Membrane targeting and secretion of mutant uromodulin in familial juvenile hyperuricemic nephropathy.
Familial juvenile hyperuricemic nephropathy (FJHN) is an autosomal dominant genetic disorder that is characterized by hyperuricemia, gout, and tubulointerstitial nephritis. FJHN is caused by mutations in the UMOD gene, which encodes for uromodulin, the most abundant urinary protein. Herein is demonstrated that patients with FJHN and renal insufficiency exhibit a profound reduction in urinary uromodulin together with either elevated or decreased plasma uromodulin. One young patient with FJHN, however, had normal serum creatinine and normal urinary uromodulin with elevated plasma uromodulin. These observations suggest that there are different urinary and plasma uromodulin profiles in early and late disease and that there may be an altered direction of uromodulin secretion in the course of FJHN as a result of improper intracellular sorting of the mutated protein in the thick ascending limb. With the use of immunohistochemistry and a quantitative immunoassay, targeting and secretion of wild-type and mutant (C77Y and N128S) uromodulin were investigated in the polarized renal epithelial cell line LLC-PK1. In transfected cells, uromodulin mutants were targeted properly to the apical membrane but were secreted less efficiently to the apical compartment than wild-type protein. The expression of mutant uromodulin had no effect on caspase 3 activity. These results indicate that the mutations studied do not impair glycosyl-phosphatidylinositol-mediated apical targeting of the protein but do affect apical secretion. Because the mutant proteins are secreted as efficiently as wild type to the basolateral compartment, the possibility arises that interactions with the immune system at the site of secretion are a contributing factor to the development of tubulointerstitial nephritis in FJHN.


Phytotoxic Activity and Identification of Phytotoxic Substances from Schumannianthus dichotomus.
The phytotoxic potential of plants and their constituents against other plants is being increasingly investigated as a possible alternative to synthetic herbicides to control weeds in crop fields. In this study, we explored the phytotoxicity and phytotoxic substances of Schumannianthus dichotomus, a perennial wetland shrub native to Bangladesh, India, and Myanmar. Leaf extracts of S. dichotomus exerted strong phytotoxicity against two dicot species, alfalfa and cress, and two monocot species, barnyard grass and Italian ryegrass. A bioassay-driven purification process yielded two phenolic derivatives, syringic acid and methyl syringate. Both constituents significantly inhibited the growth of cress and Italian ryegrass in a concentration-dependent manner. The concentrations required for 50% growth inhibition (I50 value) of the shoot and root growth of cress were 75.8 and 61.3 μM, respectively, for syringic acid, compared with 43.2 and 31.5 μM, respectively, for methyl syringate. Similarly, to suppress the shoot and root growth of Italian rye grass, a greater amount of syringic acid (I50 = 213.7 and 175.9 μM) was needed than methyl syringate (I50 = 140.4 to 130.8 μM). Methyl syringate showed higher phytotoxic potential than syringic acid, and cress showed higher sensitivity to both substances. This study is the first to report on the phytotoxic potential of S. dichotomus and to identify phytotoxic substances from this plant material.


Academic Primer Series: Key Papers About Competency-Based Medical Education.
Competency-based medical education (CBME) presents a paradigm shift in medical training. This outcome-based education movement has triggered substantive changes across the globe. Since this transition is only beginning, many faculty members may not have experience with CBME nor a solid foundation in the grounding literature. We identify and summarize key papers to help faculty members learn more about CBME. Based on the online discussions of the 2016-2017 ALiEM Faculty Incubator program, a series of papers on the topic of CBME was developed. Augmenting this list with suggestions by a guest expert and by an open call on Twitter for other important papers, we were able to generate a list of 21 papers in total. Subsequently, we used a modified Delphi study methodology to narrow the list to key papers that describe the importance and significance for educators interested in learning about CBME. To determine the most impactful papers, the mixed junior and senior faculty authorship group used three-round voting methodology based upon the Delphi method. Summaries of the five most highly rated papers on the topic of CBME, as determined by this modified Delphi approach, are presented in this paper. Major themes include a definition of core CBME themes, CBME principles to consider in the design of curricula, a history of the development of the CBME movement, and a rationale for changes to accreditation with CBME. The application of the study findings to junior faculty and faculty developers is discussed. We present five key papers on CBME that junior faculty members and faculty experts identified as essential to faculty development. These papers are a mix of foundational and explanatory papers that may provide a basis from which junior faculty members may build upon as they help to implement CBME programs.


Diagnosis and management of urinary tract infections in the outpatient setting: a review.
Urinary tract infection is among the most common reasons for an outpatient visit and antibiotic use in adult populations. The increasing prevalence of antibacterial resistance among community uropathogens affects the diagnosis and management of this clinical syndrome. To define the optimal approach for treating acute cystitis in young healthy women and in women with diabetes and men and to define the optimal approach for diagnosing acute cystitis in the outpatient setting. Evidence for optimal treatment regimens was obtained by searching PubMed and the Cochrane database for English-language studies published up to July 21, 2014. Twenty-seven randomized clinical trials (6463 patients), 6 systematic reviews, and 11 observational studies (252,934 patients) were included in our review. Acute uncomplicated cystitis in women can be diagnosed without an office visit or urine culture. Trimethoprim-sulfamethoxazole (160/800 mg twice daily for 3 days), nitrofurantoin monohydrate/macrocrystals (100 mg twice daily for 5-7 days), and fosfomycin trometamol (3 g in a single dose) are all appropriate first-line therapies for uncomplicated cystitis. Fluoroquinolones are effective for clinical outcomes but should be reserved for more invasive infections. β-Lactam agents (amoxicillin-clavulanate and cefpodoxime-proxetil) are not as effective as empirical first-line therapies. Immediate antimicrobial therapy is recommended rather than delayed treatment or symptom management with ibuprofen alone. Limited observational studies support 7 to 14 days of therapy for acute urinary tract infection in men. Based on 1 observational study and our expert opinion, women with diabetes without voiding abnormalities presenting with acute cystitis should be treated similarly to women without diabetes. Immediate antimicrobial therapy with trimethoprim-sulfamethoxazole, nitrofurantoin, or fosfomycin is indicated for acute cystitis in adult women. Increasing resistance rates among uropathogens have complicated treatment of acute cystitis. Individualized assessment of risk factors for resistance and regimen tolerability is needed to choose the optimum empirical regimen.


Habitat alteration and the conservation of African primates: case study of Kibale National Park, Uganda.
Tropical forests and the animals they support are being threatened by accelerating rates of forest conversion and degradation. In a continually fluctuating sociopolitical world, it is often impossible to protect areas from such conversion until the political environment is suitable to pursue conservation goals, by which time, the forests have often been converted to other uses. This reality suggests a need for inquiry into which primate species can persist after different types of disturbances and how quickly primate communities can recover from disturbance. Here we examine the persistence of primate populations in disturbed habitats by providing a case study of patterns of primate abundance in areas of Kibale National Park (766 km2), Uganda, that have been modified by different types and intensities of human activities, primarily commercial logging and agricultural clearing. Distributional surveys at 24 sites and detailed line-transect censuses at six sites demonstrate that primate populations in Kibale are often high and suggest that patterns of population change associated with disturbance are complex. Analysis of the land use coverage of Kibale reveals that abandoned farms (10.3%) and degraded forest (8.7%) now cover 146 km2. Unfortunately, we do not know what proportion of the farms were established on areas that were forest versus grassland. However, if the areas that are now abandoned farms were all once forested, this means that 79 km2 of forest has been lost. Based on density estimates from nearby sites, this would represent a loss of 52,612 monkeys and 200 chimpanzees. Populations would also have been affected by the degradation of the 66 km2 (8.7%) of forest. These estimates of the potential reductions in the primate populations that could have resulted from forest clearing and degradation illustrate the importance of protecting land. A review of the literature illustrates that the biomass of primates found within Kibale is very high in comparison to other locations and thus illustrates the importance of Kibale to regional conservation.


Robust estimates of overall immune-repertoire diversity from high-throughput measurements on samples.
The diversity of an organism's B- and T-cell repertoires is both clinically important and a key measure of immunological complexity. However, diversity is hard to estimate by current methods, because of inherent uncertainty in the number of B- and T-cell clones that will be missing from a blood or tissue sample by chance (the missing-species problem), inevitable sampling bias, and experimental noise. To solve this problem, we developed Recon, a modified maximum-likelihood method that outputs the overall diversity of a repertoire from measurements on a sample. Recon outputs accurate, robust estimates by any of a vast set of complementary diversity measures, including species richness and entropy, at fractional repertoire coverage. It also outputs error bars and power tables, allowing robust comparisons of diversity between individuals and over time. We apply Recon to in silico and experimental immune-repertoire sequencing data sets as proof of principle for measuring diversity in large, complex systems.


SspA up-regulates gene expression of the LEE pathogenicity island by decreasing H-NS levels in enterohemorrhagic Escherichia coli.
Enterohemorrhagic Escherichia coli (EHEC) colonizes the intestinal epithelium and causes attaching and effacing (A/E) lesions. Expression of virulence genes, particularly those from the locus of the enterocyte effacement (LEE) pathogenicity island is required for the formation of a type three secretion system, which induces A/E lesion formation. Like other horizontally acquired genetic elements, expression of the LEE is negatively regulated by H-NS. In the non-pathogenic Escherichia coli K-12 strain the stringent starvation protein A (SspA) inhibits accumulation of H-NS, and thereby allows de-repression of the H-NS regulon during the stationary phase of growth. However, the effect of SspA on the expression of H-NS-controlled virulence genes in EHEC is unknown. Here we assess the effect of SspA on virulence gene expression in EHEC. We show that transcription of virulence genes including those of the LEE is decreased in an sspA mutant, rendering the mutant strain defective in forming A/E lesions. A surface exposed pocket of SspA is functionally important for the regulation of the LEE and for the A/E phenotype. Increased expression of ler alleviates LEE expression in an sspA mutant, suggesting that the level of Ler in the mutant is insufficient to counteract H-NS-mediated repression. We demonstrate that the H-NS level is two-fold higher in an sspA mutant compared to wild type, and that the defects of the sspA mutant are suppressed by an hns null mutation, indicating that hns is epistatic to sspA in regulating H-NS repressed virulence genes. SspA positively regulates the expression of EHEC virulence factors by restricting the intracellular level of H-NS. Since SspA is conserved in many bacterial pathogens containing horizontally acquired pathogenicity islands controlled by H-NS, our study suggests a common mechanism whereby SspA potentially regulates the expression of virulence genes in these pathogens.


