Sample size considerations in the design of cluster randomized trials of combination HIV prevention.
Background Cluster randomized trials have been utilized to evaluate the effectiveness of HIV prevention strategies on reducing incidence. Design of such studies must take into account possible correlation of outcomes within randomized units. Purpose To discuss power and sample size considerations for cluster randomized trials of combination HIV prevention, using an HIV prevention study in Botswana as an illustration. Methods We introduce a new agent-based model to simulate the community-level impact of a combination prevention strategy and investigate how correlation structure within a community affects the coefficient of variation - an essential parameter in designing a cluster randomized trial. Results We construct collections of sexual networks and then propagate HIV on them to simulate the disease epidemic. Increasing level of sexual mixing between intervention and standard-of-care (SOC) communities reduces the difference in cumulative incidence in the two sets of communities. Fifteen clusters per arm and 500 incidence cohort members per community provide 95% power to detect the projected difference in cumulative HIV incidence between SOC and intervention communities (3.93% and 2.34%) at the end of the third study year, using a coefficient of variation 0.25. Although available formulas for calculating sample size for cluster randomized trials can be derived by assuming an exchangeable correlation structure within clusters, we show that deviations from this assumption do not generally affect the validity of such formulas. Limitations We construct sexual networks based on data from Likoma Island, Malawi, and base disease progression on longitudinal estimates from an incidence cohort in Botswana and in Durban as well as a household survey in Mochudi, Botswana. Network data from Botswana and larger sample sizes to estimate rates of disease progression would be useful in assessing the robustness of our model results. Conclusion Epidemic modeling plays a critical role in planning and evaluating interventions for prevention. Simulation studies allow us to take into consideration available information on sexual network characteristics, such as mixing within and between communities as well as coverage levels for different prevention modalities in the combination prevention package.


Innate immune reconstitution with suppression of HIV-1.
Progressive HIV-1 infection leads to both profound immune suppression and pathologic inflammation in the majority of infected individuals. While adaptive immune dysfunction, as evidenced by CD4+ T cell depletion and exhaustion, has been extensively studied, less is known about the functional capacity of innate immune cell populations in the context of HIV-1 infection. Given the broad susceptibility to opportunistic infections and the dysregulated inflammation observed in progressive disease, we hypothesized that there would be significant changes in the innate cellular responses. Using a cohort of patients with multiple samplings before and after antiretroviral therapy (ART) initiation, we demonstrated increased responses to innate immune stimuli following viral suppression, as measured by the production of inflammatory cytokines. Plasma viral load itself had the strongest association with this change in innate functional capacity. We further identified epigenetic modifications in the TNFA promoter locus in monocytes that are associated with viremia, suggesting a molecular mechanism for the observed changes in innate immune function following initiation of ART. These data indicate that suppression of HIV-1 viremia is associated with changes in innate cellular function that may in part determine the restoration of protective immune responses.


Usability and Acceptability of a Smartphone App to Assess Partner Communication, Closeness, Mood, and Relationship Satisfaction: Mixed Methods Study.
Interpersonal communication is critical for a healthy romantic relationship. Emotional disclosure, coupled with perceived partner responsiveness, fosters closeness and adjustment (better mood and relationship satisfaction). On the contrary, holding back from disclosure is associated with increased distress and decreased relationship satisfaction. Prior studies assessing these constructs have been cross-sectional and have utilized global retrospective reports of communication. In addition, studies assessing holding back or perceived partner responsiveness have not taken advantage of smartphone ownership for data collection and have instead required website access or use of a study-provided device. This study aimed to examine the (1) usability and acceptability of a smartphone app designed to assess partner communication, closeness, mood, and relationship satisfaction over 14 days and (2) between-person versus within-person variability of key constructs to inform the utility of their capture via ecological momentary assessment using the participants' own handheld devices. Adult community volunteers in a married or cohabiting partnered relationship received 2 smartphone prompts per day, one in the afternoon and one in the evening, for 14 days. In each prompt, participants were asked whether they had conversed with their partner either since awakening (afternoon prompt) or since the last assessment (evening prompt). If yes, a series of items assessed enacted communication, perceived partner communication, closeness, mood, and relationship satisfaction (evening only). Participants were interviewed by phone, 1 week after the end of the 14-day phase, to assess perceptions of the app. Content analysis was employed to identify key themes. Participants (N=27; mean age 36, SD 12 years; 24/27, 89% female; 25/27, 93% white and 2/27, 7% Hispanic) responded to 79.2% (555/701) of the total prompts sent and completed 553 (78.9%) of those assessments. Of the responded prompts, 79.3% (440/555) were characterized by a report of having conversed with one's partner. The app was seen as highly convenient (mean 4.15, SD 0.78, scale: 1-5) and easy to use (mean 4.39, SD 0.70, scale: 1-5). Qualitative analyses indicated that participants found the app generally easy to navigate, but the response window too short (45 min) and the random nature of receiving notifications vexing. With regard to the variability of the app-delivered items, intraclass correlation coefficients were generally <0.40, indicating that the majority of the variability in each measure was at the within-person level. Notable exceptions were enacted disclosure and relationship satisfaction. The findings of this study support the usability and acceptability of the app, with valuable user input to modify timing windows in future work. The findings also underscore the utility of an intensive repeated-measures approach, given the meaningful day-to-day variation (greater within-person vs between-person variability) in communication and mood.


