, even though the cells were often encased in the larger amyloid deposits. Based on these observations, we hypothesized that h-IAPP cytotoxicity is mediated by membrane damage induced by early h-IAPP aggregates. Consistent with this hypothesis, application of freshly dissolved h-IAPP to voltage-clamped planar bilayer membranes (a cell-free in vitro system) also caused membrane instability manifested as a marked increase in conductance, increased membrane electrical noise, and accelerated membrane breakage, effects that were absent using matured h-IAPP or rat IAPP solutions. Light-scattering techniques showed that membrane toxicity corresponded to h-IAPP aggregates containing approximately 25-6,000 IAPP molecules, an intermediate-sized amyloid particle that we term intermediate-sized toxic amyloid particles (ISTAPs). We conclude that freshly dissolved h-IAPP is cytotoxic and that this cytotoxicity is mediated through an interaction of ISTAPs with cellular membranes. Once ISTAPs mature into amyloid deposits comprising >10(6) molecules, the capacity of h-IAPP to cause membrane instability and islet cell death is significantly reduced or abolished. These data may have implications for the mechanism of cell death in other diseases characterized by local amyloid formation (such as Alzheimer's disease).


Review of extralabel use of isoxazolines for treatment of demodicosis in dogs and cats.
Amitraz is presently the only FDA-approved treatment for demodicosis in dogs in the United States. Amitraz treatment involves a protracted course of administration and risks of severe adverse effects such as sedation, bradycardia, and respiratory depression, which are caused by activation of α2-adrenergic receptors. Other treatment options include macrocyclic lactones and lime sulfur, but these products have varied efficacy and high risks of adverse effects. Several recent studies have indicated that isoxazolines are capable of reducing Demodex mite counts in canine and feline patients with demodicosis by ≥ 99% in as little as 1 month with few adverse effects. This article reviews the status of isoxazolines in regard to labeled uses in dogs and cats in the United States, extralabel clinical use for treatment of demodicosis in these species, and safety of orally administered formulations of these drugs.


[Contribution of brain function analysis to the evolution of neurorehabilitation].
Recent studies of functional neuroimaging and clinical neurophysiology have implied that functional recovery after stroke is associated with use-dependent plasticity of the damaged brain. However the property of the reorganized neural network depends on site and size of the lesion, which makes it difficult to assess what the adaptive plasticity is. From clinical point of view there is accumulating randomized controlled trials for the benefit of task-oriented rehabilitative intervention including constraint-induced movement therapy, robotics, and body-weight supported treadmill training. However dose-matched control intervention is usually as effective as a specific intervention. This raises a question regarding the specificity of a task-oriented intervention. Second question is whether such intervention goes beyond the biological destiny of human. Specifically there is no known strategy enhancing recovery of severely impaired hand. To augment functional gain, several methods of neuro-modulation may bring break-through on the assumption that they induce greater adaptive plasticity. Such neuro-modulative methods include neuropharmacological modulation, brain stimulation using transcranial magnetic stimulation and direct current stimulation, peripheral nerve stimulation, neurofeedback using real-time fMRI and real-time fNIRS, and brain-machine interface. A preliminary randomized controlled trial regarding real-time feedback of premotor activities revealed promising results for recovery of paretic hand in patients with stroke.


A global multi-hazard risk analysis of road and railway infrastructure assets.
Transport infrastructure is exposed to natural hazards all around the world. Here we present the first global estimates of multi-hazard exposure and risk to road and rail infrastructure. Results reveal that ~27% of all global road and railway assets are exposed to at least one hazard and ~7.5% of all assets are exposed to a 1/100 year flood event. Global Expected Annual Damages (EAD) due to direct damage to road and railway assets range from 3.1 to 22 billion US dollars, of which ~73% is caused by surface and river flooding. Global EAD are small relative to global GDP (~0.02%). However, in some countries EAD reach 0.5 to 1% of GDP annually, which is the same order of magnitude as national transport infrastructure budgets. A cost-benefit analysis suggests that increasing flood protection would have positive returns on ~60% of roads exposed to a 1/100 year flood event.


Population-based assessment of gastrointestinal symptoms and diseases: Cappadocia Cohort, Turkey.
The aim of this study was to determine the prevalence of symptoms and diseases of the lower and upper gastrointestinal system (GIS) in a population-based sample. The cross-sectional cohort study was conducted in Cappadocia cohort comprising the Gülşehir and Avanos districts. The "Gastrointestinal Symptom Questionnaire" was applied to persons over the age of 18 years. The GI Symptom Questionnaire was applied to 3369 subjects, and height and body weight were measured in 2797 consenting subjects. Of the participants, 61% were female and the mean patient age was 50±15 years. At least one GI symptom was present in 70.6% of the cohort. The most common upper GI symptoms were gastric bloating (31.0%) and heartburn (29.1%). The most common lower GI symptom was abnormal defecation (33.5). The prevalence of upper GIS and lower GIS diseases was 32.7% and 12.9%, respectively, and the prevalence of togetherness of upper and lower GIS diseases was 9.9%. Prevalence of GIS disease was approximately 3 times higher in females (p<0.001). All of the upper and lower GI symptoms and the prevalence of upper GIS disease increased in line with Body mass index (BMI). This first population-based, cross-sectional cohort study revealed that the prevalence of GIS diseases is critically high for optimal public health. Special attention must be paid to these diseases while planning health policies and reimbursements.


Neutron scanning reveals unexpected complexity in the enamel thickness of an herbivorous Jurassic reptile.
Eilenodontines are one of the oldest radiation of herbivorous lepidosaurs (snakes, lizards and tuatara) characterized by batteries of wide teeth with thick enamel that bear mammal-like wear facets. Unlike most reptiles, eilenodontines have limited tooth replacement, making dental longevity particularly important to them. We use both X-ray and neutron computed tomography to examine a fossil tooth from the eilenodontine Eilenodon (Late Jurassic, USA). Of the two approaches, neutron tomography was more successful and facilitated measurements of enamel thickness and distribution. We find the enamel thickness to be regionally variable, thin near the cusp tip (0.10 mm) but thicker around the base (0.15-0.30 mm) and notably greater than that of other rhynchocephalians such as the extant Sphenodon (0.08-0.14 mm). The thick enamel in Eilenodon would permit greater loading, extend tooth lifespan and facilitate the establishment of wear facets that have sharp edges for orally processing plant material such as horsetails (Equisetum). The shape of the enamel dentine junction indicates that tooth development in Eilenodon and Sphenodon involved similar folding of the epithelium but different ameloblast activity.


A Review of the New Antiepileptic Drugs for Focal-Onset Seizures in Pediatrics: Role of Extrapolation.
Most antiepileptic drugs (AEDs) receive regulatory approval for children years after the drug is available in adults, encouraging off-label use of the drug in children and hindering attempts to obtain quality pediatric data in controlled trials. Extrapolating adult efficacy data to pediatrics can reduce the time between approval in adults and that in children. To extrapolate efficacy from adults to children, several assumptions must be supported, such as (1) a similar disease progression and response to interventions in adults and children, and (2) similar exposure response in adults and children. The Pediatric Epilepsy Academic Consortium for Extrapolation (PEACE) addressed these assumptions in focal-onset seizures (FOS), the most common seizure type in both adults and children. PEACE reviewed the biological and clinical evidence that supported the assumptions that children with FOS have a similar disease progression and response to intervention as adults with FOS. After age 2 years, the pathophysiological underpinnings of FOS and the biological milieu in which seizures are initiated and propagated in children, seizure semiology, electroencephalographic features, etiology and AED response to FOS in children are similar to those in adults with FOS. PEACE concluded that extrapolation of efficacy data in adults to pediatrics in FOS is supported by strong scientific and clinical evidence. However, safety and pharmacokinetic (PK) data cannot be extrapolated from adults to children. Based on extrapolation, eslicarbazepine is now approved for children with FOS, down to age 4 years. Perampanel, lacosamide and brivaracetam are now undergoing PK and safety studies for the purposes of extrapolation down to age 2 or 4 years. When done in conjunction with PK and safety investigations in children, extrapolation of adult data from adults to children can reduce the time delay between approval of effective and safe AEDs in adults and approval in children.


How Many Older Informal Caregivers Are There in Europe? Comparison of Estimates of Their Prevalence from Three European Surveys.
Informal caregivers are people providing some type of unpaid, ongoing assistance to a person with a chronic illness or disability. Long-term care measures and policies cannot take place without taking into account the quantitatively crucial role played by informal caregivers. We use the European Health Interview Survey (EHIS), the European Quality of Life Survey (EQLS), and the Study on Health and Ageing in Europe (SHARE) to measure the prevalence of informal caregivers in the European population, and analyze associated socio-demographic factors. This rate ranges between about 13 percent in Portugal and Spain, and more than 22 percent in Luxembourg, Belgium, and Denmark. It declines in older age groups and, on average, is lower in men than in women in all countries studied, and lower among the poorly educated compared to those with higher levels of education. However, large variance was observed in the average share of informal caregivers for most countries between the three surveys. Our findings, estimated through the three surveys, reveal common trends, but also a series of disparities. Additional research will be needed to enable policy makers to access a richer and more harmonized body of data, allowing them to adopt truly evidence-based and targeted policies and interventions in this field.