Egr-1: A Candidate Transcription Factor Involved in Molecular Processes Underlying Time-Memory.
In honey bees, continuous foraging is accompanied by a sustained up-regulation of the immediate early gene Egr-1 (early growth response protein-1) and candidate downstream genes involved in learning and memory. Here, we present a series of feeder training experiments indicating that Egr-1 expression is highly correlated with the time and duration of training even in the absence of the food reward. Foragers that were trained to visit a feeder over the whole day and then collected on a day without food presentation showed Egr-1 up-regulation over the whole day with a peak expression around 14:00. When exposed to a time-restricted feeder presentation, either 2 h in the morning or 2 h in the evening, Egr-1 expression in the brain was up-regulated only during the hours of training. Foragers that visited a feeder in the morning as well as in the evening showed two peaks of Egr-1 expression. Finally, when we prevented time-trained foragers from leaving the colony using artificial rain, Egr-1 expression in the brains was still slightly but significantly up-regulated around the time of feeder training. In situ hybridization studies showed that active foraging and time-training induced Egr-1 up-regulation occurred in the same brain areas, preferentially the small Kenyon cells of the mushroom bodies and the antennal and optic lobes. Based on these findings we propose that foraging induced Egr-1 expression can get regulated by the circadian clock after time-training over several days and Egr-1 is a candidate transcription factor involved in molecular processes underlying time-memory.


Melioidosis: Case reports of confirmed Burkholderia pseudomallei in Saudi Arabia.
Melioidosis is an infectious disease of tropical climates. The disease is caused by the bacterium Burkholderia pseudomallei. Most cases are diagnosed in southeast Asia and northern Australia. Some imported cases diagnosed in returning tourists, soldiers, and immigrants from endemic areas. It caught much attention since the Centers for Disease Control and Prevention (CDC) designated B. pseudomallei as an agent for biological warfare and terrorism. We describe two cases of a 26-year-old Saudi woman who had fulminant sepsis soon after returning from Thailand & a 48-year-old woman with a long history of fever. B. pseudomallei was isolated from both patients blood cultures, and they had different consequences. A confirmed case of melioidosis was not reported before in Saudi Arabia.


Isolation and characterization of renal cancer stem cells from patient-derived xenografts.
As rapidly developing patient-derived xenografts (PDX) could represent potential sources of cancer stem cells (CSC), we selected and characterized non-cultured PDX cell suspensions from four different renal carcinomas (RCC). Only the cell suspensions from the serial xenografts (PDX-1 and PDX-2) of an undifferentiated RCC (RCC-41) adapted to the selective CSC medium. The cell suspension derived from the original tumor specimen (RCC-41-P-0) did not adapt to the selective medium and strongly expressed CSC-like markers (CD133 and CD105) together with the non-CSC tumor marker E-cadherin. In comparison, PDX-1 and PDX-2 cells exhibited evolution in their phenotype since PDX-1 cells were CD133high/CD105-/Ecadlow and PDX-2 cells were CD133low/CD105-/Ecad-. Both PDX subsets expressed additional stem cell markers (CD146/CD29/OCT4/NANOG/Nestin) but still contained non-CSC tumor cells. Therefore, using different cell sorting strategies, we characterized 3 different putative CSC subsets (RCC-41-PDX-1/CD132+, RCC-41-PDX-2/CD133-/EpCAMlow and RCC-41-PDX-2/CD133+/EpCAMbright). In addition, transcriptomic analysis showed that RCC-41-PDX-2/CD133- over-expressed the pluripotency gene ERBB4, while RCC-41-PDX-2/CD133+ over-expressed several tumor suppressor genes. These three CSC subsets displayed ALDH activity, formed serial spheroids and developed serial tumors in SCID mice, although RCC-41-PDX-1/CD132+ and RCC-41-PDX-2/CD133+ displayed less efficiently the above CSC properties. RCC-41-PDX-1/CD132+ tumors showed vessels of human origin with CSC displaying peri-vascular distribution. By contrast, RCC-41-PDX-2 originated tumors exhibiting only vessels of mouse origin without CSC peri-vascular distribution.Altogether, our results indicate that PDX murine microenvironment promotes a continuous redesign of CSC phenotype, unmasking CSC subsets potentially present in a single RCC or generating ex novo different CSC-like subsets.


An objective sign in painful crisis in sickle cell anemia: the concomitant reduction of high density red cells.
The etiopathologic basis of painful crisis in sickle cell anemia is largely unknown, and no objective criteria for its diagnosis and follow-up exist at present. We have studied 11 patients through 14 painful crises and observed a significant decrease of the densest fraction of red cells in 12 of the 14 crises as determined by isopycnic Percoll-Stractan continuous density gradients. If the first observation is normalized to 100%, the average decrease in dense cells was 77% with a range of 36% to 94%. The time needed for the percentage of dense cells to return to the steady-state level varied from seven to more than 30 days. These findings were in sharp contrast to the stability of the density pattern observed in another group of sickle cell patients, who were studied during crisis-free periods. The mechanism of the disappearance of dense cells could involve selective destruction by the reticuloendothelial (RE) system, selective sequestration in the areas of vasoocclusion, or a combination of both factors.


Retail Deli Slicer Inspection Practices: An EHS-Net Study.
The Centers for Disease Control and Prevention (CDC) estimates that 3,000 people die in the United States each year from foodborne illness, and Listeria monocytogenes causes the third highest number of deaths. Risk assessment data indicate that L. monocytogenes contamination of particularly delicatessen meats sliced at retail is a significant contributor to human listeriosis. Mechanical deli slicers are a major source of L. monocytogenes cross-contamination and growth. In an attempt to prevent pathogen cross-contamination and growth, the U.S. Food and Drug Administration (FDA) created guidance to promote good slicer cleaning and inspection practices. The CDC's Environmental Health Specialists Network conducted a study to learn more about retail deli practices concerning these prevention strategies. The present article includes data from this study on the frequency with which retail delis met the FDA recommendation that slicers should be inspected each time they are properly cleaned (defined as disassembling, cleaning, and sanitizing the slicer every 4 h). Data from food worker interviews in 197 randomly selected delis indicate that only 26.9% of workers ( n = 53) cleaned and inspected their slicers at this frequency. Chain delis and delis that serve more than 300 customers on their busiest day were more likely to have properly cleaned and inspected slicers. Data also were collected on the frequency with which delis met the FDA Food Code provision that slicers should be undamaged. Data from observations of 685 slicers in 298 delis indicate that only 37.9% of delis ( n = 113) had slicers that were undamaged. Chain delis and delis that provide worker training were more likely to have slicers with no damage. To improve slicer practices, food safety programs and the retail food industry may wish to focus on worker training and to focus interventions on independent and smaller delis, given that these delis were less likely to properly inspect their slicers and to have undamaged slicers.


A smartphone application of alcohol resilience treatment for behavioral self-control training.
High relapse rate is one of the most prominent problems in addiction treatment. Alcohol Resilience Treatment (ART), an alcohol addiction therapy, is based on Cue Exposure Treatment, which has shown promising results in preliminary studies. ART aims at optimizing the core area of relapse prevention, and intends to improve patients' capability to withstand craving of alcohol. This method emphasizes the interplay of resilience and resourcefulness. It contains 6 sessions with different topics according to the stage of treatment circuit, and each session consists of 6 steps. Due to the purity and structure of the treatment rationale, it is realistic, reasonable and manageable to transform the method into a smartphone application. An ART app in Android system and an accessory of bilateral tactile stimulation were developed and will be used in a study with behavioral self-control training. This paper presents the design and realization of the smartphone based ART application. The design of a pilot study, which is to examine the benefits of a smartphone application providing behavioral self-control training, is also reported in this paper.


The paradoxical effects of splenectomy on tumor growth.
There is a vast and contradictory literature concerning the effect of the spleen and particularly of splenectomy on tumor growth. Sometimes splenectomy seems to inhibit tumor growth, but in other cases it seems, paradoxically, to facilitate both oncogenesis and the growth of established tumors. In this essay I have selected from this large literature a few papers that seem particularly instructive, in the hope of extracting some understanding of the rules governing this paradoxical behavior. In general, whether splenectomy enhances or inhibits tumor growth seems to depend primarily upon the ratio of spleen to tumor. Small proportions of spleen cells usually stimulate tumor growth, in which case splenectomy is inhibitory. Larger proportions of the same cells, especially if they are from immunized animals, usually inhibit tumor growth, in which case splenectomy results in tumor stimulation.


Aluminum alkoxide, amide and halide complexes supported by a bulky dipyrromethene ligand: synthesis, characterization, and preliminary ε-caprolactone polymerization activity.
Aluminum halide, alkoxide and amide complexes 2-6 of the form (N,N)AlX2-nYn (n = 0, 1 and (N,N) = 1,9-dimesityl-5-phenyldipyrromethene (1)) were synthesized and characterized by NMR spectroscopy and X-ray crystallography. The in situ generated lithium salt of dipyrromethene 1 was reacted with AlX3 to afford aluminum halide complexes (N,N)AlX2 (X = Cl (2), I (3)) which were isolated as dichroic crystals. Salt metathesis reactions were employed to produce alkoxide complexes (N,N)Al(Cl)(O(t)Bu) (4) and (N,N)Al(O(t)Bu)2 (5) from compound 2. The dimethylamide complex (N,N)Al(NMe2)2 (6) was prepared by reaction of dipyrromethene 1 with [Al(NMe2)3]2. Crystallographic data revealed that the dipyrromethene is non-planar when bulky coligands are present as in compounds 3-6, while in the dichloride complex 2 the dipyrromethene is planar. Halide complexes 2 and 3 reacted with adventitious moisture in toluene to afford crystalline acid-base adducts (N,N)H·HX, (X = Cl (7), I (8)), which adopted structures reminiscent of anion receptors. Alkoxide and dimethylamide complexes 5 and 6 were also applied as precatalysts for the ring-opening polymerization of ε-caprolactone and preliminary results are reported.