Hyperconnectivity during screen-based stories listening is associated with lower narrative comprehension in preschool children exposed to screens vs dialogic reading: An EEG study.
Dialogic reading (DR) is a shared storybook reading intervention previously shown to have a positive effect on both literacy and general language skills. The aim of this study was to examine the effect of DR compared to screen-based intervention on electrophysiological markers supporting narrative comprehension using EEG. Thirty-two typically developing preschoolers, ages 4 to 6 years, were assigned to one of two intervention groups: Dialogic Reading Group (DRG, n = 16) or Screen Story Group (SSG, n = 16). We examined the effect of intervention type using behavioral assessment and a narrative comprehension task with EEG. The DRG showed improved vocabulary and decreased functional connectivity during the stories-listening task, whereas the SSG group showed no changes in vocabulary or connectivity. Significantly decreased network strength and transitivity and increased network efficiency were observed in the DRG following intervention. Greater network strength and transitivity at follow-up were correlated with increased vocabulary. The results suggest the beneficial effect of DR in preschool-age children on vocabulary and EEG-bands related to attention in the ventral stream during narrative comprehension. Decreased functional connectivity may serve as a marker for language gains following reading intervention. DR intervention for preschool-age children may reduce interfering connections related to attention, which is related to better narrative comprehension.


Soil water content and soil disaggregation by disking affects PM10 emissions.
Row crop agriculture in California's San Joaquin Valley is a major contributor of particulate matter <10 microm in aerodynamic diameter (PM10). The California Air Resources Board uses fixed PM10 emission values for various tillage operations to monitor and design attainment strategies. However, fixed emission values do not reflect emissions produced by a single implement operating under different soil conditions. This 2-yr study evaluated how PM10 mass concentrations (microg L(-1)) from disking change as a function of gravimetric soil water content (GWC), number of sequential diskings (D1, D2, D3), and the soil's weighted mean ped diameter (WMPD). Results showed PM10 increased logarithmically as the soil dried from a GWC of 14 to 4%. Average PM10 values at the lower GWCs were six to eight times greater than at the higher GWCs. Number of diskings also increased PM10, especially in drier soil. Below a GWC of 7%, PM10 for D3 was about twice that for D1. Despite strong correlations between more disking and lower WMPD, a lower WMPD did not always result in an increase in PM10. This underscored the role soil water plays in reducing PM10 at high GWCs despite low WMPDs from multiple diskings. Three-way interactions between GWC, disking, and PM10 showed, on average, that the magnitude of PM10 produced by D1 was 1.3 to 1.6 times lower than by D3, despite having insignificantly different GWC. Therefore, a disking operation can yield two different PM10 values under similar GWCs if the amount of soil disaggregation is different. Our results show that inclusion of soil parameters in PM10 emission estimates is essential to describing agriculture's role in air quality violations and to assess the value of proposed mitigation measures, such as conservation tillage.


Optimality in the zonation of ammonia detoxification in rodent liver.
The rodent liver eliminates toxic ammonia. In mammals, three enzymes (or enzyme systems) are involved in this process: glutaminase, glutamine synthetase and the urea cycle enzymes, represented by carbamoyl phosphate synthetase. The distribution of these enzymes for optimal ammonia detoxification was determined by numerical optimization. This in silico approach predicted that the enzymes have to be zonated in order to achieve maximal removal of toxic ammonia and minimal changes in glutamine concentration. Using 13 compartments, representing hepatocytes, the following predictions were generated: glutamine synthetase is active only within a narrow pericentral zone. Glutaminase and carbamoyl phosphate synthetase are located in the periportal zone in a non-homogeneous distribution. This correlates well with the paradoxical observation that in a first step glutamine-bound ammonia is released (by glutaminase) although one of the functions of the liver is detoxification by ammonia fixation. The in silico approach correctly predicted the in vivo enzyme distributions also for non-physiological conditions (e.g. starvation) and during regeneration after tetrachloromethane (CCl4) intoxication. Metabolite concentrations of glutamine, ammonia and urea in each compartment, representing individual hepatocytes, were predicted. Finally, a sensitivity analysis showed a striking robustness of the results. These bioinformatics predictions were validated experimentally by immunohistochemistry and are supported by the literature. In summary, optimization approaches like the one applied can provide valuable explanations and high-quality predictions for in vivo enzyme and metabolite distributions in tissues and can reveal unknown metabolic functions.


Politics of vulnerability: Impacts of COVID-19 and Cyclone Harold on Indo-Fijians engaged in small-scale fisheries.
The global COVID-19 pandemic has exposed the shortcomings of our health, social, and economic systems. While responding to the health crisis, governments are scrambling to understand and address the knock-on economic effects from market disruptions, and respond to other major disturbances (e.g. natural disasters). We conducted 61 key informant interviews with Indo-Fijian small-scale fisheries (SSF) actors (i.e. fishers, boat owners (that may or may not fish), crew members, and traders) in May 2020, two months after Fiji got its first case of COVID-19 and a month after Cyclone Harold hit the country. We examined how these SSF groups whose access to resources depends on their ability to navigate existing social relations of power, have lived through, experienced, and responded to the two stresses. We found the main impact of COVID-19 on SSF actors was the reduction in sales of fish (73.8 % of respondents) likely a result of reduction in local consumption and/or the loss of tourism markets. Loss of purchasing power meant almost a fifth of Indo-Fijian SSF actors interviewed (comprising 44.4 % of crew members, 16.4 % fishers, 11.5 % boat owners, 8.3 % traders) were unable to obtain sufficient food to meet their families' daily needs. Many of these SSF actors do not have access to social security or similar safety nets leaving them vulnerable to the current crisis as well as to other shocks and changes. Furthermore, social inequities and power relations surrounding access to fisheries resources and government aid contributed to their vulnerability to economic stresses from COVID-19 and a severe cyclone. An understanding of early impacts of COVID-19 on SSF through an intersectional lens can assist decision-makers to quickly mobilise assistance to help people who are most vulnerable, and avoid widening inequities among social groups.


Gender differences in distress: mental health consequences of environmental stress among jail inmates.
The present study examines the influence of gender and environmental stress on the mental health of a sample of 198 male and female jail inmates. Environmental stress is conceptualized as the degree of congruence between inmates' demand and the jail's supply of several environmental features. A state of incongruence was hypothesized to increase the mental distress of jail inmates. In addition, it was hypothesized that female inmates would have higher levels of distress than male inmates, with environmental stress as a possible explanation for gender differences in distress. Female inmates were found to have significantly higher levels of mental distress than males. However, environmental stress was found to be equally detrimental to the mental health of both male and female inmates. Thus, although congruence between environmental demand and supply is a significant predictor of mental health, it is not an explanation for the alarmingly high levels of mental distress found among female inmates.