Evaluating congenital spine deformities for intraspinal anomalies with magnetic resonance imaging.
The incidence of intraspinal abnormalities associated with congenital spinal anomalies as detected by magnetic resonance imaging (MRI) is becoming better defined. In this study, 41 nonrandomized children with congenital spinal deformities (excluding myelomeningocele) who underwent complete MR evaluation were reviewed. Of the 41 congenital spinal deformities, 37 demonstrated congenital scoliosis, with failure of formation in 19, failure of segmentation in 4, and mixed defects in 14. The remaining four deformities were cases of congenital kyphosis. Thirteen patients with congenital spine anomalies were noted to have intraspinal abnormalities identified by MRI: tethered cord in 12 patients, syringomyelia in 3 patients, and diastematomyelia in 5 patients. Of the 12 patients with tethered cord, 2 patients had neurologic deficits. Urorectal anomaly was one of the most common associated findings (15%). Considering an incidence of intraspinal anomalies of 31% and as clinical manifestations may not be initially detectable, MRI is recommended in patients with congenital spinal deformity as part of the initial evaluation even in the absence of clinical findings.


Exercise-induced changes in renal function and their relation to plasma noradrenaline in insulin-dependent diabetic children and adolescents.
Exercise-induced changes in renal haemodynamics, water homoeostasis, urinary albumin excretion and their possible relation to plasma noradrenaline were studied in seven insulin-dependent diabetic children and adolescents without signs of autonomic or somatic neuropathy. Six age-matched clinically healthy patients in remission from the minimal change nephrotic syndrome served as controls. The diabetic patients had a higher glomerular filtration rate and effective renal plasma flow and a lower venous plasma noradrenaline level in the basal state. They also had a higher systolic blood pressure compared with the controls. During exercise the diabetic patients showed a less marked reduction in glomerular filtration rate and urinary flow and a greater increase in filtration fraction. Their heart rate increase during exercise was lower. An inverse correlation between the percentage changes in noradrenaline and glomerular filtration rate during exercise was noted solely in the diabetic patients. They also showed a less marked increase in the ratio between potassium and sodium clearances during and 20 min after exercise. Albumin excretion did not differ between the two groups. We suggest that the sympatho-adrenergic effects of exercise on heart rate, renal haemodynamics and water homoeostasis are blunted in our diabetic patients. Hence, signs of impaired sympathetic activity might be disclosed in young diabetic patients with a disease of fairly short duration.


Validation of new microvolume Couette flow linear dichroism cells.
Long molecules such as fibrous proteins are particularly difficult to characterise structurally. We have recently designed a microvolume Couette flow linear dichroism (LD) cell whose sample volume is only 20-40 microL in contrast to previous cells where the volume of sample required has typically been of the order of 1000-2000 microL. This brings the sample requirements of LD to a level where it can be used for biological samples. Since LD is the difference in absorption of light polarised parallel to an orientation direction and perpendicular to that direction, it is the ideal technique for determining relative orientations of subunits of e.g. fibrous proteins, DNA-drug systems, etc. For solution phase samples, Couette flow orientation, whereby the sample is sandwiched between two cylinders, one of which rotates, has proved to be the optimal technique for LD experiments in many laboratories. Our capillary microvolume LD cell has been designed using extruded quartz rods and capillaries and focusing and collecting lenses. We have developed applications with PCR products, fibrous proteins, liposome-bound membrane proteins, as well as DNA-dye systems. Despite this range of applications, to date there is nothing reported in the literature to enable one to validate the performance of Couette flow LD cells. In this paper we establish validation criteria and show that the data from the microvolume cells are reproducible, vary by less than 1% with sample reloading, follow the Beer-Lambert law, and have signals linear in voltage over a wide voltage range. The microvolume cell data are consistent with those from the large-volume cells for DNA samples. Surprisingly, upon extending the wavelength range by adding the intercalator ethidium bromide, the spectra in the microvolume and large-volume cells differ by a wavelength dependent orientation parameter. This wavelength variation was concluded to be the result of Taylor-vortices in the large-volume cells which have inner rotating cylinders in our laboratory. Thus the microvolume LD cells can be concluded to provide better data than our large-volume LD cells, though the latter are still to be preferred for titration series as it is extremely difficult to add sample to the capillary cells without introducing artefacts.


The therapeutic effect of tissue cultured root of wild Panax ginseng C.A. Mayer on spermatogenetic disorder.
This study examined the possibility of using a tissue cultured root of wild Panax ginseng (tcwPG) as a fertility agent. The effect of tcwPG on spermatogenesis was studied using male rats. The tcwPG crude powder was administered orally to 7-week-old rats over a 6-week period. The number of sperm in the testes and epididymides was significantly higher than the control. A histological examination did not reveal any morphological changes in the testes from the tcwPG powder treated rats. Moreover, there were no significant differences in the weights of the heart, spleen, liver, kidney, brain, testes and epididymides. Oligospermia was also induced by administering 2,3,7,8-tetrachlorodaibenzo-p-dioxin (TCDD) to the rats in order to estimate the feasibility of using tcwPG as treatment for infertility caused by spermatogenic disorders. After exposing the rats to TCDD, the tcwPG saponin fraction treated rats showed some improvement in the body weight, sperm number and testis morphology. It was estimated that tcwPG had feasibility as a therapeutic agent on spermatogenic disorder.


Pattern of congestive heart failure in a Kenyan paediatric population.
Heart failure in children is a common cause of morbidity and mortality, with high socio-economic burden. Its pattern varies between countries but reports from Africa are few. The data are important to inform management and prevention strategies. To describe the pattern of congestive heart failure in a Kenyan paediatric population. This was a retrospective study done at Kenyatta National Hospital, Nairobi Kenya. Records of patients aged 12 years and younger admitted with a diagnosis of heart failure between January 2006 and December 2010 were examined for mode of diagnosis, age, gender, cause, treatment and outcome. Data were analysed using the Statistical Programme for Social Scientists version 16.0 for windows, and presented in tables, bar and pie charts. One hundred and fifty-eight cases (91 male, 67 female) patients' records were analysed. The mean age was 4.7 years, with a peak at 1-3 years. The male: female ratio was 1.4:1. All the cases were in New York Heart Association (NYHA) class II-IV. Evaluation of infants was based on the classification proposed by Ross et al. (1992). Diagnosis was made based on symptoms and signs combined with echocardiography (echo) and electrocardiography (ECG) (38%); echo alone (12.7%); ECG, echo and chest X-ray (CXR) (11.4%); and ECG alone (10.8%). The underlying cause was established on the basis of symptoms, signs, blood tests, CXR, echo and ECG results. Common causes were infection (22.8%), anaemia (17.1%), rheumatic heart disease (14.6%), congenital heart disease (13.3%), cardiomyopathy (7.6%), tuberculosis and human immunodeficiency virus (6.9% each); 77.9% of patients recovered, 13.9% after successful surgery, and 7.6% died. Congestive heart failure is not uncommon in the Kenyan paediatric population. It occurs mainly before five years of age, and affects boys more than girls. The majority are due to infection, anaemia, and rheumatic and congenital heart diseases. This differs from those in developed countries, where congenital heart disease and cardiomyopathy predominate. The majority of children usually recover. Prudent control of infection and correction of anaemia are recommended.


Making Champs and Super-Champs-Current Views, Contradictions, and Future Directions.
In our 2016 paper (Collins et al., 2016a), we proposed that superchamps (athletes who have achieved the highest level in their sport) were differentiated from their less successful counterparts by their use of positive proactive coping and a "learn from it" approach to challenge. This skill-based focus to talent development (TD) is supported extensively in the literature (e.g., MacNamara et al., 2010a,b) and suggests that the differences between levels of adult achievement relate more to what performers bring to the challenges than what they experience (Collins et al., 2016a). In this focused review we present and discuss a number of key concepts related to this paper and other parallel research in TD. We begin by presenting our pragmatic objectives and the importance of considering how we evaluate the research with an emphasis on its application to the applied setting. We then consider commonalities and differences in recent psychological approaches to TD, namely the experiential, attitudinal, and skill-based. The paper then provides further exploration of the Psychological Characteristics of Developing Excellence and their role in TD processes. We conclude with a consideration of future research and the application of research in TD. Reflecting our pragmatic stance as researchers, we hope that this focused review provides suggestions for progress and stimulates critical debate amongst practitioners, researchers and policy makers.


What Is the Association of Elite Sporting Activities With the Development of Hip Osteoarthritis?
Male athletes participating in certain elite sporting activities may be at an increased risk for development of hip osteoarthritis (OA) later in life. However, the strength of the association of participation in sporting activities with the increased risk of hip OA has not been well summarized. To investigate the association of certain high-impact sporting activities with the risk of development of hip OA in elite athletes by conducting a systematic review of the available literature. Systematic review; Level of evidence, 3. PubMed, Cochrane, and EMBASE databases were searched to identify all potential studies. Eleven studies met the inclusion criteria, which included participation in elite-level sporting activities, greater than 50% male athletes in the study population, diagnosis of hip OA by radiograph, hospital admission or total hip arthroplasty (THA), and greater than 80% follow-up. Exclusion criteria were recreational sporting activities, primarily female cohorts as there was a dearth of available literature on the topic, and self-reported symptoms without radiographic confirmation of diagnosis. Most studies were with European athletes, where elite-level was defined as involvement in national- or professional-level competition. Participation across elite-level impact sports was associated with increased risk of development of hip OA when compared with matched controls (odds ratio, 1.8-8.7). Twelve of 15 studies reviewed demonstrated an odds ratio of 1.8 or greater of developing hip OA in elite-level athletes. Handball was associated with the highest rate of OA of any sport, nearly 5 times that of matched controls. Soccer players demonstrated between 2 and 9 times increased risk of hip OA as defined by radiography or THA. Hockey players demonstrated 2 to 3 times increased risk of hip OA (THA or hospital admission). Five studies investigating the association of competitive long-distance running with hip OA demonstrated inconsistent results. Currently available literature suggests that male athletes participating in elite impact sports (soccer, handball, track and field, or hockey) are at an increased risk of developing hip OA, while those participating in high-level long-distance running do not have a clearly elevated risk. Further research is warranted to elucidate the pathomechanics of development of hip OA in these patients.


