Oral delivery systems for encapsulated bacteriophages targeted at Escherichia coli O157:H7 in feedlot cattle.
Bacteriophages are natural predators of bacteria and may mitigate Escherichia coli O157:H7 in cattle and their environment. As bacteriophages targeted to E. coli O157:H7 (phages) lose activity at low pH, protection from gastric acidity may enhance efficacy of orally administered phages. Polymer encapsulation of four phages, wV8, rV5, wV7, and wV11, and exposure to pH 3.0 for 20 min resulted in an average 13.6% recovery of phages after release from encapsulation at pH 7.2. In contrast, untreated phages under similar conditions had a complete loss of activity. Steers (n = 24) received 10(11) CFU of naladixic acid-resistant E. coli O157:H7 on day 0 and were housed in six pens of four steers. Two pens were control (naladixic acid-resistant E. coli O157:H7 only), and the remaining pens received polymer-encapsulated phages (Ephage) on days -1, 1, 3, 6, and 8. Two pens received Ephage orally in gelatin capsules (bolus; 10(10) PFU per steer per day), and the remaining two pens received Ephage top-dressed on their feed (feed; estimated 10(11) PFU per steer per day). Shedding of E. coli O157:H7 was monitored for 10 weeks by collecting fecal grab and hide swab samples. Acceptable activity of mixed phages at delivery to steers was found for bolus and feed, averaging 1.82 and 1.13 x 10(9) PFU/g, respectively. However, Ephage did not reduce shedding of naladixic acid-resistant E. coli O157:H7, although duration of shedding was reduced by 14 days (P < 0.1) in bolus-fed steers as compared with control steers. Two successful systems for delivery of Ephage were developed, but a better understanding of phage-E. coli O157:H7 ecology is required to make phage therapy a viable strategy for mitigation of this organism in feedlot cattle.


The epidermal growth factor receptor (EGFR) in head and neck cancer: its role and treatment implications.
Epidermal growth factor receptor (EGFR) is a member of the ErbB family of receptors. Its stimulation by endogenous ligands, EGF or transforming growth factor-alpha (TGF-alpha) results in activation of intracellular tyrosine kinase, therefore, cell cycle progression. High levels of EGFR expression are correlated with poor prognosis and resistance to radiation therapy in a variety of cancers, mostly in squamous-cell carcinoma of the head and neck (SCCHN). Blocking the EGFR by a monoclonal antibody results in inhibition of the stimulation of the receptor, therefore, in inhibition of cell proliferation, enhanced apoptosis, and reduced angiogenesis, invasiveness and metastases. The EGFR is a prime target for new anticancer therapy in SCCHN, and other agents in development include small molecular tyrosine kinase inhibitors and antisense therapies.


Epidemiology of astrovirus infection in Zaria, Nigeria.
Astrovirus has been shown to be an important aetiological agent associated with gastroenteritis in children, although few studies have been conducted in Africa. In this study, stool specimens were obtained from 375 young children less than 5 years of age with acute gastroenteritis presenting at Ahmadu Bello University Hospital, and from a control group of 122 children without diarrhoeal illness. The specimens were examined for the presence of human astroviruses using a monoclonal antibody-based ELISA (Astrovirus IDEIATM, Dako, UK). Negative staining electron microscopy was performed on specimens to confirm the presence of astrovirus particles. Astrovirus was detected in 6.7 per cent (25/375) of the diarrhoeal stools compared to 5.7 per cent (7/122) of the control specimens. Astrovirus seemed to infect older children and more than half the children were between 1 and 4 years of age (15/25). Only four children were less than 6 months old. A winter peak of shedding was observed.


Non-communicable disease training for public health workers in low- and middle-income countries: lessons learned from a pilot training in Tanzania.
Non-communicable diseases (NCDs) are increasing worldwide. A lack of training and experience in NCDs among public health workers is evident in low- and middle- income countries. We describe the design and outcomes of applied training in NCD epidemiology and control piloted in Tanzania that included a 2-week interactive course and a 6-month NCD field project. Trainees (n=14 initiated; n=13 completed) were epidemiology-trained Ministry of Health or hospital staff. We evaluated the training using Kirkpatrick's evaluation model for measuring reactions, learning, behavior and results using pre- and post-tests and closed-ended and open-ended questions. Significant improvements in knowledge and self-reported competencies were observed. Trainees reported applying competencies at work and supervisors reported improvements in trainees' performance. Six field projects were completed; one led to staffing changes and education materials for patients with diabetes and another to the initiation of an injury surveillance system. Workplace support and mentoring were factors that facilitated the completion of projects. Follow-up of participants was difficult, limiting our evaluation of the training's outcomes. The applied NCD epidemiology and control training piloted in Tanzania was well received and showed improvements in knowledge, skill and self-efficacy and changes in workplace behavior and institutional and organizational changes. Further evaluations are needed to better understand the impact of similar NCD trainings and future trainers should ensure that trainees have mentoring and workplace support prior to participating in an applied NCD training.


Earth-abundant oxygen evolution catalysts coupled onto ZnO nanowire arrays for efficient photoelectrochemical water cleavage.
ZnO has long been considered as a model UV-driven photoanode for photoelectrochemical water splitting, but its performance has been limited by fast charge-carrier recombination, extremely poor stability in aqueous solution, and slow kinetics of water oxidation. These issues were addressed by applying a strategy of optimization and passivation of hydrothermally grown 1D ZnO nanowire arrays. The length and diameter of bare ZnO nanowires were optimized by varying the growth time and precursor concentration to achieve optimal photoelectrochemical performance. The addition of earth-abundant cobalt phosphate (Co-Pi) and nickel borate (Ni-B) oxygen evolution catalysts onto ZnO nanowires resulted in substantial cathodic shifts in onset potential to as low as about 0.3 V versus the reversible hydrogen electrode (RHE) for Ni-B/ZnO, for which a maximum photocurrent density of 1.1 mA cm(-2) at 0.9 V (vs. RHE) with applied bias photon-to-current efficiency of 0.4 % and an unprecedented near-unity incident photon-to-current efficiency at 370 nm. In addition the potential required for saturated photocurrent was dramatically reduced from 1.6 to 0.9 V versus RHE. Furthermore, the stability of these ZnO nanowires was significantly enhanced by using Ni-B compared to Co-Pi due to its superior chemical robustness, and it thus has additional functionality as a stable protecting layer on the ZnO surface. These remarkable enhancements in both photocatalytic activity and stability directly address the current severe limitations in the use of ZnO-based photoelectrodes for water-splitting applications, and can be applied to other photoanodes for efficient solar-driven fuel synthesis.


Combination of uterine natural killer cell immunoglobulin receptor haplotype and trophoblastic HLA-C ligand influences the risk of pregnancy loss: a retrospective cohort analysis of direct embryo genotyping data from euploid transfers.
To compare maternal uterine natural killer cell immunoglobulin receptor (KIR) genotype and haplotype frequencies between patients whose euploid single-embryo transfer resulted in pregnancy loss and those that resulted in delivery and to determine if the risk of pregnancy loss was affected by the HLA-C genotype content in the embryo. Retrospective cohort. Academic research center. Autologous fresh IVF cycles resulting in positive serum β-hCG during 2009-2014. None. 1) Relative risk of pregnancy loss according to maternal KIR genotypes and haplotypes. 2) Comparison of pregnancy loss rates within each KIR haplotype according to HLA-C ligand present in trophectoderm biopsy samples. A total of 668 euploid single-embryo transfers with stored maternal DNA and available preamplification DNA from prior trophectoderm biopsy samples were studied. KIR2DS1, KIR3DS1, and KIR2DS5 were more common in patients who experienced pregnancy loss. Carriers of KIR A haplotype exhibited a decreased risk of pregnancy loss compared with KIR B haplotype carriers. However, among KIR A haplotype carriers, the risk of loss was significantly influenced by whether the transferred embryo carried a C1 allele versus no C1 alleles. KIR A haplotype carriers experienced fewer pregnancy losses than KIR B haplotype carriers after euploid single-embryo transfer. However, this risk was modified by HLA-C alleles present in the embryo. High-risk combinations (KIR A/homozygous C2 and KIR B/homozygous C1) resulted in a 51% increased risk of loss over all other combinations.


Superficial digital flexor tendonitis in cutting horses: 19 cases (2007-2011).
To characterize superficial digital flexor (SDF) tendon lesions in the forelimbs of cutting horses and determine recurrence rate and prognosis for this condition. Retrospective case series. 19 cutting horses with SDF tendonitis. Medical records for horses evaluated for SDF tendonitis in 2007 through 2011 were reviewed. Data regarding age, sex, lameness grade, affected limb, and treatment were collected. Ultrasonographic images were reviewed, and lesion characteristics were recorded. Follow-up telephone interviews with owners or trainers were conducted to determine recurrence of SDF tendonitis, return of horse to its previous level of activity, and duration of the convalescent period. All 19 horses initially evaluated for SDF tendonitis had similar lesions in the lateral aspect of the tendon. The right forelimb was affected in 11 horses, and the left forelimb was affected in 7 horses; 1 horse was affected in both forelimbs. Mean lameness grade was 1.26 (range, 0 to 3). Of 17 horses for which follow-up information was available, 3 had recurrence of tendon lesions and 1 developed a lesion in the contralateral forelimb SDF tendon; 16 horses returned to their previous level of activity. The location of SDF tendonitis in cutting horses appeared to be unique in that no central core lesions were detected ultrasonographically. Lesions at the periphery of the tendon may have an increased ability to heal, compared with lesions at the central core. Results suggested that cutting horses with SDF tendonitis have a better prognosis than that reported for affected racehorses.