What are the molecular ties that maintain genomic loops?
The formation of genomic loops by proteins bound at sites scattered along a chromosome has a central role in many cellular processes, such as transcription, recombination and replication. Until recently, few such loops had been analyzed in any detail, and there was little agreement about the nature of the molecular ties maintaining these loops. Recent evidence suggests that loops are found in both prokaryotes and eukaryotes, and that the transcription machinery is a molecular tie. In addition, results obtained using site-specific recombination in bacteria and chromosome conformation capture in eukaryotes support the idea that active transcription units are in close contact. These data are consistent with a model for genome organization in which active polymerases cluster into transcription 'factories', which, inevitably, loops the intervening DNA. They are also consistent with the ties functioning as barriers, silencers, enhancers or locus control regions, depending on their positions relative to other genes.


Moderate blast exposure results in increased IL-6 and TNFα in peripheral blood.
A unique cohort of military personnel exposed to isolated blast was studied to explore acute peripheral cytokine levels, with the aim of identifying blast-specific biomarkers. Several cytokines, including interleukin (IL) 6, IL-10 and tumor necrosis factor alpha (TNFα) have been linked to pre-clinical blast exposure, but remained unstudied in clinical blast exposure. To address this gap, blood samples from 62 military personnel were obtained at baseline, and daily, during a 10-day blast-related training program; changes in the peripheral concentrations of IL-6, IL-10 and TNFα were evaluated using an ultrasensitive assay. Two groups of trainees were matched on age, duration of military service, and previous history of blast exposure(s), resulting in moderate blast cases and no/low blast controls. Blast exposures were measured using helmet sensors that determined the average peak pressure in pounds per square inch (psi). Moderate blast cases had significantly elevated concentrations of IL-6 (F1,60=18.81, p<0.01) and TNFα (F1,60=12.03, p<0.01) compared to no/low blast controls; levels rebounded to baseline levels the day after blast. On the day of the moderate blast exposure, the extent of the overpressure (psi) in those exposed correlated with IL-6 (r=0.46, p<0.05) concentrations. These findings indicate that moderate primary blast exposure results in changes, specifically acute and transient increases in peripheral inflammatory markers which may have implications for neuronal health.


Immunization, urbanization and slums - a systematic review of factors and interventions.
In 2014, over half (54%) of the world's population lived in urban areas and this proportion will increase to 66% by 2050. This urbanizing trend has been accompanied by an increasing number of people living in urban poor communities and slums. Lower immunization coverage is found in poorer urban dwellers in many contexts. This study aims to identify factors associated with immunization coverage in poor urban areas and slums, and to identify interventions to improve coverage. We conducted a systematic review, searching Medline, Embase, Global Health, CINAHL, Web of Science and The Cochrane Database with broad search terms for studies published between 2000 and 2016. Of 4872 unique articles, 327 abstracts were screened, leading to 63 included studies: 44 considering factors and 20 evaluating interventions (one in both categories) in 16 low or middle-income countries. A wide range of socio-economic characteristics were associated with coverage in different contexts. Recent rural-urban migration had a universally negative effect. Parents commonly reported lack of awareness of immunization importance and difficulty accessing services as reasons for under-immunization of their children. Physical distance to clinics and aspects of service quality also impacted uptake. We found evidence of effectiveness for interventions involving multiple components, especially if they have been designed with community involvement. Outreach programmes were effective where physical distance was identified as a barrier. Some evidence was found for the effective use of SMS (text) messaging services, community-based education programmes and financial incentives, which warrant further evaluation. No interventions were identified that provided services to migrants from rural areas. Different factors affect immunization coverage in different urban poor and slum contexts. Immunization services should be designed in collaboration with slum-dwelling communities, considering the local context. Interventions should be designed and tested to increase immunization in migrants from rural areas.


About the Gaussian-Schell pulse train spectrum.
The Gaussian-Schell (G-S) pulse model describes variations in Gaussian shaped light pulses. Recent studies proposed random functions fitted to G-S pulses in a non-stationary context. More specifically, this paper provides a cyclo-stationary model. The associated power spectrum is derived. A correlation between pulses is shown to remove the power spectrum Gaussian character.


The highest background radiation school in the world and the health status of its students and their offspring.
Although the average effective human dose from natural background radiation is about 2.4 mSv per year, the students of the Saeid Nafisi school in Ramsar received effective doses of about 250 mSv while studying there for over 5 years. The goal of this project was a retrospective study of the health status of former students of this school and their offspring. The list of the students of the Saeid Nafisi school (high background radiation) and Taleghani and Kashani schools (ordinary background radiation) was provided by the Department of Education. After matching sex, age and socioeconomic level and obtaining their consent, part 1 of the specifically designed questionnaire was filled out by interview, and clinical examinations were recorded in part 2 of the questionnaire by a physician. The data were analysed using Statistical Package for the Social Sciences 16. Our study shows that 88.1 % of general examinations of high background radiation school students were normal as compared with 85.7 % for control group. There were no significant differences. This study is interesting and unique. It reveals that there is no health emergency related to these high radiation doses. We recommend continuing the health supervision of this population in the future.


Stochastic modelling of African swine fever in wild boar and domestic pigs: Epidemic forecasting and comparison of disease management strategies.
African swine fever (ASF), caused by the African swine fever virus (ASFV), is highly virulent in domestic pigs and wild boar (Sus scrofa), causing up to 100% mortality. The recent epidemic of ASF in Europe has had a serious economic impact and poses a threat to global food security. Unfortunately, there is no effective treatment or vaccine against ASFV, limiting the available disease management strategies. Mathematical models allow us to further our understanding of infectious disease dynamics and evaluate the efficacy of disease management strategies. The ASF Challenge, organised by the French National Research Institute for Agriculture, Food, and the Environment, aimed to expand the development of ASF transmission models to inform policy makers in a timely manner. Here, we present the model and associated projections produced by our team during the challenge. We developed a stochastic model combining transmission between wild boar and domestic pigs, which was calibrated to synthetic data corresponding to different phases describing the epidemic progression. The model was then used to produce forward projections describing the likely temporal evolution of the epidemic under various disease management scenarios. Despite the interventions implemented, long-term projections forecasted persistence of ASFV in wild boar, and hence repeated outbreaks in domestic pigs. A key finding was that it is important to consider the timescale over which different measures are evaluated: interventions that have only limited effectiveness in the short term may yield substantial long-term benefits. Our model has several limitations, partly because it was developed in real-time. Nonetheless, it can inform understanding of the likely development of ASF epidemics and the efficacy of disease management strategies, should the virus continue its spread in Europe.


Potential effects of nutrient profiles on nutrient intakes in the Netherlands, Greece, Spain, USA, Israel, China and South-Africa.
Nutrient profiling is defined as the science of categorising foods based on their nutrient composition. The Choices Programme is a nutrient profile system with criteria that determine whether foods are eligible to carry a "healthier option" stamp. The Daily Menu Method which has been developed to evaluate these criteria is described here. This method simulates the change in calculated nutrient intakes which would be the result of consumers changing their diets in favour of food products that comply with the criteria. Average intakes of energy, trans fatty acids (TFA), saturated fatty acids (SAFA), sodium, added sugar and fibre were derived from dietary intake studies and food consumption surveys of 7 countries: The Netherlands, Greece, Spain, the USA, Israel, China and South Africa. For each of the key nutrients, these average intakes were translated into three Typical Daily Menus per country. Average intakes based on these three menus were compared with average intakes from three Choices Daily Menus. To compose the Choices Menus, foods from the Typical Menus that did not comply with the Choices criteria were replaced with foods that did comply and are available on the market. Comparison of intakes from the Choices Menus with the survey data showed that calculated intakes of energy, SAFA, TFA, sodium and added sugar were reduced. Fibre intakes were increased. The size of the effect differed per country. The Daily Menu Method is a useful means to predict the potential effects of nutrient profiles such as the Choices criteria, on daily nutrient intakes. The method can be applied internationally and confirms that the criteria of the Choices Programme are in line with the aim of the programme: to improve nutrient intakes in the direction of the recommendations.


Why people do, or do not, immediately contact emergency medical services following the onset of acute stroke: qualitative interview study.
To identify the reasons why individuals contact, or delay contacting, emergency medical services in response to stroke symptoms. Qualitative interview study with a purposive sample of stroke patients and witnesses, selected according to method of accessing medical care and the time taken to do so. Data were analysed using the Framework approach. Area covered by three acute stroke units in the north east of England. Nineteen stroke patients and 26 witnesses who had called for help following the onset of stroke symptoms. Factors influencing who called emergency medical services and when they called included stroke severity, how people made sense of symptoms and their level of motivation to seek help. Fear of the consequences of stroke, including future dependence or disruption to family life, previous negative experience of hospitals, or involving a friend or relations in the decision to access medical services, all resulted in delayed admission. Lack of knowledge of stroke symptoms was also an important determinant. Perceptions of the remit of medical services were a major cause of delays in admission, with many people believing the most appropriate action was to telephone their GP. Variations in the response of primary care teams to acute stroke symptoms were also evident. The factors influencing help-seeking decisions are complex. There remains a need to improve recognition by patients, witnesses and health care staff of the need to treat stroke as a medical emergency by calling emergency medical services, as well as increasing knowledge of symptoms of stroke among patients and potential witnesses. Fear, denial and reticence to impose on others hinders the process of seeking help and will need addressing specifically with appropriate interventions. Variability in how primary care services respond to stroke needs further investigation to inform interventions to promote best practice. UK Clinical Research Network UKCRN 6590.


Should vascular surgery patients be screened preoperatively for methicillin-resistant Staphylococcus aureus?
Methicillin-resistant Staphylococcus aureus (MRSA) can cause severe infection in patients who are undergoing vascular surgical operations. Testing all vascular surgery patients preoperatively for MRSA and attempting to decolonize those who have positive results may be a strategy to prevent MRSA infection. The economic value of such a strategy has not yet been determined. We developed a decision-analytic computer simulation model to determine the economic value of using such a strategy before all vascular surgical procedures from the societal and third-party payer perspectives at different MRSA prevalence and decolonization success rates. The model showed preoperative MRSA testing to be cost-effective (incremental cost-effectiveness ratio, <$50,000 per quality-adjusted life year) when the MRSA prevalence is > or = 0.01 and the decolonization success rate is > or = 0.25. In fact, this strategy was dominant (ie, less costly and more effective) at the following thresholds: MRSA prevalence > or = 0.01 and decolonization success rate > or = 0.5, and MRSA prevalence > or = 0.025 and decolonization success rate > or = 0.25. Testing and decolonizing patients for MRSA before vascular surgery may be a cost-effective strategy over a wide range of MRSA prevalence and decolonization success rates.