Determinants of adherence to exercise in women treated for breast cancer.
We describe adherence to a three group randomised controlled trial that involved aerobic exercise therapy (N=34) and exercise-placebo intervention (N=36) or usual care (N=38) in women treated for breast cancer. We also investigated relationships between routes of trial recruitment, socio-economic characteristics, health behaviours, cancer treatment regimen(s) and subsequent adherence to the interventions. Women who had completed breast cancer treatment 12-36 months previously were randomised to one of the groups. The intervention groups attended an exercise facility three times per week for eight weeks. A total of 77% of the aerobic exercise therapy and 88.9% of the exercise-placebo groups attended at least 70% of prescribed sessions. The percentage of women achieving 30min of aerobic exercise per session steadily increased during the intervention but many women were not able to achieve this goal. Routes of trial recruitment, socio-economic characteristics, health behaviours and cancer treatment regimen(s) were not related to intervention. Exercise therapy participants were able to comply with the progressive nature of the aerobic exercise intervention, although some women were not able to achieve 30min of aerobic exercise per session by intervention completion. Research is still required to tease out all the potential determinants of exercise in this population.


The next generation of epidermal growth factor receptor tyrosine kinase inhibitors in the treatment of lung cancer.
The discovery of "driver" genomic alterations in patients with non-small cell lung cancer (NSCLC) has dramatically changed the field of thoracic oncology in recent years. The best understood of these molecular drivers are those involving the epidermal growth factor receptor (EGFR), which when aberrantly activated are integral to the development of a subset of NSCLC tumors. First-generation and second-generation tyrosine kinase inhibitors (TKIs) specific to the activated EGFR have shown significant efficacy and have brought about the era of targeted therapy for NSCLC. The most common resistance mechanism is a threonine-to-methionine substitution (T790M) in exon 20 of the EGFR gene. Although the previous standard of care in patients with EGFR-mutated NSCLC that progressed on initial TKI therapy was chemotherapy, third-generation EGFR TKIs have now been developed and have yielded promising results for this population of patients with NSCLC. This article reviews the emerging data regarding third-generation agents in the treatment of patients with advanced NSCLC.


Clinical management and humoral immune responses to rabies post-exposure prophylaxis among three patients who received solid organs from a donor with rabies.
The rabies virus causes a fatal encephalitis and can be transmitted through organ transplantation. In 2013, a man developed rabies 18 months after receiving a kidney from a donor with rabies, who was not known to have been infected when the organs were procured. Three additional persons who received organs from the same donor (liver, kidney, heart), all of whom were not vaccinated for rabies before transplantation, received rabies post-exposure prophylaxis (PEP) with rabies immune globulin and 5 doses of rabies vaccine as soon as the diagnosis of rabies was made in the donor (18 months after their transplant surgeries). We describe their clinical management. As the 3 recipients were all on immunosuppressive medications, post-vaccination serologic testing was performed using the rapid fluorescent focus inhibition test to measure rabies virus neutralizing antibodies (RVNAs). An acceptable antibody response to administration of rabies vaccine was defined as detection of RVNAs at a concentration ≥0.1 IU/mL from a serum specimen collected ≥7 days after the fifth vaccine dose. All 3 recipients demonstrated an acceptable antibody response despite their immunosuppressed states. More than 36 months have passed since their transplant surgeries, and all 3 recipients have no evidence of rabies. The survival of 3 previously unvaccinated recipients of solid organs from a donor with rabies is unexpected. Although the precise factors that led to their survival remain unclear, our data suggest that PEP can possibly enhance transplant safety in settings in which donors are retrospectively diagnosed with rabies.


Editorial: Global action to reduce HIV stigma and discrimination.
There is no question that the stigma and discrimination associated with HIV and AIDS can be reduced through intervention. The inclusion of stigma and discrimination reduction as a critical component of achieving an AIDS-free generation in recent UNAIDS, UN and PEPFAR political initiatives is promising. Yet national governments need evidence on effective interventions at the individual, community and societal levels in order to strategically incorporate stigma and discrimination reduction into national AIDS plans. Currently, the heterogeneity of stigma and discrimination reduction approaches and measurement makes it challenging to compare and contrast evaluated interventions. Moving forward, it is critical for the research community to: (1) clearly link intervention activities to the domains of stigma to be shifted; (2) assess the stigma domains in a consistent manner; and (3) link stigma and discrimination reduction with HIV prevention, care and treatment outcomes (e.g., uptake, adherence and retention of ART). These steps would further advance the scientific evidence base of stigma and discrimination reduction and allow for the identification of effective interventions that could be scaled up by national governments.


Exact and approximate graph matching using random walks.
In this paper, we propose a general framework for graph matching which is suitable for different problems of pattern recognition. The pattern representation we assume is at the same time highly structured, like for classic syntactic and structural approaches, and of subsymbolic nature with real-valued features, like for connectionist and statistic approaches. We show that random walk based models, inspired by Google's PageRank, give rise to a spectral theory that nicely enhances the graph topological features at node level. As a straightforward consequence, we derive a polynomial algorithm for the classic graph isomorphism problem, under the restriction of dealing with Markovian spectrally distinguishable graphs (MSD), a class of graphs that does not seem to be easily reducible to others proposed in the literature. The experimental results that we found on different test-beds of the TC-15 graph database show that the defined MSD class "almost always" covers the database, and that the proposed algorithm is significantly more efficient than top scoring VF algorithm on the same data. Most interestingly, the proposed approach is very well-suited for dealing with partial and approximate graph matching problems, derived for instance from image retrieval tasks. We consider the objects of the COIL-100 visual collection and provide a graph-based representation, whose node's labels contain appropriate visual features. We show that the adoption of classic bipartite graph matching algorithms offers a straightforward generalization of the algorithm given for graph isomorphism and, finally, we report very promising experimental results on the COIL-100 visual collection.


A model for a G-protein-mediated mechanism for synaptic channel modulation.
Neurons communicate with other neurons via specialized structures called synapses, at which the digital voltage signal encoded in an action potential is converted into an analog chemical signal. An action potential that arrives at the presynaptic face triggers release of neurotransmitter from vesicles in a calcium-dependent manner, and the neurotransmitter diffuses across the synaptic cleft and binds to receptors on the post-synaptic face, where it may trigger a postsynaptic action potential. Calcium is a critical component of the release process, and its spatio-temporal dynamics can control the release and can lead to facilitation or augmentation. However, how cells regulate cytoplasmic calcium so that exocytosis can be triggered successfully is still not completely understood. We propose a mechanism, based upon the experimental findings of Barrett and Rittenhouse [C.F. Barrett, A.R. Rittenhouse, Modulation of N-type calcium channel activity by G-proteins and protein kinase C, J. Gen. Physiol. 115 (3) (2000) 277], for the regulation of calcium influx through N-type channels in the presynaptic terminal by PKC and downstream effectors of G-protein activation. This proposed modulatory mechanism consists of a feedback loop involving cytoplasmic calcium, neurotransmitters and G-protein-coupled receptors. We study the dynamics of each component separately and then we address how kinetic properties of the components and the frequency of the stimuli affect the regulatory mechanisms presented here.


Accuracy of dual-source CT coronary angiography: First experience in a high pre-test probability population without heart rate control.
The aim of this study was to assess the diagnostic accuracy of dual-source computed tomography (DSCT) for evaluation of coronary artery disease (CAD) in a population with extensive coronary calcifications without heart rate control. Thirty patients (24 male, 6 female, mean age 63.1+/-11.3 years) with a high pre-test probability of CAD underwent DSCT coronary angiography and invasive coronary angiography (ICA) within 14+/-9 days. No beta-blockers were administered prior to the scan. Two readers independently assessed image quality of all coronary segments with a diameter > or =1.5 mm using a four-point score (1: excellent to 4: not assessable) and qualitatively assessed significant stenoses as narrowing of the luminal diameter >50%. Causes of false-positive (FP) and false-negative (FN) ratings were assigned to calcifications or motion artifacts. ICA was considered the standard of reference. Mean body mass index was 28.3+/-3.9 kg/m2 (range 22.4-36.3 kg/m2), mean heart rate during CT was 70.3+/-14.2 bpm (range 47-102 bpm), and mean Agatston score was 821+/-904 (range 0-3,110). Image quality was diagnostic (scores 1-3) in 98.6% (414/420) of segments (mean image quality score 1.68+/-0.75); six segments in three patients were considered not assessable (1.4%). DSCT correctly identified 54 of 56 significant coronary stenoses. Severe calcifications accounted for false ratings in nine segments (eight FP/one FN) and motion artifacts in two segments (one FP/one FN). Overall sensitivity, specificity, positive and negative predictive value for evaluating CAD were 96.4, 97.5, 85.7, and 99.4%, respectively. First experience indicates that DSCT coronary angiography provides high diagnostic accuracy for assessment of CAD in a high pre-test probability population with extensive coronary calcifications and without heart rate control.