Caspase-3 probes for PET imaging of apoptotic tumor response to anticancer therapy.
Apoptosis is a highly regulated process involved in the normal organism development and homeostasis. In the context of anticancer therapy, apoptosis is also studied intensively in an attempt to induce cell death in cancer cells. Caspase activation is a known key event in the apoptotic process. In particular, active caspase-3 and -7 are the common effectors in several apoptotic pathways, therefore effector caspase activation may be a promising biomarker for response evaluation to anticancer therapy. Quantitative imaging of apoptosis in vivo could provide early assessment of therapeutic effectiveness and could also be used in drug development to evaluate the efficacy as well as potential toxicity of novel treatments. Positron Emission Tomography (PET) is a highly sensitive molecular imaging modality that allows non-invasive in vivo imaging of biological processes such as apoptosis by using radiolabeled probes. Here we describe the development and evaluation of fluorine-18-labeled caspase-3 activity-based probes (ABPs) for PET imaging of apoptosis. ABPs were selected by screening of a small library of fluorine-19-labeled DEVD peptides containing different electrophilic warhead groups. An acyloxymethyl ketone was identified with low nanomolar affinity for caspase-3 and was radiolabeled with fluorine-18. The resulting radiotracer, [18F]MICA-302, showed good labeling of active caspase-3 in vitro and favorable pharmacokinetic properties. A μPET imaging experiment in colorectal tumor xenografts demonstrated an increased tumor accumulation of [18F]MICA-302 in drug-treated versus control animals. Therefore, our data suggest this radiotracer may be useful for clinical PET imaging of response to anticancer therapy.


Endoscopic and Pathologic Characterization of Papillary Architecture in Struvite Stone Formers.
To describe the endoscopic characteristics of renal papillae in struvite stone formers (SFs). From 2009 to 2014, patients undergoing percutaneous nephrolithotomy were prospectively enrolled in our study. Endoscopic analysis and biopsy of papillae were performed to demonstrate the presence and percentage surface area (SA) of Randall's plaque or ductal plug. Comparison with idiopathic calcium oxalate (CaOx) SF and non-SF controls was performed. We identified 29 struvite SFs to compare with 90 idiopathic CaOx SFs and 17 controls. On endoscopic mapping, 28 struvite SFs (97%) demonstrated Randall's plaque and 9 (31%) had plugging. The average mean SA of Randall's plaque in struvite SF (1.5 ± 1.4%) was less than CaOx SFs (3.7 ± 4.3%, P = .0018) and similar to controls (1.7 ± 2.7%, P = .76). Average mean plug SA was similar between struvite SFs, CaOx SFs, and controls. On metabolic assessment, 83% of struvite SFs had at least one urine abnormality, with urinary uric acid and oxalate levels significantly higher among struvite SFs compared to controls (P = .002). Despite lack of active urinary tract infection, interstitial inflammation was more prevalent in struvite SFs compared to CaOx SFs (43.5% vs 7.3%, P = .0001). Our findings suggest a limited role for Randall's plaque in struvite stone formation. Struvite SFs have less plaque formation than CaOx SFs, but demonstrate evidence of severe parenchymal inflammation compared to other SFs. The role of this prominent interstitial inflammation requires further study.


Potential for diagnosis of infectious disease from the 100,000 Genomes Project Metagenomic Dataset: Recommendations for reporting results.
The identification of microbiological infection is usually a diagnostic investigation, a complex process that is firstly initiated by clinical suspicion. With the emergence of high-throughput sequencing (HTS) technologies, metagenomic analysis has unveiled the power to identify microbial DNA/RNA from a diverse range of clinical samples (1). Metagenomic analysis of whole human genomes at the clinical/research interface bypasses the steps of clinical scrutiny and targeted testing and has the potential to generate unexpected findings relating to infectious and sometimes transmissible disease. There is no doubt that microbial findings that may have a significant impact on a patient's treatment and their close contacts should be reported to those with clinical responsibility for the sample-donating patient. There are no clear recommendations on how such findings that are incidental, or outside the original investigation, should be handled. Here we aim to provide an informed protocol for the management of incidental microbial findings as part of the 100,000 Genomes Project which may have broader application in this emerging field. As with any other clinical information, we aim to prioritise the reporting of data that are most likely to be of benefit to the patient and their close contacts. We also set out to minimize risks, costs and potential anxiety associated with the reporting of results that are unlikely to be of clinical significance. Our recommendations aim to support the practice of microbial metagenomics by providing a simplified pathway that can be applied to reporting the identification of potential pathogens from metagenomic datasets. Given that the ambition for UK sequenced human genomes over the next 5 years has been set to reach 5 million and the field of metagenomics is rapidly evolving, the guidance will be regularly reviewed and will likely adapt over time as experience develops.


A Case of False-Positive Mycobacterium tuberculosis Caused by Mycobacterium celatum.
Mycobacterium celatum is a nontuberculous mycobacterium shown to cause symptoms similar to pulmonary M. tuberculosis. Certain strains have been shown to cross-react with the probes used to detect M. tuberculosis, making this a diagnostic challenge. We present a 56-year-old gentleman who developed signs and symptoms of lung infection with computed tomography scan of the chest showing right lung apex cavitation. Serial sputum samples were positive for acid-fast bacilli and nucleic acid amplification testing identified M. tuberculosis ribosomal RNA, resulting in treatment initiation. Further testing with high performance liquid chromatography showed a pattern consistent with M. celatum. This case illustrates the potential for M. celatum to mimic M. tuberculosis in both its clinical history and laboratory testing due to the identical oligonucleotide sequence contained in both. An increasing number of case reports suggest that early reliable differentiation could reduce unnecessary treatment and public health intervention associated with misdiagnosed tuberculosis.


Four malaria success stories: how malaria burden was successfully reduced in Brazil, Eritrea, India, and Vietnam.
While many countries struggle to control malaria, four countries, Brazil, Eritrea, India, and Vietnam, have successfully reduced malaria burden. To determine what led these countries to achieve impact, published and unpublished reports were reviewed and selected program and partner staff were interviewed to identify common factors that contributed to these successes. Common success factors included conducive country conditions, a targeted technical approach using a package of effective tools, data-driven decision-making, active leadership at all levels of government, involvement of communities, decentralized implementation and control of finances, skilled technical and managerial capacity at national and sub-national levels, hands-on technical and programmatic support from partner agencies, and sufficient and flexible financing. All these factors were essential in achieving success. If the goals of Roll Back Malaria are to be achieved, governments and their partners must take the lessons learned from these program successes and apply them in other affected countries.


Panhypogammaglobulinemia in systemic lupus erythematosus: in vitro demonstration of multiple cellular defects.
Classically, systemic lupus erythematosus (SLE) is a disease of antibody overproduction, whereas the hallmark of acquired immune deficiency is antibody underproduction. Two patients are presented in whom panhypogammaglobulinemia developed during the course of SLE. In both patients, the levels of the major immunoglobulin (Ig) classes did not fall simultaneously. Anti-DNA antibodies were present, and exacerbations of SLE nephritis occurred in both cases 6 to 8 yr after Ig levels became subnormal. One patient still requires immunosuppressive therapy for renal disease; both patients are experiencing recurrent sinopulmonary bacterial infections. In the pokeweed mitogen--stimulated Ig biosynthesis assay, both patients showed abnormal Ig production due to defective function of three cell types: hyporesponsive B cells, excessive T suppression, and subnormal T help. The latter defect is rare in common variable hypogammaglobulinemia. One patient also showed extreme suppression of Ig production by phagocytic mononuclear cells. Thus, despite the similarity in the histories, the cellular function of these two patients was not identical in vitro.


Recovery patterns over 4 years after distal radius fracture: Descriptive changes in fracture-specific pain/disability, fall risk factors, bone mineral density, and general health status.
Descriptive/Longitudinal cohort. Distal radius fracture (DRF) is a common fall related fragility fracture that is known to be an early and independent predictor of secondary osteoporotic (OP) fractures. Changes in falls risk status, bone status and general health has not been evaluated prospectively in a population that has sustained a DRF. The purpose of our study was to describe the status of fracture-specific pain/disability, fall risk factors such as physical activity (PA) and fear of falling (FOF), bone mineral density (BMD) and general health status (HS) in people with a DRF and how these variables change over four years with respect to sex, age, incidence of secondary falls and secondary OP fractures. Patients (n = 94) self-reported their fracture-specific pain and disability (Patient-Rated Wrist Evaluation), PA (Rapid Assessment of Physical Activity), FOF (Modified Fall Efficacy Scale), HS (12-item Short Form Health Survey) and completed dual-energy X-ray absorptiometry scan based BMD assessment (lumbar spine and total hip) at baseline (1-2 weeks post-fracture), six months and four years after DRF. Descriptive statistics and general linear models were used to describe changes in recovery patterns over four years. There was significant (p<0.001) improvement in fracture-specific pain/disability (60 points), FOF (1 point) and physical HS (11 points) between baseline and 4 year follow-up. There were no significant changes in PA and BMD. When stratified with respect to age, sex, presence of subsequent falls and OP fractures, there were no significant differences in fracture-specific pain/disability, PA, FOF, and BMD at baseline, six months or four years after DRF. The physical HS was significantly (p<0.05) less/poorer among those with secondary falls (lower by 2-6 points) and fractures (lower by 5-6 points) compared to those without. Similarly, mental HS was significantly (p<0.05) poorer among people with secondary falls (lower by 2-6 points) and in 50-64 year age group (lower by 3-5 points) than those without secondary falls and in 65-80 year age group, respectively. Post DRF, the majority of the improvement in fracture-specific pain/disability, FOF and HS was completed at six months and very small changes were observed between the six month and four year follow-up. NA.


A case of Raoultella planticola causing a urinary tract infection in a pediatric patient.
Raoultella planticola (R. planticola) is an abundant environmental bacterium that rarely infects humans. There is only one known case of R. planticola causing a urinary tract infection (UTI) in a pediatric patient. This is a second case of R. planticola potentially causing a symptomatic UTI in a 2-month-old female child with no known medical problems.


Temperature stability of nanocellulose dispersions.
Cellulose nanofibrils (CNF) have potential as rheology modifiers of water based fluids, e.g. drilling fluids for use in oil wells or as additives in injection water for enhanced oil recovery (EOR). The temperature in oil wells can be high (>100°C), and the retention time long; days for drilling fluids and months for EOR fluids. Hence, it is important to assess the temperature stability over time of nanocellulose dispersions to clarify their suitability as rheology modifiers of water based fluids at such harsh conditions. Dispersions of CNF produced mechanically, by using TEMPO mediated oxidation and by using carboxymethylation as pretreatment, in addition to cellulose nanocrystals (CNC), have been subjected to heat aging. Temperature stability was best for CNC and for mechanically produced CNF that were stable after heating to 140°C for three days. The effect of additives was evaluated; cesium formate and sodium formate increased the temperature stability of the dispersions, while there was no effect of using phosphate buffer.