TGFβ receptor I transactivation mediates stretch-induced Pak1 activation and CTGF upregulation in mesangial cells.
Increased intraglomerular pressure is an important pathogenic determinant of kidney fibrosis in the progression of chronic kidney disease, and can be modeled by exposing glomerular mesangial cells (MC) to mechanical stretch. MC produce extracellular matrix and profibrotic cytokines, including connective tissue growth factor (CTGF) when stretched. We show that p21-activated kinase 1 (Pak1) is activated by stretch in MC in culture and in vivo in a process marked by elevated intraglomerular pressures. Its activation is essential for CTGF upregulation. Rac1 is an upstream regulator of Pak1 activation. Stretch induces transactivation of the type I transforming growth factor β1 receptor (TβRI) independently of ligand binding. TβRI transactivation is required not only for Rac1/Pak1 activation, but also for activation of the canonical TGFβ signaling intermediate Smad3. We show that Smad3 activation is an essential requirement for CTGF upregulation in MC under mechanical stress. Pak1 regulates Smad3 C-terminal phosphorylation and transcriptional activation. However, a second signaling pathway, that of RhoA/Rho-kinase and downstream Erk activation, is also required for stretch-induced CTGF upregulation in MC. Importantly, this is also regulated by Pak1. Thus, Pak1 serves as a novel central mediator in the stretch-induced upregulation of CTGF in MC.


Classification of dengue cases in Southwest India based on the WHO systems-a retrospective analysis.
The WHO dengue classification 1997 was revised in 2009 and the revised classification system has now been in use for the past decade. This study was carried out to compare the 1997 and revised classifications in assessing the severity of dengue infection among all age groups during a dengue outbreak in southwest India. This retrospective cross-sectional study was carried out including serologically confirmed dengue cases. A total of 1033 dengue cases were classified on the basis of the 1997 WHO classification and 2009 revised classification. The statistical analysis was carried out using SPSS 15.0 for Windows (SPSSTM Inc, Chicago, IL, USA). Both the 1997 and revised WHO classifications were applied to 1033 confirmed dengue cases, including 692 males (67%) and 341 females (33%). The median age of the study participants was 23 years (IQR 10-33), including 112 (10.8%) children at and below the age of 5 years. The level of agreement between the two systems of classification was poor (kappa=0.143, 0.055-0.198, p-value <0.001). A greater sensitivity and specificity of the revised classification was observed in comparison with the 1997 WHO classification. In the context of changing dengue epidemiology and geographical expansion, the revised classification is helpful in the identification of severe cases, facilitating timely management.


Embolotherapy for Neuroendocrine Tumor Liver Metastases: Prognostic Factors for Hepatic Progression-Free Survival and Overall Survival.
The purpose of the study was to evaluate prognostic factors for survival outcomes following embolotherapy for neuroendocrine tumor (NET) liver metastases. This was a multicenter retrospective study of 155 patients (60 years mean age, 57 % male) with NET liver metastases from pancreas (n = 71), gut (n = 68), lung (n = 8), or other/unknown (n = 8) primary sites treated with conventional transarterial chemoembolization (TACE, n = 50), transarterial radioembolization (TARE, n = 64), or transarterial embolization (TAE, n = 41) between 2004 and 2015. Patient-, tumor-, and treatment-related factors were evaluated for prognostic effect on hepatic progression-free survival (HPFS) and overall survival (OS) using unadjusted and propensity score-weighted univariate and multivariate Cox proportional hazards models. Median HPFS and OS were 18.5 and 125.1 months for G1 (n = 75), 12.2 and 33.9 months for G2 (n = 60), and 4.9 and 9.3 months for G3 tumors (n = 20), respectively (p < 0.05). Tumor burden >50 % hepatic volume demonstrated 5.5- and 26.8-month shorter median HPFS and OS, respectively, versus burden ≤50 % (p < 0.05). There were no significant differences in HPFS or OS between gut or pancreas primaries. In multivariate HPFS analysis, there were no significant differences among embolotherapy modalities. In multivariate OS analysis, TARE had a higher hazard ratio than TACE (unadjusted Cox model: HR 2.1, p = 0.02; propensity score adjusted model: HR 1.8, p = 0.11), while TAE did not differ significantly from TACE. Higher tumor grade and tumor burden prognosticated shorter HPFS and OS. TARE had a higher hazard ratio for OS than TACE. There were no significant differences in HPFS among embolotherapy modalities.


Outdoor fecal deposition by free-roaming cats and attitudes of cat owners and nonowners toward stray pets, wildlife, and water pollution.
To estimate cat population size, management, and outside fecal deposition and evaluate attitudes of cat owners and nonowners to stray animal control, water pollution, and wildlife protection. Cross-sectional survey. 294 adult residents of Cayucos, Los Osos, and Morro Bay, Calif. Telephone survey. The region's cat population was estimated at 7,284 owned and 2,046 feral cats, and 38% of surveyed households owned a mean of 1.9 cats/household. Forty-four percent of cats defecated outside >75% of the time. Annual fecal deposition (wet weight) by owned cats in the 3 communities was estimated to be 77.6 tonnes (76.4 tons). Cat owners were more likely to oppose cat licensing and impounding stray cats and support trap-neuter-return for stray cats and less likely to be concerned about water pollution, than were noncat owners. Feral cats represented a sizeable proportion (22%) of the free roaming cats in this area and could be contributing 30.0 tonnes (29.5 tons) of feces to the environment per year. However, feral cats are not the principal source of fecal loading because owned cats defecating outdoors contribute an estimated 77.6 tonnes (76.4 tons) or 72% of the annual outdoor fecal deposition.


Variation in ancient Egyptian stature and body proportions.
Stature and the pattern of body proportions were investigated in a series of six time-successive Egyptian populations in order to investigate the biological effects on human growth of the development and intensification of agriculture, and the formation of state-level social organization. Univariate analyses of variance were performed to assess differences between the sexes and among various time periods. Significant differences were found both in stature and in raw long bone length measurements between the early semipastoral population and the later intensive agricultural population. The size differences were greater in males than in females. This disparity is suggested to be due to greater male response to poor nutrition in the earlier populations, and with the increasing development of social hierarchy, males were being provisioned preferentially over females. Little change in body shape was found through time, suggesting that all body segments were varying in size in response to environmental and social conditions. The change found in body plan is suggested to be the result of the later groups having a more tropical (Nilotic) form than the preceding populations.


The role of ATF4 stabilization and autophagy in resistance of breast cancer cells treated with Bortezomib.
The ubiquitin-proteasome system plays a key regulatory role in cellular homeostasis. The inhibition of the 26S proteasome by Bortezomib leads to the accumulation of misfolded proteins, resulting in endoplasmic reticulum stress followed by a coordinated cellular response called unfolded protein response (UPR). Endoplasmic reticulum stress is also a potent inducer of macroautophagy. Bortezomib is a selective and potent inhibitor of the 26S proteasome and is approved for the treatment of multiple myeloma. Clinical trials with Bortezomib have shown promising results for some types of cancers, but not for some others, including those of the breast. In this study, we show that Bortezomib induces the UPR and autophagy in MCF7 breast cancer cells. Surprisingly, Bortezomib did not induce phosphorylation of PERK, a key initial step of the UPR. We show that induction of autophagy by Bortezomib is dependent on the proteasomal stabilisation of ATF4 and up-regulation of LC3B by ATF4. We show that ATF4 and LC3B play a critical role in activating autophagy and protecting cells from Bortezomib-induced cell death. Our experiments also reveal that HDAC6 knockdown results in decreased LC3B protein and reduced autophagy. Our work shows that the induction of autophagy through ATF4 may be an important resistance mechanism to Bortezomib treatment in breast cancer, and targeting autophagy may represent a novel approach to sensitize breast cancers to Bortezomib.


Excess electron localization sites in neutral water clusters.
We present approximate pseudopotential quantum-mechanical calculations of the excess electron states of equilibrated neutral water clusters sampled by classical molecular dynamics simulations. The internal energy of the clusters are representative of those present at temperatures of 200 and 300 K. Correlated electronic structure calculations are used to validate the pseudopotential for this purpose. We find that the neutral clusters support localized, bound excess electron ground states in about 50% of the configurations for the smallest cluster size studied (n = 20), and in almost all configurations for larger clusters (n > 66). The state is always exterior to the molecular frame, forming typically a diffuse surface state. Both cluster size and temperature dependence of energetic and structural properties of the clusters and the electron distribution are explored. We show that the stabilization of the electron is strongly correlated with the preexisting instantaneous dipole moment of the neutral clusters, and its ground state energy is reflected in the electronic radius. The findings are consistent with electron attachment via an initial surface state. The hypothetical spectral dynamics following such attachment is also discussed.


Mass of pluto.
Analysis of the observations of Neptune indicates a reciprocal mass of Pluto of 1,812,000 (0.18 Earth masses). If the density is the same as that of Earth, the diameter would be 7200 kilometers. If 6400 kilometers is accepted (from other sources) as the upper limit of the diameter, then Pluto must be at least 1.4 times as dense as Earth.


Resident motivations and willingness-to-pay for urban biodiversity conservation in Guangzhou (China).
The monetary assessment of biodiversity measures the welfare damages brought by biodiversity losses and the cost-benefit analysis of conservation projects in a socio-economic context. The contingent valuation method could include motivational factors to strengthen economic analysis of nature conservation. This study analyzed Guangzhou residents' motivations and willingness-to-pay (WTP) for an urban biodiversity conservation program in the National Baiyun Mountain Scenic Area (BMSA). The peri-urban natural site, offering refuge to some endemic species, is under increasing development pressures for recreational and residential use. A questionnaire survey was conducted in the Guangzhou metropolitan area during June to October 2007. We interviewed face-to-face 720 stratified sampled households to probe residents' attitudes towards the city's environmental issues, motivations for urban nature conservation, and WTP for biodiversity conservation. Principal component analysis identified five motivational factors, including environmental benefit, ecological diversity, nature-culture interaction, landscape-recreation function, and intergenerational sustainability, which illustrated the general economic values of urban nature. Logistic regression was applied to predict the probability of people being willing to pay for the urban biodiversity conservation in BMSA. The significant predictors of WTP included household income and the factor nature-culture interaction. The median WTP estimated RMB149/household (about US$19.5/household) per year and an aggregate of RMB291 million (approximately US$38.2 million) annually to support the urban conservation project. Including public motivations into contingent valuation presents a promising approach to conduct cost-benefit analysis of public projects in China.