Power output in traditional and ballistic bench press in elite athletes: Influence of training background.
This study aimed to compare the power production in traditional bench-press (TBP) and ballistic bench-throw (BBT) exercises. Furthermore, we assessed the differences in velocity, force, and power outputs between TBP and BBT. Finally, we tested the differences between the loads used to optimize power (optimum power load; OPL) in both exercises, using three distinct power-variables: mean power (MP), mean propulsive power (MPP), and peak power (PP). Sixty athletes from different sports were divided into two groups, according to their training characteristics: hypertrophy-based trained athletes (HTA), thirty-one athletes performing hypertrophy training programmes for (at least) 12-weeks; and power-based trained athletes (PTA), twenty-nine athletes performing power-oriented training sessions for (at least) 12-weeks. Magnitude-based inferences were used to test for differences between groups. Independent of the variable analyzed (MP, MPP, or PP), the PTA produced greater power values in BBT, whereas the HTA generated higher outputs during TBP. The OPL in the HTA was likely heavier in TBP than in BBT, whereas no differences related to this variable were found in the PTA. Despite the apparent superiority of ballistics to produce power, it seems that in elite athletes, the strength-power training routine might affect the ability to apply high forces at very-high velocities.


The effect of amblyopia therapy on ocular alignment.
We sought to describe the change in ocular alignment at 2 years after treatment of amblyopia in children younger than 7 years of age at enrollment. A randomized clinical trial of patching versus atropine for 6 months followed by standard clinical care for 18 months was conducted in 357 children with anisometropic, strabismic, or combined amblyopia (20/40-20/100) whose ages ranged from 3 to younger than 7 years at enrollment. Ocular alignment was evaluated at enrollment and after 2 years of follow-up. At enrollment when tested at distance fixation, 161 (45%) children were orthotropic, 91 (25%) had a microtropia (1-8 Delta), and 105 (29%) had a heterotropia >8 Delta. Of the 161 patients with no strabismus, similar proportions of patients initially assigned to the patching and atropine groups developed new strabismus by 2 years (18% vs. 16%, P = 0.84). Of these cases of new strabismus, only 2 patients in the patching group and 3 patients in the atropine group developed a deviation that was greater than 8 Delta. Microtropia at enrollment progressed to a deviation greater than 8 Delta with similar frequency in both treatment groups (13% vs. 15%, P = 1.00). Of the 105 patients with strabismus greater than 8 Delta at enrollment, 13% of those in the patching group and 16% of those in the atropine group improved to orthotropia without strabismus surgery. Strabismus surgery was performed in 32 patients during the 2-year study period. Patients who had amblyopia treatment with patching or atropine for 6 months followed by standard clinical care were found to have similar rates of deterioration and improvement of ocular alignment. When parents begin amblyopia treatment for children without strabismus, they should be warned of the possibility of development of strabismus, although it is most often a small angle deviation. Strabismus resolved after amblyopia therapy in some cases.


Reducing hospital admissions from nursing homes: a systematic review.
The geriatric nursing home population is vulnerable to acute and deteriorating illness due to advanced age, multiple chronic illnesses and high levels of dependency. Although the detriments of hospitalising the frail and old are widely recognised, hospital admissions from nursing homes remain common. Little is known about what alternatives exist to prevent and reduce hospital admissions from this setting. The objective of this study, therefore, is to summarise the effects of interventions to reduce acute hospitalisations from nursing homes. A systematic literature search was performed in Cochrane Library, PubMed, MEDLINE, EMBASE and ISI Web of Science in April 2013. Studies were eligible if they had a geriatric nursing home study population and were evaluating any type of intervention aiming at reducing acute hospital admission. Systematic reviews, randomised controlled trials, quasi randomised controlled trials, controlled before-after studies and interrupted time series were eligible study designs. The process of selecting studies, assessing them, extracting data and grading the total evidence was done by two researchers individually, with any disagreement solved by a third. We made use of meta-analyses from included systematic reviews, the remaining synthesis is descriptive. Based on the type of intervention, the included studies were categorised in: 1) Interventions to structure and standardise clinical practice, 2) Geriatric specialist services and 3) Influenza vaccination. Five systematic reviews and five primary studies were included, evaluating a total of 11 different interventions. Fewer hospital admissions were found in four out of seven evaluations of structuring and standardising clinical practice; in both evaluations of geriatric specialist services, and in influenza vaccination of residents. The quality of the evidence for all comparisons was of low or very low quality, using the GRADE approach. Overall, eleven interventions to reduce hospital admissions from nursing homes were identified. None of them were tested more than once and the quality of the evidence was low for every comparison. Still, several interventions had effects on reducing hospital admissions and may represent important aspects of nursing home care to reduce hospital admissions.


Achondroplasia in Turkey is defined by recurrent G380R mutation of the FGFR3 gene.
Achondroplasia, the most common form of skeletal dysplasia in man, has autosomal dominant inheritance and causes severe dwarfism. More than 90% of patients with achondroplasia have a G to A transversion or G to C transversion at position 1138 of the fibroblast growth factor receptor-3 (FGFR3) gene resulting in the substitution of an arginine for a glycine residue at position 380 (G380R) of the FGFR3 protein. In this study, 12 unrelated Turkish patients with achondroplasia were evaluated for the G to A and G to C transversion at position 1138 of the FGFR3 gene. Eleven of 12 patients carried the G to A mutation heterozygously. None of the patients had the G to C mutation at the same position. In conclusion, the vast majority of Turkish achondroplasia patients have the same mutation that has been most often defined in patients with achondroplasia from other countries. Our results give further support to the fact that the G380R mutation of FGFR-3 is the most common mutation causing achondroplasia in different populations.


Zebrafish model in drug safety assessment.
Over the past decade, zebrafish are being increasingly used in assessing the effects of chemical compounds. Especially, the embryos and larvae, due to their microscopically small size and optical transparency, are compatible with multi-well microtiter plates for high throughput screening. Being transparent, they allow for non-invasive visualization of internal organs during early development. The organization of the genome, the genetic pathways controlling signal transduction and the developmental pattern appear to be significantly conserved between zebrafish and humans. Major organ systems including the nervous, cardiovascular, digestive and visual systems of zebrafish are also similar to their mammalian counterparts at the anatomical, physiological and molecular levels. Therefore, zebrafish assays are ideal for evaluating multiple organ toxicities simultaneously that contrast in vitro assays performed on cultured cells or tissue explants and organ slices. Although research on zebrafish as a model system began a few decades ago, later studies on zebrafish developmental biology and developmental genetics resulted in the characterization of a large number of genes involved in vertebrate development and biological pathways thus establishing zebrafish as a relevant human disease model for research. Recently, zebrafish have become an attractive vertebrate model for pharmaceutical and toxicological studies. We have outlined in this review some of the toxicological screens and tools that used zebrafish early life stages, and the efforts made to validate zebrafish assays against mammalian drug screens.


Sexual conflict and the function of genitalic claws in guppies (Poecilia reticulata).
Poeciliid fish, freshwater fish with internal fertilization, are known for the diversity of structures on the male intromittent organ, the gonopodium. Prominent among these, in some species, is a pair of claws at its tip. We conducted a manipulative study of these claws in the guppy, Poecilia reticulata, to determine if these aid in transferring sperm to resistant females. We compared the sperm transfer rates of clawed versus surgically declawed males attempting to mate with either receptive or unreceptive (i.e. resistant) females. Our analyses demonstrate that the gonopodial claws function to increase sperm transfer to unreceptive females during uncooperative matings but not during receptive matings. Up to threefold more sperm were transferred to unreceptive females by clawed than declawed males. These data suggest that the claw is a sexually antagonistic trait, functioning to aid in transferring sperm to resistant females, and implicate sexual conflict as a selective force in the diversification of the gonopodium in the Poeciliidae.


A single institution experience with living donor liver transplantation for acute-on-chronic hepatitis B liver failure.
This study reports our preliminary experience of living donor liver transplantation (LDLT) for patients with acute-on-chronic liver failure (AoCLF) caused by hepatitis B. 47 patients who demonstrated Ao- CLF caused by hepatitis B with mean (±SD) Model for End-Stage Liver Disease scores of 39.2±5.1 were divided by the transplantation group (n=19) and the non-transplantation group (n=28) according to whether or not undergoing LDLT. At the same time, 30 hepatitis B cirrhosis recipients who underwent LDLT and did not reach the criteria of AoCLF were selected as the control group (n=30). In the transplantation group, veno-venous bypass, molecular adsorbent recirculating system (MARS) and continuous renal replacement therapy (CRRT) were introduced. The intraoperative data, post-transplant complications and mortality were analyzed retrospectively. There were no significant differences in survival rates of 1, 6 and 12 months and the postoperative complications except for pneumonia and diabetes, between the control group and the transplantation group (p>0.05). Recurrence of hepatitis B was not found in the recipients of the control group and the transplantation group. Right-lobe LDLT may be an effective therapeutic option for patients with acute-on-chronic hepatitis B liver failure.


