Doxycycline for community treatment of suspected COVID-19 in people at high risk of adverse outcomes in the UK (PRINCIPLE): a randomised, controlled, open-label, adaptive platform trial.
Doxycycline is often used for treating COVID-19 respiratory symptoms in the community despite an absence of evidence from clinical trials to support its use. We aimed to assess the efficacy of doxycycline to treat suspected COVID-19 in the community among people at high risk of adverse outcomes. We did a national, open-label, multi-arm, adaptive platform randomised trial of interventions against COVID-19 in older people (PRINCIPLE) across primary care centres in the UK. We included people aged 65 years or older, or 50 years or older with comorbidities (weakened immune system, heart disease, hypertension, asthma or lung disease, diabetes, mild hepatic impairment, stroke or neurological problem, and self-reported obesity or body-mass index of 35 kg/m2 or greater), who had been unwell (for ≤14 days) with suspected COVID-19 or a positive PCR test for SARS-CoV-2 infection in the community. Participants were randomly assigned using response adaptive randomisation to usual care only, usual care plus oral doxycycline (200 mg on day 1, then 100 mg once daily for the following 6 days), or usual care plus other interventions. The interventions reported in this manuscript are usual care plus doxycycline and usual care only; evaluations of other interventions in this platform trial are ongoing. The coprimary endpoints were time to first self-reported recovery, and hospitalisation or death related to COVID-19, both measured over 28 days from randomisation and analysed by intention to treat. This trial is ongoing and is registered with ISRCTN, 86534580. The trial opened on April 2, 2020. Randomisation to doxycycline began on July 24, 2020, and was stopped on Dec 14, 2020, because the prespecified futility criterion was met; 2689 participants were enrolled and randomised between these dates. Of these, 2508 (93·3%) participants contributed follow-up data and were included in the primary analysis: 780 (31·1%) in the usual care plus doxycycline group, 948 in the usual care only group (37·8%), and 780 (31·1%) in the usual care plus other interventions group. Among the 1792 participants randomly assigned to the usual care plus doxycycline and usual care only groups, the mean age was 61·1 years (SD 7·9); 999 (55·7%) participants were female and 790 (44·1%) were male. In the primary analysis model, there was little evidence of difference in median time to first self-reported recovery between the usual care plus doxycycline group and the usual care only group (9·6 [95% Bayesian Credible Interval [BCI] 8·3 to 11·0] days vs 10·1 [8·7 to 11·7] days, hazard ratio 1·04 [95% BCI 0·93 to 1·17]). The estimated benefit in median time to first self-reported recovery was 0·5 days [95% BCI -0·99 to 2·04] and the probability of a clinically meaningful benefit (defined as ≥1·5 days) was 0·10. Hospitalisation or death related to COVID-19 occurred in 41 (crude percentage 5·3%) participants in the usual care plus doxycycline group and 43 (4·5%) in the usual care only group (estimated absolute percentage difference -0·5% [95% BCI -2·6 to 1·4]); there were five deaths (0·6%) in the usual care plus doxycycline group and two (0·2%) in the usual care only group. In patients with suspected COVID-19 in the community in the UK, who were at high risk of adverse outcomes, treatment with doxycycline was not associated with clinically meaningful reductions in time to recovery or hospital admissions or deaths related to COVID-19, and should not be used as a routine treatment for COVID-19. UK Research and Innovation, Department of Health and Social Care, National Institute for Health Research.


Health disparities in cardiometabolic risk among Black and Hispanic youth in the United States.
Cardiometabolic risk factors in children and adolescents track into adulthood and are associated with increased risk of atherosclerotic cardiovascular disease. The purpose of this review is to examine the pervasive race and ethnic disparities in cardiometabolic risk factors among Black and Hispanic youth in the United States. We focus on three traditional cardiometabolic risk factors (obesity, type 2 diabetes mellitus, and dyslipidemia) as well as on the emerging cardiometabolic risk factor of non-alcoholic fatty liver disease. Additionally, we highlight interventions aimed at improving cardiometabolic health among these minority pediatric populations. Finally, we advocate for continued research on effective prevention strategies to reduce cardiometabolic risk and avert further disparities in cardiovascular morbidity and mortality.


Mouse mast cell protease 4 is the major chymase in murine airways and has a protective role in allergic airway inflammation.
It is widely established that mast cells (MCs) have a harmful role in asthma, for example by secreting various proinflammatory substances stored within their secretory granule. However, in this study, we show that one of the substances stored within MC granule, chymase, in fact has a protective role in allergic airway inflammation, indicating that MCs may possess both harmful and protective activities in connection with this type of disease. Wild-type (WT) mice and mice lacking mouse MC protease 4 (mMCP-4), a chymase that is functionally homologous to human chymase, were sensitized and challenged with OVA, followed by the assessment of airway physiology and inflammatory parameters. Our results show that the airway hyperresponsiveness was significantly higher in mMCP-4(-/-) as compared with WT mice. Moreover, the degree of lung tissue inflammation was markedly higher in mice lacking mMCP-4 than in WT controls. Histological analysis revealed that OVA sensitization/challenge resulted in a marked increased in the thickness of the smooth muscle cell (SMC) layer and, notably, that the degree of SMC layer thickening was more pronounced in mMCP-4(-/-) animals than in WT controls, thus indicating that chymase may have an effect on airway SMCs. In support of this, mMCP-4-positive MCs were located in the close vicinity of the SMC layer, mainly in the upper airways, and mMCP-4 was shown to be the major chymase expressed in these MCs. Taken together, our results indicate that chymase present in the upper airways protects against allergic airway responses, possibly by regulating SMCs.


Demonstration of optical switching by means of solitary wave collisions in a fiber ring reflector.
We have demonstrated the use of solitary wave collisions in optical pulse switching. Our apparatus consisted of a fiber ring with 11 sections of polarization-maintaining fiber, with successive sections fusion spliced with the axes rotated 90 deg. The configuration yielded enhanced transmission (autocorrelation contrast ratio 2.82:1), in agreement with expectation for this number of sections and the unoptimized fiber coupler that was used. Design criteria for complete switching are presented.


Fatigue in multiple sclerosis is not due to sleep apnoea.
Fatigue is a frequent and disabling phenomenon among patients with multiple sclerosis (MS). Daytime sleepiness is a typical symptom of the sleep apnoea/hypopnoea syndrome due to nocturnal hypoxia and recurrent arousals causing sleep fragmentation. Since MS plaques are often found in the midbrain, brain stem and upper cervical cord on magnetic resonance imaging (MRI) we hypothesized that fatigue in MS patients might be caused by a central respiratory dysfunction. We investigated 10 patients with definite MS by oligography, two questionnaires assessing fatigue (Fatigue Severity Scale, FSS) and daytime sleepiness (Epworth Sleepiness Scale, ESS), MRI and pulmonary function tests. A total of six patients had either an elevated FSS and/or an elevated ESS. None of the six patients with an elevated FSS and/or ESS has an apnoea index > 5/hour. CT90 was normal in nine patients. We conclude that fatigue and daytime sleepiness in MS cannot be explained by nocturnal apnoeas or oxygen desaturations. The Fatigue Severity Scale should be integrated to the extended Barthel index, which is a new instrument for disability assessment in MS patients.


The effects of a rotator cuff tear on activities of daily living in older adults: A kinematic analysis.
Rotator cuff tears (RCT) in older individuals may compound age-associated physiological changes and impact their ability to perform daily functional tasks. Our objective was to quantify thoracohumeral kinematics for functional tasks in 18 older adults (mean age=63.3±2.2), and compare findings from nine with a RCT to nine matched controls. Motion capture was used to record kinematics for 7 tasks (axilla wash, forward reach, functional pull, hair comb, perineal care, upward reach to 90°, upward reach to 105°) spanning the upper limb workspace. Maximum and minimum joint angles and motion excursion for the three thoracohumeral degrees of freedom (elevation plane, elevation, axial rotation) were identified for each task and compared between groups. The RCT group used greater minimum elevation angles for axilla wash and functional pull (p≤0.0124) and a smaller motion excursion for functional pull (p=0.0032) compared to the control group. The RCT group also used a more internally rotated maximum axial rotation angle than controls for functional reach, functional pull, hair comb, and upward reach to 105° (p≤0.0494). The most differences between groups were observed for axial rotation, with the RCT group using greater internal rotation to complete functional tasks, and significant differences between groups were identified for all three thoracohumeral degrees of freedom for functional pull. We conclude that older adults with RCT used more internal rotation to perform functional tasks than controls. The kinematic differences identified in this study may have consequences for progression of shoulder damage and further functional impairment in older adults with RCT.


A medical review approach to Medicare outpatient documentation.
Blue Cross of California has recognized the problem of reimbursement delays due to Medicare claims that have been returned or denied. The information in this article applies to the medical review process and suggests an interpretation of the new Medicare outpatient guidelines used by the Blue Cross of California Medicare reviewers. As a solution to the problem of delayed reimbursement, medical reviewers--who are also practicing occupational therapists--offer an explanation of the Medicare review process and suggestions for correcting technical billing errors. Methods for keeping complete and timely medical records are discussed, with the suggestion that clinicians follow these methods from the start of care. A process for documentation is presented that will clearly prove to the medical reviewer the need for the special skills of an occupational therapist. This article also proposes reasons for noncoverage because of insufficient medical necessity. It is important to note, however, that variance between fiscal intermediaries' interpretations and requirements exists nationwide. This article, therefore, represents only one fiscal intermediary's approach to reviewing claims and medical records.