Can brain-injured patients participate in an aerobic exercise programme during early inpatient rehabilitation?
We investigated the capacity of brain-injured patients to participate in an aerobic exercise programme early after injury. Retrospective analysis of exercise achievements in patients participating in a randomized controlled trial. Ninety patients participated in an exercise training programme on a cycle ergometer at four inpatient neurological rehabilitation units for younger patients. At intake, impairments and function were rated on: Motricity Index, Ashworth Scale, Berg Balance Scale, Barthel Index and Functional Independence Measure. Patients cycled for up to 30 minutes three times weekly for 24-36 sessions over 12 weeks. Exercise performance was measured by: (a) number of sessions to achieve a cycling time of 30 minutes, (b) overall mean cycling time per session over 24 sessions and (c) mean time per session cycling at >60% of age predicted maximum heart rate (HR max) over 24 sessions. Fifty-five patients completed 24 sessions. Thirty-five withdrew, largely for logistic reasons, before completing training; they were significantly less disabled than the 55 who remained. Forty-four of the 55 patients trained for an average of at least 20 minutes per session, 18 training at >60% HR max for this time. There were no differences in performance on the three exercise parameters between two groups of patients with baseline Barthel scores of < or = 12 and > or = 13. Brain-injured patients with a range of disabilities have the capacity to participate in an exercise programme during early inpatient rehabilitation, though some may take longer to achieve adequate intensity of aerobic exercise.


The association between higher maternal pre-pregnancy body mass index and increased birth weight, adiposity and inflammation in the newborn.
Excess adiposity and adiposity-related inflammation are known risk factors for cardiovascular disease in adults; however, little is known regarding the determinants of adiposity-related inflammation at birth. The aim of this study was to investigate the association between maternal pre-pregnancy BMI and newborn adiposity and inflammation. Paired maternal (28-week gestation) and infant (umbilical cord) blood samples were collected from a population-derived birth cohort (Barwon Infant Study, n = 1074). Data on maternal comorbidities and infant birth anthropomorphic measures were compiled, and infant aortic intima-media thickness was measured by trans-abdominal ultrasound. In a selected subgroup of term infants (n = 161), matched maternal and cord lipids, high-sensitivity C-reactive protein (hsCRP) and maternal soluble CD14 were measured. Analysis was completed by using pairwise correlation and linear regression. Because of their non-normal distribution, pathology blood measures were log transformed prior to analysis. Maternal pre-pregnancy BMI was positively associated with increased birth weight (mean difference 17.8 g per kg m-2 , 95% CI 6.6 to 28.9; p = 0.002), newborn mean skin-fold thickness (mean difference 0.1 mm per kg m-2 , 95% CI 0.0 to 0.1; p < 0.001) and cord blood hsCRP (mean difference of 4.2% increase in hsCRP per kg m-2 increase in pre-pregnancy BMI, 95% CI 0.6 to 7.7%, p = 0.02), but not cord blood soluble CD14. Inclusion of maternal hsCRP as a covariate attenuated the associations between pre-pregnancy BMI and both newborn skin-fold thickness and cord blood hsCRP. Higher maternal pre-pregnancy BMI is associated with increased newborn adiposity and inflammation. These associations may be partially mediated by maternal inflammation during pregnancy.


Hepatic steatosis in chronic hepatitis C is a significant risk factor for developing hepatocellular carcinoma independent of age, sex, obesity, fibrosis stage and response to interferon therapy.
Hepatic steatosis is linked to development of hepatocellular carcinoma (HCC) in non-viral liver disease such as non-alcoholic steatohepatitis. The present study aimed to assess whether hepatic steatosis is associated with the development of HCC in chronic hepatitis C. We studied a retrospective cohort of 1279 patients with chronic hepatitis C who received interferon (IFN) therapy between 1994 and 2005 at a single regional hospital in Japan. Of these patients, 393 had a sustained virological response (SVR) and 886 had non-SVR to IFN therapy. After IFN therapy, these patients were screened for development of HCC every 6 months. The average period of observation was 4.5 years. HCC developed in 68 patients. The annual incidence of HCC was 2.73% for patients with a steatosis grade of 10% or greater and 0.69% for patients with a steatosis grade of 0-9%. On multivariate analysis, higher grade of steatosis was a significant risk factor for HCC independent of older age, male sex, higher body mass index (BMI), advanced fibrosis stage and non-SVR to IFN therapy. The adjusted risk ratio of hepatic steatosis was 3.04 (confidence interval 1.82-5.06, P < 0.0001), which was higher than that of older age (1.09), male sex (2.12), non-SVR to IFN (2.43) and higher BMI (1.69). Hepatic steatosis is a significant risk factor for development of HCC in chronic hepatitis C independent of other known risk factors, which suggest the possibility that amelioration of hepatic steatosis may prevent hepatocarcinogenesis.


Coccidiosis in the European badger, Meles meles in Wytham Woods: infection and consequences for growth and survival.
In total 1502 faecal samples were collected from a population of European badgers (Meles meles) between 1992 and 1995 at Wytham Woods, Oxfordshire, UK. Two coccidia species, Eimeria melis and Isospora melis, were identified. Cubs showed a marked seasonal pattern of infection with E. melis, with infection occurring at significantly higher intensity and prevalence than in adults. There was preliminary evidence to suggest that infantile coccidiosis in badgers may be associated with impaired growth and increased mortality.