Effect of the Affordable Care Act on Breastfeeding Outcomes.
To assess how the 2012 Affordable Care Act (ACA) policy change, which required most private health insurance plans to cover lactation-support services and breastfeeding equipment (without cost-sharing), affected breastfeeding outcomes. We used a regression-adjusted difference-in-differences approach with cross-sectional observational data from the US National Immunization Survey from 2008 to 2014 to estimate the effect of the ACA policy change on breastfeeding outcomes, including initiation, duration, and age at first formula feeding. The sample included children aged 19 to 23 months covered by private health insurance or Medicaid. The ACA policy change was associated with an increase in breastfeeding duration by 10% (0.57 months; P = .007) and duration of exclusive breastfeeding by 21% (0.74 months; P = .001) among the eligible population. Results indicate no significant effects on breastfeeding initiation and age at first formula feeding. Reducing barriers to receiving support services and breastfeeding equipment shows promise as part of a broader effort to encourage breastfeeding, particularly the duration of breastfeeding and the amount of time before formula supplementation.


Developing operational algorithms using linear and non-linear squares estimation in Python for the identification of Culex pipiens and Culex restuans in a mosquito abatement district (Cook County, Illinois, USA).
In this research, community level spatial models were developed for determining mosquito abundance and environmental factors that could aid in the risk prediction of West Nile virus (WNv) outbreaks. Adult Culex pipiens and Culex restuan mosquitoes and multiple habitat covariates were collected from nine sites within Cook County, Illinois, USA, to provide spatio-temporal information on the abundance of WNv vectors from 2002 to 2005. Regression analyses of the sampled covariates revealed that the adult Culex population was positively associated with temperature throughout the sampling frame. The model output also indicated that precipitation was negatively associated to mosquito abundance in 2002, 2003 and 2005 (P <0.05), but positively associated in 2004 (P <0.05). A land use land cover classification, based on QuickBird visible and near infra-red data, acquired at 0.61 m resolution, was used to investigate possible associations between geographical features and the abundance of sampled Culex oviposition surveillance sites. A maximum likelihood unsupervised classification in ArcInfo 9.2(R) revealed that the highest overall mosquito abundance was found in sites having a low-to-moderate range of built environment (40%) and high forest composition. A set of propagation equations were then designed to model the calibration uncertainties, which revealed that normalized difference vegetation index (NDVI), and two NDVI variants, were informative markers for the sampled mosquito data. Spatial dependence of the covariates of Cx. restuans and Cx. pipiens oviposition sites were indexed using semivariograms, which suggested that all main effects of the explanatory variables were statistically significant in the model. Additionally, a multispectral classification and digital elevation model-based geographical information system method were able to evaluate stream flow direction and accumulation for identification of terrain covariates associated with the sampled habitat data. These results demonstrate that remotely sensed operational indices can be used to identify parameters associated with field-sampled Cx. pipiens and Cx. restuans aquatic habitats.


Using several pair-wise informant sequences for de novo prediction of alternatively spliced transcripts.
As part of the ENCODE Genome Annotation Assessment Project (EGASP), we developed the MARS extension to the Twinscan algorithm. MARS is designed to find human alternatively spliced transcripts that are conserved in only one or a limited number of extant species. MARS is able to use an arbitrary number of informant sequences and predicts a number of alternative transcripts at each gene locus. MARS uses the mouse, rat, dog, opossum, chicken, and frog genome sequences as pairwise informant sources for Twinscan and combines the resulting transcript predictions into genes based on coding (CDS) region overlap. Based on the EGASP assessment, MARS is one of the more accurate dual-genome prediction programs. Compared to the GENCODE annotation, we find that predictive sensitivity increases, while specificity decreases, as more informant species are used. MARS correctly predicts alternatively spliced transcripts for 11 of the 236 multi-exon GENCODE genes that are alternatively spliced in the coding region of their transcripts. For these genes a total of 24 correct transcripts are predicted. The MARS algorithm is able to predict alternatively spliced transcripts without the use of expressed sequence information, although the number of loci in which multiple predicted transcripts match multiple alternatively spliced transcripts in the GENCODE annotation is relatively small.


Toxicity of hydroxyurea in rats and dogs.
The toxicity of hydroxyurea, a treatment for specific neoplasms, sickle-cell disease, polycythemia, and thrombocytosis that kills cells in mitosis, was assessed in repeat-dose, oral gavage studies in rats and dogs and a cardiovascular study in telemetered dogs. Hydroxyurea produced hematopoietic, lymphoid, cardiovascular, and gastrointestinal toxicity with steep dose response curves. In rats dosed for 10 days, 50 mg/kg/day was tolerated; 500 mg/kg/day produced decreased body weight gain; decreased circulating leukocytes, erythrocytes, and platelets; decreased cellularity of thymus, lymph nodes, and bone marrow; and epithelial degeneration and/or dysplasia of the stomach and small intestine; 1,500 mg/kg/day resulted in deaths on day 5. In dogs, a single dose at ≥ 250 mg/kg caused prostration leading to unscheduled euthanasia. Dogs administered 50 mg/kg/day for 1 month had decreased circulating leukocytes, erythrocytes, and platelets; increased bone marrow cellularity with decreased maturing granulocytes; increased creatinine kinase activity; and increased iron pigment in bone marrow and hepatic sinusoidal cells. In telemetered dogs, doses ≥ 15 mg/kg decreased systolic blood pressure (BP); 50 mg/kg increased diastolic BP, heart rate, and change in blood pressure over time (+dP/dt), and decreased QT and PR intervals and maximum left ventricular systolic and end diastolic pressures with measures returning to control levels within 24 hr.


Subclinical cardiac dysfunction increases the risk of stroke and dementia: the Rotterdam Study.
To investigate the association between cardiac function and the risk of stroke and dementia in elderly free of clinical cardiac disease. Additionally, we investigated the relation between cardiac function and MRI markers of subclinical cerebrovascular disease. This study was conducted within the population-based Rotterdam Study. A total of 3,291 participants (60.8% female, age-range 58-98 years) free of coronary heart disease, heart failure, atrial fibrillation, stroke, and dementia underwent echocardiography in 2002-2005 to measure cardiac function. Follow-up finished in 2012. In 2005-2006, a random subset of 577 stroke-free people without dementia underwent brain MRI on which infarcts and white matter lesion volume were assessed. During 21,785 person-years of follow-up, 164 people had a stroke and during 19,462 person-years of follow-up, 208 people developed dementia. Measures of better diastolic function, such as higher E/A ratio, were associated with a lower risk of stroke (hazard ratio [HR] 0.82; 95% confidence interval [CI] 0.69; 0.98) and dementia (HR 0.82; 95% CI 0.70; 0.96). Better systolic function, measured as higher fractional shortening, was only associated with a lower risk of stroke (HR 0.84; 95% CI 0.72; 0.98). Better diastolic function was related to a lower prevalence of silent infarcts on MRI, especially lacunar infarcts. In elderly free of clinical cardiac disease, worse diastolic function is associated with clinical stroke, dementia, and silent infarcts on MRI, whereas worse systolic function is related only to clinical stroke. These findings can form the basis for future research on the utility of cardiac function as potential intervention target for prevention of neurologic diseases.


Ribose Supplementation Alone or with Elevated Creatine Does Not Preserve High Energy Nucleotides or Cardiac Function in the Failing Mouse Heart.
Reduced levels of creatine and total adenine nucleotides (sum of ATP, ADP and AMP) are hallmarks of chronic heart failure and restoring these pools is predicted to be beneficial by maintaining the diseased heart in a more favourable energy state. Ribose supplementation is thought to support both salvage and re-synthesis of adenine nucleotides by bypassing the rate-limiting step. We therefore tested whether ribose would be beneficial in chronic heart failure in control mice and in mice with elevated myocardial creatine due to overexpression of the creatine transporter (CrT-OE). FOUR GROUPS WERE STUDIED: sham; myocardial infarction (MI); MI+ribose; MI+CrT-OE+ribose. In a pilot study, ribose given in drinking water was bioavailable, resulting in a two-fold increase in myocardial ribose-5-phosphate levels. However, 8 weeks post-surgery, total adenine nucleotide (TAN) pool was decreased to a similar amount (8-14%) in all infarcted groups irrespective of the treatment received. All infarcted groups also presented with a similar and substantial degree of left ventricular (LV) dysfunction (3-fold reduction in ejection fraction) and LV hypertrophy (32-47% increased mass). Ejection fraction closely correlated with infarct size independently of treatment (r(2) = 0.63, p<0.0001), but did not correlate with myocardial creatine or TAN levels. Elevating myocardial ribose and creatine levels failed to maintain TAN pool or improve post-infarction LV remodeling and function. This suggests that ribose is not rate-limiting for purine nucleotide biosynthesis in the chronically failing mouse heart and that alternative strategies to preserve TAN pool should be investigated.


Neighborhood predictors of plant performance.
We developed models of inter-individual interference to predict the fecundity of individuals in populations of the annual plant species, Arabidopsis thaliana. An individual plant is modeled as having a neighborhood which is a circular area of fixed radius with the plant at its center. Other plants which share the circle with the focal plant are termed neighbors of the focal plant. We developed an index of neighborhood interference which is the independent variable in a non-linear regression model that predicts individual plant fecundity. We present methods of exploratory data analysis that are useful in determining a best neighborhood radius, defined as that radius which minimizes residual sum of squares, and in deciding on the functional form of the interference index. In developing the interference index for Arabidopsis, we focus on aspects of the spatial distribution of neighbors: their number, distance and angular dispersion.We found that a best (or optimal) neighborhood radius can be resolved, which provides the best predictor of plant performance. Fecundity predictors based on adult neighbors were noticeably better than those based on neighbors at the seedling stage. Rosettes of Arabidopsis may change location during development (they fall over) and the new "fallen" positions do provide some improvement in the predictor. Taking into account distance to neighbors within the neighborhood provided only negligible improvement in the model. Finally, the incorporation of angular dispersion in the crowding index produced a considerably better fit. The fecundity predictor that included number of neighbors and angular dispersion in the crowding index explained about 70% of the variation in individual seed set.