Defining the metabolome: size, flux, and regulation.
Renewed interest in metabolic research over the last two decades has inspired an explosion of technological developments for studying metabolism. At the forefront of methodological innovation is an approach referred to as "untargeted" or "discovery" metabolomics. The experimental objective of this technique is to comprehensively measure the entire metabolome, which constitutes a largely undefined set of molecules. Given its potential comprehensive coverage, untargeted metabolomics is often the first choice of experiments for investigators pursuing a metabolic research question. It is important to recognize, however, that untargeted metabolomics may not always be the optimal experimental approach. Conventionally, untargeted metabolomics only provides information about relative differences in metabolite pool sizes. Therefore, depending on the specific scientific question at hand, a complementary approach involving stable isotopes (such as metabolic flux analysis) may be better suited to provide biological insights. Unlike untargeted metabolomics, stable-isotope methods can provide information about differences in reaction rates.


Antioxidant properties of blirubin in the model organism, Caenorhabditis elegans.
We evaluated the effects of bilirubin on the nematode, C. elegans, specifically addressing the ability of bilirubin to induce oxidative stress and alter glutathione (GSH) content as measures of injury. Bilirubin exposure caused a doubling of the spectrophotometric absorption at 440 nm in wild-type C. elegans irrespective of the bilirubin concentration used, suggesting that bilirubin is readily taken up by the worms. No changes were noted in growth, phenotype or reproductive cycle at any bilirubin concentration at 24, 48, and 72 hrs. The oxidative stress inducible green fluorescent protein (GFP) expression in the gst-4::GFP strain was decreased upon exposure to bilirubin at a concentration of 0.5 mM at 24, 48 and 72 hrs. This trend was not statistically significant at 24 or 72 hrs, but did reach statistical significance at 48 hours (p < 0.05). Glutathione (GSH) content in the gst-4::GFP strain showed a significant increase as early as 20 hours post treatment with 0.5 mM bilirubin (p < 0.05). Microarray analysis showed that in bilirubin-exposed worms, 27 genes were up-regulated, and 90 genes were down-regulated (by >1.3 fold vs. controls). The transcription factor asc-1 was induced, whereas genes involved in transcription, trafficking and mitochondrial function were down-regulated. Our findings corroborate earlier findings of bilirubin's ability to act as an antioxidant, most likely by reducing the metabolic requirement in C. elegans.


Inhibition of morphine-induced cAMP overshoot: a cell-based assay model in a high-throughput format.
Opiates are not only potent analgesics but also drugs of abuse mainly because they produce euphoria. Chronic use of opiates results in the development of tolerance and dependence. Dr Marshall Nirenberg's group at the National Institutes of Health (NIH) was the first to use a cellular model system of Neuroblastoma × Glioma hybrid cells (NG108-15) to study morphine addiction. They showed that opiates affect adenylyl cyclase (AC) by two opposing mechanisms mediated by the opiate receptor. Although the cellular mechanisms that cause addiction are not yet completely understood, the most observed correlative biochemical adaptation is the upregulation of AC. This model also provides the opportunity to look for compounds which could dissociate the acute effect of opiates from the delayed response, upregulation of AC, and thus lead to the discovery of non-addictive drugs. To identify small molecule compounds that can inhibit morphine-induced cAMP overshoot, we have validated and optimized a cell-based assay in a high throughput format that measures cellular cAMP production after morphine withdrawal. The assay performed well in the 1536-well plate format. The LOPAC library of 1,280 compounds was screened in this assay on a quantitative high-throughput screening (qHTS) platform. A group of compounds that can inhibit morphine-induced cAMP overshoot were identified. The most potent compounds are eight naloxone-related compounds, including levallorphan tartrate, naloxonazine dihydrochloride, naloxone hydrochloride, naltrexone hydrochloride, and naltriben methanesulfonate. The qHTS approach we used in this study will be useful in identifying novel inhibitors of morphine induced addiction from a larger scale screening.


GENTRIFICATION WITHOUT SEGREGATION? RACE, IMMIGRATION, AND RENEWAL IN A DIVERSIFYING CITY.
Research on how neighborhood racial composition affects where gentrification unfolds yields mixed conclusions, but these studies either capture broad national trends or highly segregated cities. Drawing on the case of Seattle-a majority-white city with low segregation levels and growing ethnoracial diversity, this study uncovers an underexplored mechanism shaping patterns of uneven development and residential selection in the contemporary city: immigrant replenishment. The share of all minorities is negatively associated with gentrification during the 1970s and 1980s, and, in contrast to expectations, shares of blacks positively predicts recent gentrification while shares of Asians negatively predicts it. Increased concentrations of recent immigrants in neighborhoods with greater shares of Asians explain these relationships. These findings suggest that where arriving immigrants move limits residential selection in gentrification and shifts pressures to low-cost black neighborhoods. This study highlights how immigration and points of entry are important factors for understanding uneven development in the contemporary city and has implications for the future of racial stratification as cities transform.


Extracellular Myocardial Volume in Patients With Aortic Stenosis.
Myocardial fibrosis is a key mechanism of left ventricular decompensation in aortic stenosis and can be quantified using cardiovascular magnetic resonance (CMR) measures such as extracellular volume fraction (ECV%). Outcomes following aortic valve intervention may be linked to the presence and extent of myocardial fibrosis. This study sought to determine associations between ECV% and markers of left ventricular decompensation and post-intervention clinical outcomes. Patients with severe aortic stenosis underwent CMR, including ECV% quantification using modified Look-Locker inversion recovery-based T1 mapping and late gadolinium enhancement before aortic valve intervention. A central core laboratory quantified CMR parameters. Four-hundred forty patients (age 70 ± 10 years, 59% male) from 10 international centers underwent CMR a median of 15 days (IQR: 4 to 58 days) before aortic valve intervention. ECV% did not vary by scanner manufacturer, magnetic field strength, or T1 mapping sequence (all p > 0.20). ECV% correlated with markers of left ventricular decompensation including left ventricular mass, left atrial volume, New York Heart Association functional class III/IV, late gadolinium enhancement, and lower left ventricular ejection fraction (p < 0.05 for all), the latter 2 associations being independent of all other clinical variables (p = 0.035 and p < 0.001). After a median of 3.8 years (IQR: 2.8 to 4.6 years) of follow-up, 52 patients had died, 14 from adjudicated cardiovascular causes. A progressive increase in all-cause mortality was seen across tertiles of ECV% (17.3, 31.6, and 52.7 deaths per 1,000 patient-years; log-rank test; p = 0.009). Not only was ECV% associated with cardiovascular mortality (p = 0.003), but it was also independently associated with all-cause mortality following adjustment for age, sex, ejection fraction, and late gadolinium enhancement (hazard ratio per percent increase in ECV%: 1.10; 95% confidence interval [1.02 to 1.19]; p = 0.013). In patients with severe aortic stenosis scheduled for aortic valve intervention, an increased ECV% is a measure of left ventricular decompensation and a powerful independent predictor of mortality.


Migration and Pacific mortality: estimating migration effects on Pacific mortality rates using Bayesian models.
Pacific people living in New Zealand have higher mortality rates than New Zealand residents of European/Other ethnicity. The aim of this paper is to see whether Pacific mortality rates vary by natality and duration of residence. We used linked census-mortality information for 25- to 74-year-olds in the 2001 census followed for up to three years. Hierarchical Bayesian modeling provided a means of handling sparse data. Posterior mortality rates were directly age-standardized. We found little evidence of mortality differences between the overseas-born and the New Zealand-born for all-cause, cancer, and cardiovascular disease (CVD) mortality. However, we found evidence for lower all-cause (and possibly cancer and CVD) mortality rates for Pacific migrants resident in New Zealand for less than 25 years relative to those resident for more than 25 years. This result may arise from a combination of processes operating over time, including health selection effects from variations in New Zealand's immigration policy, the location of Pacific migrants within the social, political, and cultural environment of the host community, and health impacts of the host culture. We could not determine the relative importance of these processes, but identifying the (modifiable) drivers of the inferred long-term decline in health of the overseas-born Pacific population relative to more-recent Pacific migrants is important to Pacific communities and from a national health and policy perspective.


Quality of life in adult patients after stem cell transplantation.
Many articles pertaining to quality of life (QOL) following stem cell transplantation have been published in the US and western Europe. However, since the actions of health insurance systems and overall cultural aspects are strongly associated with QOL, investigations into QOL should be carried out within all countries. Therefore, we have investigated the QOL of adult patients following stem cell transplantation at 31 hospitals in Japan. The survivors, who were surveyed by mail questionnaire, were 20 years or older at the time of this study. The underlying diseases were acute lymphoblastic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, myelodysplastic syndrome, and multiple myeloma. Median age at the time of the study was 36 years, and median interval after transplantation was 35.3 months. Of 383 patients surveyed, 282 (73.6%) responded to the questionnaire. One hundred and ninety-two patients were treated with an allogeneic-related transplantation, 52 with allogeneic-unrelated, and 38 with an autologous transplantation. Our data revealed that the length of time since transplantation and the diagnosis of chronic GVHD were associated with QOL. When unrelated and related transplantation recipients were compared, ratings on relief from pain, stability in weight, and confidence in dealing with daily life were lower among unrelated transplantation patients.