DNA single strand breakage, DNA adducts, and sister chromatid exchange in lymphocytes and phenanthrene and pyrene metabolites in urine of coke oven workers.
To investigate the specificity of biological monitoring variables (excretion of phenanthrene and pyrene metabolites in urine) and the usefulness of some biomarkers of effect (alkaline filter elution, 32P postlabelling assay, measurement of sister chromatid exchange) in workers exposed to polycyclic aromatic hydrocarbons (PAHs). 29 coke oven workers and a standardised control group were investigated for frequencies of DNA single strand breakage, DNA protein cross links (alkaline filter elution assay), sister chromatid exchange, and DNA adducts (32P postlabelling assay) in lymphocytes. Phenanthrene and pyrene metabolites were measured in 24 hour urine samples. 19 different PAHs (including benzo(a)pyrene, pyrene, and phenanthrene) were measured at the workplace by personal air monitoring. The GSTT1 activity in erythrocytes and lymphocyte subpopulations in blood was also measured. Concentrations of phenanthrene, pyrene, and benzo(a)pyrene in air correlated well with the concentration of total PAHs in air; they could be used for comparisons of different workplaces if the emission compositions were known. The measurement of phenanthrene metabolites in urine proved to be a better biological monitoring variable than the measurement of 1-hydroxypyrene. Significantly more DNA strand breaks in lymphocytes of coke oven workers were found (alkaline filter elution assay); the DNA adduct rate was not significantly increased in workers, but correlated with exposure to PAHs in a semiquantitative manner. The number of sister chromatid exchanges was lower in coke oven workers but this was not significant; thus counting sister chromatid exchanges was not a good variable for biomonitoring of coke oven workers. Also, indications for immunotoxic influences (changes in lymphocyte subpopulations) were found. The measurement of phenanthrene metabolites in urine seems to be a better biological monitoring variable for exposure to PAHs than measurement of hydroxypyrene. The alkaline filter elution assay proved to be the most sensitive biomarker for genotoxic damage, whereas the postlabelling assay was the only one with some specificity for DNA alterations caused by known compounds.


A review of floc strength and breakage.
The main focus of the paper is to review current understanding of floc structure and strength. This has been done by reviewing current theoretical understanding of floc growth and breakage and an analysis of different techniques used for measuring floc strength. An overview has also been made of the general trends seen in floc strength analysis. The rate of floc formation is a balance between breakage and aggregation with flocs eventually reaching a steady-state size for a given shear rate. The steady-state floc size for a particular shear rate can, therefore, be a good indicator of floc strength. This has resulted in the development of a range of techniques to measure floc size at different applied shear levels using a combination of one or more of the following tools: light scattering and transmission; microscopy; photography; video and image analysis software. Floc strength may be simply quantified using the initial floc size for a given shear rate and the floc strength factor. More complex techniques have used theoretical modelling to determine whether flocs break by large-scale fragmentation or smaller-scale surface erosion effects, although this interpretation is open to debate. Impeller-based mixing, ultrasound and vibrating columns have all been used to provide a uniform, accurate and controllable dissipation of energy onto a floc suspension to determine floc strength. Other more recent techniques have used sensitive micromanipulators to measure the force required to break or compress individual flocs, although these techniques have been limited to the measurement of only a few hundred flocs. General trends emerge showing that smaller flocs tend to have greater strength than larger flocs, whilst the use of polymer seems to give increased strength to only some types of floc. Finally, a comparison of the strength of different types of floc (activated sludge flocs, organic matter flocs, sweep flocs and charge neutralised flocs) has been made highlighting differences in relative floc strength.


Clinical audit of COPD patients requiring hospital admissions in Spain: AUDIPOC study.
AUDIPOC is a nationwide clinical audit that describes the characteristics, interventions and outcomes of patients admitted to Spanish hospitals because of an exacerbation of chronic obstructive pulmonary disease (ECOPD), assessing the compliance of these parameters with current international guidelines. The present study describes hospital resources, hospital factors related to case recruitment variability, patients' characteristics, and adherence to guidelines. An organisational database was completed by all participant hospitals recording resources and organisation. Over an 8-week period 11,564 consecutive ECOPD admissions to 129 Spanish hospitals covering 70% of the Spanish population were prospectively identified. At hospital discharge, 5,178 patients (45% of eligible) were finally included, and thus constituted the audited population. Audited patients were reassessed 90 days after admission for survival and readmission rates. A wide variability was observed in relation to most variables, hospital adherence to guidelines, and readmissions and death. Median inpatient mortality was 5% (across-hospital range 0-35%). Among discharged patients, 37% required readmission (0-62%) and 6.5% died (0-35%). The overall mortality rate was 11.6% (0-50%). Hospital size and complexity and aspects related to hospital COPD awareness were significantly associated with case recruitment. Clinical management most often complied with diagnosis and treatment recommendations but rarely (<50%) addressed guidance on healthy life-styles. The AUDIPOC study highlights the large across-hospital variability in resources and organization of hospitals, patient characteristics, process of care, and outcomes. The study also identifies resources and organizational characteristics associated with the admission of COPD cases, as well as aspects of daily clinical care amenable to improvement.


Delayed internal fixation of femoral shaft fracture reduces mortality among patients with multisystem trauma.
Fractures of the femoral shaft are common and have potentially serious consequences in patients with multiple injuries. The appropriate timing of fracture repair is controversial. The purpose of the present study was to assess the effect of timing of internal fixation on mortality in patients with multisystem trauma. We performed a retrospective cohort study with use of data from public and private trauma centers throughout the United States that were reported to the National Trauma Data Bank (version 5.0 for 2000 through 2004). The study included 3069 patients with multisystem trauma (Injury Severity Score, > or =15) who underwent internal fixation of a femoral shaft fracture. The time to treatment was defined in categories as the time from admission to internal fixation: t(0) (twelve hours or less), t(1) (more than twelve hours to twenty-four hours), t(2) (more than twenty-four hours to forty-eight hours), t(3) (more than forty-eight hours to 120 hours), and t(4) (more than 120 hours). The relative risk of in-hospital mortality when the four later periods were compared with the earliest one was estimated with inverse probability of treatment-weighted analysis. Subgroups with serious head or neck, chest, abdominal, and additional extremity injury were investigated. When compared with that during the first twelve hours after admission, the estimated mortality risk was significantly lower in three time categories: t(1) (relative risk, 0.45; 95% confidence interval, 0.15 to 0.98; p = 0.03), t(3) (relative risk, 0.58; 95% confidence interval, 0.28 to 0.93; p = 0.03), and t(4) (relative risk, 0.43; 95% confidence interval, 0.10 to 0.94; p = 0.03). Patients with serious abdominal trauma (Abbreviated Injury Score, > or =3) experienced the greatest benefit from a delay of internal fixation beyond twelve hours (relative risk, 0.82 [95% confidence interval, 0.54 to 1.35] for patients with an Abbreviated Injury Score of <3, compared with 0.36 [95% confidence interval, 0.13 to 0.87] for those with an Abbreviated Injury Score of > or =3) (p value for effect modification, 0.09). Delayed repair of femoral shaft fracture beyond twelve hours in patients with multisystem trauma, which may allow time for appropriate resuscitation, reduces mortality by approximately 50%. Patients with serious abdominal injury benefit most from delayed treatment. These results support delaying definitive treatment of long-bone injuries in patients with multisystem trauma as a means of so-called damage-control in order to reduce adverse outcomes.


Plyometric training in female athletes. Decreased impact forces and increased hamstring torques.
The purpose of this study was to test the effect of a jump-training program on landing mechanics and lower extremity strength in female athletes involved in jumping sports. These parameters were compared before and after training with those of male athletes. The program was designed to decrease landing forces by teaching neuromuscular control of the lower limb during landing and to increase vertical jump height. After training, peak landing forces from a volleyball block jump decreased 22%, and knee adduction and abduction moments (medially and laterally directed torques) decreased approximately 50%. Multiple regression analysis revealed that these moments were significant predictors of peak landing forces. Female athletes demonstrated lower landing forces than male athletes and lower adduction and abduction moments after training. External knee extension moments (hamstring muscle-dominant) of male athletes were threefold higher than those of female athletes. Hamstring-to-quadriceps muscle peak torque ratios increased 26% on the nondominant side and 13% on the dominant side, correcting side-to-side imbalances. Hamstring muscle power increased 44% with training on the dominant side and 21% on the nondominant. Peak torque ratios of male athletes were significantly greater than those of untrained female athletes, but similar to those of trained females. Mean vertical jump height increased approximately 10%. This training may have a significant effect on knee stabilization and prevention of serious knee injury among female athletes.


Increased Resting Hippocampal and Basal Ganglia Perfusion in People at Ultra High Risk for Psychosis: Replication in a Second Cohort.
We recently reported that resting hippocampal, basal ganglia and midbrain perfusion is elevated in people at ultra high risk (UHR) for psychosis. The present study sought to replicate our previous finding in an independent UHR cohort, and examined the relationship between resting perfusion in these regions, psychosis and depression symptoms, and traumatic experiences in childhood. Pseudo-Continuous Arterial Spin Labelling (p-CASL) imaging was used to measure resting cerebral blood flow (rCBF) in 77 UHR for psychosis individuals and 25 healthy volunteers in a case-control design. UHR participants were recruited from clinical early detection services at 3 sites in the South of England. Symptoms levels were assessed using the Comprehensive Assessment of At Risk Mental States (CAARMS), the Hamilton Depression Scale (HAM-D), and childhood trauma was assessed retrospectively using the Childhood Trauma Questionnaire (CTQ). Right hippocampal and basal ganglia rCBF were significantly increased in UHR subjects compared to controls, partially replicating our previous finding in an independent cohort. In UHR participants, positive symptoms were positively correlated with rCBF in the right pallidum. CTQ scores were positively correlated with rCBF values in the bilateral hippocampus and negatively associated with rCBF in the left prefrontal cortex. Elevated resting hippocampal and basal ganglia activity appears to be a consistent finding in individuals at high risk for psychosis, consistent with data from preclinical models of the disorder. The association with childhood trauma suggests that its influence on the risk of psychosis may be mediated through an effect on hippocampal function.


Longitudinal study of circulating miR-122 in a rat model of non-alcoholic fatty liver disease.
Circulating microRNAs (miRs) may be promising biomarkers for several diseases. We previously found that miR-122 can function as a biomarker for non-alcoholic fatty liver disease (NAFLD). However, little is known regarding the time course of circulating miR-122 levels during the development of NAFLD. Here, we examined circulating miR-122 levels using a rat model of NAFLD. To clarify changes in serum levels of miR-122 during development of NAFLD, experimental rats were fed a high-fat diet (HFD) for 2-10 weeks, while control rats received standard chow. Serum and liver tissue was collected from all animals at 2, 6, and 10 weeks of feeding. Clinical laboratory parameters (cholesterol, TG, AST, ALT, NEFA) were determined by biochemistry analyzer. Hepatic lipid accumulation was estimated by Oil red O staining. Circulating miR-122 levels were then measured by real-time polymerase chain reaction. Over the 10 weeks of feeding, body weight, total liver lipids, and liver and serum triacylglycerol were increased in the HFD group compared to the control group. However, no significant changes in serum alanine aminotransferase activity were observed, suggesting that NAFLD status was mild. In contrast, we observed drastic up-regulation of circulating miR-122 levels. Our findings suggest that serum miR-122 level is indeed useful for assessing early NAFLD and might be superior to clinical markers traditionally used to monitor hepatic disease.