Self-Rated Health Status and Risk of Incident Stroke in 0.5 Million Chinese Adults: The China Kadoorie Biobank Study.
Self-rated health (SRH) is a consistent and strong predictor of all-cause and cardiovascular mortality in various populations. However, the associations between SRH measures and risk of first-ever or recurrent stroke were rarely explored. We thus aim to prospectively investigate the associations between SRH measures and risk of total and subtypes of stroke in Chinese population. A total of 494,113 participants from the China Kadoorie Biobank without prior heart diseases or cancer (486,541 without stroke and 7,572 with stroke) were followed from baseline (2004 to 2008) until December 31, 2013. General and age-comparative SRH were obtained from baseline questionnaires. First-ever stroke or recurrent events were ascertained through linkage to disease registry system and health insurance data. We identified 27,662 first-ever stroke and 2,909 recurrent events during an average of 7.0 years of follow-up. Compared with excellent general SRH, the hazard ratios (HRs) and 95% confidence intervals (CIs) for first-ever stroke associated with good, fair, and poor general SRH were 1.04 (1.00 to 1.08), 1.19 (1.15 to 1.23), and 1.49 (1.42 to 1.56) in the multivariate model, respectively. Compared with better age-comparative SRH, the HRs (95% CIs) of same and worse age-comparative SRH were 1.13 (1.10 to 1.17) and 1.51 (1.45 to 1.58), respectively. The relations of SRH measures with ischemic stroke, hemorrhagic stroke, and recurrent stroke were similar to that with total first-ever stroke. However, the magnitude of associations was much stronger for fatal stroke than for non-fatal stroke. This large-scale prospective cohort suggests that self-perceived health status is associated with incident stroke, regardless of stroke subtype.


Postural orientation in subjects with anterior cruciate ligament injury: development and first evaluation of a new observational test battery.
Anterior cruciate ligament (ACL) injury is associated with mechanical instability and defective neuromuscular function, and can lead to further injury, increased joint loading and osteoarthritis. Patients with ACL injury demonstrate altered postural orientation, manifested as observable "substitution patterns" (SPs) but no one has applied a clinically useful method to systematically study postural orientation in these patients. Here, we investigated the presence of such patterns in 24 adults with ACL injury and in 49 controls, in parallel with the development and a first evaluation of a new test battery, test for SPs. The rationale behind the test for SPs was to characterize postural orientation as the ability to maintain appropriate relationships between body segments and environment during weight-bearing movements. In this first study, patients displayed SPs more frequently and/or more clearly on their injured, but also their uninjured side than did controls. Inter-rater and intra-rater reproducibility was good at a group level. Future studies of validity, responsiveness and including other subgroups of patients with ACL injury will have to prove if the test for SPs can be used in the diagnostics of defective neuromuscular function following knee injury, when planning and carrying out training and rehabilitation and when deciding appropriate time to return to activity and sports after ACL injury.


Breaking the relentless course of Hallopeau's acrodermatitis by dapsone.
We report the case of a 74-year-old man who suffered for four years from recurrent generalized pustular eruptions associated with acrodermatitis continua of Hallopeau (ACH). Initial therapy using topical corticosteroids, tars and PUVA failed to improve the acral lesions. The disease was also unresponsive to systemic therapy using corticosteroids, methotrexate, etretinate, acitretine and cyclosporin A. Serious side-effects were experienced with these drugs. A marked improvement of the generalized pustules was observed under treatment combining etretinate and methotrexate, without having any effect on the acropustulosis. During hospitalization, a topical tar therapy cleared the pustular rash. Again no effect was observed on the acral lesions. Disulone (DDS) therapy, initiated at a dose of 200 mg/day, completely cleared ACH after 4 weeks. The disease relapsed when the dose was reduced to 100 mg/day. Increasing the DDS to 150 mg/day was rapidly followed by a new clearance of the acral lesions. After a 3-month treatment, the patient has remained lesion free and no DDS-related side-effects have occurred. In conclusion, sulfones may be considered as an alternative therapeutic option in drug-resistant ACH.


Adaptive divergence in experimental populations of Pseudomonas fluorescens. III. Mutational origins of wrinkly spreader diversity.
Understanding the connections among genotype, phenotype, and fitness through evolutionary time is a central goal of evolutionary genetics. Wrinkly spreader (WS) genotypes evolve repeatedly in model Pseudomonas populations and show substantial morphological and fitness differences. Previous work identified genes contributing to the evolutionary success of WS, in particular the di-guanylate cyclase response regulator, WspR. Here we scrutinize the Wsp signal transduction pathway of which WspR is the primary output component. The pathway has the hallmarks of a chemosensory pathway and genetic analyses show that regulation and function of Wsp is analogous to the Che chemotaxis pathway from Escherichia coli. Of significance is the methyltransferase (WspC) and methylesterase (WspF) whose opposing activities form an integral feedback loop that controls the activity of the kinase (WspE). Deductions based on the regulatory model suggested that mutations within wspF were a likely cause of WS. Analyses of independent WS genotypes revealed numerous simple mutations in this single open reading frame. Remarkably, different mutations have different phenotypic and fitness effects. We suggest that the negative feedback loop inherent in Wsp regulation allows the pathway to be tuned by mutation in a rheostat-like manner.


Rotavirus vaccine trial in Jamaica.
Worldwide, rotaviruses have been a significant cause of dehydrating gastroenteritis. This contributed to increased infant morbidity and mortality in Jamaica. We enrolled 1804 Jamaican infants in the international randomized, placebo-controlled, pentavalent (G1, G2, G3, G4 and P1) rotavirus vaccine trial. This pentavalent vaccine was found to significantly reduce rotavirus gastroenteritis attributable emergency room visits and hospitalizations, without increasing the rates of intussusception, or other serious adverse events in Jamaican infants. It is recommended that the rotavirus vaccine be included in Jamaica's National Immunization Programme in accordance with recommendations from the World Health Organization.


Refractory period studies in a human neuromuscular preparation.
A subtraction technique has been employed to study refractoriness in the median nerve and thenar muscles in man. The absolutely refractory periods of the distal motor nerve branches and of the muscle fibres were found to be similar, the majority being within the 2.5--3.0 ms range; the values were distributed in a unimodal manner. The relatively refractory periods of the motor nerve terminals were about 5 ms. In some experiments it was possible to record non-propagated potentials at the endplate zones of the muscle fibres.


Impact of a new alcohol policy on homemade alcohol consumption and sales in Russia.
To describe the effects of Russian policy since 2006 affecting price and availability on the consumption of recorded and unrecorded alcohol, with specific reference to homemade alcohol, and to investigate other factors affecting homemade alcohol consumption and purchasing. Consumption and preferred beverage data were collected from RLMS-HSE nationwide panel surveys from 1994 to 2013, with a detailed analysis of 2012 data (18,221 respondents aged 16+ years). Official statistics on manufactured alcohol sales, regional price increase and real disposable income were used. Homemade distilled spirits (samogon) consumption decreased together with that of recorded and unrecorded manufactured spirits since 2000. The consumption of spirits was partially replaced by the consumption of beer and wine. These trends in alcohol consumption were interrupted in 2008-2013. The interruption was more likely affected by the economic crisis and recession than by the new alcohol policy. Social networks and availability of unrecorded alcohol were more important predictors of homemade alcohol consumption and purchasing than was a recorded alcohol price increase. Homemade alcohol consumption does not replace the declining market for recorded spirits in Russia. The effects of economic and social factors on homemade alcohol consumption are greater than are the short-term effects of the new alcohol policy. The very recent (2015) reduction of the minimum unit price of vodka may be premature.


100% Complete response rate in patients with cutaneous metastatic melanoma treated with intralesional interleukin (IL)-2, imiquimod, and topical retinoid combination therapy: results of a case series.
Patients with cutaneous melanoma metastases have experienced excellent responses to intralesional interleukin (IL)-2. This has led to its recent inclusion into the US National Comprehensive Cancer Network guidelines for management of cutaneous melanoma metastases. Despite this, intralesional IL-2 has not been highlighted in the US literature nor have US physicians adopted it. We sought to evaluate the effectiveness of intralesional IL-2 combined with topical imiquimod and retinoid for treatment of cutaneous metastatic melanoma. A retrospective case series of 11 patients with cutaneous metastatic melanoma were treated with intralesional IL-2 combined with topical imiquimod and retinoid. A 100% complete local response rate with long-term follow-up (average of 24 months) was seen in all 11 patients treated with this proposed regimen. Biopsy specimens of treated sites confirmed absence of malignant cells. The most common treatment-related adverse event was rigors. Small number of patients, retrospective review of charts, and lack of a comparison group were limitations. Intralesional IL-2 administered concomitantly with topical imiquimod and a retinoid cream is a promising therapeutic option for managing cutaneous melanoma metastases. The regimen was well tolerated and should be considered as a reasonable alternative to surgical excision.


Heart Failure Society of South Africa (HeFSSA) perspective on the European Society of Cardiology (ESC) 2012 chronic heart failure guideline.
The South African Heart Association (SA Heart) is an affiliate of the European Society of Cardiology (ESC). SA Heart endorses ESC treatment guidelines with modification to suit local circumstances. The Heart Failure Society of South Africa (HeFSSA) is a special interest group of SA Heart. This guideline has been compiled on behalf of the HeFSSA and is based on the ESC guidelines for the diagnosis and treatment of acute and chronic heart failure 2012. The focus is on heart failure with reduced ejection fraction (HF-REF) (i.e. ejection fraction <50%). We have recommended interventions in symptomatic patients with HF-REF in general to clarify the 'grey area' between the ESC guidelines definition of REF (<50%) and the predefined ejection fraction used in randomised heart failure trials (<35%). To highlight new changes in the diagnosis and treatment of chronic heart failure with particular emphasis on areas that are relevant to SA. Randomised clinical trials are a crucial, but not the only, guide in treating HF-REF patients. There always remain questions that are unanswered and groups of patients not studied, so prudent clinical decisions are required.