Transmission of Streptococcus pneumoniae in rural Gambian villages: a longitudinal study.
BACKGROUND. To prepare for national introduction of a pneumococcal vaccine of restricted valency, we studied the pattern of nasopharyngeal carriage of Streptococcus pneumoniae and its transmission in Gambian villages over time. METHODS. We collected nasopharyngeal swab specimens every 2 weeks from 158 villagers in 19 households in 2 villages over 1 year. We studied the prevalence and duration of S. pneumoniae carriage, the effect of household size and composition on carriage, and sequence type-specific carriage within and between households. RESULTS. Ninety-seven percent of children and 85% of adults carried S. pneumoniae at some time. Fifty-three serotypes were represented among 1522 isolates. Carriage was more common among children than adults for all serotypes studied except 9V. There was an overall trend toward shorter carriage with increasing age (P = .043) and significant differences in carriage duration between serotypes. For most serotypes, the odds of being a carrier were greater if there were other carriers in the household. The prevalence of carriage varied by serotype. Most notably, serotype 5 carriage occurred in only 1 village and was transient. Multilocus sequence typing of serotype 6B isolates from 1 village revealed 8 different sequence types and strong evidence of nonrandom distribution among households (P < .001). Study by sequence type suggested household spread starting most commonly in children, followed by spread to adults. CONCLUSIONS. This longitudinal carriage study in Gambian villages provides unique information on the pattern of spread of S. pneumoniae in rural Africa and a baseline for evaluating the impact of the introduction of pneumococcal conjugate vaccine into the region.


Safety and toxicological evaluation of a novel anti-obesity formulation LI85008F in animals.
LI85008F is a novel synergistic composition of Moringa oleifera, Murraya koenigi, and Curcuma longa. These herbs are well recognized and widely used in ayurvedic system of medicine for treating a variety of diseases and are also have been used for culinary purposes for thousands of years. LI85008F inhibits preadipocyte differentiation and potentiates lipid breakdown in mature adipocytes. In diet-induced obese rats, LI85008F significantly reduced weight gain and improved serum adiponectin levels. These findings motivated the authors to determine the broad-spectrum safety of LI85008F. Acute oral toxicity, acute dermal toxicity, primary skin irritation, primary eye irritation, and dose-dependent 28-day sub-acute toxicity studies were conducted. The acute oral LD50 of LI85008F was greater than 5000 mg/kg in female SD rats and no changes in body weight or adverse effects were observed following necropsy. Acute dermal LD50 of LI85008F was greater than 2000 mg/kg. LI85008F was classified as non-irritating to skin in a primary dermal irritation study conducted using New Zealand Albino rabbits. LI85008F caused minimal irritation to eyes in a primary eye irritation test conducted on New Zealand Albino rabbits. A dose-dependent 28-day sub-acute toxicity study demonstrated no significant changes in selected organ weights. Evaluations on hematology, clinical chemistry, and histopathology did not show any significant adverse changes. The NOAEL of LI85008F was found to be greater than 2500 mg/kg body weight. These results demonstrate the broad spectrum safety of LI85008F in animal models.


Trends and determinants of stillbirth in developing countries: results from the Global Network's Population-Based Birth Registry.
Stillbirth rates remain high, especially in low and middle-income countries, where rates are 25 per 1000, ten-fold higher than in high-income countries. The United Nations' Every Newborn Action Plan has set a goal of 12 stillbirths per 1000 births by 2030 for all countries. From a population-based pregnancy outcome registry, including data from 2010 to 2016 from two sites each in Africa (Zambia and Kenya) and India (Nagpur and Belagavi), as well as sites in Pakistan and Guatemala, we evaluated the stillbirth rates and rates of annual decline as well as risk factors for 427,111 births of which 12,181 were stillbirths. The mean stillbirth rates for the sites were 21.3 per 1000 births for Africa, 25.3 per 1000 births for India, 56.9 per 1000 births for Pakistan and 19.9 per 1000 births for Guatemala. From 2010 to 2016, across all sites, the mean stillbirth rate declined from 31.7 per 1000 births to 26.4 per 1000 births for an average annual decline of 3.0%. Risk factors for stillbirth were similar across the sites and included maternal age < 20 years and age > 35 years. Compared to parity 1-2, zero parity and parity > 3 were both associated with increased stillbirth risk and compared to women with any prenatal care, women with no prenatal care had significantly increased risk of stillbirth in all sites. At the current rates of decline, stillbirth rates in these sites will not reach the Every Newborn Action Plan goal of 12 per 1000 births by 2030. More attention to the risk factors and treating the causes of stillbirths will be required to reach the Every Newborn Action Plan goal of stillbirth reduction. NCT01073475 .


Pain is common after sexual assault and posttraumatic arousal/reactivity symptoms mediate the development of new or worsening persistent pain.
Clinically significant new or worsening pain (CSNWP) is a common, yet often overlooked, sequelae of sexual assault. Little is known regarding factors influencing the development of CSNWP in sexual assault survivors. The current study used data from a recently completed prospective study to evaluate whether posttraumatic alterations in arousal and reactivity in the early aftermath of sexual assault influence the transition from acute to clinically significant new or worsening persistent pain. Women ≥ 18 years of age (n = 706) presenting for emergency care after sexual assault to 13 emergency care sites were enrolled in the study. Women completed assessments at the time of presentation as well as at 1 week (n = 706, 100%) and 6 weeks (n = 630, 91%). Nearly 70% of women reported CSNWP at the time of emergency care (n = 475, 69%), which persisted to 6 weeks in approximately 2 in 5 survivors (n = 248, 41%). A structural equation model adjusted for age, race, past trauma exposure, and preassault pain levels suggested that posttraumatic alterations in arousal/reactivity symptoms 1 week after assault partially mediated the transition from acute to persistent CSNWP. A significant portion (41%) of women sexual assault survivors develop CSNWP 6 weeks postassault. Posttraumatic arousal/reactivity symptoms in the early aftermath of assault contribute to CSNWP development; such symptoms are potential targets for secondary preventive interventions to reduce chronic postassault pain.


Self-mutilation and suicide attempt: distinguishing features in prisoners.
Nonlethal forms of self-injury are often discussed together with suicide attempts as though they belonged on a continuum of self-harm. Both types of self-injury are common in prisons, which have a predominantly male population; however, most studies of nonlethal self-injury have been done with female subjects. This exploratory study tested the hypothesis that prisoners who injured themselves without intending to die would differ clinically from prisoners who had attempted suicide. Inmates admitted to the prison unit of a public hospital for treatment of self-inflicted wounds or who had a history of previous self-injury were administered a standardized intake protocol by the first author, which included asking about their intent at the time they injured themselves. Patients were classified as self-mutilators or suicide attempters on the basis of intent. Fifteen patients reported that they had attempted to take their own lives, while 16 reported other reasons for harming themselves. Suicide attempt was associated with adult affective disorder 13/15 versus 2/16 mutilators); self-mutilation with a history of childhood hyperactivity (12/16 versus 1/15 suicide attempters) and a mixed dysthymia/anxiety syndrome that began in childhood or early adolescence (9/16). Prison self-mutilators and suicide attempters had very different clinical presentations and histories. The history of childhood hyperactivity in self-mutilators deserves further study in both correctional and noncorrectional populations.


Development of a high throughput and low cost model for the study of semi-dry biofilms.
The persistence of microorganisms as biofilms on dry surfaces resistant to the usual terminal cleaning methods may pose an additional risk of transmission of infections. In this study, the Centre for Disease Control (CDC) dry biofilm model (DBM) was adapted into a microtiter plate format (Model 1) and replicated to create a novel in vitro model that replicates conditions commonly encountered in the healthcare environment (Model 2). Biofilms of Staphylococcus aureus grown in the two models were comparable to the biofilms of the CDC DBM in terms of recovered log10 CFU well-1. Assessment of the antimicrobial tolerance of biofilms grown in the two models showed Model 2 a better model for biofilm formation. Confirmation of the biofilms' phenotype with an extracellular matrix deficient S. aureus suggested stress tolerance through a non-matrix defined mechanism in microorganisms. This study highlights the importance of conditions maintained in bacterial growth as they affect biofilm phenotype and behaviour.


Comparison of Hybribio GenoArray and Roche human papillomavirus (HPV) linear array for HPV genotyping in anal swab samples.
Human papillomavirus (HPV) is causally associated with anal cancer, as HPV DNA is detected in up to 90% of anal intraepithelial neoplasias and anal cancers. With the gradual increase of anal cancer rates, there is a growing need to establish reliable and clinically relevant methods to detect anal cancer precursors. In resource-limited settings, HPV DNA detection is a potentially relevant tool for anal cancer screening. Here, we evaluated the performance of the Hybribio GenoArray (GA) for genotyping HPV in anal samples, against the reference standard Roche Linear Array (LA). Anal swab samples were obtained from sexually active men who have sex with men. Following DNA extraction, each sample was genotyped using GA and LA. The overall interassay agreement, type-specific, and single and multiple genotype agreements were evaluated by kappa statistics and McNemar's χ(2) tests. Using GA and LA, 68% and 76% of samples were HPV DNA positive, respectively. There was substantial interassay agreements for the detection of all HPV genotypes (κ = 0.70, 86% agreement). Although LA was able to detect more genotypes per sample, the interassay agreement was acceptable (κ = 0.53, 63% agreement). GA had poorer specific detection of HPV genotypes 35, 42, and 51 (κ < 0.60). In conclusion, GA and LA showed good interassay agreement for the detection of most HPV genotypes in anal samples. However, the detection of HPV DNA in up to 76% of anal samples warrants further evaluation of its clinical significance.