Universal amplification, next-generation sequencing, and assembly of HIV-1 genomes.
Whole HIV-1 genome sequences are pivotal for large-scale studies of inter- and intrahost evolution, including the acquisition of drug resistance mutations. The ability to rapidly and cost-effectively generate large numbers of HIV-1 genome sequences from different populations and geographical locations and determine the effect of minority genetic variants is, however, a limiting factor. Next-generation sequencing promises to bridge this gap but is hindered by the lack of methods for the enrichment of virus genomes across the phylogenetic breadth of HIV-1 and methods for the robust assembly of the virus genomes from short-read data. Here we report a method for the amplification, next-generation sequencing, and unbiased de novo assembly of HIV-1 genomes of groups M, N, and O, as well as recombinants, that does not require prior knowledge of the sequence or subtype. A sensitivity of at least 3,000 copies/ml was determined by using plasma virus samples of known copy numbers. We applied our novel method to compare the genome diversities of HIV-1 groups, subtypes, and genes. The highest level of diversity was found in the env, nef, vpr, tat, and rev genes and parts of the gag gene. Furthermore, we used our method to investigate mutations associated with HIV-1 drug resistance in clinical samples at the level of the complete genome. Drug resistance mutations were detected as both major variant and minor species. In conclusion, we demonstrate the feasibility of our method for large-scale HIV-1 genome sequencing. This will enable the phylogenetic and phylodynamic resolution of the ongoing pandemic and efficient monitoring of complex HIV-1 drug resistance genotypes.


Isolation and longterm culture of human intestinal microvascular endothelial cells.
Microvascular endothelial cells play an important part in inflammation as well as in organ specific leucocyte traffic, and may be functionally different from large vessel endothelium in this respect. This study therefore established a method for isolation and longterm culture of human intestinal microvascular endothelial cells (HIMEC). After dissociation by collagenase/dispase/DNase of mucosal and submucosal tissue obtained from normal adult jejunum, cells were plated and cultured to subconfluence in endothelial serum free medium containing 2.5% fetal calf serum, hydrocortisone, and N6, O2-dibutyryladenosine cyclic monophosphate. Primary cultures were trypsinised and endothelial cells were isolated by paramagnetic beads armed with monoclonal antibody to CD31. Optimal growth conditions for HIMEC cultures were established, allowing up to nine passages (three months in vitro). The cells contained Weibel-Palade bodies, expressed von Willebrand factor, CD31, and VE-cadherin; and bound Ulex Europaeus lectin I. A method to establish longterm cell cultures of HIMEC will facilitate further investigation of the function of intestinal endothelial cells and their participation in physiological and pathological events in the gut.


First-order symmetry-adapted perturbation theory for multiplet splittings.
We present a symmetry-adapted perturbation theory (SAPT) for the interaction of two high-spin open-shell molecules (described by their restricted open-shell Hartree-Fock determinants) resulting in low-spin states of the complex. The previously available SAPT formalisms, except for some system-specific studies for few-electron complexes, were restricted to the high-spin state of the interacting system. Thus, the new approach provides, for the first time, a SAPT-based estimate of the splittings between different spin states of the complex. We have derived and implemented the lowest-order SAPT term responsible for these splittings, that is, the first-order exchange energy. We show that within the so-called S2 approximation commonly used in SAPT (neglecting effects that vanish as fourth or higher powers of intermolecular overlap integrals), the first-order exchange energies for all multiplets are linear combinations of two matrix elements: a diagonal exchange term that determines the spin-averaged effect and a spin-flip term responsible for the splittings between the states. The numerical factors in this linear combination are determined solely by the Clebsch-Gordan coefficients: accordingly, the S2 approximation implies a Heisenberg Hamiltonian picture with a single coupling strength parameter determining all the splittings. The new approach is cast into both molecular-orbital and atomic-orbital expressions: the latter enable an efficient density-fitted implementation. We test the newly developed formalism on several open-shell complexes ranging from diatomic systems (Li⋯H, Mn⋯Mn, …) to the phenalenyl dimer.


An intragenic tandem duplication of genomic DNA is responsible for the f3N mutation of Drosophila melanogaster.
Among the numerous X chromosome-linked forked bristle (f) mutations described in Drosophila melanogaster, one designated f3N exhibits the unusual property of reverting spontaneously to wild type at an inordinate frequency, a frequency that can be increased with x-ray irradiation. In contrast to the f mutants described thus far, all of which are associated with the insertion of mobile DNA elements, f3N is associated with an intragenic duplication of 2.8 kb of genomic DNA that resolves to the normal sequence when reversions occur. Consideration is given to intrachromosomal recombination as the mechanism of reversion and truncation of the forked protein as a cause for the mutant phenotype.


FLT3 mutations in canine acute lymphocytic leukemia.
FMS-like tyrosine kinase 3 (FLT3) is a commonly mutated protein in a variety of human acute leukemias. Mutations leading to constitutively active FLT3, including internal tandem duplications of the juxtamembrane domain (ITD), result in continuous cellular proliferation, resistance to apoptotic cell death, and a poorer prognosis. A better understanding of the molecular consequences of FLT3 activation would allow improved therapeutic strategies in these patients. Canine lymphoproliferative diseases, including lymphoma and acute leukemias, share evolutionarily conserved chromosomal aberrations and exhibit conserved mutations within key oncogenes when compared to their human counterparts. A small percentage of canine acute lymphocytic leukemias (ALL) also exhibit FLT3 ITD mutations. We molecularly characterized FLT3 mutations in two dogs and one cell line, by DNA sequencing, gene expression analysis via quantitative real-time PCR, and sensitivity to the FLT3 inhibitor lestaurtinib via in vitro proliferation assays. FLT 3 and downstream mediators of FLT3 activation were assessed by Western blotting. The canine B-cell leukemia cell line, GL-1, and neoplastic cells from 2/7 dogs diagnosed cytologically with ALL were found to have FLT3 ITD mutations and FLT3 mRNA up-regulation. Lestaurtinib, a small molecule FLT3 inhibitor, significantly inhibited the growth of GL-1 cells, while not affecting the growth of two other canine lymphoid cell lines without the FLT3 mutation. Finally, western blots were used to confirm the conserved downstream mediators of FLT3 activating mutations. These results show that ALL and FLT3 biology is conserved between canine and human patients, supporting the notion that canine ALL, in conjunction with the GL-1 cell line, will be useful in the development of a relevant large animal model to aid in the study of human FLT3 mutant leukemias.


WE-C-217A-01: Risk Estimation versus Risk Perception.
The recent attention in the scientific and popular press to the potential risks associated with ionizing radiation from medical imaging has resulted in some patient becoming unwilling to undergo medically appropriate procedures that use ionizing radiation. Scientific data on the magnitude of both potential risks and benefits are not always successful in allaying patient concerns, even though the potential risks are very small and the potential benefits are very large. An understanding of the psychological factors affecting the perception of risk helps to explain this behavior. For patients to feel comfortable with medical procedures involving ionizing radiation, they must be believe that their medical providers have their best interest in mind, that the ordered procedure is medically appropriate and may help their doctor in managing their care, and that those providing the imaging services are actively engaged in efforts to ensure patient safety. 1. Differentiate between risk estimation and risk perception 2. List several factors that affect a person's perception of risk 3. Describe some of the consequences that may occur when patients do not feel that they are safe.


Characterization of the role of very late expression factor 1 in baculovirus capsid structure and DNA processing.
Very late expression factor 1 (VLF-1) of Autographa californica multiple nucleopolyhedrovirus is a putative tyrosine recombinase and is required for both very late gene expression and budded virus production. In this report, we show that a vlf-1 knockout bacmid was able to synthesize viral DNA at levels similar to that detected for a gp64 knockout bacmid that served as a noninfectious control virus. Additionally, analysis of replicated bacmid DNA by field-inversion gel electrophoresis indicated that VLF-1 is not required for synthesizing high-molecular-weight intermediates that could be resolved into unit-length genomes when cut at a unique restriction site. However, immunoelectron microscopic analysis revealed that in cells transfected with a vlf-1 knockout bacmid, aberrant tubular structures containing the capsid protein vp39 were observed, suggesting that this virus construct was defective in producing mature capsids. In contrast, rescuing the vlf-1 knockout bacmid construct with a copy of VLF-1 that carries a mutation of a highly conserved tyrosine (Y355F) was sufficient to restore the production of nucleocapsids with a normal appearance, but not infectious virus production. Furthermore, the results of a DNase I protection assay indicated that the DNA packaging efficiency of the VLF-1(Y355F) virus construct was similar to that of the gp64 knockout control. Finally, a recombinant virus containing a functional hemagglutinin epitope-tagged version of VLF-1 was constructed to investigate the association of VLF-1 with the nucleocapsid. Analysis by immunoelectron microscopy of Sf-9 cells infected with this virus showed that VLF-1 localized to an end region of the nucleocapsid. Collectively, these results indicate that VLF-1 is required for normal capsid assembly and serves an essential function during the final stages of the DNA packaging process.


Cost-effective large-scale fabrication of diffractive optical elements by using conventional semiconducting processes.
In this article, we introduce a simple fabrication method for SiO(2)-based thin diffractive optical elements (DOEs) that uses the conventional processes widely used in the semiconductor industry. Photolithography and an inductively coupled plasma etching technique are easy and cost-effective methods for fabricating subnanometer-scale and thin DOEs with a refractive index of 1.45, based on SiO(2). After fabricating DOEs, we confirmed the shape of the output light emitted from the laser diode light source and applied to a light-emitting diode (LED) module. The results represent a new approach to mass-produce DOEs and realize a high-brightness LED module.