Sexuality and the Internet: A Study of the Perspectives of Turkish University Students.
The purpose of this study was to determine Turkish university students' uses of and attitudes toward the Internet concerning sexuality. The study was conducted in two public universities in the Central Anatolia and Eastern Anatolia regions of Turkey. Among the students invited to take part in the study, 1,330 students agreed to do so. The study data were collected using a questionnaire designed by researchers to determine participants' personal characteristics, computer and Internet uses in general, and for sexual matters. The findings suggest that approximately half of the students (51%) reported using the Internet to obtain information about sexuality. Among the students, 30.5% said that they visit erotic and pornographic Web sites, 21.1% said that they chat on the Internet about sexuality, and 9.3% said that they bought sexual products online. Compared to the female students, the male students, statistically, more frequently show behaviors such as obtaining sexual information on the Internet, online sexual shopping, chatting on the Internet about sexuality, and visiting pornographic and erotic Web sites. In addition, female students have more negative attitudes toward using the Internet for sexual purposes. This study discusses its results along with the literature from Turkey and other countries.


Immune Response and Intraocular Inflammation in Patients With Leber Hereditary Optic Neuropathy Treated With Intravitreal Injection of Recombinant Adeno-Associated Virus 2 Carrying the ND4 Gene: A Secondary Analysis of a Phase 1/2 Clinical Trial.
Intravitreal gene therapy is regarded as generally safe with limited mild adverse events, but its systemic effects remain to be investigated. To examine the association between immune response and intraocular inflammation after ocular gene therapy with recombinant adeno-associated virus 2 carrying the ND4 gene (rAAV2/2-ND4). This secondary analysis of an open-label, dose-escalation phase 1/2 randomized clinical trial of rAAV2/2-ND4 included data from February 13, 2014 (first patient visit), to March 30, 2017 (last patient visit at week 96), the first 2 years after injection. Patients older than 15 years with diagnosed ND4 Leber hereditary optic neuropathy (LHON) and visual acuity of at least counting fingers were enrolled in 1 of 5 cohorts. Four dose cohorts of 3 patients each were treated sequentially. An extension cohort of 3 patients received the dose of 9 × 1010 viral genomes per eye. Patients received increasing doses of rAAV2/2-ND4 (9 × 109, 3 × 1010, 9 × 1010, and 1.8 × 1011 viral genomes per eye) as a single unilateral intravitreal injection. Patients were monitored for 96 weeks after injection; ocular examinations were performed regularly, and blood samples were collected for immunologic testing. A composite ocular inflammation score (OIS) was calculated based on grades of anterior chamber cells and flare, vitreous cells, and haze according to the Standardization of Uveitis Nomenclature. The systemic immune response was quantified by enzyme-linked immunospot (cellular immune response), enzyme-linked immunosorbent assay (IgG titers), and luciferase assay (neutralizing antibody [NAb] titers). The present analysis included 15 patients (mean [SD] age, 47.9 [17.2] years; 13 men and 2 women) enrolled in the 5 cohorts of the clinical trial. Thirteen patients experienced intraocular inflammation after rAAV2/2-ND4 administration. Mild anterior chamber inflammation and vitritis were reported at all doses, and all cases were responsive to treatment. A maximum OIS of 9.5 was observed in a patient with history of idiopathic uveitis. Overall, OIS was not associated with the viral dose administered. No NAbs against AAV2 were detected in aqueous humor before treatment. Two patients tested positive for cellular immune response against AAV2 at baseline and after treatment. Humoral immune response was not apparently associated with the dose administered or with the immune status of patients at baseline. No association was found between OISs and serum NAb titers. In this study, intravitreal administration of rAAV2/2-ND4 in patients with LHON was safe and well tolerated. Further investigations may shed light into the local immune response to rAAV2/2-ND4 as a potential explanation for the observed intraocular inflammation.


Autologous transplantation of adipose-derived mesenchymal stem cells attenuates cerebral ischemia and reperfusion injury through suppressing apoptosis and inducible nitric oxide synthase.
The purpose of this study was to investigate whether autologous transplantation of adipose-derived mesenchymal stem cells (ADMSCs) has a neuroprotective effect against cerebral ischemia/reperfusion (I/R) injury in rats and to explore the possible underlying mechanisms. Adult male Sprague-Dawley rats were randomly assigned into the sham, I/R injury model (I/R), and model plus autologous transplantation of ADMSCs (ADMSC) groups. Cerebral I/R injury was induced by 2 h middle cerebral artery occlusion (MCAO) followed by reperfusion for 24 h. Rats in the I/R and ADMSC groups were intravenously injected with culture medium and ADMSCs (2.0x10(6)), respectively, at the onset of reperfusion and 12 h after reperfusion. Cerebral infarct volume was detected by triphenyltetrazolium chloride (TTC) staining. The histopathological changes and neuronal apoptosis in the ischemic penumbra were evaluated with H&E staining and the TUNEL assay, respectively. The nitric oxide (NO) content, caspase-3 activity and the Bax/Bcl-2 protein ratio were also measured. Moreover, the inducible nitric oxide synthase (iNOS) expression in the ischemic regions of rats was determined by immunohistochemical staining, quantitative real-time RT-PCR and western blot analysis. We found that autologous transplantation of ADMSCs significantly reduced the cerebral infarct volume, improved the I/R injury-induced brain damages and inhibited the neuronal apoptosis. ADMSC implantation also decreased caspase-3 activity and the Bax/Bcl-2 protein ratio, and markedly downregulated the expression of iNOS and thus prevented NO release in response to cerebral I/R injury. Taken together, our results demonstrated that autologous transplantation of ADMSCs can protect the brain against cerebral I/R injury via the inhibition of neuronal apoptosis and iNOS expression.


Psychedelics not linked to mental health problems or suicidal behavior: a population study.
A recent large population study of 130,000 adults in the United States failed to find evidence for a link between psychedelic use (lysergic acid diethylamide, psilocybin or mescaline) and mental health problems. Using a new data set consisting of 135,095 randomly selected United States adults, including 19,299 psychedelic users, we examine the associations between psychedelic use and mental health. After adjusting for sociodemographics, other drug use and childhood depression, we found no significant associations between lifetime use of psychedelics and increased likelihood of past year serious psychological distress, mental health treatment, suicidal thoughts, suicidal plans and suicide attempt, depression and anxiety. We failed to find evidence that psychedelic use is an independent risk factor for mental health problems. Psychedelics are not known to harm the brain or other body organs or to cause addiction or compulsive use; serious adverse events involving psychedelics are extremely rare. Overall, it is difficult to see how prohibition of psychedelics can be justified as a public health measure.


Glucose-6-phosphate dehydrogenase deficiency. WHO Working Group.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the commonest enzyme disorder of human beings and a globally important cause of neonatal jaundice, which can lead to kernicterus and death or spastic cerebral palsy. It can also lead to life-threatening haemolytic crises in childhood and at later ages, by interacting with specific drugs and with fava beans in the diet. The complications of G6PD deficiency can largely be prevented by education and information, and neonatal jaundice can be successfully treated by phototherapy, a cheap and simple approach suitable for use in primary health care. This update describes developments in the methodology for characterizing G6PD deficiency, recent knowledge of the factors that can cause haemolysis, community approaches for prevention of haemolytic crises and neonatal jaundice, and the implications of recent advances at the DNA level.


Highly efficient production of the Alzheimer's γ-secretase integral membrane protease complex by a multi-gene stable integration approach.
Inefficient production of membrane-embedded multi-protein complexes by conventional methods has largely prevented the generation of high-resolution structural information and the performance of high-throughput drug discovery screens for this class of proteins. Not exempt from this rule is γ-secretase, an intramembrane-cleaving protease complex regulating a multitude of signaling pathways and biological processes by influencing gene transcription. γ-Secretase is also implicated in the pathogenesis of Alzheimer's disease and several types of cancer. As an additional challenge, the reconstitution of the protease complex in its active form requires an intricate assembly and maturation process, including a highly regulated endoproteolytic processing of its catalytic component. In this article we report the application of a transposon-mediated multigene stable integration technology to produce active γ-secretase in mammalian cells in amounts adequate for crystallization studies and drug screening. Our strategy is expected to help elucidate the molecular mechanisms of intramembrane proteolysis. It is further expected to be widely used for the production of other multi-protein complexes for applications in structural biology and drug development.


Risk factors of suicidal ideation in a population of UK military veterans seeking support for mental health difficulties.
Little has been reported regarding the risk factors of suicidal ideation in individuals once they have left the military in the UK. The aim of this paper was to explore the risk factors associated with suicidal ideation in a sample of treatment-seeking veterans. Using a cross-sectional design, participants included veterans (n=144) seeking treatment from a national mental health charity in the UK. Individuals completed questionnaires regarding their military experiences, pre-enlistment factors and health. Data were then linked to risk assessments extracted from clinical records. After controlling for relevant variables, suicidal ideation was significantly higher in veterans who were unemployed (OR 8.01; 95% CI 1.79 to 35.80), were early service leavers (OR 8.46; 95% CI 2.21 to 32.35) and those with a history of childhood adversity (OR 6.92; 95% CI 2.10 to 22.82). In addition, taking longer than 5 years to seek help was associated with a reduced risk of suicidal ideation (OR 0.10; 95% CI 0.06 to 0.87). There was no association between health outcomes and suicidal ideation. Risk factors associated with suicidal ideation in this sample of veterans included: being unemployed, an early service leaver, taking less than 5 years to seek help and experiencing preservice adversity.


SU-E-J-210: Lung Tumor Target Volume Contours on EPID Cine Mode Images.
While real time imaging of treatment through an electronic portal imaging device (EPID) is a powerful tool to monitor treatment, limited field of view and lower contrast from an MV beam can make assessment difficult for physicians. This work will develops a method to register and project contour outlines for the internal target volume (ITV) and planning target volume (PTV) of lung tumor cases onto cine mode EPID images to help physicians in interpretation during treatment. A sequence of EPID images, acquired during treatment, was registered to treatment planning computed tomography (CT) by machine geometry and patient setup with cone-beam computed tomography (CBCT). The planning CT was converted from Hounsfield scale to electron density by calibration curves of our CT simulator and digitally reconstructed radiographs (DRRs) were produced to match the EPID geometry, pixel for pixel. ITV and PTV structures as defined on the planning CT were then projected onto the DRRs. The DRRs were registered to the EPID images using cross correlation of a single template defined within the treatment aperture of each image. Once registered, the contours from the DRR were transferred to the EPID. We were able to successfully register MV DRRs to EPID images and display the projected target volumes. Without introduced motion, geometric registration and CBCT guided patient setup up were sufficient to register the contours within a single pixel, as normalized cross correlations produced no additional shift. We expect the DRR/EPID registration to be an important step when looking at cases with substantial tumor movement. The visualization of target volumes provides a tool for physicians to interpret EPID images and assess treatment, especially in cases with tumor movement. The methods developed will serve as the basis for a clinical tool providing real time contours.