Prediction of bone marrow iron findings from tests performed on peripheral blood.
After evaluating multiple tests, the authors have devised a scheme to predict bone marrow iron findings from tests performed on peripheral blood. They examined bone marrows from 97 consecutive patients with anemia who were divided into five marrow morphologic groups: (1) iron deficiency; (2) anemia of chronic disease; (3) abnormal sideroblasts; (4) ring sideroblasts; and (5) other. Tests of peripheral blood included hemoglobin, hematocrit, red blood cell count and red blood cell indices, reticulocyte count, sedimentation rate or zetacrit, ferritin, iron, iron binding capacity, free erythrocyte protoporphyrin, and tests of hepatic and renal function. Cluster analysis, multidimensional scaling, and logistic discriminant analysis were used to derive a graph of serum ferritin with the sedimentation rate, allowing accurate confirmation or exclusion of iron deficiency in most patients. Percent saturation of serum transferrin and serum ferritin allowed identification of only 50 percent of patients with abnormal or ring sideroblasts while excluding 100 percent of patients without abnormal or ring sideroblasts. In three years of follow-up, two of 19 patients with abnormal or ring sideroblast have developed the dysmyelopoietic syndrome or ANLL, respectively. With the aid of the two parameter graphs described, the authors believe the differential diagnosis of the hypoproliferative anemias relating to iron metabolism can frequently be made without examination of the bone marrow.


Neuroprotective benefits of grape seed and skin extract in a mouse model of Parkinson's disease.
Parkinson's disease is a neurodegenerative disorder characterized by the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra pars compacta, and it involves oxidative stress. Our goal was to evaluate the neuroprotective effect of Vitis vinifera red grape seed and skin extract (GSSE) in a model of Parkinson's disease. GSSE is very rich in phenolic compounds, such as flavonoids, anthocyanins, catechins and stilbenes, which are present in the pulp, seeds, and leaves of the fruit. GSSE is known for its antioxidant properties and has shown beneficial effects against oxidative injury in different organs, such as the kidneys, liver, heart and brain. In this study, we revealed the neuroprotective effect of GSSE on midbrain dopaminergic neurons both in vitro and in vivo. We used the neurotoxin 6-hydroxydopamine (6-OHDA), which induces oxidative damage and mimics the degeneration of dopaminergic neurons observed in Parkinson's disease. We found that GSSE was effective in protecting dopamine neurons from 6-OHDA toxicity by reducing apoptosis, the level of reactive oxygen species (ROS) and inflammation. Furthermore, we found that GSSE treatment efficiently protected against neuronal loss and improved motor function in an in vivo 6-OHDA model of Parkinson's disease (PD). Altogether, our results show that GSSE acts at multiple levels to protect dopamine neurons from degeneration in a model of PD.


Ion-Induced Formation of Nanocrystalline Cellulose Colloidal Glasses Containing Nematic Domains.
Controlling the assembly of colloids in dispersion is a fundamental approach toward the production of functional materials. Nanocrystalline cellulose (NCC) is a charged nanoparticle whose colloidal interactions can be modulated from repulsive to attractive by increasing ionic strength. Here, we combine polarized optical microscopy, rheology, and small-angle scattering techniques to investigate (i) the concentration-driven transition from isotropic dispersion to cholesteric liquid crystals and (ii) salt-induced NCC phase transitions. In particular, we report on the formation of NCC attractive glasses containing nematic domains. At increasing NCC concentration, a structure peak was observed in small-angle X-ray scattering (SAXS) patterns. The evolution of the structure peak demonstrates the decrease in NCC interparticle distance, favoring orientational order during the isotropic-cholesteric phase transition. Small amounts of salt reduce the cholesteric volume fraction and pitch by a decrease in excluded volume. Beyond a critical salt concentration, NCC forms attractive glasses due to particle caging and reduced motility. This results in a sharp increase in viscosity and formation of viscoelastic glasses. The presence of nematic domains is suggested by the appearance of interference colors and the Cox-Merz rule failure and was confirmed by an anisotropic SAXS scattering pattern at q ranges associated with the presence of nematic domains. Thus, salt addition allows the formation of NCC attractive glasses with mechanical properties similar to those of gels while remaining optically active owed to entrapped nematic domains.


Efflux pump inhibitors (EPIs) as new antimicrobial agents against Pseudomonas aeruginosa.
Pseudomonas aeruginosa is an opportunistic human pathogen and one of the leading causes of nosocomial infections worldwide. The difficulty in treatment of pseudomonas infections arises from being multidrug resistant (MDR) and exhibits resistance to most antimicrobial agents due to the expression of different mechanisms overcoming their effects. Of these resistance mechanisms, the active efflux pumps in Pseudomonas aeruginosa that belong to the resistance nodulation division (RND) plays a very important role in extruding the antibiotics outside the bacterial cells providing a protective means against their antibacterial activity. Beside its role against the antimicrobial agents, these pumps can extrude biocides, detergents, and other metabolic inhibitors. It is clear that efflux pumps can be targets for new antimicrobial agents. Peptidomimetic compounds such as phenylalanine arginyl β-naphthylamide (PAβN) have been introduced as efflux pump inhibitors (EPIs); their mechanism of action is through competitive inhibition with antibiotics on the efflux pump resulting in increased intracellular concentration of antibiotic, hence, restoring its antibacterial activity. The advantage of EPIs is the difficulty to develop bacterial resistance against them, but the disadvantage is their toxic property hindering their clinical application. The structure activity relationship of these compounds showed other derivatives from PAβN that are higher in their activity with higher solubility in biological fluids and decreased toxicity level. This raises further questions on how can we compact Pseudomonas infections. Of particular importance, the recent resurgence in the use of older antibiotics such as polymyxins and probably applying stricter control measures in order to prevent their spread in clinical sittings.


Biological and Molecular Variability Among High Plains virus Isolates.
The High Plains virus (HPV), vectored by the wheat curl mite (WCM) (Aceria tosichella), causes a severe disease of maize (Zea mays) in the U. S. High Plains. In the present study, five HPV isolates from five states were separated from co-infecting Wheat streak mosaic virus and their molecular and biological variability studied. Molecular studies involved time-of-flight mass spectrometry (TOFMS) to determine amino acid sequence variability of the 32-kDa nucleoprotein (32 np) of the isolates. Biological studies involved testing the ability of the five HPV isolates to infect a maize line previously shown to have resistance. Inoculations of the HPV isolates were conducted using vascular puncture inoculation (VPI) and viruliferous WCM. TOFMS analyses demonstrated an 18-amino acid sequence in the isolates at the N-terminus of the 32 np, the presence of amino acid sequence differences among the isolates, and variability among amino acid sequences of the 32 np of some isolates. Three of the five HPV isolates infected the resistant maize inbred, B73, using VPI, and two of the same three HPV isolates infected this line using WCM inoculation, albeit low numbers of plants were infected by each technique.


Total and state-specific medical and absenteeism costs of COPD among adults aged ≥ 18 years in the United States for 2010 and projections through 2020.
COPD remains a leading cause of morbidity and mortality. The objectives of this study were to estimate (1) national US COPD-attributable annual medical costs by payer (direct) and absenteeism (indirect) in 2010 and projected medical costs through 2020 and (2) state-specific COPD-attributable medical and absenteeism costs in 2010. We used the 2006-2010 Medical Expenditure Panel Survey, the 2004 National Nursing Home Survey, and 2010 Centers for Medicare and Medicaid Services data to generate cost estimates and 2010 census data to project medical costs through 2020. In 2010, total national medical costs attributable to COPD and its sequelae were estimated at $32.1 billion, and total absenteeism costs were $3.9 billion, for a total burden of COPD-attributable costs of $36 billion. An estimated 16.4 million days of work were lost because of COPD. Of the medical costs, 18% was paid for by private insurance, 51% by Medicare, and 25% by Medicaid. National medical costs are projected to increase from $32.1 billion in 2010 to $49.0 billion in 2020. Total state-specific costs in 2010 ranged from $49.1 million in Wyoming to $2.8 billion in California: medical costs ranged from $42.5 million in Alaska to $2.5 billion in Florida and absenteeism costs ranged from $8.4 million in Wyoming to $434.0 million in California. Costs attributable to COPD and its sequelae are substantial and are projected to increase through 2020. Evidence-based interventions that prevent tobacco use and reduce the clinical complications of COPD may result in potential decreased COPD-attributable costs.


Construction of a computable cell proliferation network focused on non-diseased lung cells.
Critical to advancing the systems-level evaluation of complex biological processes is the development of comprehensive networks and computational methods to apply to the analysis of systems biology data (transcriptomics, proteomics/phosphoproteomics, metabolomics, etc.). Ideally, these networks will be specifically designed to capture the normal, non-diseased biology of the tissue or cell types under investigation, and can be used with experimentally generated systems biology data to assess the biological impact of perturbations like xenobiotics and other cellular stresses. Lung cell proliferation is a key biological process to capture in such a network model, given the pivotal role that proliferation plays in lung diseases including cancer, chronic obstructive pulmonary disease (COPD), and fibrosis. Unfortunately, no such network has been available prior to this work. To further a systems-level assessment of the biological impact of perturbations on non-diseased mammalian lung cells, we constructed a lung-focused network for cell proliferation. The network encompasses diverse biological areas that lead to the regulation of normal lung cell proliferation (Cell Cycle, Growth Factors, Cell Interaction, Intra- and Extracellular Signaling, and Epigenetics), and contains a total of 848 nodes (biological entities) and 1597 edges (relationships between biological entities). The network was verified using four published gene expression profiling data sets associated with measured cell proliferation endpoints in lung and lung-related cell types. Predicted changes in the activity of core machinery involved in cell cycle regulation (RB1, CDKN1A, and MYC/MYCN) are statistically supported across multiple data sets, underscoring the general applicability of this approach for a network-wide biological impact assessment using systems biology data. To the best of our knowledge, this lung-focused Cell Proliferation Network provides the most comprehensive connectivity map in existence of the molecular mechanisms regulating cell proliferation in the lung. The network is based on fully referenced causal relationships obtained from extensive evaluation of the literature. The computable structure of the network enables its application to the qualitative and quantitative evaluation of cell proliferation using systems biology data sets. The network is available for public use.