Risk of Giardia intestinalis infection in children from an artificially recharged groundwater area in Mexico City.
The objective of this study was to assess the risk of infection with Giardia intestinalis in children living in an area with artificial groundwater recharge and potable water reuse in Mexico City. Eligible wells and surrounding homesteads were defined by using a geographic information system. Five wells were tested for G. intestinalis cysts per 400 liters of water. A total of 750 eligible households were visited during two cross-sectional surveys. Stool samples were provided by 986 children in the rainy season study and 928 children during the dry season survey for parasitologic tests. Their guardians provided information on water, sanitation, hygiene, and socioeconomic variables. The prevalence rates of G. intestinalis infection were 9.4% in the rainy season and 4.4% in the dry season. Higher rates of infection were observed in older individuals (9.5% and 10.6%) and girls had a lower risk of infection than boys (odds ratio [OR] =0.55, 95% confidence interval [CI] = 0.34, 0.88 in the rainy season and OR = 0.47, 95% CI = 0.25, 0.90 in the dry season). During the wet season survey, a health risk was detected among those storing water in unprotected receptacles (OR = 4.00, 4.69, and 5.34 for those using uncovered jars, cisterns or tanks, and buckets, respectively), and bathing outside the dwelling, i.e., using a tap (OR = 1.93, 95% CI = 1.10, 3.39). A health risk was also detected among children from households with unsafe food hygiene practices (OR =2.41, 95% CI =1.10, 5.30) and those with no hand-washing habits (OR = 2.27, 95% CI = 1.00, 5.20). Groundwater reserves are at risk of fecal pollution, as indicated by the presence of G. intestinalis cysts. However, the endemic pattern of intestinal infection reflects low standards of personal hygiene and unsafe drinking water storage and food-related practices at household level. Prevention activities must address health education and environmental protection policies.


Continued Investigation Into 17-OHPC: Results From the Preclinical RUPP Rat Model of Preeclampsia.
Preeclampsia is characterized by elevated TNF-α (tumor necrosis factor-α), antiangiogenic factors, such as sFlt-1 (soluble vascular endothelial growth factor receptor 1), increased uterine artery resistance index, and decreased of NO during pregnancy. Previously we showed that 17-hydroxyprogesterone caproate (17-OHPC) administered into reduced uterine perfusion pressure (RUPP) rats on day 18 of gestation improved hypertension without improving pup weight. We hypothesized that earlier administration of 17-OHPC on day 15 of gestation could improve pathophysiology of preeclampsia and fetal outcomes in response to placental ischemia. Carotid catheters were inserted on day 18, and mean arterial blood pressure and samples were collected on day 19. Mean arterial blood pressure in normal pregnant rats was 102±2, 105±2 in normal pregnant+day 15 of gestation (GD15) 17-OHPC, 127±2 in RUPP and 112±1 mm Hg in RUPP+GD15 17-OHPC, P<0.05. Pup weight and litter size were improved from 1.9±0.05, 10.1±1.4 in RUPP to 2.1±0.07 g and 13.2±0.6 in RUPP+GD15 17-OHPC, P<0.05. Uterine artery resistance index was 0.8±0.03 in RUPP, which was decreased to 0.6±0.04 in RUPP+GD15 17-OHPC, P<0.05. Plasma TNF-α levels were 164±34 in RUPP and blunted to 29±9 pg/mL in RUPP+GD15 17-OHPC, P<0.05. Plasma nitrate-nitrite levels were 10.8±2.3 in RUPP rats and significantly increased to 25.5±5.2 µmol/L in RUPP+GD15 17-OHPC, P<0.05. sFlt-1 levels were 386±141 in RUPP rats, which were reduced to 110.2±11 in RUPP+17-OHPC, P<0.05. These data indicate that GD15 17-OHPC improves pathophysiology in RUPP rats, possibly via improving sFlt-1 reduced NO during pregnancy.


Identification of symbol digit modality test score extremes in Huntington's disease.
Studying individuals with extreme phenotypes could facilitate the understanding of disease modification by genetic or environmental factors. Our aim was to identify Huntington's disease (HD) patients with extreme symbol digit modality test (SDMT) scores. We first examined in HD the contribution of cognitive measures of the Unified Huntington's Disease Rating Scale (UHDRS) in predicting clinical endpoints. The language-independent SDMT was used to identify patients performing very well or very poorly relative to their CAG and age cohort. We used data from REGISTRY and COHORT observational study participants (5,603 HD participants with CAG repeats above 39 with 13,868 visits) and of 1,006 healthy volunteers (with 2,241 visits), included to identify natural aging and education effects on cognitive measures. Separate Cox proportional hazards models with CAG, age at study entry, education, sex, UHDRS total motor score and cognitive (SDMT, verbal fluency, Stroop tests) scores as covariates were used to predict clinical endpoints. Quantile regression for longitudinal language-independent SDMT data was used for boundary (2.5% and 97.5% quantiles) estimation and extreme score analyses stratified by age, education, and CAG repeat length. Ten percent of HD participants had an extreme SDMT phenotype for at least one visit. In contrast, only about 3% of participants were consistent SDMT extremes at two or more visits. The thresholds for the one-visit and two-visit extremes can be used to classify existing and new individuals. The identification of these phenotype extremes can be useful in the search for disease modifiers.