[Epidemiology of infectious hematopoietic necrosis (IHN) of salmonid fish in France: study of the course of natural infection by combined use of viral and seroneutralization test and eradication attempts].
Infectious haematopoietic necrosis (IHN), a rhabdoviral infection of salmonid fish, was considered to be an exotic disease in Europe until it was recognized in France and Italy in 1987. In France, the existence of this new condition led the authorities in charge of animal health to order epidemiological studies to be undertaken. These studies were based upon virological, serological and experimental diagnostic methods and also encompassed disease eradication attempts. Studies were conducted at 7 fish farming sites, involved 1,545 salmonid fish, of which 848 were sacrificed, and represented 262 virological examinations and 1,782 serum neutralization tests. The presence of the IHN virus was detected in the 7 trout farm fish populations that were located in 5 regions, one of which was situated 600 km from the place where the first isolation of IHN virus was made. Moreover, 6 out of 7 rainbow trout populations reared in these farms also harboured viral haemorrhagic septicaemia virus (VHSV) often resulting in overt disease. Rainbow trout was the only salmonid fish species found infected with IHN. Overt infection, which was observed in fish ageing less than 2,200 degrees-days, always occurred at water temperatures below 14 degrees C, and the younger fish were more susceptible (mortality rate > or = 80%). Although the IHN virus is easily isolable from fish undergoing overt infection, it was hardly detectable in survivors until they were adults, at which stage the virus was shedded via sexual products which constituted suitable materials for virological examination and disease transmission assays. Survivors of overt and dormant IHN infection developed consistent immune response and special attention was paid to neutralizing antibodies (NAb) to IHN virus. The detection of such NAb in fish from infected farming sites or other NAb from presumably IHN-free sites, correlated fairly well with the presence and further detection of IHN virus among such fish populations. Our data provide arguments for considering the serological technique as a suitable means of completing fish health surveillance programmes for IHN. Although our results are in agreement with part of the existing knowledge on IHN, they differed in several points: rainbow trout was the only susceptible fish species; overt IHN was always recorded in juveniles and at water temperatures below 14 degrees C; IHN virus could not be recovered from the mucus of infected broodfish; IHN infection usually coexisted with VHS infection in same fish population; and serology was widely and successfully used for the diagnosis of IHN.


Recent improvements to the SMART domain-based sequence annotation resource.
SMART (Simple Modular Architecture Research Tool, http://smart.embl-heidelberg.de) is a web-based resource used for the annotation of protein domains and the analysis of domain architectures, with particular emphasis on mobile eukaryotic domains. Extensive annotation for each domain family is available, providing information relating to function, subcellular localization, phyletic distribution and tertiary structure. The January 2002 release has added more than 200 hand-curated domain models. This brings the total to over 600 domain families that are widely represented among nuclear, signalling and extracellular proteins. Annotation now includes links to the Online Mendelian Inheritance in Man (OMIM) database in cases where a human disease is associated with one or more mutations in a particular domain. We have implemented new analysis methods and updated others. New advanced queries provide direct access to the SMART relational database using SQL. This database now contains information on intrinsic sequence features such as transmembrane regions, coiled-coils, signal peptides and internal repeats. SMART output can now be easily included in users' documents. A SMART mirror has been created at http://smart.ox.ac.uk.


Mesenchymal stromal cells for cardiovascular disease.
The fields of regenerative medicine and cellular therapy have been the subject of tremendous hype and hope. In particular, the perceived usage of somatic cells like mesenchymal stromal cells (MSCs) has captured the imagination of many. MSCs are a rare population of cells found in multiple regions within the body that can be readily expanded ex vivo and utilized clinically. Originally, it was hypothesized that transplantation of MSCs to sites of injury would lead to de novo tissue-specific differentiation and thereby replace damaged tissue. Now, it is generally agreed that MSC home to sites of injury and direct positive remodeling via the secretion of paracrine factors. Consequently, their clinical utilization has largely revolved around their abilities to promote neovascularization for ischemic disorders and modulate overly exuberant inflammatory responses for autoimmune and alloimmune conditions. One of the major issues surrounding the development of somatic cell therapies like MSCs is that despite evoking a positive response, long-term engraftment and persistence of these cells is rare. Consequently, very large cell doses need be administered for raising production, delivery, and efficacy issues. In this review, we will outline the field of MSC in the context of ischemia and discuss causes for their lack of persistence. In addition, some of the methodologies be used to enhance their therapeutic potential will be highlighted.


Association of Early Stress Testing with Outcomes for Emergency Department Evaluation of Suspected Acute Coronary Syndrome.
Professional society guidelines suggest early stress testing (within 72 hours) after an emergency department (ED) evaluation for suspected acute coronary syndrome (ACS). However, there is increasing concern that current practice results in over-testing without evidence of benefit. We test the hypothesis that early stress testing improves outcomes. We analyzed prospectively collected data from 9 EDs on patients with suspected ACS, 1999-2001. We excluded patients with an ED diagnosis of ACS. The primary outcome was 30-day major adverse cardiac events (MACEs), including all-cause death, acute myocardial infarction, and revascularization. We used the HEART score to determine pretest ACS risk (low, intermediate, and high). To mitigate potential confounding, patients with and without early stress testing were matched within pretest risk strata in a 1:2 ratio using propensity scores. Of 7127 potentially eligible patients, 895 (13%) received early stress testing. The analytic cohort included 895 patients with early stress testing matched to 1790 without early stress testing. The overall 30-day MACE rate in both the source and analytic population was 3%. There were no baseline imbalances after propensity score matching (P > 0.1 for more than 30 variables). There was no association between early stress testing and 30-day MACE [odds ratio, 1.0; 95% confidence interval (CI), 0.6-1.7]. There was no effect modification by pretest risk (low: odds ratio, 1.0; 95% CI, 0.2-3.7; intermediate: 1.2; 95% CI, 0.6-2.6; high: 0.4; 95% CI, 0.1-1.6). Early stress testing is not associated with reduced MACE in patients evaluated for suspected ACS. Early stress testing may have limited value in populations with low MACE rate.


Selecting sites for converting farmlands to wetlands in the Sanjiang Plain, Northeast China, based on remote sensing and GIS.
Wetlands in the Sanjiang Plain are rich in biodiversity and natural resources in the northeast of China. However, this wetland area has decreased in size and deteriorated in quality owing to expanded agricultural activities since the 1950s. Converting farmlands to wetlands is necessary to improve these conditions. Using Remote Sensing (RS) and Geographic Information Systems (GIS) technologies, we derived farmland productivity data and hydrology data for the Sanjiang Plain. The farmland productivity data were derived from land use and net primary productivity (NPP) data of the MODIS products. We obtained three productivity farmland classes (low, medium, and high) through the NPP anomaly percentage method. We were only concerned with the low-productivity farmland. Hydrology data were modeled with a wetness index, which was derived from Digital Elevation Model (DEM) data. Based on these two data layers, we identified and prioritized sites for the conversion of farmlands to wetlands. The areas with low farmland productivity and medium or high wetness values have potential to support the conversion of farmlands to wetlands. Potential sites were prioritized in terms of patch size and proximity to natural wetlands and water bodies. We obtained three priority classes, among which the high-priority class would be used as the areas for the recent conversion of farmlands to wetlands. The area of this class was 75,888 ha and accounted for 1.3% of the total farmland area.


Improving shape retrieval by spectral matching and meta similarity.
We propose two computational approaches for improving the retrieval of planar shapes. First, we suggest a geometrically motivated quadratic similarity measure, that is optimized by way of spectral relaxation of a quadratic assignment. By utilizing state-of-the-art shape descriptors and a pairwise serialization constraint, we derive a formulation that is resilient to boundary noise, articulations and nonrigid deformations. This allows both shape matching and retrieval. We also introduce a shape meta-similarity measure that agglomerates pairwise shape similarities and improves the retrieval accuracy. When applied to the MPEG-7 shape dataset in conjunction with the proposed geometric matching scheme, we obtained a retrieval rate of 92.5%.


Emotional Experiences of COVID-19 Patients in China: A Qualitative Study.
This study explored the emotional experiences of COVID-19 patients in China. Thirty-four patients diagnosed with COVID-19 participated in semi-structured telephone interviews. We used qualitative methods to investigate the distribution patterns and characteristics of patients' emotional experiences. The results indicated that emotional experiences showed different characteristics at different stages during isolation and treatment. COVID-19 patients' emotional discourse encompassed eight main themes, namely, feelings of shock at the diagnosis, yearning for future life, attachment to one's family, depression during the treatment, self-restriction due to probable contagiousness, powerlessness about the disease, open-mindedness about death, and faith in the joint efforts to fight COVID-19. These themes related to experiences concerning infection, isolation, outlook on life and death, stigma, and macro-identity. The findings suggest that the unexpected experience of COVID-19 infection exacerbated patients' negative emotions. COVID-19 patients' emotional stress stemmed from isolated environments, physiological effects of the disease, panic about the unknown, and realistic economic pressure. The government, medical staff, family members of patients, and the media should therefore work together to ensure proper emotional care for COVID-19 patients.


Forecasting seasonal outbreaks of influenza.
Influenza recurs seasonally in temperate regions of the world; however, our ability to predict the timing, duration, and magnitude of local seasonal outbreaks of influenza remains limited. Here we develop a framework for initializing real-time forecasts of seasonal influenza outbreaks, using a data assimilation technique commonly applied in numerical weather prediction. The availability of real-time, web-based estimates of local influenza infection rates makes this type of quantitative forecasting possible. Retrospective ensemble forecasts are generated on a weekly basis following assimilation of these web-based estimates for the 2003-2008 influenza seasons in New York City. The findings indicate that real-time skillful predictions of peak timing can be made more than 7 wk in advance of the actual peak. In addition, confidence in those predictions can be inferred from the spread of the forecast ensemble. This work represents an initial step in the development of a statistically rigorous system for real-time forecast of seasonal influenza.