Survival after diagnosis of localized T1a kidney cancer: current population-based practice of surgery and nonsurgical management.
To compare overall and cancer-specific survival (CSS) of patients who undergo nonsurgical management (NSM), partial nephrectomy (PN), and radical nephrectomy (RN). NSM is being increasingly used for older patients with early-stage kidney cancer and competing risks of death. However, survival is poorly characterized for this approach compared with surgery with PN or RN. The Surveillance, Epidemiology and End Results-Medicare database from 1995 to 2007 was used to identify patients aged 65 years or older diagnosed with localized T1a kidney cancer treated with PN, RN, or NSM. We used Cox proportional hazards regression, Fine and Gray competing risks regression, and propensity score matching to adjust for patient and tumor characteristics. Of 7177 Medicare beneficiaries meeting the inclusion criteria, 754 (10.5%) underwent NSM, 1849 (25.8%) PN, and 4574 (63.7%) RN, with 436 (57.8%), 389 (21.0%), and 1598 (34.9%) patients dying from any cause, respectively, at a median follow-up of 56 months. Overall survival favored PN and RN compared with NSM (hazard ratio [95% CI]: 0.40 [0.34-0.46] and 0.50 [0.45-0.56], respectively) as did CSS (hazard ratio [95% CI]: 0.42 [0.27-0.64] and 0.62 [0.46-0.85], respectively). However, there was no difference in CSS between any 2 treatment groups for younger patients (<75 years), whereas there was an excess of kidney cancer deaths for NSM patients aged 75-79 years and an attenuated difference for patients aged 80 years or older. NSM was associated with an increased risk of kidney cancer death among Medicare beneficiaries aged 75-79 years. Evolving active surveillance protocols will need to develop robust selection criteria to maximize cancer survival for older patients with kidney cancer.


A method to discriminate between closely related bovine major histocompatibility complex class I alleles by combining established PCR-SSP assays with RFLPs.
We have developed a polymerase chain reaction-sequence-specific primers-restriction fragment length polymorphism (PCR-SSP-RFLP) method to rapidly differentiate between the A18 and A18 variant (v) BoLA haplotypes and between A14 and A15/A15v BoLA haplotypes in Holstein/Friesian cattle. We used published SSP to PCR amplify BoLA alleles expressed in animals of known haplotype and exposed the amplicons to the restriction enzyme PvuII that was predicted to cut at a unique site in the middle of BoLA-6*01302 (A18v) and BoLA-1*00901 (A15) but not in BoLA-6*01301 (A18) or BoLA-1*02301 (A14) alleles. Whereas the method does not discriminate between the A15 and A15v haplotypes, as the BoLA-1*00902 allele associated with A15v also contains a PvuII site, we are interested in cattle of A18 and A14 haplotype for vaccine related studies. Our results also indicated that the BoLA-6*01302 (A18v) allele is much more abundant than BoLA-6*01301 (A18) in the cattle that we sampled.


Age and Muscle Function Are More Closely Associated With Intracellular Magnesium, as Assessed by 31P Magnetic Resonance Spectroscopy, Than With Serum Magnesium.
Total serum magnesium is a common clinical measurement for assessing magnesium status; however, magnesium in blood represents less than 1% of the body's total magnesium content. We measured intramuscular ionized magnesium by phosphorus magnetic resonance spectroscopy (31P-MRS) and tested the hypothesis that this measure better correlates with skeletal muscle function and captures more closely the effect of aging than the traditional measure of total serum magnesium. Data were collected from 441 participants (age 24-98 years) in the Baltimore Longitudinal Study of Aging (BLSA), a study of normative aging that encompasses a broad age range. Results showed that intramuscular ionized magnesium was negatively associated with age (β = -0.29, p < 0.001, R 2 = 0.08) and positively associated with knee-extension strength (β = 0.31, p < 0.001, and R 2 = 0.1 in women; and β = 0.2, p = 0.003, and R 2 = 0.04 in men), while total serum magnesium showed no association with age or strength (p = 0.27 and 0.1, respectively). Intramuscular ionized magnesium was significantly lower in women that in men (p < 0.001), perhaps due to chronic latent Mg deficiency in women that is not otherwise detected by serum magnesium levels. Based on these findings, we suggest that intramuscular ionized magnesium from 31P-MRS is a better clinical measure of magnesium status than total serum magnesium, and could be measured when muscle weakness of unidentified etiology is detected. It may also be used to monitor the effectiveness of oral magnesium interventions, including supplementation.


Bloody Zebrafish: Novel Methods in Normal and Malignant Hematopoiesis.
Hematopoiesis is an optimal system for studying stem cell maintenance and lineage differentiation under physiological and pathological conditions. In vertebrate organisms, billions of differentiated hematopoietic cells need to be continuously produced to replenish the blood cell pool. Disruptions in this process have immediate consequences for oxygen transport, responses against pathogens, maintenance of hemostasis and vascular integrity. Zebrafish is a widely used and well-established model for studying the hematopoietic system. Several new hematopoietic regulators were identified in genetic and chemical screens using the zebrafish model. Moreover, zebrafish enables in vivo imaging of hematopoietic stem cell generation and differentiation during embryogenesis, and adulthood. Finally, zebrafish has been used to model hematopoietic diseases. Recent technological advances in single-cell transcriptome analysis, epigenetic regulation, proteomics, metabolomics, and processing of large data sets promise to transform the current understanding of normal, abnormal, and malignant hematopoiesis. In this perspective, we discuss how the zebrafish model has proven beneficial for studying physiological and pathological hematopoiesis and how these novel technologies are transforming the field.


Role of receptor protein tyrosine phosphatase alpha (RPTPalpha) and tyrosine phosphorylation in the serotonergic inhibition of voltage-dependent potassium channels.
The activity of voltage-gated potassium (Kv) channels can be dynamically modulated by several events, including neurotransmitter-stimulated biochemical cascades mediated by G-protein-coupled receptors. By using a heterologous expression system, we show that activating the 5-HT2C receptor inhibits both Kv1.1 and Kv1.2 channels through a tyrosine phosphorylation mechanism. The major molecular determinants of channel inhibition were identified as two tyrosine residues located in the N-terminal region of the Kv channel subunit. Furthermore, we demonstrate that receptor protein tyrosine phosphatase alpha (RPTPalpha), a receptor protein tyrosine phosphatase, co-ordinates the inhibition process mediated via 5-HT2C receptors. We therefore propose that the serotonergic regulation of human Kv1.1 and Kv1.2 channel activity by the 5-HT2C receptor involves the dual coordination of both RPTPalpha and specific tyrosine kinases coupled to this receptor.


Focus on cutaneous and uveal melanoma specificities.
Cutaneous melanoma (CM) and uveal melanoma (UM) derive from cutaneous and uveal melanocytes that share the same embryonic origin and display the same cellular function. However, the etiopathogenesis and biological behaviors of these melanomas are very different. CM and UM display distinct landscapes of genetic alterations and show different metastatic routes and tropisms. Hence, therapeutic improvements achieved in the last few years for the treatment of CM have failed to ameliorate the clinical outcomes of patients with UM. The scope of this review is to discuss the differences in tumorigenic processes (etiologic factors and genetic alterations) and tumor biology (gene expression and signaling pathways) between CM and UM. We develop hypotheses to explain these differences, which might provide important clues for research avenues and the identification of actionable vulnerabilities suitable for the development of new therapeutic strategies for metastatic UM.


Identification and characterization of novel TRPM1 autoantibodies from serum of patients with melanoma-associated retinopathy.
Melanoma-associated retinopathy (MAR) is a rare paraneoplastic retinal disorder usually occurring in the context of metastatic melanoma. Patients present with night blindness, photopsias and a constriction of the visual field. MAR is an auto-immune disorder characterized by the production of autoantibodies targeting retinal proteins, especially autoantibodies reacting to the cation channel TRPM1 produced in melanocytes and ON-bipolar cells. TRPM1 has at least three different isoforms which vary in the N-terminal region of the protein. In this study, we report the case of three new MAR patients presenting different anti-TRPM1 autoantibodies reacting to the three isoforms of TRPM1 with variable binding affinity. Two sera recognized all isoforms of TRPM1, while one recognized only the two longest isoforms upon immunolocalization studies on overexpressing cells. Similarly, the former two sera reacted with all TRPM1 isoforms on western blot, but an immunoprecipitation enrichment step was necessary to detect all isoforms with the latter serum. In contrast, all sera labelled ON-bipolar cells on Tprm1+/+ but not on Trpm1-/- mouse retina as shown by co-immunolocalization. This confirms that the MAR sera specifically detect TRPM1. Most likely, the anti-TRPM1 autoantibodies of different patients vary in affinity and concentration. In addition, the binding of autoantibodies to TRPM1 may be conformation-dependent, with epitopes being inaccessible in some constructs (truncated polypeptides versus full-length TRPM1) or applications (western blotting versus immunohistochemistry). Therefore, we propose that a combination of different methods should be used to test for the presence of anti-TRPM1 autoantibodies in the sera of MAR patients.