Silicene growth through island migration and coalescence.
We perform massively-parallel classical molecular dynamics (MD) simulations to study the long timescale monolayer silicene growth on an Ir (111) surface. We observe an intricate multi-stage growth process driven by atomic and cluster migration on the surface. Initial growth involves formation of sub-nanometer clusters via adatom surface diffusion. Subsequently, these clusters rearrange spontaneously with each additional Si atom, forming clusters containing 4-7 member rings. Growth of each cluster through adatom adhesion is accompanied by the formation of larger islands through cluster migration and coalescence. Coalescence of smaller, more mobile islands into larger clusters is aided by the internal rearrangement of rings within each cluster. This flexibility, both of clusters and their constituent atoms, allows the impinging clusters to reorient after first contact and form a more perfect union. We also report on the effect of temperature and flux on the growth process and the final nanostructure. Our study provides atomistic insights into the early stage growth mechanisms of silicene which can be significant for controlled synthesis of its 2D monolayers.


Polymorphisms and promoter overactivity of the p22(phox) gene in vascular smooth muscle cells from spontaneously hypertensive rats.
In a previous study, we found that the p22(phox) subunit of the NADH/NADPH oxidase is overexpressed in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHRs) with enhanced vascular production of superoxide anion ((.)O(2)(-)). Thus, we have investigated whether changes in the sequence or activity of the promoter region of p22(phox) gene are present in SHRs. To carry out this analysis, first of all, we characterized the rat gene structure and promoter region for the p22(phox) subunit. The p22(phox) gene spans approximately 10 kb and contains 6 exons and 5 introns. Primer extension analysis indicated the transcriptional start site 100 bp upstream from the translational start site. The immediate promoter region of the p22(phox) gene does not contain a TATA box, but there are a CCAC box and putative recognition sites for nuclear factors, such as SP1, gamma-interferon, and nuclear factor-kappaB. Using reporter-gene transfection analysis, we found that this promoter was functional in VSMCs. Furthermore, we observed that p22(phox) promoter activity was significantly higher in VSMCs from SHRs than from normotensive Wistar-Kyoto rats. In addition, we found that there were 5 polymorphisms in the sequence of p22(phox) promoter between Wistar-Kyoto rats and SHRs and that they were functional. The results obtained in this study provide a tool to explore the mechanisms that regulate the expression of p22(phox) gene in rat VSMCs. Furthermore, our findings show that changes in the sequence of p22(phox) gene promoter and in the degree of activation of VSMCs are responsible for upregulated expression of p22(phox) in SHRs.


Mind the gap: diversity and reactivity relationships among multihaem cytochromes of the MtrA/DmsE family.
Shewanella oneidensis MR-1 has the ability to use many external terminal electron acceptors during anaerobic respiration, such as DMSO. The pathway that facilitates this electron transfer includes the decahaem cytochrome DmsE, a paralogue of the MtrA family of decahaem cytochromes. Although both DmsE and MtrA are decahaem cytochromes implicated in the long-range electron transfer across a ~300 Å (1 Å=0.1 nm) wide periplasmic 'gap', MtrA has been shown to be only 105 Å in maximal length. In the present paper, DmsE is further characterized via protein film voltammetry, revealing that the electrochemistry of the DmsE haem cofactors display macroscopic potentials lower than those of MtrA by 100 mV. It is possible this tuning of the redox potential of DmsE is required to shuttle electrons to the outer-membrane proteins specific to DMSO reduction. Other decahaem cytochromes found in S. oneidensis, such as the outer-membrane proteins MtrC, MtrF and OmcA, have been shown to have electrochemical properties similar to those of MtrA, yet possess a different evolutionary relationship.


OARSI Clinical Trials Recommendations: Design, conduct, and reporting of clinical trials for knee osteoarthritis.
The goal of this document is to update the original OARSI recommendations specifically for the design, conduct, and reporting of clinical trials that target symptom or structure modification among individuals with knee osteoarthritis (OA). To develop recommendations for the design, conduct, and reporting of clinical trials for knee OA we initially drafted recommendations through an iterative process. Members of the working group included representatives from industry and academia. After the working group members reviewed a final draft, they scored the appropriateness for recommendations. After the members voted we calculated the median score among the nine members of the working group who completed the score. The document includes 25 recommendations regarding randomization, blocking and stratification, blinding, enhancing accuracy of patient-reported outcomes (PRO), selecting a study population and index knee, describing interventions, patient-reported and physical performance measures, structural outcome measures, biochemical biomarkers, and reporting recommendations. In summary, the working group identified 25 recommendations that represent the current best practices regarding clinical trials that target symptom or structure modification among individuals with knee OA. These updated recommendations incorporate novel technologies (e.g., magnetic resonance imaging (MRI)) and strategies to address the heterogeneity of knee OA.


A comparative study between cone-beam computed tomography and periapical radiographs in the diagnosis of simulated endodontic complications.
To compare cone-beam computed tomography (CBCT) with periapical radiography for the identification of simulated endodontic complications. Sixteen human teeth, in three mandibles, were submitted to the following simulated endodontic complications: G1) fractured endodontic file; G2) root perforation; G3) cast post with deviation; G4) external root resorption. Periapical radiographs were taken of each tooth at three different angles, and CBCT scan was taken. One calibrated examiner who was specialized in dental radiology interpreted the images. The results were analysed using the following scoring system: 0 - unidentified alteration; 1 - alteration identified with inaccurate diagnosis; and 2 - alteration identified with accurate diagnosis. Data were analysed using McNemar and Wilcoxon tests (alfa=0.05). In the overall assessment, CBCT was superior when compared with periapical radiographs (P<0.05). When individual results on each complication were evaluated, CBCT was superior only in the identification of external root resorption (100% Score 2) (P<0.05). Cone-beam computed tomography could be an alternative to periapical radiographs especially in the detection and assessment of external root resorption.


Abundant genetic overlap between blood lipids and immune-mediated diseases indicates shared molecular genetic mechanisms.
Epidemiological studies suggest a relationship between blood lipids and immune-mediated diseases, but the nature of these associations is not well understood. We used genome-wide association studies (GWAS) to investigate shared single nucleotide polymorphisms (SNPs) between blood lipids and immune-mediated diseases. We analyzed data from GWAS (n~200,000 individuals), applying new False Discovery Rate (FDR) methods, to investigate genetic overlap between blood lipid levels [triglycerides (TG), low density lipoproteins (LDL), high density lipoproteins (HDL)] and a selection of archetypal immune-mediated diseases (Crohn's disease, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, celiac disease, psoriasis and sarcoidosis). We found significant polygenic pleiotropy between the blood lipids and all the investigated immune-mediated diseases. We discovered several shared risk loci between the immune-mediated diseases and TG (n = 88), LDL (n = 87) and HDL (n = 52). Three-way analyses differentiated the pattern of pleiotropy among the immune-mediated diseases. The new pleiotropic loci increased the number of functional gene network nodes representing blood lipid loci by 40%. Pathway analyses implicated several novel shared mechanisms for immune pathogenesis and lipid biology, including glycosphingolipid synthesis (e.g. FUT2) and intestinal host-microbe interactions (e.g. ATG16L1). We demonstrate a shared genetic basis for blood lipids and immune-mediated diseases independent of environmental factors. Our findings provide novel mechanistic insights into dyslipidemia and immune-mediated diseases and may have implications for therapeutic trials involving lipid-lowering and anti-inflammatory agents.


Competing phytoplankton undermines allelopathy of a bloom-forming dinoflagellate.
Biotic interactions in the plankton can be both complex and dynamic. Competition among phytoplankton is often chemically mediated, but no studies have considered whether allelopathic compounds are modified by biotic interactions. Here, we show that compounds exuded during Karenia brevis blooms were allelopathic to the cosmopolitan diatom Skeletonema costatum, but that bloom allelopathy varied dramatically among collections and years. We investigated several possible causes of this variability and found that neither bloom density nor concentrations of water-borne brevetoxins correlated with allelopathic potency. However, when we directly tested whether the presence of competing phytoplankton influenced bloom allelopathy, we found that S. costatum reduced the growth-inhibiting effects of bloom exudates, suggesting that S. costatum has a mechanism for undermining K. brevis allelopathy. Additional laboratory experiments indicated that inducible changes to K. brevis allelopathy were restricted to two diatoms among five sensitive phytoplankton species, whereas five other species were constitutively resistant to K. brevis allelopathy. Our results suggest that competitors differ in their responses to phytoplankton allelopathy, with S. costatum exhibiting a previously undescribed method of resistance that may influence community structure and alter bloom dynamics.


Rapid changes in the incidence of urinary system cancers in Turkey.
Estimation of national cancer incidence for major cancer sites in Turkey has been carried out by analyzing the data obtained from active cancer registry, and published regularly by Institute of Public Health of Ministry of Health. In the light of these statistics, the incidence of urinary cancers in both sexes and their age related distributions have been discussed, paying special attention to prostate, kidney and bladder cancers. The annual incidence of all cancer cases increased gradually, reaching to 221.5 per 100,000 population in 2009, the latest confirmed figure available at present. Among males the most frequent cancers were those of the lung, prostate and bladder. The incidence rates of urinary cancers among males were 36.1, 21.4 and 6.3 per 100,000 for prostate, bladder and kidney respectively. The reliability of current data concerning the incidence of cancer has been discussed by comparing them with the previously reported national cancer data.


Mouse early extra-embryonic lineages activate compensatory endocytosis in response to poor maternal nutrition.
Mammalian extra-embryonic lineages perform the crucial role of nutrient provision during gestation to support embryonic and fetal growth. These lineages derive from outer trophectoderm (TE) and internal primitive endoderm (PE) in the blastocyst and subsequently give rise to chorio-allantoic and visceral yolk sac placentae, respectively. We have shown maternal low protein diet exclusively during mouse preimplantation development (Emb-LPD) is sufficient to cause a compensatory increase in fetal and perinatal growth that correlates positively with increased adult-onset cardiovascular, metabolic and behavioural disease. Here, to investigate early mechanisms of compensatory nutrient provision, we assessed the influence of maternal Emb-LPD on endocytosis within extra-embryonic lineages using quantitative imaging and expression of markers and proteins involved. Blastocysts collected from Emb-LPD mothers within standard culture medium displayed enhanced TE endocytosis compared with embryos from control mothers with respect to the number and collective volume per cell of vesicles with endocytosed ligand and fluid and lysosomes, plus protein expression of megalin (Lrp2) LDL-family receptor. Endocytosis was also stimulated using similar criteria in the outer PE-like lineage of embryoid bodies formed from embryonic stem cell lines generated from Emb-LPD blastocysts. Using an in vitro model replicating the depleted amino acid (AA) composition found within the Emb-LPD uterine luminal fluid, we show TE endocytosis response is activated through reduced branched-chain AAs (leucine, isoleucine, valine). Moreover, activation appears mediated through RhoA GTPase signalling. Our data indicate early embryos regulate and stabilise endocytosis as a mechanism to compensate for poor maternal nutrient provision.