Mutations in the clk-1 gene of Caenorhabditis elegans affect developmental and behavioral timing.
We have identified three allelic, maternal-effect mutations that affect developmental and behavioral timing in Caenorhabditis elegans. They result in a mean lengthening of embryonic and postembryonic development, the cell cycle period and life span, as well as the periods of the defecation, swimming and pumping cycles. These mutants also display a number of additional phenotypes related to timing. For example, the variability in the length of embryonic development is several times larger in the mutants than in the wild type, resulting in the occasional production of mutant embryos developing more rapidly than the most rapidly developing wild-type embryos. In addition, the duration of embryonic development of the mutants, but not of the wild type, depends on the temperature at which their parents were raised. Finally, individual variations in the severity of distinct mutant phenotypes are correlated in a counterintuitive way. For example, the animals with the shortest embryonic development have the longest defecation cycle and those with the longest embryonic development have the shortest defecation cycle. Most of the features affected by these mutations are believed to be controlled by biological clocks, and we therefore call the gene defined by these mutations clk-1, for "abnormal function of biological clocks."


A 5-hydroxytryptamine receptor antagonist, sarpogrelate, reduces renal tubulointerstitial fibrosis by suppressing PAI-1.
A selective 5-hydroxytryptamine (5-HT) 2A receptor antagonist sarpogrelate (SG) blocks serotonin-induced platelet aggregation. It has been used clinically for the treatment of peripheral arterial disease. SG might be able to improve chronic ischemia, which contributes to renal fibrosis progression by maintaining renal microcirculation. This study investigated whether SG suppresses renal fibrosis. C57BL/6 mice fed a 0.2% adenine-containing diet for 6 wk developed severe tubulointerstitial fibrosis with kidney dysfunction. Subsequent SG treatment (30 mg·kg(-1)·day(-1)) for 4 wk improved these changes significantly by increasing peritubular blood flow in the fibrotic area, as evaluated by intravital microscopy and decreasing fibrin deposition. Urinary L-type fatty acid-binding protein, up-regulated by renal hypoxia, was also reduced by SG. Additionally, results showed that mRNA expression of plasminogen activator inhibitor-1 (PAI-1), which is known to promote fibrosis by mediating and enhancing transforming growth factor (TGF)-β1 signaling, was suppressed by SG treatment in the kidney. In vitro experiments using cultured murine proximal tubular epithelial (mProx) cells revealed that incubation with TGF-β1 and 5-HT increased PAI-1 mRNA expression; SG significantly reduced it. In conclusion, SG reduces renal fibrosis not only by the antithrombotic effect of maintaining peritubular blood flow but also by suppressing PAI-1 expression in renal tubular cells.


Elective spinal surgery in asymptomatic HIV-seropositive persons: perioperative complications and outcomes.
This is a retrospective study consisting of medical records review and evaluation of pertinent radiographs. In this study, complications and outcomes are reviewed in a series of HIV-positive patients undergoing spinal surgery. Surgery was performed for conditions that were unrelated to HIV infection. From the period 1996 to 2000, we identified 10 seropositive patients who underwent spinal procedures for conditions that did not arise as a complication of HIV infection. A retrospective review was done, which included inpatient and out patient records. A total of 6 women and 5 men who were HIV-positive underwent spinal surgery during the time period reviewed. Surgery was performed for a variety of conditions, including lumbar disc herniaition, degenerative disc disease, cervical disc herniation, spinal fractures, and Arnold-Chiari malformation. The mean CD4 count was 279 cells/mm3 before surgery. All patients had at least 6 months of conservative management before surgery, except in the case of those who had spinal fractures. Two patients sustained postoperative complications: 1 patient developed a superficial wound infection, which was successfully treated with antibiotics, and 1 patient experienced prolonged fever (7 days) after surgery, which resolved without a clear diagnosis. All patients were improved after surgery (mean follow-up, 29.3 months) Many physicians have held a nihilistic approach, when it comes to the treatment of HIV-positive persons. However, our results in this small series of patients suggest that spinal surgery may be appropriate and can be performed with acceptable outcomes in selected patients.


The re-enactment of childhood sexual abuse in maternity care: a qualitative study.
The process of pregnancy and birth are profound events that can be particularly challenging for women with a history of childhood sexual abuse. The silence that surrounds childhood sexual abuse means that few women disclose it and those caring for them will often not be aware of their history. It is known from anecdotal accounts that distressing memories may be triggered by childbirth and maternity care but research data on the subject are rare. This paper explores aspects of a study on the maternity care experiences of women who were sexually abused in childhood that demonstrate ways that maternity care can be reminiscent of abuse. Its purpose is to inform those providing care for these women. The experiences of women were explored through in-depth interviews in this feminist narrative study. The Voice-Centred Relational Method and thematic analysis were employed to examine interview data. Women sometimes experienced re-enactment of abuse through intimate procedures but these were not necessarily problematic in themselves. How they were conducted was important. Women also experienced re-enactment of abuse through pain, loss of control, encounters with strangers and unexpected triggers. Many of these experiences were specific to the woman, often unpredictable and not necessarily avoidable. Maternity care was reminiscent of abuse for women irrespective of whether they had disclosed to midwives and was not necessarily prevented by sensitive care. 'Re-enactment of abuse' occurred both as a result of events that involved the crossing of a woman's body boundaries and more subjective internal factors that related to her sense of agency. As staff may not know of a woman's history, they must be alert to unspoken messages and employ 'universal precautions' to mitigate hidden trauma. Demonstrating respect and enabling women to retain control is crucial. Getting to know women is important in the building of trusting relationships that will facilitate the delivery of sensitive care and enable women to feel safe so that the re-enactment of abuse in maternity care is minimised.


Temperature dependence of thermo-optic effects of single-crystal and ceramic TGG.
The temperature dependence of the thermo-optic effects in single crystal and ceramic TGG were evaluated by using the Fizou interferometer method. The temperature dependence of the refractive index and thermal expansion are significantly improved at low temperature for both ceramics and single crystals. Our estimation using a figure of merit indicated that a TGG ceramics cooled to liquid nitrogen temperature can reduce thermal wave-front distortion by a factor of up to 4.7 with respect to that at 300 K, and can reduce thermal birefringence effects by up to a factor of 12 with respect to those at 300 K.


Experiences of Australian podiatrists working through the 2020 coronavirus (COVID-19) pandemic: an online survey.
On the 19th of January, 2020, the Chief Medical Officer of Australia issued a statement about a novel coronavirus, or SARS-CoV-2. Since this date, there have been variable jurisdictional responses, including lockdowns, and restrictions on podiatry practice. This study aimed to describe impacts of the SARS-CoV-2 pandemic on the podiatry profession in Australia. This was a cross sectional study of Australian podiatrists using demographic data collected between 2017 and 2020, and pandemic-related question responses collected between 30th March and 31st August, 2020. Data were collected online and participants described their work settings, patient funding types, business decisions and impacts, and information sources used to guide practice decisions during this time-period. Inductive thematic analysis was used to analyse open-ended questions about their practice impact of SARS-CoV-2. There were 732 survey responses, with 465 Australian podiatrists or podiatric surgeons providing responses describing pandemic impact. From these responses, 223 (49% of 453) podiatrists reported no supply issues, or having adequate supplies for the foreseeable future with personal protective equipment (PPE) or consumables to support effective infection prevention and control. The most frequent responses about employment, or hours of work, impact were reported in the various categories of "business as usual" (n = 312, 67%). Participants described most frequently using the local state and territory Department of Health websites (n = 347, 75%), and the Australian Podiatry Association (n = 334, 72%) to make decisions about their business. Overarching themes which resounded through open-ended comments was that working through the pandemic was likened to a marathon, and not a sprint. Themes were: (i) commitment to do this, (ii) it's all in the plan, but not everything goes to plan, (iii) my support team must be part of getting through it, (iv) road blocks happen, and (v) nothing is easy, what's next? Podiatrists in Australia reported variable pandemic impact on their business decisions, PPE stores, and their valued sources of information. Podiatrists also described their "marathon" journey through the pandemic to date, with quotes describing their challenges and highlights. Describing these experiences should provide key learnings for future workforce challenges, should further restrictions come into place.


Smoking cessation intervention in Australian general practice: a secondary analysis of a cluster randomised controlled trial.
GPs have limited capacity to routinely provide smoking cessation support. New strategies are needed to reach all smokers within this setting. To evaluate the effect of a pharmacist-coordinated interdisciplinary smoking cessation intervention delivered in Australian general practice. Secondary analysis of a cluster randomised controlled trial (RCT) conducted in 41 Australian general practices. In all, 690 current smokers were included in this study: 373 from intervention clinics (n = 21) and 317 from control clinics (n = 18). A total of 166 current smokers had spirometry-confirmed chronic obstructive pulmonary disease (COPD). In the intervention clinics, trained pharmacists provided smoking cessation support plus Quitline referral. Control clinics provided usual care plus Quitline referral. Those with COPD in the intervention group (n = 84) were referred for home medicines review (HMR) and home-based pulmonary rehabilitation (HomeBase), which included further smoking cessation support. Outcomes included carbon monoxide (CO)-validated smoking abstinence, self-reported use of smoking cessation aids, and differences between groups in readiness-to-quit score at 6 months. Intention-to-treat analysis showed similar CO-validated abstinence rates at 6 months in the intervention (4.0%) and control clinics (3.5%). No differences were observed in readiness-to-quit scores between groups at 6 months. CO-validated abstinence rates were similar in those who completed HMR and at least six sessions of HomeBase to those with COPD in usual care. A pharmacist-coordinated interdisciplinary smoking cessation intervention when integrated in a general practice setting had no advantages over usual care. Further research is needed to evaluate the effect of HMR and home-based pulmonary rehabilitation on smoking abstinence in smokers with COPD.