Chamber-specific differences in human cardiac fibroblast proliferation and responsiveness toward simvastatin.
Fibroblasts, the most abundant cells in the heart, contribute to cardiac fibrosis, the substrate for the development of arrythmogenesis, and therefore are potential targets for preventing arrhythmic cardiac remodeling. A chamber-specific difference in the responsiveness of fibroblasts from the atria and ventricles toward cytokine and growth factors has been described in animal models, but it is unclear whether similar differences exist in human cardiac fibroblasts (HCFs) and whether drugs affect their proliferation differentially. Using cardiac fibroblasts from humans, differences between atrial and ventricular fibroblasts in serum-induced proliferation, DNA synthesis, cell cycle progression, cyclin gene expression, and their inhibition by simvastatin were determined. The serum-induced proliferation rate of human atrial fibroblasts was more than threefold greater than ventricular fibroblasts with faster DNA synthesis and higher mRNA levels of cyclin genes. Simvastatin predominantly decreased the rate of proliferation of atrial fibroblasts, with inhibition of cell cycle progression and an increase in the G0/G1 phase in atrial fibroblasts with a higher sensitivity toward inhibition compared with ventricular fibroblasts. The DNA synthesis and mRNA levels of cyclin A, D, and E were significantly reduced by simvastatin in atrial but not in ventricular fibroblasts. The inhibitory effect of simvastatin on atrial fibroblasts was abrogated by mevalonic acid (500 μM) that bypasses 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibition. Chamber-specific differences exist in the human heart because atrial fibroblasts have a higher proliferative capacity and are more sensitive to simvastatin-mediated inhibition through HMG-CoA reductase pathway. This mechanism may be useful in selectively preventing excessive atrial fibrosis without inhibiting adaptive ventricular remodeling during cardiac injury.


HIV-1 infection of nondividing cells through the recognition of integrase by the importin/karyopherin pathway.
The karyophilic properties of the HIV-1 nucleoprotein complex facilitate infection of nondividing cells such as macrophages and quiescent T lymphocytes, and allow the in vivo delivery of transgenes by HIV-derived retroviral vectors into terminally differentiated cells such as neurons. Although the viral matrix (MA) and Vpr proteins have previously been shown to play important roles in this process, we demonstrate here that integrase, the enzyme responsible for mediating the integration of the viral genome in the host cell chromosome, can suffice to connect the HIV-1 preintegration complex with the cell nuclear import machinery. This novel function of integrase reflects the recognition of an atypical bipartite nuclear localization signal by the importin/karyopherin pathway.


Heparin is an activating ligand of the orphan receptor tyrosine kinase ALK.
Anaplastic lymphoma kinase (ALK) is one of the few remaining "orphan" receptor tyrosine kinases (RTKs) in which the ligands are unknown. Ligand-mediated activation of RTKs is important throughout development. ALK is particularly relevant to the development of the nervous system. Increased activation of RTKs by mutation, genetic amplification, or signals from the stroma contributes to disease progression and acquired drug resistance in cancer. Aberrant activation of ALK occurs in subsets of lung adenocarcinoma, neuroblastoma, and other cancers. We found that heparin is a ligand that binds specifically to the ALK extracellular domain. Whereas heparins with short chain lengths bound to ALK in a monovalent manner and did not activate the receptor, longer heparin chains induced ALK dimerization and activation in cultured neuroblastoma cells. Heparin lacking N- and O-linked sulfate groups or other glycosaminoglycans with sulfation patterns different than heparin failed to activate ALK. Moreover, antibodies that bound to the extracellular domain of ALK interfered with heparin binding and prevented heparin-mediated activation of ALK. Thus, heparin and perhaps related glycosaminoglycans function as ligands for ALK, revealing a potential mechanism for the regulation of ALK activity in vivo and suggesting an approach for developing ALK-targeted therapies for cancer.


Microsecond Rearrangements of Hydrophobic Clusters in an Initially Collapsed Globule Prime Structure Formation during the Folding of a Small Protein.
Determining how polypeptide chain collapse initiates structure formation during protein folding is a long standing goal. It has been challenging to characterize experimentally the dynamics of the polypeptide chain, which lead to the formation of a compact kinetic molten globule (MG) in about a millisecond. In this study, the sub-millisecond events that occur early during the folding of monellin from the guanidine hydrochloride-unfolded state have been characterized using multiple fluorescence and fluorescence resonance energy transfer probes. The kinetic MG is shown to form in a noncooperative manner from the unfolded (U) state as a result of at least three different processes happening during the first millisecond of folding. Initial chain compaction completes within the first 37μs, and further compaction occurs only after structure formation commences at a few milliseconds of folding. The transient nonnative and native-like hydrophobic clusters with side chains of certain residues buried form during the initial chain collapse and the nonnative clusters quickly disassemble. Subsequently, partial chain desolvation occurs, leading to the formation of a kinetic MG. The initial chain compaction and subsequent chain rearrangement appear to be barrierless processes. The two structural rearrangements within the collapsed globule appear to prime the protein for the actual folding transition.


PD-L1 and thyroid cytology: A possible diagnostic and prognostic marker.
Programmed death-ligand 1 (PD-L1) expression is emerging as an important predictive biomarker in anti-PD-L1 cancer immunotherapy. Its role has been clearly defined in various human cancers and is linked to a poor prognosis and resistance to anticancer therapies. The role of PD-L1 in thyroid cancers has not been well defined in fine-needle aspiration cytology (FNAC). The authors examined the performance of PD-L1 immunostaining in liquid-based cytology (LBC) to determine whether it could be a biomarker of malignancy or aggressive disease. From January 2018 to March 2019, 236 thyroid lesions, which had been diagnosed by FNAC as indeterminate lesions, suspicious for malignancy (SFM), and malignant, were enrolled. PD-L1 immunostaining was performed on both LBC and corresponding histology samples. The FNAC cohort included 50 benign negative controls, 42 samples of atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS), 33 samples of follicular neoplasm/suspicious for follicular neoplasm (FN/SFN), 53 samples that were suspicious for malignancy (SFM), and 58 malignant samples. AUS/FLUS samples included 3 goiters, 32 follicular adenomas (FAs), 1 noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP), 5 invasive follicular variants of papillary thyroid carcinoma (I-FVPTCs), and 1 follicular carcinoma; whereas FN/SFN samples included 24 FAs and 9 malignancies (4 I-FVPTCs, 1 NIFTP, 3 papillary thyroid carcinomas [PTCs], and 1 oncocytic follicular carcinoma). The 53 SFM samples were diagnosed on histopathology as 2 FAs, 5 NIFTPs, 15 I-FVPTCs, and 31 PTCs; whereas the 58 malignant specimens included 5 NIFTPs, 5 I-FVPTCs, and 48 PTCs. Increased plasma membrane and cytoplasmic PD-L1 expression was found in 79 cases (38.5%), including 61 PTCs (conventional and variants). Negative PD-L1 expression was found in NIFTPs and FAs. A BRAF V600E mutation was identified in 15% of PD-L1-positive malignancies. The current data suggest that PD-L1 expression in the thyroid gland might represent a marker of malignancy that correlates with PTC, but not with NIFTP. Thyroid neoplasms with PD-L1 expression also ae enriched with BRAF V600E mutations, suggesting that they are associated with more aggressive behavior.


Control of cell division in the root epidermis of Arabidopsis thaliana.
The formation of the root epidermis in Arabidopsis thaliana provides a simple model to study mechanisms underlying patterning in plants. In this paper we have analyzed the relationships between cell fate specification and the pattern of cell division that occur in the root epidermis. Using clonal analysis, the two cell types of the developing root epidermis, trichoblasts and atrichoblasts, were distinguished by different rates of cell division, highest in trichoblasts. This character appears to be dependent on TTG which controls epidermal cell fate specification. The ability of epidermal cells to undergo longitudinal divisions which are involved in the control of the radial symmetry was shown to be controlled in a cell-specific manner by TTG. The control of the rate and the orientation of cell division in the root meristem epidermal layer thus appear to be under the control of cell fate specification mechanisms.


Effects of low-dose alcohol consumption on postural control with a particular focus on the role of the visual system.
There is clear evidence that vision contributes to stabilizing posture and that large quantities of alcohol affect balance. It has, however, not yet been investigated whether and how the consumption of low doses of ethanol affects postural control and the visual system. The purpose of this study was therefore to assess the influence of low-dose alcohol intake on balance. After having performed stability tests in a sober condition, 26 healthy males were instructed to consume 0.32 g of ethanol/kg body weight. At predefined time points, blood samples were collected and stability index scores were calculated using computerized dynamic posturography. Thirty minutes after ethanol intake, blood alcohol levels reached a mean peak of 0.037%. Whereas the ability to maintain balance significantly deteriorated during eyes-open testing, it surprisingly did not decrease during eyes-closed testing. Apparently, the visual system is particularly affected by ethanol and plays a major role in maintaining postural stability.


Survey of radiologists' knowledge regarding the management of severe contrast material-induced allergic reactions.
To evaluate radiologists' knowledge of the appropriate management of severe contrast material-induced allergic reactions by means of a telephone survey. Institutional research ethics board approval was obtained. Following verbal consent, a telephone survey of radiologists working in Canada's 13 English-speaking and 13 U.S. university-affiliated radiology departments was performed. Participants were selected by using a multistage sampling scheme and simple random sampling within departments. Given a severe contrast material-induced allergic reaction case scenario, radiologists were first asked their initial medication of choice, then questioned specifically on the use of epinephrine. The Canadian and U.S. cohorts were compared by using the chi(2) and Fisher exact tests, as appropriate, and proportions and 95% confidence intervals (CIs) were computed. A total of 253 (81%) of 311 radiologists from a 30% target population were surveyed. Ninety-one percent (231 of 253; 95% CI: 88%, 94%) of radiologists chose epinephrine as the most important initial medication. No radiologist gave the ideal response, but 41% (94 of 231; 95% CI: 35%, 47%) provided an acceptable administration route, concentration, and dose; 17% (n = 39; 95% CI: 12%, 22%) of radiologists provided an overdose. Only 11% (27 of 253; 95% CI: 7%, 15%) of radiologists knew what concentration of epinephrine was available in their drug kit and/or crash cart and what equipment would be required to administer it to a patient. Radiologists' knowledge of epinephrine for the management of severe contrast material-induced allergic reactions is deficient.