Tau phosphorylation and cleavage in ethanol-induced neurodegeneration in the developing mouse brain.
Previous studies indicated that ethanol-induced neurodegeneration in postnatal day 7 (P7) mice, widely used as a model for the fetal alcohol spectrum disorders, was accompanied by glycogen synthase kinase-3beta (GSK-3beta) and caspase-3 activation. Presently, we examined whether tau, a microtubule associated protein, is modified by GSK-3beta and caspase-3 in ethanol-treated P7 mouse forebrains. We found that ethanol increased phosphorylated tau recognized by the paired helical filament (PHF)-1 antibody and by the antibody against tau phosphorylated at Ser199. Ethanol also generated tau fragments recognized by an antibody against caspase-cleaved tau (C-tau). C-tau was localized in neurons bearing activated caspase-3 and fragmented nuclei. Over time, cell debris and degenerated projections containing C-tau appeared to be engulfed by activated microglia. A caspase-3 inhibitor partially blocked C-tau formation. Lithium, a GSK-3beta inhibitor, blocked ethanol-induced caspase-3 activation, phosphorylated tau elevation, C-tau formation, and microglial activation. These results indicate that tau is phosphorylated by GSK-3beta and cleaved by caspase-3 during ethanol-induced neurodegeneration in the developing brain.


First Report of Boeremia Blight Caused by Boeremia exigua var. exigua on Pyrethrum in Australia.
Pyrethrum (Tanacetum cinerariifolium) is produced for extraction of insecticidal compounds from the flower achenes. In 2004 and 2006, isolations from necrotic lesions on stems and leaves in three fields in northern Tasmania, Australia yielded four unidentified fungal isolates. Leaf lesions were medium brown and circular (2 to 4 mm in diameter) or irregular in shape (2 to 5 mm long). Stem lesions were irregular, necrotic spots, 5 to 15 mm below the flower peduncle, medium brown, 2 to 4 mm long, and 1 to 2 mm wide. Isolations were conducted on water agar following surface sterilization. Isolates were identified by colony characteristics and the presence of metabolite 'E' (1). On oatmeal agar (OA), colonies had irregular margins, were greenish olivaceous-to-olivaceous gray with sparse, white, floccose, aerial mycelia. On malt extract agar (MEA), cultures were variable in color with olivaceous black centers with soft, dense, aerial mycelia. Conidia were hyaline, ellipsoidal to oblong, mainly aseptate, but occasionally 1-septate with dimensions ranging from 2.5 to 7.5 × 1.8 to 3.8 μm (length/width ratio = 1.7 to 2.1). All isolates had moderate reactions to the NaOH test for metabolite 'E'. DNA was extracted from all four isolates with a DNeasy Plant Mini Kit (QIAGEN Inc., Valencia, CA). For identification, the internal transcribed spacer region (ITS1, 5.8s, and ITS2) and part of the translation elongation factor (TEF) region were amplified and sequenced. Primers ITS1 and ITS4 (2) were used for the ITS region and primers EFCF1 (5'-AGTGCGGTGGTATCGACAAG) and EFCF6 (3'-CATGTCACGGACGGCGAAAC) were used for the TEF. Amplicons were sequenced in both directions and consensus sequences assembled. The ITS sequence was 100% identical to Boeremia exigua var. exigua (GenBank Accession No. GU237715). Base pairs 413 to 1,214 of the TEF sequence from the pyrethrum isolates matched base pairs 1 to 802 (799 of 802 identities) of B. exigua var. exigua (GenBank Accession No. GU349080). All isolates were confirmed as B. exigua var. exigua using morphology and sequencing. Pathogenicity tests were conducted three times in separate glasshouse trials for two of the four isolates. For each isolate, conidial suspensions in water (3 ml/plant) from MEA, adjusted to 5 × 105/ml were applied with Tween 20 (1 drop per 100 ml of water) to 8-week-old pyrethrum plants (five pots per isolate with four plants per pot) using a hand-held spray bottle. Twenty plants were sprayed with water and Tween 20 as nontreated controls. Plants were covered with plastic bags for 48 h after inoculation and examined for symptoms after 15 days at 20°C. Disease incidence (number of symptomatic leaves affected per total number of leaves) of the inoculated plants varied from 7.5 to 9.4%. Noninoculated plants did not develop symptoms. Isolations resulted in cultures morphologically identical on MEA and OA to those inoculated. To our knowledge, this is the first report of B. exigua var. exigua causing disease in pyrethrum. Cultures were deposited in the New South Wales Department of Agriculture collection (DAR79101 to 79104) and TEF and ITS sequences for DAR79101 in GenBank (Accession Nos. JF925328 and JF925329, respectively). Boeremia blight is likely to contribute to the fungal disease complex causing reductions in green leaf area in Australian pyrethrum production. References: (1) M. M. Aveskamp et al. Stud. Mycol. 65:1, 2010. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.


Development and validation of a clinically applicable arm use monitor for people after stroke.
To develop and validate a clinically applicable and easy-to-use accelerometry-based device to measure arm use in people after stroke; the Activ8 arm use monitor (Activ8-AUM). Development and validation study. A total of 25 people at different stages of rehabilitation after stroke were included in this study. The Activ8-AUM consists of 3 single-sensor Activ8s: one on the unaffected thigh and one on each wrist. Arm use was calculated by combining movement intensity of the arms with data from body posture and movements on the leg sensor. Data were divided into 2 sets: one for determining situation-specific movement intensity thresholds for arm use, and the other to validate the Activ8-AUM using video recordings. Overall agreement between the Activ8-AUM and video recordings was 75%, sensitivity was 73% and specificity was 77%. Agreement between the different categories of arm use ranged from 42% to 93% for the affected arm and from 24% to 82% for the unaffected arm. By combining the movement intensity threshold with body posture and movements, good agreement was reached between the Activ8-AUM and video recordings. This result, together with the easy-to-use configuration, makes the Activ8-AUM a promising device to measure arm use in people after stroke.


Beam quality measurements using digitized laser beam images.
A method is described for measuring various laser beam characteristics with modest experimental complexity by digital processing of the near and far field images. Gaussian spot sizes, peak intensities, and spatial distributions of the images are easily found. Far field beam focusability is determined by computationally applying apertures of circular or elliptical diameters to the digitized image. Visualization of the magnitude of phase and intensity distortions is accomplished by comparing the 2-D fast Fourier transform of both smoothed and unsmoothed near field data to the actual far field data. The digital processing may be performed on current personal computers to give the experimenter unprecedented capabilities for rapid beam characterization at relatively low cost.


An enhanced cerium(IV)-rhodamine 6G chemiluminescence system using guest-host interactions in a lab-on-a-chip platform for estimating the total phenolic content in food samples.
Two chemiluminescence-microfluidic (CL-MF) systems, e.g., Ce(IV)-rhodamine B (RB) and Ce(IV)-rhodamine 6G (R6G), for the determination of the total phenolic content in teas and some sweeteners were evaluated. The results indicated that the Ce(IV)-R6G system was more sensitive in comparison to the Ce(IV)-RB CL system. Therefore, a simple (CL-MF) method based on the CL of Ce(IV)-R6G was developed, and the sensitivity, selectivity and stability of this system were evaluated. Selected phenolic compounds (PCs), such as quercetin (QRC), catechin (CAT), rutin (RUT), gallic acid (GA), caffeic acid (CA) and syringic acid (SA), produced analytically useful chemiluminescence signals with low detection limits ranging from 0.35 nmol L(-1) for QRC to 11.31 nmol L(-1) for SA. The mixing sequence and the chip design were crucial, as the sensitivity and reproducibility could be substantially affected by these two factors. In addition, the anionic surfactant (i.e., sodium dodecyl sulfate (SDS)) can significantly enhance the CL signal intensity by as much as 300% for the QRC solution. Spectroscopic studies indicated that the enhancement was due to a strong guest-host interaction between the cationic R6G molecules and the anionic amphiphilic environment. Other parameters that could affect the CL intensities of the PCs were carefully optimized. Finally, the method was successfully applied to tea and sweetener samples. Six different tea samples exhibited total phenolic/antioxidant levels from 7.32 to 13.5 g per 100g of sample with respect to GA. Four different sweetener samples were also analyzed and exhibited total phenolic/antioxidant levels from 500.9 to 3422.9 mg kg(-1) with respect to GA. The method was selective, rapid and sensitive when used to estimate the total phenolic/antioxidant level, and the results were in good agreement with those reported for honey and tea samples.


Randomized Trial of Medical versus Surgical Treatment for Refractory Heartburn.
Heartburn that persists despite proton-pump inhibitor (PPI) treatment is a frequent clinical problem with multiple potential causes. Treatments for PPI-refractory heartburn are of unproven efficacy and focus on controlling gastroesophageal reflux with reflux-reducing medication (e.g., baclofen) or antireflux surgery or on dampening visceral hypersensitivity with neuromodulators (e.g., desipramine). Patients who were referred to Veterans Affairs (VA) gastroenterology clinics for PPI-refractory heartburn received 20 mg of omeprazole twice daily for 2 weeks, and those with persistent heartburn underwent endoscopy, esophageal biopsy, esophageal manometry, and multichannel intraluminal impedance-pH monitoring. If patients were found to have reflux-related heartburn, we randomly assigned them to receive surgical treatment (laparoscopic Nissen fundoplication), active medical treatment (omeprazole plus baclofen, with desipramine added depending on symptoms), or control medical treatment (omeprazole plus placebo). The primary outcome was treatment success, defined as a decrease of 50% or more in the Gastroesophageal Reflux Disease (GERD)-Health Related Quality of Life score (range, 0 to 50, with higher scores indicating worse symptoms) at 1 year. A total of 366 patients (mean age, 48.5 years; 280 men) were enrolled. Prerandomization procedures excluded 288 patients: 42 had relief of their heartburn during the 2-week omeprazole trial, 70 did not complete trial procedures, 54 were excluded for other reasons, 23 had non-GERD esophageal disorders, and 99 had functional heartburn (not due to GERD or other histopathologic, motility, or structural abnormality). The remaining 78 patients underwent randomization. The incidence of treatment success with surgery (18 of 27 patients, 67%) was significantly superior to that with active medical treatment (7 of 25 patients, 28%; P = 0.007) or control medical treatment (3 of 26 patients, 12%; P<0.001). The difference in the incidence of treatment success between the active medical group and the control medical group was 16 percentage points (95% confidence interval, -5 to 38; P = 0.17). Among patients referred to VA gastroenterology clinics for PPI-refractory heartburn, systematic workup revealed truly PPI-refractory and reflux-related heartburn in a minority of patients. For that highly selected subgroup, surgery was superior to medical treatment. (Funded by the Department of Veterans Affairs Cooperative Studies Program; ClinicalTrials.gov number, NCT01265550.).