Effectiveness of alternative measures to reduce antimicrobial usage in pig production in four European countries.
The reduction of antimicrobial usage (AMU) is in the focus in modern pig production. The objective of this study was to assess the effectiveness of alternatives to reduce AMU at herd level. In a prospective study, 68 farrow-to-finish pig herds located in Belgium, France, Germany and Sweden were recruited on a voluntary basis to implement tailor-made intervention plans to reduce their AMU. Alternative measures included improvement of biosecurity (n = 29 herds), vaccination (n = 30), changes of feeding schemes or drinking water quality (n = 45), improved pig health and welfare care (n = 21) as well as changes in stable climate and zootechnical measures (n = 14). Herds were followed for 1 year after implementation of measures. Annual antimicrobial expenditures or treatment records, as well as disease incidence scores were collected and compared to those of the year before intervention. AMU was measured as the treatment incidence and calculated by age category, antimicrobial class and administration route. Compliance with the intervention plans was high (median 93%). AMU was significantly reduced following the implementation of alternative measures: in the median herd of the four countries, pigs were treated before intervention 25% of their expected lifespan (200 days from birth to slaughter) and after intervention 16%. AMU of suckling and weaned pigs were significantly reduced by 37 and 54%, respectively. The usage of polymyxins and tetracyclines was significantly reduced by 69 and 49%, respectively. AMU via feed and water, as well as parenteral AMU were significantly reduced by 46 and 36%, respectively. Herds with a higher AMU level before intervention achieved a bigger reduction. The majority of disease incidence were similar before and after intervention, with a few exceptions of disorders related to the gastro-intestinal tract in suckling pigs (decreased) and in breeding pigs (increased). Following tailor-made implementation of alternative measures, a substantial reduction of AMU in pig production was achievable without jeopardizing animal health. The AMU reduction in the youngest age categories (suckling and weaned pigs) and the reduction of group treatments via feed and water was in line with the recent European Guidelines on the prudent use of antimicrobials in veterinary medicine.


The role of redundant information in cultural transmission and cultural stabilization.
Redundant copying has been proposed as a manner to achieve the high-fidelity necessary to pass on and preserve complex traits in human cultural transmission. There are at least 2 ways to define redundant copying. One refers to the possibility of copying repeatedly the same trait over time, and another to the ability to exploit multiple layers of information pointing to the same trait during a single copying event. Using an individual-based model, we explore how redundant copying (defined as in the latter way) helps to achieve successful transmission. The authors show that increasing redundant copying increases the likelihood of accurately transmitting a behavior more than either augmenting the number of copying occasions across time or boosting the general accuracy of social learning. They also investigate how different cost functions, deriving, for example, from the need to invest more energy in cognitive processing, impact the evolution of redundant copying. The authors show that populations converge either to high-fitness/high-costs states (with high redundant copying and complex culturally transmitted behaviors; resembling human culture) or to low-fitness/low-costs states (with low redundant copying and simple transmitted behaviors; resembling social learning forms typical of nonhuman animals). This outcome may help to explain why cumulative culture is rare in the animal kingdom.


Scenario approach for assessing the utility of dispersal information in decision support for aerially spread plant pathogens, applied to Phytophthora infestans.
Opportunities exist to improve decision support systems through the use of dispersal information gained from epidemiological research. However, dispersal and demographic information is often fragmentary in plant pathology, and this uncertainty creates a risk of inappropriate action whenever such information is used as a basis for decision making. In this article, a scenario-based simulation approach is used to evaluate crop and economic risks and benefits in the use of dispersal information for decision making using the potato late blight pathosystem (Phytophthora infestans-Solanum tuberosum) as a case study. A recently validated spatiotemporal potato late blight model was coupled to submodels for crop growth, tuber dry matter production, and fungicide efficacy. The yield response of a range of management scenarios to a single influx of primary inoculum (the initial spore load) was calculated. Damage curves (relative yield loss versus initial spore load) from a range of combinations of varietal susceptibility and fungicide treatments were used to classify the various management scenarios as either sensitive to initial spore load or tolerant to initial spore load, thus identifying where a high degree of accuracy would be required in dispersal information for appropriate decision making, and where a greater degree of uncertainty could be tolerated. General epidemics, resulting from spatially homogeneous initial spore loads, responded more strongly to the size of the initial spore load than focal epidemics, resulting from an initial spot infection. Susceptible cultivars responded with sizeable yield losses even at low levels of initial spore load, regardless of the fungicide management regime used. These results indicated that, for susceptible cultivars (late cultivars in particular), the degree of accuracy that would be required in dispersal information for appropriate decision making is unlikely to be practically attainable. The results also indicated that, contrary to "folk wisdom," spore loads of a few hundred spores per square meter do not lead to appreciable crop loss in resistant cultivars and are therefore acceptable. We conclude that scope exists for including dispersal information in decision making for potato late blight with resistant potato cultivars but not for susceptible cultivars. The modeling framework used in this study can be extended to investigate the scope for inclusion of dispersal information in decision support for other aerially transmitted pathogens.


SU-E-T-291: Dosimetry of Double Scattered Proton Beam Fields Used for Cranio-Spinal Irradiation.
To investigate the effect of source to surface distance on treatment field lateral penumbra width and the consequence of setup error on the dose distribution in the junction between two spinal fields of double scattered proton beams. The CT images of the Spine Phantom from Radiological Physics Center was used to design a double scattered proton beam treatment plan using Varian Eclipse treatment planning system. PTV included spinal cord, vertebral body and part of spinous process. The PTV was divided into superior and inferior parts and two posterior fields were used to cover the targets with the prescribed dose. 2D dose was measured using IBA MatriXX and EBT2 film at a depth close to the center of SOBPs of both the fields located both at the nominal source to axis distance (SAD) of 270 cm and at an extended SAD. The field separation was changed by ±1 mm to study the effect of setup error. The measured and TPS calculated dose distributions in verification plans in a water phantom were compared. The measured 2-D doses agreed very well with planned ones for individual fields. 99% of pixels pass 3%/3 mm dose/distance agreement criteria. The CAX dose differences are within 2%. The 80% to 20% penumbra widths at nominal SAD are 7.4/7.7/7.8 mm for planned/MatriXX/EBT2 film respectively, and about 1 mm wider for the extended SAD. The measured maximum dose was about 10% higher than that from the plan, and decreased/increased about 7% when the fields were separated by ±1 mm. The penumbra width is modestly affected by the extended SSD often used for patient treatment. Dose in the junction of two fields is very sensitive to the setup error and the accuracy of the TPS dose calculation in this region may be limited.