Visceral primitive peripheral neuroectodermal tumors: a clinicopathologic and molecular study.
Ewing sarcoma-primitive neuroectodermal tumor (EWS/PNET) belongs to the group of pediatric small round blue cell tumors; although EWS/PNET is classically a tumor of the soft tissue or bone in children and young adults, individual cases have been described in patients of all ages. A group of chromosomal translocations involving the EWS gene and a member of the Ets transcription factor family of genes has been detected in EWS/PNET, and heterogeneity in the precise breakpoint of the translocation has been shown to generate a group of related fusion transcripts that may have prognostic significance. Within the last decade, the clinicopathologic spectrum of EWS/PNET has been markedly expanded by recognition that the tumor may also have a visceral origin. To determine whether visceral EWS/PNET has the same pattern of genetic alterations and range of fusion transcripts as EWS/PNET of bone and soft tissue, we performed reverse-transcription polymerase chain reaction-based testing of formalin-fixed, paraffin-embedded tissue from a series of visceral tumors for which the diagnosis of EWS/PNET was well established. Together with additional cases compiled from the literature, EWS-Fli1 (or a related fusion transcript) was present in 18 of 19 visceral EWS/PNET, with a distribution of transcript types not statistically different from EWS/PNET of soft tissue and bone (P >.05, chi(2) test). These results firmly establish the genetic relationship between EWS/PNET of visceral sites, soft tissue, and bone.


Transcellular degradation of axonal mitochondria.
It is generally accepted that healthy cells degrade their own mitochondria. Here, we report that retinal ganglion cell axons of WT mice shed mitochondria at the optic nerve head (ONH), and that these mitochondria are internalized and degraded by adjacent astrocytes. EM demonstrates that mitochondria are shed through formation of large protrusions that originate from otherwise healthy axons. A virally introduced tandem fluorophore protein reporter of acidified mitochondria reveals that acidified axonal mitochondria originating from the retinal ganglion cell are associated with lysosomes within columns of astrocytes in the ONH. According to this reporter, a greater proportion of retinal ganglion cell mitochondria are degraded at the ONH than in the ganglion cell soma. Consistently, analyses of degrading DNA reveal extensive mtDNA degradation within the optic nerve astrocytes, some of which comes from retinal ganglion cell axons. Together, these results demonstrate that surprisingly large proportions of retinal ganglion cell axonal mitochondria are normally degraded by the astrocytes of the ONH. This transcellular degradation of mitochondria, or transmitophagy, likely occurs elsewhere in the CNS, because structurally similar accumulations of degrading mitochondria are also found along neurites in superficial layers of the cerebral cortex. Thus, the general assumption that neurons or other cells necessarily degrade their own mitochondria should be reconsidered.


Folate receptor-α (FOLR1) expression and function in triple negative tumors.
Folate receptor alpha (FOLR1) has been identified as a potential prognostic and therapeutic target in a number of cancers. A correlation has been shown between intense overexpression of FOLR1 in breast tumors and poor prognosis, yet there is limited examination of the distribution of FOLR1 across clinically relevant breast cancer subtypes. To explore this further, we used RNA-seq data from multiple patient cohorts to analyze the distribution of FOLR1 mRNA across breast cancer subtypes comprised of estrogen receptor positive (ER+), human epidermal growth factor receptor positive (HER2+), and triple negative (TNBC) tumors. FOLR1 expression varied within breast tumor subtypes; triple negative/basal tumors were significantly associated with increased expression of FOLR1 mRNA, compared to ER+ and HER2+ tumors. However, subsets of high level FOLR1 expressing tumors were observed in all clinical subtypes. These observations were supported by immunohistochemical analysis of tissue microarrays, with the largest number of 3+ positive tumors and highest H-scores of any subtype represented by triple negatives, and lowest by ER+ tumors. FOLR1 expression did not correlate to common clinicopathological parameters such as tumor stage and nodal status. To delineate the importance of FOLR1 overexpression in triple negative cancers, RNA-interference was used to deplete FOLR1 in overexpressing triple negative cell breast lines. Loss of FOLR1 resulted in growth inhibition, whereas FOLR1 overexpression promoted folate uptake and growth advantage in low folate conditions. Taken together, our data suggests patients with triple negative cancers expressing high FOLR1 expression represent an important population of patients that may benefit from targeted anti-FOLR1 therapy. This may prove particularly helpful for a large number of patients who would typically be classified as triple negative and who to this point have been left without any targeted treatment options.


How to optimize the evaluation and use of antibiotics in neonates.
Early and late-onset neonatal sepsis has specific pathogen distribution and infection rates in neonates with different gestational and postnatal ages. Despite the fact that early-onset sepsis is relatively rare (<1% of total deliveries), it is a major cause of mortality and morbidity. The known immunological immaturity of the neonate combined with non-specific clinical symptoms of infection has resulted in the frequent overuse of antibiotics in neonatal intensive care units (NICUs). In addition to this overuse there is a huge variability in the choice of antibacterial agents and dosing regimens used in NICUs across the world. Therefore, a more rational approach in the neonatal use of antibiotics is needed because of two major reasons: the emergence of multi-resistant bacteria in NICUs, and short- and long-term side effects of frequently used antibacterial agents in the neonatal population. This paper will focus on the optimal use of aminoglycosides (both used in early and late onset sepsis) and vancomycin (primarily used in late onset infections) in NICUs, and will underscore the need for specialists in neonatal medicine and pediatric pharmacology to work closely together to reach the most effective and safe way of using medicines in the NICU.


An inter-island comparison of Darwin's finches reveals the impact of habitat, host phylogeny, and island on the gut microbiome.
Darwin's finch species in the Galapagos Archipelago are an iconic adaptive radiation that offer a natural experiment to test for the various factors that influence gut microbiome composition. The island of Floreana has the longest history of human settlement within the archipelago and offers an opportunity to compare island and habitat effects on Darwin's finch microbiomes. In this study, we compare gut microbiomes in Darwin's finch species on Floreana Island to test for effects of host phylogeny, habitat (lowlands, highlands), and island (Floreana, Santa Cruz). We used 16S rRNA Illumina sequencing of fecal samples to assess the gut microbiome composition of Darwin's finches, complemented by analyses of stable isotope values and foraging data to provide ecological context to the patterns observed. Overall bacterial composition of the gut microbiome demonstrated co-phylogeny with Floreana hosts, recapitulated the effect of habitat and diet, and showed differences across islands. The finch phylogeny uniquely explained more variation in the microbiome than did foraging data. Finally, there were interaction effects for island × habitat, whereby the same Darwin's finch species sampled on two islands differed in microbiome for highland samples (highland finches also had different diets across islands) but not lowland samples (lowland finches across islands had comparable diet). Together, these results corroborate the influence of phylogeny, age, diet, and sampling location on microbiome composition and emphasize the necessity for comprehensive sampling given the multiple factors that influence the gut microbiome in Darwin's finches, and by extension, in animals broadly.


Editing of an Alpha-Kafirin Gene Family Increases, Digestibility and Protein Quality in Sorghum.
Kafirins are the major storage proteins in sorghum (Sorghum bicolor) grains and form protein bodies with poor digestibility. Since kafirins are devoid of the essential amino acid lysine, they also impart poor protein quality to the kernel. The α-kafirins, which make up most of the total kafirins, are largely encoded by the k1C family of highly similar genes. We used a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing approach to target the k1C genes to create variants with reduced kafirin levels and improved protein quality and digestibility. A single guide RNA was designed to introduce mutations in a conserved region encoding the endoplasmic reticulum signal peptide of α-kafirins. Sequencing of kafirin PCR products revealed extensive edits in 25 of 26 events in one or multiple k1C family members. T1 and T2 seeds showed reduced α-kafirin levels, and selected T2 events showed significantly increased grain protein digestibility and lysine content. Thus, a single consensus single guide RNA carrying target sequence mismatches is sufficient for extensive editing of all k1C genes. The resulting quality improvements can be deployed rapidly for breeding and the generation of transgene-free, improved cultivars of sorghum, a major crop worldwide.


Longer term impact of the mass media campaign to promote the Get Healthy Information and Coaching Service®: increasing the saliency of a new public health program.
The Get Healthy Information and Coaching Service® (GHS) was introduced in New South Wales in February 2009. It used mass reach media advertising and direct mail and/or proactive marketing to recruit participants. This article reports on the long-term impact of the campaign on GHS participation from July 2011 to June 2012. A stand-alone population survey collected awareness, knowledge, and behavioral variables before the first advertising phase, (n = 1,544, August-September 2010), during the advertising period (n = 1,500, February-March 2011; n = 1,500, June-July 2011; n = 1,500, February 2012), and after the advertising period (n = 1,500, June-July 2012). GHS usage data (n = 6,095) were collated during July 2011-June 2012. Unprompted and prompted awareness of GHS mass media significantly increased (0% to 8.0%, p < .001; and 14.1% to 43.9%, p < .001, respectively) as well as knowledge and perceived effectiveness of the GHS. Those from the lowest three quintiles of socioeconomic disadvantage and respondents who were overweight or obese were significantly more likely to report prompted campaign awareness. The majority (84.4%) of new GHS calls occurred when television advertising was present. Participants who cited mass media as their referral source were significantly more likely to enroll in the intensive coaching program. Mass media campaigns remain an effective method of promoting a telephone-based statewide lifestyle program.