Elective induction of labor at 39 weeks compared with expectant management: a meta-analysis of cohort studies.
Elective induction of labor at 39 weeks among low-risk nulliparous women has reduced the chance of cesarean and other adverse maternal and perinatal outcomes in a randomized trial, although its clinical effectiveness in nonresearch settings remains uncertain. To perform a systematic review of observational studies that compared elective induction of labor at 39 weeks among nulliparous women with expectant management and to use meta-analytic techniques to estimate the association of elective induction with cesarean delivery, as well as other maternal and perinatal outcomes. Studies were eligible for this meta-analysis only if they: (1) were observational; (2) compared women undergoing labor induction at 39 weeks with women undergoing expectant management beyond that gestational age; (3) included women in the induction group only if they had no other indication for labor induction at 39 weeks; and (4) provided data specifically for nulliparous women. The predefined primary outcome was cesarean delivery, and secondary outcomes representing other maternal and perinatal morbidities also were evaluated. Outcome data from different studies were combined to estimate pooled relative risks with 95% confidence intervals using random-effects models. Of 375 studies identified by the initial search, 6 cohort studies, which included 66,019 women undergoing elective labor induction at 39 weeks and 584,390 undergoing expectant management, met inclusion criteria. Elective induction of labor at 39 weeks was associated with a significantly lower frequency of cesarean delivery (26.4% vs 29.1%; relative risk, 0.83; 95% confidence interval, 0.74-0.93), as well as of peripartum infection (2.8% vs 5.2%; relative risk, 0.53; 95% confidence interval, 0.39-0.72). Neonates of women in the induction group were less likely to have respiratory morbidity (0.7% vs 1.5%; relative risk, 0.71; 95% confidence interval, 0.59-0.85); meconium aspiration syndrome (0.7% vs 3.0%; relative risk, 0.49; 95% confidence interval, 0.26-0.92); and neonatal intensive care unit admission (3.5% vs 5.5%; relative risk, 0.80; 95% confidence interval, 0.72-0.88). There also was a lower risk of perinatal mortality (0.04% vs 0.2%; relative risk, 0.27; 95% confidence interval, 0.09-0.76). This meta-analysis of 6 cohort studies demonstrates that elective induction of labor at 39 weeks, compared with expectant management beyond that gestational age, was associated with a significantly lower risk of cesarean delivery, maternal peripartum infection, and perinatal adverse outcomes, including respiratory morbidity, intensive care unit admission, and mortality.


Fluorescent in situ folding control for rapid optimization of cell-free membrane protein synthesis.
Cell-free synthesis is an open and powerful tool for high-yield protein production in small reaction volumes predestined for high-throughput structural and functional analysis. Membrane proteins require addition of detergents for solubilization, liposomes, or nanodiscs. Hence, the number of parameters to be tested is significantly higher than with soluble proteins. Optimization is commonly done with respect to protein yield, yet without knowledge of the protein folding status. This approach contains a large inherent risk of ending up with non-functional protein. We show that fluorophore formation in C-terminal fusions with green fluorescent protein (GFP) indicates the folding state of a membrane protein in situ, i.e. within the cell-free reaction mixture, as confirmed by circular dichroism (CD), proteoliposome reconstitution and functional assays. Quantification of protein yield and in-gel fluorescence intensity imply suitability of the method for membrane proteins of bacterial, protozoan, plant, and mammalian origin, representing vacuolar and plasma membrane localization, as well as intra- and extracellular positioning of the C-terminus. We conclude that GFP-fusions provide an extension to cell-free protein synthesis systems eliminating the need for experimental folding control and, thus, enabling rapid optimization towards membrane protein quality.


Co-Salient Object Detection Based on Deep Saliency Networks and Seed Propagation Over an Integrated Graph.
This paper presents a co-salient object detection method to find common salient regions in a set of images. We utilize deep saliency networks to transfer co-saliency prior knowledge and better capture high-level semantic information. The resulting initial co-saliency maps are enhanced by seed propagation steps over an integrated graph. The deep saliency networks are trained in a supervised manner to avoid weakly supervised online learning and exploit them not only to extract high-level features but also to produce both intra- and inter-image saliency maps. Through a refinement step, the initial co-saliency maps can uniformly highlight co-salient regions and locate accurate object boundaries. To handle input image groups inconsistent in size, we propose to pool multi-regional descriptors including both within-segment and within-group information. In addition, the integrated multilayer graph is constructed to find the regions that the previous steps may not detect by seed propagation with low-level descriptors. In this paper, we utilize the useful complementary components of high- and low-level information and several learning-based steps. Our experiments have demonstrated that the proposed approach outperforms comparable co-saliency detection methods on widely used public databases and can also be directly applied to co-segmentation tasks.


A comprehensive genetic study of autosomal recessive ocular albinism in Caucasian patients.
Autosomal recessive ocular albinism (AROA) is a group of genetic disorders in which reduced pigmentation of the eye is associated with decreased visual acuity, nystagmus, strabismus, and photophobia, although pigmentation of skin and hair is relatively normal. Previous studies have shown that AROA in some cases constitutes a clinically mild presentation of oculocutaneous albinism (OCA), due to mutations in either the TYR (OCA1) or OCA2 (P) genes. The purpose of this study was to characterize the relative prevalence of different genetic forms of AROA, and to characterize a sample repertoire of gene mutations in a large series of Caucasian patients with AROA. Thirty-six unrelated Caucasian patients carrying the clinical diagnosis of AROA were studied by DNA sequence analysis of the four classic OCA genes: TYR, OCA2 (P), TYRP1, and SLC45A2 (MATP), as appropriate. In all patients with no apparent pathologic mutations in these genes, DNA sequence analysis was performed of a candidate OCA gene, SILV, and the two genes most often involved in Hermansky-Pudlak syndrome, HPS1 and HPS4, the most frequent syndromic form of OCA. TYR gene mutations were identified in 20 (56%) patients, OCA2 mutations in 3 (8%), mutations in both TYR and OCA2 in 2 (6%), and possible TYRP1 mutations in 2 (6%). In at least nine patients, no mutations were found in any of the genes studied. Almost all patients with OCA1-related AROA were compound heterozygous for severe OCA1 mutant alleles and the common R402Q variant. Most patients with AROA represent phenotypically mild variants of OCA, well over half of which is OCA1.


The Tailored Activity Program to reduce behavioral symptoms in individuals with dementia: feasibility, acceptability, and replication potential.
The Tailored Activity Program (TAP) is a home-based occupational therapy intervention shown to reduce behavioral symptoms and caregiver burden in a randomized trial. This article describes TAP, its assessments, acceptability, and replication potential. TAP involves 8 sessions for a period of 4 months. Interventionists identify preserved capabilities, previous roles, habits, and interests of individuals with dementia; develop activities customized to individual profiles; and train families in activity use. Interventionists documented time spent and ease conducting assessments, and observed receptivity of TAP. For each implemented prescribed activity, caregivers reported the amount of time their relative spent in activity and perceived benefits. The TAP assessment, a combination of neuropsychological tests, standardized performance-based observations, and clinical interviewing, yielded information on capabilities from which to identify and tailor activities. Assessments were easy to administer, taking an average of two 1-hr sessions. Of 170 prescribed activities, 81.5% were used, for an average of 4 times for 23 min by families between treatment sessions for a period of months. Caregivers reported high confidence in using activities, being less upset with behavioral symptoms (86%), and enhanced skills (93%) and personal control (95%). Interventionists observed enhanced engagement (100%) and pleasure (98%) in individuals with dementia during sessions. TAP offers families knowledge of their relative's capabilities and easy-to-use activities. The program was well received by caregivers. Prescribed activities appeared to be pleasurable and engaging to individuals with dementia. TAP merits further evaluation to establish efficacy with larger more diverse populations and consideration as a nonpharmacological approach to manage behavioral symptoms.


Activation of CB2 receptors as a potential therapeutic target for migraine: evaluation in an animal model.
Experimental animal models of migraine have suggested the existence of interactions between the endocannabinoid system and pain mediation in migraine. Extensive evidence has demonstrated a role for the cannabinoid-1 (CB1) receptor in antinociception. However, recent research suggests that also CB2 receptors, especially located outside the central nervous system, play a role in the perception of pain. Systemic administration of nitroglycerin (NTG) consistently induces spontaneous-like headache attacks in migraneurs; in the rat, systemic NTG induces a condition of hyperalgesia, probably through the activation of cerebral/spinal structures involved in nociceptive transmission. In this study we evaluated the role of CB2 receptors in two animal models of pain that may be relevant for migraine: the tail flick test and the formalin test performed during NTG-induced hyperalgesia. The study was performed in male Sprague-Dawley rats pre-treated with NTG (10 mg/kg, i.p.) or vehicle (4 hours before) and treated with the CB2 agonist AM1241 o dimethylsulfoxide (DMSO) 60 minutes before both the tail flick test and the formalin test. AM1241 showed a significant analgesic effect in baseline conditions in both tests. Furthermore, when administered 3 hours after NTG administration, AM1241 at both doses significantly reduced the total number of flinches/shakes during phase II of the test. These findings suggest that the pharmacological manipulation of the CB2 receptor may represent a potential therapeutic tool for the treatment of migraine.