Natural isoforms of the Photosystem II D1 subunit differ in photoassembly efficiency of the water-oxidizing complex.
Oxygenic photosynthesis efficiency at increasing solar flux is limited by light-induced damage (photoinhibition) of Photosystem II (PSII), primarily targeting the D1 reaction center subunit. Some cyanobacteria contain two natural isoforms of D1 that function better under low light (D1:1) or high light (D1:2). Herein, rates and yields of photoassembly of the Mn4CaO5 water-oxidizing complex (WOC) from the free inorganic cofactors (Mn(2+), Ca(2+), water, electron acceptor) and apo-WOC-PSII are shown to differ significantly: D1:1 apo-WOC-PSII exhibits a 2.3-fold faster rate-limiting step of photoassembly and up to seven-fold faster rate to the first light-stable Mn(3+) intermediate, IM1*, but with a much higher rate of photoinhibition than D1:2. Conversely, D1:2 apo-WOC-PSII assembles slower but has up to seven-fold higher yield, achieved by a higher quantum yield of charge separation and slower photoinhibition rate. These results confirm and extend previous observations of the two holoenzymes: D1:2-PSII has a greater quantum yield of primary charge separation, faster [P680 (+) Q A (-) ] charge recombination and less photoinhibition that results in a slower rate and higher yield of photoassembly of its apo-WOC-PSII complex. In contrast, D1:1-PSII has a lower quantum yield of primary charge separation, a slower [P680 (+) Q A (-) ] charge recombination rate, and faster photoinhibition that together result in higher rate but lower yield of photoassembly at higher light intensities. Cyanobacterial PSII reaction centers that contain the high- and low-light D1 isoforms can tailor performance to optimize photosynthesis at varying light conditions, with similar consequences on their photoassembly kinetics and yield. These different efficiencies of photoassembly versus photoinhibition impose differential costs for biosynthesis as a function of light intensity.


Photosynthetic responses of a dominant C4 grass to an experimental heat wave are mediated by soil moisture.
Extreme heat waves and drought are predicted to increase in frequency and magnitude with climate change. These extreme events often co-occur, making it difficult to separate their direct and indirect effects on important ecophysiological and carbon cycling processes such as photosynthesis. Here, we assessed the independent and interactive effects of experimental heat waves and drought on photosynthesis in Andropogon gerardii, a dominant C4 grass in a native mesic grassland. We experimentally imposed a two-week heat wave at four intensity levels under two contrasting soil moisture regimes: a well-watered control and an extreme drought. There were three main findings from this study. First, the soil moisture regimes had large effects on canopy temperature, leading to extremely high temperatures under drought and low temperatures under well-watered conditions. Second, soil moisture mediated the photosynthetic response to heat; heat reduced photosynthesis under the well-watered control, but not under the extreme drought treatment. Third, the effects of heat on photosynthesis appeared to be driven by a direct thermal effect, not indirectly through other environmental or ecophysiological variables. These results suggest that while photosynthesis in this dominant C4 grass is sensitive to heat stress, this sensitivity can be overwhelmed by extreme drought stress.


Residential cognitive therapy versus residential interpersonal therapy for social phobia: a randomized clinical trial.
Eighty patients meeting DSM-IV criteria for social phobia were randomly assigned to 10 weeks of residential cognitive therapy (RCT) or residential interpersonal therapy (RIPT). Subjects were assessed at pretreatment, midtreatment, posttreatment, and 1 year after end of treatment. The patients reported chronic, highly comorbid social phobia. Most had tried other treatments without success. Existing individual treatment protocols for cognitive therapy and interpersonal therapy were extensively modified for an integrated group, individual, and residential format. The RCT and RIPT patients improved significantly on the primary outcome measures from pre- to posttreatment. No significant differences were observed between treatments. Patients also completed three weekly secondary outcome measures; on one, social role security, RCT was superior to RIPT. The entire sample reported continued improvement from posttreatment to 1-year follow-up, indicating that improvements were robust. RCT in the present trial was associated with less improvement compared to individual CT in other recent trials.


Hazard of Arm Injury in Professional Starting and Relief Pitchers.
How different pitching roles affect the risk of arm injury in professional pitchers is currently unclear. To investigate differences between professional baseball starting and relief pitchers in the hazard of (1) arm injury and (2) elbow and shoulder injury. Prospective cohort study. Minor League Baseball (MiLB) from 2013 to 2019. Pitchers in MiLB. Pitchers were followed for the entire MiLB season, and athlete-exposures and injuries were recorded. Risk ratios and risk difference were calculated between starting and relieving MiLB pitchers. A Cox survival analysis was then performed in relation to time to arm injury between starting and relieving MiLB pitchers. Subgroup analyses were conducted for elbow and shoulder injuries. A total of 297 pitchers were included, with 85 270 player-days recorded. The incidence of arm injuries was 11.4 per 10 000 athlete-exposures. Starting pitchers demonstrated a greater risk ratio (1.2 [95% CI = 1.1, 1.3]), risk difference (13.6 [95% CI = 5.6, 21.6]), and hazard of arm injury (2.4 [95% CI = 1.5, 4.0]) than relief pitchers. No differences were observed for the hazard of elbow injury between starting and relief pitchers (1.9; 95% CI = 0.8, 4.2). Starting pitchers had a greater hazard of shoulder injury than relief pitchers (3.8 [95% CI = 2.0, 7.1]). Starting pitchers displayed a 2.4 times greater hazard of arm injury than relief pitchers. Subgroup analyses indicated that starters exhibited a greater hazard of shoulder injury than relievers, but no differences occurred for the hazard of elbow injury. However, due to the wide CIs, these subgroup analyses should be interpreted with caution. Clinicians may need to consider cumulative exposure and fatigue and how these factors relate to different pitching roles when assessing the risk of pitching arm injury.


The extraordinary dielectric properties of biological materials and the action of enzymes.
From very general theoretical considerations it is concluded that many biomolecules (i) should have metastable excited states with very high dipole moment, and (ii) should be capable of strongly excited giant dipole vibrations with frequencies near 10(11) Hz. Experimental evidence available so far seems to support these postulates. It is suggested that the two postulates should be of importance for the action of enzymes, and relevant experiments are proposed.