Thermodynamics of HMGB1 interaction with duplex DNA.
The high mobility group protein HMGB1 is a small, highly abundant protein that binds to DNA in a non-sequence-specific manner. HMGB1 consists of 2 DNA binding domains, the HMG boxes A and B, followed by a short basic region and a continuous stretch of 30 glutamate or aspartate residues. Isothermal titration calorimetry was used to characterize the binding of HMGB1 to the double-stranded model DNAs poly(dAdT).(dTdA) and poly(dGdC).(dCdG). To elucidate the contribution of the different structural motifs to DNA binding, calorimetric measurements were performed comparing the single boxes A and B, the two boxes plus or minus the basic sequence stretch (AB(bt) and AB), and the full-length HMGB1 protein. Thermodynamically, binding of HMGB1 and all truncated constructs to duplex DNA was characterized by a positive enthalpy change at 15 degrees C. From the slopes of the temperature dependence of the binding enthalpies, heat capacity changes of -0.129 +/- 0.02 and -0.105 +/- 0.05 kcal mol(-1) K(-1) were determined for box A and full-length HMGB1, respectively. Significant differences in the binding characteristics were observed using full-length HMGB1, suggesting an important role for the acid tail in modulating DNA binding. Moreover, full-length HMGB1 binds differently these two DNA templates: binding to poly(dAdT).(dTdA) was cooperative, had a larger apparent binding site size, and proceeded with a much larger unfavorable binding enthalpy than binding to poly(dGdC).(dCdG).


Mutations in the homologous ZDS1 and ZDS2 genes affect cell cycle progression.
The Saccharomyces cerevisiae ZDS1 and ZDS2 genes were identified as multicopy suppressors in distinct genetic screens but were found to encode highly similar proteins. We show that at semipermissive temperatures, a yeast strain with a cdc28-1N allele was uniquely deficient in plasmid maintenance in comparison with strains harboring other cdc28 thermolabile alleles. Quantitative analysis of plasmid loss rates in cdc28-1N strains carrying plasmids with multiple replication origins suggests that a defect in initiating DNA replication probably causes this plasmid loss phenotype. The ZDS1 gene was isolated as a multicopy suppressor of the cdc28-1N plasmid loss defect. A zds1 deletion exhibits genetic interactions with cdc28-1N but not with other cdc28 alleles. SIN4 encodes a protein which is part of the RNA polymerase II holoenzyme-mediator complex, and a sin4 null mutation has pleiotropic effects suggesting roles in transcriptional regulation and chromatin structure. The ZDS2 gene was isolated as a multicopy suppressor of the temperature-sensitive growth defect caused by the sin4 null mutation. Disruption of either ZDS1 or ZDS2 causes only modest phenotypes. However, a strain with both ZDS1 and ZDS2 disrupted is extremely slowly growing, has marked defects in bud morphology, and shows defects in completing S phase or entering mitosis.


Fluoride intake by infants.
Many infants are fully or partially breast fed during the early months of life; however, the percentage of such infants decreases to about 30 percent by 4 months of age. The majority of US infants are fed formulas for most of the first 10 months of life. Although fluoride (F) intakes by fully breast-fed infants are low, F intakes by partially breast-fed infants and by formula-fed infants are highly variable, depending primarily on the F content of the water used to dilute concentrated liquid or powdered infant formula products. In communities with F content of the drinking water less than 0.3 ppm, F consumption by many infants will be 30 to 40 micrograms.kg-1.d-1. The addition of a F supplement of 0.25 mg/d for a 4 kg infant would increase the F intake by 63 micrograms.kg-1.d-1, resulting in a total intake of about 100 micrograms.kg-1.d-1, an intake in the range believed to be associated with development of fluorosis of the permanent teeth. However, for the US infant population generally, many fewer infants are exposed to high F intakes from formula plus a supplement (recommended only for communities with water providing less than 0.3 ppm F) than from formula alone in communities with F content of 1 ppm in the drinking water. In assessing the possible effects of F intake during infancy on development of fluorosis, it is important to recognize that infant feeding practices have changed greatly during the past 30 years. In the 1960s, most infants over 4 months of age were fed fresh cow's milk and intakes of F were therefore low. By the mid 1970s a trend toward more extended feeding of formula was evident and this trend has continued into the 1990s. Prolonged exposure to high intakes of fluoride during infancy is much more common now than in the past.


Multi-Institutional Assessment of Adverse Health Outcomes Among North American Testicular Cancer Survivors After Modern Cisplatin-Based Chemotherapy.
Purpose To provide new information on adverse health outcomes (AHOs) in testicular cancer survivors (TCSs) after four cycles of etoposide and cisplatin (EPX4) or three or four cycles of bleomycin, etoposide, cisplatin (BEPX3/BEPX4). Methods Nine hundred fifty-two TCSs > 1 year postchemotherapy underwent physical examination and completed a questionnaire. Multinomial logistic regression estimated AHOs odds ratios (ORs) in relation to age, cumulative cisplatin and/or bleomycin dose, time since chemotherapy, sociodemographic factors, and health behaviors. Results Median age at evaluation was 37 years; median time since chemotherapy was 4.3 years. Chemotherapy consisted largely of BEPX3 (38.2%), EPX4 (30.9%), and BEPX4 (17.9%). None, one to two, three to four, or five or more AHOs were reported by 20.4%, 42.0%, 25.1%, and 12.5% of TCSs, respectively. Median number after EPX4 or BEPX3 was two (range, zero to nine and zero to 11, respectively; P > .05) and two (range, zero to 10) after BEPX4. When comparing individual AHOs for EPX4 versus BEPX3, Raynaud phenomenon (11.6% v 21.4%; P < .01), peripheral neuropathy (29.2% v 21.4%; P = .02), and obesity (25.5% v 33.0%; P = .04) differed. Larger cumulative bleomycin doses (OR, 1.44 per 90,000 IU) were significantly associated with five or more AHOs. Increasing age was a significant risk factor for one to two, three to four, or five or more AHOs versus zero AHOs (OR, 1.22, 1.50, and 1.87 per 5 years, respectively; P < .01); vigorous physical activity was protective (OR, 0.62, 0.51, and 0.41, respectively; P < .05). Significant risk factors for three to four and five or more AHOs included current (OR, 3.05 and 3.73) or former (OR, 1.61 and 1.76) smoking ( P < .05). Self-reported health was excellent/very good in 59.9% of TCSs but decreased as AHOs increased ( P < .001). Conclusion Numbers of AHOs after EPX4 or BEPX3 appear similar, with median follow-up of 4.3 years. A healthy lifestyle was associated with reduced number of AHOs.


Failure mode and effects analysis of witnessing protocols for ensuring traceability during PGD/PGS cycles.
Preimplantation genetic diagnosis and aneuploidy testing (PGD/PGS) use is constantly growing in IVF, and embryo/biopsy traceability during the additional laboratory procedures needed is pivotal. An electronic witnessing system (EWS), which showed a significant value in decreasing mismatch occurrence and increasing detection possibilities during standard care IVF, still does not guarantee the same level of efficiency during PGD/PGS cycles. Specifically, EWS cannot follow single embryos throughout the procedure. This is however critical when an unambiguous diagnosis corresponds to each embryo. Failure Mode and Effects Analysis (FMEA) is a proactive method generally adopted to define tools ensuring safety along a procedure. Due to the implementation of a large quantitative PCR (qPCR)-based blastocyst stage PGD/PGS programme in our centre, and to evaluate the potential procedural risks, a FMEA was performed in September 2014. Forty-four failure modes were identified, among which six were given a moderate risk priority number (>15) (RPN; product of estimated occurrence, severity and detection). Specific corrective measures were then introduced and implemented, and a second evaluation performed six months later. The meticulous and careful application of such measures allowed the risks to be decreased along the whole protocol, by reducing their estimated occurrence and/or increasing detection possibilities.


Worldwide variations in prevalence of symptoms of allergic rhinoconjunctivitis in children: the International Study of Asthma and Allergies in Childhood (ISAAC).
As part of the International Study of Asthma and Allergies in Childhood (ISAAC), prevalence surveys were conducted among representative samples of school children from locations in Europe, Asia, Africa, Australia, North and South America. 257,800 children aged 6-7 years from 91 centres in 38 countries, and 463,801 children aged 13-14 years from 155 centres in 56 countries. Written symptom questionnaires were translated from English into the local language for self-completion by the 13-14-year-olds and completion by the parents of the 6-7-year-olds. Rhinitis was described as a problem with sneezing, or a runny, or blocked nose when you (your child) DID NOT have a cold or the flu. Additional questions were asked about rhinitis associated with itchy-watery eyes, interference with activities and a history of hay fever ever. The prevalence of rhinitis with itchy-watery eyes ("rhinoconjunctivitis") in the past year varied across centres from 0.8% to 14.9% in the 6-7-year-olds and from 1.4% to 39.7% in the 13-14-year-olds. Within each age group, the global pattern was broadly consistent across each of the symptom categories. In centres of higher prevalence there was great variability in the proportion of rhinoconjunctivitis labelled as hay fever. The lowest prevalences of rhinoconjunctivitis were found in parts of eastern Europe, south and central Asia. High prevalences were reported from centres in several regions. These results suggest substantial worldwide variations in the prevalence and labelling of symptoms of allergic rhinoconjunctivitis which require further study. These differences, if real, may offer important clues to environmental influences on allergy.


Do ecological niche model predictions reflect the adaptive landscape of species?: a test using Myristica malabarica Lam., an endemic tree in the Western Ghats, India.
Ecological niche models (ENM) have become a popular tool to define and predict the "ecological niche" of a species. An implicit assumption of the ENMs is that the predicted ecological niche of a species actually reflects the adaptive landscape of the species. Thus in sites predicted to be highly suitable, species would have maximum fitness compared to in sites predicted to be poorly suitable. As yet there are very few attempts to address this assumption. Here we evaluate this assumption. We used Bioclim (DIVA GIS version 7.3) and Maxent (version 3.3.2) to predict the habitat suitability of Myristica malabarica Lam., an economically important tree occurring in the Western Ghats, India. We located populations of the trees naturally occurring in different habitat suitability regimes (from highly suitable to poorly suitable) and evaluated them for their regeneration ability and genetic diversity. We also evaluated them for two plant functional traits, fluctuating asymmetry--an index of genetic homeostasis, and specific leaf weight--an index of primary productivity, often assumed to be good surrogates of fitness. We show a significant positive correlation between the predicted habitat quality and plant functional traits, regeneration index and genetic diversity of populations. Populations at sites predicted to be highly suitable had a higher regeneration and gene diversity compared to populations in sites predicted to be poor or unsuitable. Further, individuals in the highly suitable sites exhibited significantly less fluctuating asymmetry and significantly higher specific leaf weight compared to individuals in the poorly suitable habitats. These results for the first time provide an explicit test of the ENM with respect to the plant functional traits, regeneration ability and genetic diversity of populations along a habitat suitability gradient. We discuss the implication of these results for designing viable species conservation and restoration programs.