Phase behavior and design rules for plastic colloidal crystals of hard polyhedra via consideration of directional entropic forces.
Plastic crystals - like liquid crystals - are mesophases that can exist between liquids and crystals and possess some of the characteristic traits of each of these states of matter. Plastic crystals exhibit translational order but orientational disorder. Here, we characterize the phase behavior in systems of hard polyhedra that self-assemble plastic face-centered cubic (pFCC) colloidal crystals. We report a first-order transition from a pFCC to a body-centered tetragonal (BCT) crystal, a smooth crossover from pFCC to an orientationally-ordered FCC crystal, and an apparent orientational glass transition wherein long-range order fails to develop from a plastic crystal upon an increase in density. Using global order parameters and local environment descriptors, we describe how particle shape influences the development of orientational order with increasing density, and we provide design rules based on the arrangement of facets for engineering plastic crystal behavior in colloidal systems.


Off-axis digital holographic microscopy with LED illumination based on polarization filtering.
A reflection mode digital holographic microscope with light emitting diode (LED) illumination and off-axis interferometry is proposed. The setup is comprised of a Linnik interferometer and a grating-based 4f imaging unit. Both object and reference waves travel coaxially and are split into multiple diffraction orders in the Fourier plane by the grating. The zeroth and first orders are filtered by a polarizing array to select orthogonally polarized object waves and reference waves. Subsequently, the object and reference waves are combined again in the output plane of the 4f system, and then the hologram with uniform contrast over the entire field of view can be acquired with the aid of a polarizer. The one-shot nature in the off-axis configuration enables an interferometric recording time on a millisecond scale. The validity of the proposed setup is illustrated by imaging nanostructured substrates, and the experimental results demonstrate that the phase noise is reduced drastically by an order of 68% when compared to a He-Ne laser-based result.


Prospective memory or prospective attention: physiological and pharmacological support for an attentional model.
Previous studies have reported that nicotine, a cholinergic agonist, could improve prospective memory (PM) - memory for a delayed intention - in healthy young adults. In the present study, we asked whether nicotine effects on PM performance were attributable to a drug-induced non-specific increase in arousal. Therefore, a double-blind, placebo-controlled study compared the effect of nicotine to the effect of an arousal manipulation on PM performance. All participants were non-smokers; half received 1 mg nicotine via a nasal spray and half received a matched placebo. Within these groups, half of the volunteers were exposed to hard anagrams and exhibited heightened tense arousal, while half of the volunteers were given easy anagrams and showed no change in arousal. These manipulations resulted in four conditions, placebo/low-arousal (n=12), placebo/high-arousal (n=10), nicotine/low-arousal (n=12), nicotine/high-arousal (n=13). All participants completed an ongoing lexical decision task while maintaining a PM intention (to make a separate, non-focal, response to certain items embedded within the ongoing task). When introduced separately, both nicotine and high tense arousal improved PM performance, but when combined, this improvement was eliminated. It is argued that both nicotine and high tense arousal increase attentional resources, specifically improving monitoring of the PM targets, but when combined they no longer produce beneficial effects. Additionally, given that nicotine exerted no effect on physiological or subjective measures of arousal, we conclude that the observed effects of nicotine and of arousal on PM performance are driven by different pharmacological mechanisms.


Shift of monocyte subsets along their continuum predicts cardiovascular outcomes.
Distribution of monocyte subsets has been shown to predict cardiovascular outcomes. However, monocytes form a continuum and categorization into discrete subsets may be an oversimplification. We herein aimed at establishing whether distribution of monocytes based on CD14 and CD16 fluorescence intensity provides incremental and complementary information on cardiovascular outcomes beyond enumeration of traditional subsets. A cohort of 227 patients at high cardiovascular risk was characterized at baseline and followed for a median of 4 years. We quantified monocytes subsets by flow cytometry based on CD14 and CD16 expression and evaluated the continuous distribution of CD14 and CD16 fluorescence within each subset. A consistent shift toward higher CD16 fluorescence intensity within each monocyte subset was observed in patients with type 2 diabetes, despite no change in their frequencies. Patients with coronary artery disease (CAD) at baseline showed a doubling of CD14++CD16+ intermediate monocytes and a shift of non-classical and classical monocytes towards intermediates ones. During follow-up, cardiovascular death or cardiovascular events occurred in 26 patients, who showed monocyte skewing similar to those of patients with baseline CAD. In fully adjusted Cox proportional hazard regression models, higher CD16 expression on classical monocytes, but not the level of intermediate monocytes or other subsets, independently predicted adverse cardiovascular outcomes. Shift of monocyte subsets along the CD14/CD16 continuum, more than their frequencies, predicted adverse cardiovascular outcomes. This finding illustrates how the concept of monocyte continuum can be used to model the cardiovascular risk.


Disassociation of muscle triglyceride content and insulin sensitivity after exercise training in patients with Type 2 diabetes.
We determined the effect of exercise training on insulin sensitivity and muscle lipids (triglyceride [TG(m)] and long-chain fatty acyl CoA [LCACoA] concentration) in patients with Type 2 diabetes. Seven patients with Type 2 diabetes and six healthy control subjects who were matched for age, BMI, % body fat and VO(2)peak participated in a 3 days per week training program for 8 weeks. Insulin sensitivity was determined pre- and post-training during a 120 min euglycaemic-hyperinsulinaemic clamp and muscle biopsies were obtained before and after each clamp. Oxidative enzyme activities [citrate synthase (CS), beta-hydroxy-acyl-CoA (beta-HAD)] and TG(m) were determined from basal muscle samples pre- and post training, while total LCACoA content was measured in samples obtained before and after insulin-stimulation, pre- and post training. The training-induced increase in VO(2)peak (approximately 20%, p<0.01) was similar in both groups. Compared with control subjects, insulin sensitivity was lower in the diabetic patients before and after training (approximately 60%; p<0.05), but was increased to the same extent in both groups with training (approximately 30%; p<0.01). TG(m) was increased in patients with Type 2 diabetes (170%; p<0.05) before, but was normalized to levels observed in control subjects after training. Basal LCACoA content was similar between groups and was unaltered by training. Insulin-stimulation had no detectable effect on LCACoA content. CS and beta-HAD activity were increased to the same extent in both groups in response to training ( p<0.001). We conclude that the enhanced insulin sensitivity observed after short-term exercise training was associated with a marked decrease in TG(m) content in patients with Type 2 diabetes. However, despite the normalization of TG(m )to levels observed in healthy individuals, insulin resistance was not completely reversed in the diabetic patients.


A case of adult α-type spinal deformity with spinal cord rotation greater than 90°.
This report describes a 42-year-old man who presented with an α-type spinal deformity with a Cobb angle of 224.9° and associated spinal cord rotation greater than 90°. Preoperative imaging revealed extensive spinal deformity, and 3D modeling confirmed the α-type nature of his deformity. Intraoperative photography demonstrated spinal cord rotation greater than 90°, which likely contributed to the patient's poor neurological status. Reports of patients with Cobb angles ≥ 100° are rare, and to the authors' knowledge, there have been no published cases of adult α-type spinal deformity. Furthermore, very few cases or case series of spinal cord rotation have been published previously, with no single patient having rotation greater than 90° to the authors' knowledge. Given these two rarities presenting in the same patient, this report can provide important insights into the operative management of this difficult form of spinal deformity.


From cells to circuits: development of the zebrafish spinal cord.
The ability of an animal to carry out its normal behavioral repertoire requires generation of an enormous diversity of neurons and glia. The relative simplicity of the spinal cord makes this an especially attractive part of the nervous system for addressing questions about the development of vertebrate neural specification and function. The last decade has witnessed an explosion in our understanding of spinal cord development and the functional interactions among spinal cord neurons and glia. Cellular, genetic, molecular, physiological and behavioral studies in zebrafish have all been important in providing insights into questions that remained unanswered by studies from other vertebrate model organisms. This is the case because many zebrafish spinal neurons can be individually identified and followed over time in living embryos and larvae. In this review, we discuss what is currently known about the cellular, genetic and molecular mechanisms involved in specifying distinct cell types in the zebrafish spinal cord and how these cells establish the functional circuitry that mediates particular behaviors. We start by describing the early signals and morphogenetic movements that form the nervous system, and in particular, the spinal cord. We then provide an overview of the cell types within the spinal cord and describe how they are specified and patterned. We begin ventrally with floor plate and proceed dorsally, through motoneurons and oligodendrocytes, interneurons, astrocytes and radial glia, spinal sensory neurons and neural crest. We next describe axon pathfinding of spinal neurons. Finally, we discuss the roles of particular spinal cord neurons in specific behaviors.


Course of renal function in type 2 diabetic patients with abnormalities of albumin excretion rate.
Heterogeneity in renal structure has been described in type 2 diabetic patients with both microalbuminuria and proteinuria; in fact, only a subset of type 2 diabetic patients have the typical diabetic glomerulopathy. However, it is currently unknown whether abnormalities in albumin excretion rate (AER) have a different renal prognostic value depending on the underlying renal structure. Aims of this study were: 1) to study the course of renal function in type 2 diabetic patients with altered AER; 2) to evaluate the relationship between the course of glomerular filtration rate (GFR) and renal structure; and 3) to evaluate the relationship between the course of GFR and baseline AER levels, metabolic control, and blood pressure levels during a follow-up period of 4 years. A total of 108 type 2 diabetic patients, 74 with microalbuminuria (MA) and 34 with proteinuria (P), were recruited into a prospective study that encompassed: 1) a baseline kidney biopsy with morphometric measurements of glomerular parameters; 2) intensified antihypertensive treatment for an average 4-year period (blood pressure target <140/90 mmHg); and 3) determinations of GFR at baseline and every 6 months. Mean (+/- SD) GFR significantly decreased from baseline in both MA (-1.3+/-9.4 [95% CI -3.51 to +0.86], P < 0.05) and P (-3.0+/-13.0 ml x min(-1) x 1.73 m(-2) per year [-7.71 to +1.61], P < 0.01). However, the changes in GFR were quite heterogeneous. Thus, on the basis of percent GFR change per year from baseline (delta%GFR), both MA and P patients were defined as progressors or nonprogressors when they were below or above the median, respectively. Baseline parameters of glomerular structure had a strong influence on the course of GFR. Indeed, the odds ratios of being progressors significantly increased across the quartiles of baseline glomerular basement membrane (GBM) width and mesangial fractional volume [Vv(mes/glom)], being 2.71 and 2.85 higher, respectively, in the fourth quartile than in the first quartile (P < 0.01 for both). Conversely, nonprogressors outnumbered progressors in the first quartile of GBM width (odds ratio: 2.14, P < 0.05) and in the first quartile of Vv(mes/glom) (odds ratio: 2.28, P < 0.01). Baseline albumin excretion rate (AER) did not influence delta%GFR; in fact, the number of progressors did not increase across quartiles of baseline AER among either MA or P. Similarly, mean blood pressure levels during follow-up (and intensified antihypertensive therapy) did not affect the course of GFR: the number of progressors and nonprogressors did not change across quartiles of mean blood pressure. In contrast, HbA1c during follow-up had an impact on delta%GFR: the odds ratio for being a progressor increased across quartiles of HbA1c, particularly for the highest quartile (HbA1c >9.0%). In conclusion, the course of renal function is heterogeneous in type 2 diabetic patients with microalbuminuria or proteinuria. In fact, a subset of patients has a rapid decline in GFR over a 4-year follow-up period; these patients have more advanced diabetic glomerulopathy and worse metabolic control than the remaining patients, whose GFR remains stable. These two cohorts are otherwise undistinguishable as regards the degree of AER at baseline and tight blood pressure control. Kidney biopsy has an important prognostic role in these patients. Thus, tight blood pressure control, when not associated with satisfactory glycemic control, is unable to prevent rapid GFR decline in type 2 diabetic patients with typical diabetic glomerulopathy.