Mortality of patients infected with HIV in the intensive care unit (2005 through 2010): significant role of chronic hepatitis C and severe sepsis.
The combination antiretroviral therapy (cART) has led to decreased opportunistic infections and hospital admissions in human immunodeficiency virus (HIV)-infected patients, but the intensive care unit (ICU) admission rate remains constant (or even increased in some instances) during the cART era. Hepatitis C virus (HCV) infection is associated with an increased risk for hospital admission and/or mortality (particularly those related to severe liver disease) compared with the general population. The aim of this study was to assess the mortality among HIV-infected patients in ICU, and to evaluate the impact of HIV/HCV coinfection and severe sepsis on ICU mortality. We carried out a retrospective study based on patients admitted to ICU who were recorded in the Minimum Basic Data Set (2005 through 2010) in Spain. HIV-infected patients (All-HIV-group (n = 1,891)) were divided into two groups: HIV-monoinfected patients (HIV group (n = 1,191)) and HIV/HCV-coinfected patients (HIV/HCV group (n = 700)). A control group (HIV(-)/HCV(-)) was also included (n = 7,496). All-HIV group had higher frequencies of severe sepsis (57.7% versus 39.4%; P < 0.001) than did the control group. Overall, ICU mortality in patients with severe sepsis was much more frequent than that in patients without severe sepsis (other causes) at days 30 and 90 in HIV-infected patients and the control group (P < 0.001). Moreover, the all-HIV group in the presence or absence of severe sepsis had a higher percentage of death than did the control group at days 7 (P < 0.001), 30 (P < 0.001) and 90 (P < 0.001). Besides, the HIV/HCV group had a higher percentage of death, both in patients with severe sepsis and in patients without severe sepsis compared with the HIV group at days 7 (P < 0.001) and 30 (P < 0.001), whereas no differences were found at day 90. In a bayesian competing-risk model, the HIV/HCV group had a higher mortality risk (adjusted hazard ratio (aHR) = 1.44 (95% CI = 1.30 to 1.59) and aHR = 1.57 (95% CI = 1.38 to 1.78) for patients with and without severe sepsis, respectively). HIV infection was related to a higher frequency of severe sepsis and death among patients admitted to the ICU. Besides, HIV/HCV coinfection contributed to an increased risk of death in both the presence and the absence of severe sepsis.


Optical transverse excitation in quaterthiophene crystals by ultraviolet-visible internal and attenuated total reflection.
We report internal and attenuated total reflection of light at the interface between glass and a quaterthiophene crystal in the spectral region of the electronic transitions. The bands corresponding to the absorption of the a(u) and b(u) Frenkel exciton states are detected for different polarization of the incident light. In particular, the wave-normal vector being almost perpendicular to the b(u) transition dipole moment allows its transverse component to be accessed, whose excitation in conventional external reflection or transmission spectroscopies is forbidden.


Weekly change in mindfulness and perceived stress in a mindfulness-based stress reduction program.
The purpose of the study was to examine weekly change in self-reported mindfulness and perceived stress in participants who completed an 8-week course in mindfulness-based stress reduction (MBSR). Participants were 87 adults with problematic levels of stress related to chronic illness, chronic pain, and other life circumstances (mean age = 49 years, 67% female) participating in MBSR in an academic medical center. They completed weekly self-report assessments of mindfulness skills and perceived stress. It was hypothesized that significant improvement in mindfulness skills would precede significant change in stress. Mindfulness skills and perceived stress both changed significantly from pretreatment to posttreatment. Significant increases in mindfulness occurred by the second week of the program, whereas significant improvements in perceived stress did not occur until week 4. Extent of change in mindfulness skills during the first three weeks predicted change in perceived stress over the course of the intervention. Evidence that changes in mindfulness precede changes in perceived stress in a standard MBSR course is consistent with previous studies suggesting that improvements in mindfulness skills may mediate the effects of mindfulness training on mental health outcomes.


Tracking picosecond strain pulses in heterostructures that exhibit giant magnetostriction.
We combine ultrafast X-ray diffraction (UXRD) and time-resolved Magneto-Optical Kerr Effect (MOKE) measurements to monitor the strain pulses in laser-excited TbFe2/Nb heterostructures. Spatial separation of the Nb detection layer from the laser excitation region allows for a background-free characterization of the laser-generated strain pulses. We clearly observe symmetric bipolar strain pulses if the excited TbFe2 surface terminates the sample and a decomposition of the strain wavepacket into an asymmetric bipolar and a unipolar pulse, if a SiO2 glass capping layer covers the excited TbFe2 layer. The inverse magnetostriction of the temporally separated unipolar strain pulses in this sample leads to a MOKE signal that linearly depends on the strain pulse amplitude measured through UXRD. Linear chain model simulations accurately predict the timing and shape of UXRD and MOKE signals that are caused by the strain reflections from multiple interfaces in the heterostructure.


Massive horizontal gene transfer, strictly vertical inheritance and ancient duplications differentially shape the evolution of Bacillus cereus enterotoxin operons hbl, cytK and nhe.
Bacillus cereus sensu lato comprises eight closely related species including the human pathogens Bacillus anthracis and Bacillus cereus. Within B. cereus sensu lato, chromosomally and plasmid-encoded toxins exist. While plasmid-mediated horizontal gene transfer of the emetic toxin, anthrax and insecticidal toxins is known, evolution of enterotoxin genes within the group has not been studied. We report draft genome assemblies of 25 strains, a phylogenetic network of 142 strains based on ANI derived from genome sequences and a phylogeny based on whole-genome SNP analysis. The data clearly support subdivision of B. cereus sensu lato into seven phylogenetic groups. While group I, V and VII represent B. pseudomycoides, B. toyonensis and B. cytotoxicus, which are distinguishable at species level (ANI border ≥ 96 %), strains ascribed to the other five species do not match phylogenic groups. The chromosomal enterotoxin operons nheABC and hblCDAB are abundant within B. cereus both isolated from infections and from the environment. While the duplicated hbl variant hbl a is present in 22 % of all strains investigated, duplication of nheABC is extremely rare (0.02 %) and appears to be phylogenetically unstable. Distribution of toxin genes was matched to a master tree based on seven concatenated housekeeping genes, which depicts species relationships in B. cereus sensu lato as accurately as whole-genome comparisons. Comparison to the phylogeny of enterotoxin genes uncovered ample evidence for horizontal transfer of hbl, cytK and plcR, as well as frequent deletion of both toxins and duplication of hbl. No evidence for nhe deletion was found and stable horizontal transfer of nhe is rare. Therefore, evolution of B. cereus enterotoxin operons is shaped unexpectedly different for yet unknown reasons. Frequent exchange of the pathogenicity factors hbl, cytK and plcR in B. cereus sensu lato appears to be an important mechanism of B. cereus virulence evolution, including so-called probiotic or non-pathogenic species, which might have consequences for risk assessment procedures. In contrast, exclusively vertical inheritance of nhe was observed, and since nhe-negative strains appear to be extremely rare, we suggest that fitness loss may be associated with deletion or horizontal transfer of the nhe operon.


Ligand photo-isomerization triggers conformational changes in iGluR2 ligand binding domain.
Neurological glutamate receptors bind a variety of artificial ligands, both agonistic and antagonistic, in addition to glutamate. Studying their small molecule binding properties increases our understanding of the central nervous system and a variety of associated pathologies. The large, oligomeric multidomain membrane protein contains a large and flexible ligand binding domains which undergoes large conformational changes upon binding different ligands. A recent application of glutamate receptors is their activation or inhibition via photo-switchable ligands, making them key systems in the emerging field of optochemical genetics. In this work, we present a theoretical study on the binding mode and complex stability of a novel photo-switchable ligand, ATA-3, which reversibly binds to glutamate receptors ligand binding domains (LBDs). We propose two possible binding modes for this ligand based on flexible ligand docking calculations and show one of them to be analogues to the binding mode of a similar ligand, 2-BnTetAMPA. In long MD simulations, it was observed that transitions between both binding poses involve breaking and reforming the T686-E402 protein hydrogen bond. Simulating the ligand photo-isomerization process shows that the two possible configurations of the ligand azo-group have markedly different complex stabilities and equilibrium binding modes. A strong but slow protein response is observed after ligand configuration changes. This provides a microscopic foundation for the observed difference in ligand activity upon light-switching.


Steroid correlates of territorial behavior in male jacky dragons, Amphibolurus muricatus.
Male jacky dragons, Amphibolurus muricatus, indicate territoriality to rivals during the mating season through the use of stereotyped motor displays. The relationship between corticosterone (B) and testosterone (T) and its effects on territorial display expression were investigated in captive lizards. Results demonstrated that territorial display production was most effectively predicted by elevated baseline T levels. This parallels the heightened T concentrations exhibited by males in the field during the spring mating period. The effect of social interaction on B and T levels was also examined by presenting territory-holding males with a size-matched male intruder. Resident males exhibited clear differences in the level of territorial response evoked by a male intruder, which were correlated with differences in physiological activity. Males that gave no territorial response had moderate B levels that did not change with social interaction, and significantly lower T levels than males that responded to the intruder with territorial displays. Among displaying males, those exhibiting low levels of territorial responses showed no change in B or T. In contrast, high-level territorial responders exhibited acute B increases when faced with an intruder, which might assist in supporting sustained metabolic activity and could possibly reflect differences in the perception of social stress. These same males also exhibited a reduction in T levels at such times, probably due to the acute rise in B. Combined, these results suggest that high circulating T is acting in a preparatory manner to increase the likelihood of producing a territorial response upon engaging in a social encounter. However, once this response has been initiated T may not need to remain elevated to allow continued expression of territorial behavior.


Mechanisms of Winner-Take-All and Group Selection in Neuronal Spiking Networks.
A major function of central nervous systems is to discriminate different categories or types of sensory input. Neuronal networks accomplish such tasks by learning different sensory maps at several stages of neural hierarchy, such that different neurons fire selectively to reflect different internal or external patterns and states. The exact mechanisms of such map formation processes in the brain are not completely understood. Here we study the mechanism by which a simple recurrent/reentrant neuronal network accomplish group selection and discrimination to different inputs in order to generate sensory maps. We describe the conditions and mechanism of transition from a rhythmic epileptic state (in which all neurons fire synchronized and indiscriminately to any input) to a winner-take-all state in which only a subset of neurons fire for a specific input. We prove an analytic condition under which a stable bump solution and a winner-take-all state can emerge from the local recurrent excitation-inhibition interactions in a three-layer spiking network with distinct excitatory and inhibitory populations, and demonstrate the importance of surround inhibitory connection topology on the stability of dynamic patterns in spiking neural network.