Genome-wide identification of genes regulated by the Rcs phosphorelay system in Erwinia amylovora.
The exopolysaccharide amylovoran is one of the major pathogenicity factors in Erwinia amylovora, the causal agent of fire blight of apples and pears. We have previously demonstrated that the RcsBCD phosphorelay system is essential for virulence by controlling amylovoran biosynthesis. We have also found that the hybrid sensor kinase RcsC differentially regulates amylovoran production in vitro and in vivo. To further understand how the Rcs system regulates E. amylovora virulence gene expression, we conducted genome-wide microarray analyses to determine the regulons of RcsB and RcsC in liquid medium and on immature pear fruit. Array analyses identified a total of 648 genes differentially regulated by RcsCB in vitro and in vivo. Consistent with our previous findings, RcsB acts as a positive regulator in both conditions, while RcsC positively controls expression of amylovoran biosynthetic genes in vivo but negatively controls expression in vitro. Besides amylovoran biosynthesis and regulatory genes, cell-wall and cell-envelope (membrane) as well as regulatory genes were identified as the major components of the RcsBC regulon, including many novel genes. We have also demonstrated that transcripts of rcsA, rcsC, and rcsD genes but not the rcsB gene were up-regulated when bacterial cells were grown in minimal medium or following infection of pear fruits compared with those grown in Luria Bertani medium. Furthermore, using the genome of E. amylovora ATCC 49946, a hidden Markov model predicted 60 genes with a candidate RcsB binding site in the intergenic region, 28 of which were identified in the microarray assay. Based on these findings as well as previous reported data, a working model has been proposed to illustrate how the Rcs phosphorelay system regulates virulence gene expression in E. amylovora.


Fate of ochratoxin A content in Argentinian red wine during a pilot scale vinification.
The aim of this work was to evaluate the fate of ochratoxin A (OTA) content from must to wine during the red wine making process in a pilot scale vinification. The study was done using musts obtained from two red grape varieties (Bonarda and Tempranillo) artificially contaminated with two OTA levels. A duplicate set of tanks of 100 I each was established for each must (Bonarda and Tempranillo). The fermentations were initiated by inoculation of two Saccharomyces spp. strains having different fermentation performance. The must from the Tempranillo variety was spiked with 6 microg/I of OTA while that from the Bonarda variety with 0.3 microg/I of the toxin. Samples were collected at different stages of the process. Performance of the alcoholic and malolactic fermentations was monitored. Titratable and volatile acidity, pH, ethanol, sugar and SO2 concentrations were determined following standard methods proposed by the Office International de la Vigne et du Vin (OIV). OTA analysis was done by HPLC. Detection and quantification limits were 0.01 and 0.1 ng/ml, respectively. The OTA levels during the vinification trials dropped to an average of about 86.5%. The type of Saccharomyces strains used showed no effect on toxin reduction.


Bouncing on Mars and the Moon-the role of gravity on neuromuscular control: correlation of muscle activity and rate of force development.
On our astronomical neighbors Mars and the Moon, bouncing movements are the preferred locomotor techniques. During bouncing, the stretch-shortening cycle describes the muscular activation pattern. This study aimed to identify gravity-dependent changes in kinematic and neuromuscular characteristics in the stretch-shortening cycle. Hence, neuromuscular control of limb muscles as well as correlations between the muscles' pre-activation, reflex components, and force output were assessed in lunar, Martian, and Earth gravity. During parabolic flights, peak force (Fmax), ground-contact-time, rate of force development (RFD), height, and impulse were measured. Electromyographic (EMG) activities in the m. soleus (SOL) and gastrocnemius medialis (GM) were assessed before (PRE) and during bounces for the reflex phases short-, medium-, and long-latency response (SLR, MLR, LLR). With gradually decreasing gravitation, Fmax, RFD, and impulse were reduced, whereas ground-contact time and height increased. Concomitantly, EMG_GM decreased for PRE, SLR, MLR, and LLR, and in EMG_SOL in SLR, MLR, and LLR. For SLR and MLR, Fmax and RFD were positively correlated to EMG_SOL. For PRE and LLR, RFD and Fmax were positively correlated to EMG_GM. Findings emphasize that biomechanically relevant kinematic adaptations in response to gravity variation were accompanied by muscle- and phase-specific modulations in neural control. Gravitational variation is anticipated and compensated for by gravity-adjusted muscle activities. Importantly, the pre-activation and reflex phases were differently affected: in SLR and MLR, SOL is assumed to contribute to the decline in force output with a decreasing load, and, complementary in PRE and LLR, GM seems to be of major importance for force generation.


Functions of S-layers.
Although S-layers are being increasingly identified on Bacteria and Archaea, it is enigmatic that in most cases S-layer function continues to elude us. In a few instances, S-layers have been shown to be virulence factors on pathogens (e.g. Campylobacter fetus ssp. fetus and Aeromonas salmonicida), protective against Bdellovibrio, a depository for surface-exposed enzymes (e.g. Bacillus stearothermophilus), shape-determining agents (e.g. Thermoproteus tenax) and nucleation factors for fine-grain mineral development (e.g. Synechococcus GL 24). Yet, for the vast majority of S-layered bacteria, the natural function of these crystalline arrays continues to be evasive. The following review up-dates the functional basis of S-layers and describes such diverse topics as the effect of S-layers on the Gram stain, bacteriophage adsorption in lactobacilli, phagocytosis by human polymorphonuclear leukocytes, the adhesion of a high-molecular-mass amylase, outer membrane porosity, and the secretion of extracellular enzymes of Thermoanaerobacterium. In addition, the functional aspect of calcium on the Caulobacter S-layer is explained.


Effects of stress on growth, cortisol and glucose levels in non-domesticated Eurasian perch (Perca fluviatilis) and domesticated rainbow trout (Oncorhynchus mykiss).
Eurasian perch (Perca fluviatilis) is a promising aquaculture candidate, but the growth performance of this non-domesticated species may be negatively affected by its stress responsiveness to intensive culture conditions. To evaluate this potential problem, juvenile Eurasian perch were exposed to a standardized handling stressor twice a week for an 8-week period. A similar study was conducted on domesticated rainbow trout (Oncorhynchus mykiss) for comparison of intra- and inter-specific differences. The stressed fish of both species showed lower body growth than the non-stressed control fish, however, the final mean body mass was 35.4% lower in the stressed Eurasian perch than in the non-stressed controls, compared to 22.8% difference between the two groups in rainbow trout. The stress responsiveness was examined by comparing the post-stress cortisol and glucose levels in repeatedly stressed fish and fish exposed to the stressor only once. The cortisol stress response in both species strongly indicated a habituation to the repeated stressor. Thus, repeatedly stressed Eurasian perch reached maximum cortisol levels of 130 ng/mL after 0.5 h compared to 200 ng/mL in the fish stressed once, while considerably smaller differences in cortisol levels were shown between the repeatedly and single stressed rainbow trout. Rainbow trout also showed lower post-stress glucose levels in the repeatedly stressed fish compared to the single stressed fish. In contrast, the glucose levels in both groups of Eurasian perch increased abruptly after stress treatment and remained elevated at approximately 19 mM for 6 h; levels were three times as high as the peak levels 3 h post-stress in rainbow trout. Together, the habituation of the stress response shown in both species did not eliminate the growth difference found in the repeatedly stressed fish versus the control fish. Further, the lower growth performance of Eurasian perch compared to rainbow trout could partly be due to the increased energy consumption in the more stress responsive Eurasian perch.


Importin 8 is a gene silencing factor that targets argonaute proteins to distinct mRNAs.
Small regulatory RNAs including small interfering RNAs (siRNAs) and microRNAs (miRNAs) guide Argonaute (Ago) proteins to specific target RNAs leading to mRNA destabilization or translational repression. Here, we report the identification of Importin 8 (Imp8) as a component of miRNA-guided regulatory pathways. We show that Imp8 interacts with Ago proteins and localizes to cytoplasmic processing bodies (P bodies), structures involved in RNA metabolism. Furthermore, we detect Ago2 in the nucleus of HeLa cells, and knockdown of Imp8 reduces the nuclear Ago2 pool. Using immunoprecipitations of Ago2-associated mRNAs followed by microarray analysis, we further demonstrate that Imp8 is required for the recruitment of Ago protein complexes to a large set of Ago2-associated target mRNAs, allowing for efficient and specific gene silencing. Therefore, we provide evidence that Imp8 is required for cytoplasmic miRNA-guided gene silencing and affects nuclear localization of Ago proteins.


Advanced glycation end products suppress osteoblastic differentiation of stromal cells by activating endoplasmic reticulum stress.
Advanced glycation end products (AGEs) are involved in bone quality deterioration in diabetes mellitus. We previously showed that AGE2 or AGE3 inhibited osteoblastic differentiation and mineralization of mouse stromal ST2 cells, and also induced apoptosis and decreased cell growth. Although quality management for synthesized proteins in endoplasmic reticulum (ER) is crucial for the maturation of osteoblasts, the effects of AGEs on ER stress in osteoblast lineage are unknown. We thus examined roles of ER stress in AGE2- or AGE3-induced suppression of osteoblastogenesis of ST2 cells. An ER stress inducer, thapsigargin (TG), induced osteoblastic differentiation of ST2 cells by increasing the levels of Osterix, type 1 collagen (Col1), alkaline phosphatase (ALP) and osteocalcin (OCN) mRNA. AGE2 or AGE3 suppressed the levels of ER stress sensors such as IRE1α, ATF6 and OASIS, while they increased the levels of PERK and its downstream molecules, ATF4. A reduction in PERK level by siRNA did not affect the AGEs-induced suppression of the levels of Osterix, Col1 and OCN mRNA. In conclusion, AGEs inhibited the osteoblastic differentiation of stromal cells by suppressing ER stress sensors and accumulating abnormal proteins in the cells. This process might accelerate AGEs-induced suppression of bone formation found in diabetes mellitus.