Seed-to-seedling transitions exhibit distance-dependent mortality but no strong spacing effects in a Neotropical forest.
Patterns of seed dispersal and seed mortality influence the spatial structure of plant communities and the local coexistence of competing species. Most seeds are dispersed in proximity to the parent tree, where mortality is also expected to be the highest, because of competition with siblings or the attraction of natural enemies. Whereas distance-dependent mortality in the seed-to-seedling transition was often observed in tropical forests, few studies have attempted to estimate the shape of the survival-distance curves, which determines whether the peak of seedling establishment occurs away from the parent tree (Janzen-Connell pattern) or if the peak attenuates but remains at the parent location (Hubbell pattern). In this study, we inferred the probability density of seed dispersal and two stages of seedling establishment (new recruits, and seedlings 20 cm or taller) with distance for 24 tree species present in the 50-ha Forest Dynamics Plot of Barro Colorado Island, Panama. Using data from seed traps, seedling survey quadrats, and tree-census records spanning the 1988-2014 period, we fit hierarchical Bayesian models including parameters for tree fecundity, the shape of the dispersal kernel, and overdispersion of seed or seedling counts. We combined predictions from multiple dispersal kernels to obtain more robust inferences. We find that Hubbell patterns are the most common and Janzen-Connell patterns are very rare among those species; that distance-dependent mortality may be stronger in the seed stage, in the early recruit stage, or comparable in both; and that species with larger seeds experience less overall mortality and less distance-dependent mortality. Finally, we describe how this modeling approach could be extended at a community scale to include less abundant species.


Identifying infection-associated genes of Candida albicans in the postgenomic era.
The human pathogenic yeast Candida albicans can cause an unusually broad range of infections reflecting a remarkable potential to adapt to various microniches within the human host. The exceptional adaptability of C. albicans is mediated by rapid alterations in gene expression in response to various environmental stimuli and this transcriptional flexibility can be monitored with tools such as microarrays. Using such technology it is possible to (1) capture a genome-wide portrait of the transcriptome that mirrors the environmental conditions, (2) identify known genes, signalling pathways and transcription factors involved in pathogenesis, (3) identify new patterns of gene expression and (4) identify previously uncharacterized genes that may be associated with infection. In this review, we describe the molecular dissection of three distinct stages of infections, covering both superficial and invasive disease, using in vitro, ex vivo and in vivo infection models and microarrays.


Therapeutic approach guided by genetic alteration: use of MTOR inhibitor in renal medullary carcinoma with loss of PTEN expression.
Renal Medullary Cancer (RMC) is a rare and aggressive type of renal cell cancer that presents predominantly in patients with sickle cell hemoglobinopathies, and is typically metastatic at the time of presentation. Although platinum based chemotherapeutic regimens have recently emerged as the best option for producing a clinically significant response as reported in various case series, the response is far from satisfactory, as most RMC patients still succumb to their disease within a year of diagnosis. There is currently no standard of care for treatment of this disease. We report, to our knowledge, the first case of RMC where in molecular characterization of the tumor was used to guide therapy. In our patient, molecular analysis identified a decreased expression of Ribonucleotide Reductase M1(RRM1) and phosphatase and tensin homolog (PTEN). Based on these results of PTEN deficiency, we started our patient on everolimus (an MTOR inhibitor) maintenance after treating him with an induction chemotherapy regimen of Paclitaxel-Cisplatin-Gemcitabine (PCG). His tumor responded to induction therapy and he went into complete remission and remained in remission for 7 months. He is now alive about 14 months from his diagnosis and is asymptomatic with minimal disease. The rarity of RMC makes it very difficult to do any meaningful clinical trials in this group of patients. The overall prognosis for RMC remains very poor and knowledge about driver mutations may help in guiding therapy to improve survival in this select group of patients, where there is dearth of available therapies.


Offline Optimization of the Relative Timing of Movements in a Sequence Is Blocked by Retroactive Behavioral Interference.
Acquisition of motor skills often involves the concatenation of single movements into sequences. Along the course of learning, sequential performance becomes progressively faster and smoother, presumably by optimization of both motor planning and motor execution. Following its encoding during training, "how-to" memory undergoes consolidation, reflecting transformations in performance and its neurobiological underpinnings over time. This offline post-training memory process is characterized by two phenomena: reduced sensitivity to interference and the emergence of delayed, typically overnight, gains in performance. Here, using a training protocol that effectively induces motor sequence memory consolidation, we tested temporal and kinematic parameters of performance within (online) and between (offline) sessions, and their sensitivity to retroactive interference. One group learned a given finger-to-thumb opposition sequence (FOS), and showed robust delayed (consolidation) gains in the number of correct sequences performed at 24 h. A second group learned an additional (interference) FOS shortly after the first and did not show delayed gains. Reduction of touch times and inter-movement intervals significantly contributed to the overall offline improvement of performance overnight. However, only the offline inter-movement interval shortening was selectively blocked by the interference experience. Velocity and amplitude, comprising movement time, also significantly changed across the consolidation period but were interference -insensitive. Moreover, they paradoxically canceled out each other. Current results suggest that shifts in the representation of the trained sequence are subserved by multiple processes: from distinct changes in kinematic characteristics of individual finger movements to high-level, temporal reorganization of the movements as a unit. Each of these processes has a distinct time course and a specific susceptibility to retroactive interference. This multiple-component view may bridge the gap in understanding the link between the behavioral changes, which define online and offline learning, and the biological mechanisms that support those changes.


Impact of spikelets on hippocampal CA1 pyramidal cell activity during spatial exploration.
In vivo intracellular recordings of hippocampal neurons reveal the occurrence of fast events of small amplitude called spikelets or fast prepotentials. Because intracellular recordings have been restricted to anesthetized or head-fixed animals, it is not known how spikelet activity contributes to hippocampal spatial representations. We addressed this question in CA1 pyramidal cells by using in vivo whole-cell recording in freely moving rats. We observed a high incidence of spikelets that occurred either in isolation or in bursts and could drive spiking as fast prepotentials of action potentials. Spikelets strongly contributed to spiking activity, driving approximately 30% of all action potentials. CA1 pyramidal cell firing and spikelet activity were comodulated as a function of the animal's location in the environment. We conclude that spikelets have a major impact on hippocampal activity during spatial exploration.


Musculoskeletal Screening Tests and Bony Hip Morphology Cannot Identify Male Professional Soccer Players at Risk of Groin Injuries: A 2-Year Prospective Cohort Study.
Musculoskeletal hip/groin screening tests are commonly performed to detect at-risk individuals. Bony hip morphology is considered a potential intrinsic risk factor but has not been examined prospectively. To evaluate the association between intrinsic risk factors identified from musculoskeletal and radiographic screening tests and hip/groin injuries leading to time loss from training and/or match play in professional male soccer players. Prospective cohort study; Level of evidence, 2. Male professional soccer players, aged ≥18 years, underwent screening specific for hip/groin pain during 2 consecutive seasons of the Qatar Stars League. The screening battery included pain provocation, range of motion, and strength tests as well as a hip radiographic examination. The radiographic examination included an anteroposterior pelvic view and 45° Dunn view, with bony hip morphology determined using quantitative methods. Time-loss (≥1 day) hip/groin injuries and individual player exposure (training and match play) were recorded prospectively, and injuries were categorized as adductor-related, inguinal-related, iliopsoas-related, pubic-related, or hip-related groin pain, or "other," as recommended in the Doha agreement. We calculated hazard ratios (HRs) from univariate and multivariate Cox regression models to assess the relationship between potential risk factors and hip/groin injuries. There were 438 players, completing 609 player seasons, and 113 hip/groin injuries that met the criteria for inclusion, with 85 injuries categorized as adductor-related. The proportion of players with bony morphological variants was the following: cam, 71%; pincer, 5%; and acetabular dysplasia, 13%. Previous hip/groin injuries (HR, 1.8; 95% CI, 1.2-2.7) and eccentric adduction strength were associated with the risk of hip/groin injuries. Higher (>1 SD above the mean) than normal eccentric adduction strength was associated with an increased risk for all hip/groin injuries (HR, 1.6; 95% CI, 1.0-2.5). Lower (<1 SD below the mean) than normal eccentric adduction strength was associated with an increased risk for adductor-related injuries (HR, 1.7; 95% CI, 1.0-3.0). No other musculoskeletal screening test or bony hip morphology variables were associated with the injury risk. Previous groin injuries and eccentric adduction strength were associated with the risk of groin injuries. However, these associations were not strong enough to identify an "at-risk" individual, and therefore, musculoskeletal screening tests were not useful to dictate individualized prevention strategies. Bony hip morphology was not associated with the risk of groin injuries.