Randomized phase II study of preoperative afatinib in untreated head and neck cancers: predictive and pharmacodynamic biomarkers of activity.
There is no strong and reliable predictive biomarker in head and neck squamous cell carcinoma (HNSCC) for EGFR inhibitors. We aimed to identify predictive and pharmacodynamic biomarkers of efficacy of afatinib, a pan-HER tyrosine kinase inhibitor, in a window-of-opportunity trial (NCT01415674). Multi-omics analyses were carried out on pre-treatment biopsy and surgical specimen for biological assessment of afatinib activity. Sixty-one treatment-naïve and operable HNSCC patients were randomised to afatinib 40 mg/day for 21-28 days versus no treatment. Afatinib produced a high rate of metabolic response. Responders had a higher expression of pERK1/2 (P = 0.02) and lower expressions of pHER4 (P = 0.03) and pRB1 (P = 0.002) in pre-treatment biopsy compared to non-responders. At the cellular level, responders displayed an enrichment of tumor-infiltrating B cells under afatinib (P = 0.02). At the molecular level, NF-kappa B signaling was over-represented among upregulated genes in non-responders (P < 0.001; FDR = 0.01). Although exploratory, phosphoproteomics-based biomarkers deserve further investigations as predictors of afatinib efficacy.


Structural and functional characterization of an intradiol ring-cleavage dioxygenase from the polyphagous spider mite herbivore Tetranychus urticae Koch.
Genome analyses of the polyphagous spider mite herbivore Tetranychus urticae (two-spotted spider mite) revealed the presence of a set of 17 genes that code for secreted proteins belonging to the "intradiol dioxygenase-like" subgroup. Phylogenetic analyses indicate that this novel enzyme family has been acquired by horizontal gene transfer. In order to better understand the role of these proteins in T. urticae, we have structurally and functionally characterized one paralog (tetur07g02040). It was demonstrated that this protein is indeed an intradiol ring-cleavage dioxygenase, as the enzyme is able to cleave catechol between two hydroxyl-groups using atmospheric dioxygen. The enzyme was characterized functionally and structurally. The active site of the T. urticae enzyme contains an Fe3+ cofactor that is coordinated by two histidine and two tyrosine residues, an arrangement that is similar to those observed in bacterial homologs. However, the active site is significantly more solvent exposed than in bacterial proteins. Moreover, the mite enzyme is monomeric, while almost all structurally characterized bacterial homologs form oligomeric assemblies. Tetur07g02040 is not only the first spider mite dioxygenase that has been characterized at the molecular level, but is also the first structurally characterized intradiol ring-cleavage dioxygenase originating from a eukaryote.


BCR/ABL regulates mammalian RecA homologs, resulting in drug resistance.
RAD51 is one of six mitotic human homologs of the E. coli RecA protein (RAD51-Paralogs) that play a central role in homologous recombination and repair of DNA double-strand breaks (DSBs). Here we demonstrate that RAD51 is important for resistance to cisplatin and mitomycin C in cells expressing the BCR/ABL oncogenic tyrosine kinase. BCR/ABL significantly enhances the expression of RAD51 and several RAD51-Paralogs. RAD51 overexpression is mediated by a STAT5-dependent transcription as well as by inhibition of caspase-3-dependent cleavage. Phosphorylation of the RAD51 Tyr-315 residue by BCR/ABL appears essential for enhanced DSB repair and drug resistance. Induction of the mammalian RecA homologs establishes a unique mechanism for DNA damage resistance in mammalian cells transformed by an oncogenic tyrosine kinase.


Distributed Implementation of Boolean Functions by Transcriptional Synthetic Circuits.
Starting in the early 2000s, sophisticated technologies have been developed for the rational construction of synthetic genetic networks that implement specified logical functionalities. Despite impressive progress, however, the scaling necessary in order to achieve greater computational power has been hampered by many constraints, including repressor toxicity and the lack of large sets of mutually orthogonal repressors. As a consequence, a typical circuit contains no more than roughly seven repressor-based gates per cell. A possible way around this scalability problem is to distribute the computation among multiple cell types, each of which implements a small subcircuit, which communicate among themselves using diffusible small molecules (DSMs). Examples of DSMs are those employed by quorum sensing systems in bacteria. This paper focuses on systematic ways to implement this distributed approach, in the context of the evaluation of arbitrary Boolean functions. The unique characteristics of genetic circuits and the properties of DSMs require the development of new Boolean synthesis methods, distinct from those classically used in electronic circuit design. In this work, we propose a fast algorithm to synthesize distributed realizations for any Boolean function, under constraints on the number of gates per cell and the number of orthogonal DSMs. The method is based on an exact synthesis algorithm to find the minimal circuit per cell, which in turn allows us to build an extensive database of Boolean functions up to a given number of inputs. For concreteness, we will specifically focus on circuits of up to 4 inputs, which might represent, for example, two chemical inducers and two light inputs at different frequencies. Our method shows that, with a constraint of no more than seven gates per cell, the use of a single DSM increases the total number of realizable circuits by at least 7.58-fold compared to centralized computation. Moreover, when allowing two DSM's, one can realize 99.995% of all possible 4-input Boolean functions, still with at most 7 gates per cell. The methodology introduced here can be readily adapted to complement recent genetic circuit design automation software. A toolbox that uses the proposed algorithm was created and made available at https://github.com/sontaglab/DBC/.


Absence of acute ocular damage in humans after prolonged exposure to intense RF EMF.
The eye is considered to be a critical organ when determining safety standards for radio frequency (RF) radiation. Experimental data obtained using animals showed that RF heating of the eye, particularly over a specific threshold, can induce cataracts. During the treatment of cancer in the head and neck by hyperthermia, the eyes receive a considerable dose of RF radiation due to stray radiation from the prolonged (60 min) and intense exposure at 434 MHz of this region. In the current study, we verified the exposure guidelines for humans by determining the association between the electromagnetic and thermal dose in the eyes with the reported ocular effects. We performed a simulation study to retrospectively assess the specific absorption rate (SAR) and temperature increase in the eyes of 16 selected patients (encompassing a total of 74 treatment sessions) whose treatment involved high power delivery as well as a minimal distance between the tumor site and the eye. Our results show that the basic restrictions on the peak 10 g spatial-averaged SAR (10 W kg(-1)) and peak tissue temperature increase (1 °C) are exceeded by up to 10.4 and 4.6 times, on average, and by at least 6.2 and 1.8 times when considering the lower limit of the 95% confidence interval. Evaluation of the acute effects according to patients' feedback (all patients), the common toxicity criteria scores (all patients) and an ophthalmology investigation (one patient with the highest exposure) revealed no indication of any serious acute ocular effect, even though the eyes were exposed to high electromagnetic fields, leading to a high thermal dose. We also found that, although there is a strong correlation (R (2) = 0.88) between the predicted induced SAR and temperature in the eye, there are large uncertainties regarding the temperature-SAR relationship. Given this large uncertainty (129%) compared with the uncertainty of 3D temperature simulations (61%), we recommend using temperature simulations as a dosimetric measure in electromagnetic exposure risk assessments.


Zebrafish Mecp2 is required for proper axonal elongation of motor neurons and synapse formation.
Rett syndrome is a severe neurodevelopmental disorder. It is caused by a mutation in methyl-CpG binding protein 2 (MecP2), a transcriptional regulator that recruits protein complexes involved in histone modification and chromatin remodeling. However, the role of Mecp2 in Rett syndrome remains unclear. In this study, we investigated the function of Mecp2 in neuronal development using zebrafish embryos. Mecp2 expression was detected ubiquitously in the central nervous system and muscles at 28 h postfertilization (hpf). We injected an antisense morpholino oligonucleotide (AMO) to induce Mecp2 knockdown phenotype. In mecp2 morphants (embryos with Mecp2 knockdown by AMO) at 28 and 72 hpf, we found an increase in abnormal axonal branches of caudal primary motor neurons and a decrease in motor activity. In mecp2 morphants at 24 hpf, we observed an increase in the expression of an mecp2 downstream candidate gene, brain derived neurotrophic factor (bdnf). In mecp2 morphants at 72 hpf, the presynaptic area stained by an anti-SV2 antibody was increased at the neuromuscular junction (NMJ). Interestingly, the size of SV2-positive presynaptic area at the NMJ was also increased following bdnf mRNA injection, while it was normalized in a double knockdown of mecp2 and bdnf. These results imply that Mecp2 is an important functional regulator of bdnf gene expression during neural circuit formation in zebrafish embryo. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1101-1113, 2017.


Role of leptin as antioxidant in obstructive sleep apnea: an in vitro study using electron paramagnetic resonance method.
As in obstructive sleep apnea (OSA), the chronic cycles of hypoxia and reoxygenation are thought to be conducive of oxidative stress (OS) with generation of reactive oxygen species, identifying effective mechanisms of protection against oxidant-mediated tissue damage becomes of outmost importance. Leptin's role had been recently extended into that of participant to OS; while its exact role in this process is yet to be defined, elevated leptin levels correlate significantly with several indices of OSA disease severity such as nocturnal hypoxemia, possibly acting as a counteractive mechanism against the chronic intermittent hypoxia-related OS and serving as a marker of future risk of atherosclerotic disease. We therefore investigated leptin's antioxidant mechanism on superoxide (O (2) (-•) ) anions using spectrophotometry and electron paramagnetic resonance (EPR). The O (2) (-•) was generated by oxidation of xanthine (XAN) by xanthine oxidase (XO) in the presence of spin trap 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide with various concentrations of leptin (0.001, 0.01, 0.1, and 1 mg/ml) and without leptin. Signal intensity between 3,440 and 3,540 G was expressed as standard means ± SD. The activity of leptin on XO was determined by monitoring the conversion of XAN to uric acid at 293 nm using a Beckman DU 800 UV-visible spectrophotometer. Leptin added to aqueous solutions at 0.1 and 1 mg/ml concentrations was associated with a statistically significant decrease in the EPR signal due to leptin's direct scavenging activity towards the O (2) (-•) . Leptin is an antioxidant agent of possible use as a marker of OS and future risk of atherosclerotic disease in OSA.


Effect of paternal age on reproductive outcomes in oocyte donation model: a systematic review.
To perform a meta-analysis of the literature examining the influence of paternal age on oocyte donation outcomes. Systematic review of the literature with no language or time restrictions. Not applicable. None. None. Pregnancy and live-birth rates. By independent screening of titles and abstracts, two investigators selected original studies examining the influence of paternal age on oocyte donation outcomes. Twelve articles were included, encompassing a total of 12,538 oocyte-donation cases. No statistically significant correlation was found by most studies between advanced paternal age and the rate of fertilization, cleavage embryo development, implantation, pregnancy, miscarriage, or live birth. A statistically significant decrease in blastocyst embryo formation was suggested in two articles. Except for volume and possibly motility, other sperm characteristics such as concentration and morphology did not alter with age. However, the overall quality of the evidence was rated as very low according to Grading of Recommendations Assessment, Development, and Evaluation criteria. The available evidence suggests that advancing paternal age is not associated with adverse oocyte donation outcomes, including pregnancy and live-birth rates. However, the suboptimal quality of the available evidence necessitates high-quality, well-adjusted prospective trials that are also aimed at evaluating additional outcomes such as congenital anomalies and various specific long-term disorders.


FokI polymorphism of the vitamin D receptor gene correlates with parameters of bone mass and turnover in a female population of the Italian island of Lampedusa.
One of the most promising genetic approaches to dissecting a multifactorial disease is represented by genetically isolated population studies. We studied a genetic marker in a cohort of women living on the Mediterranean island of Lampedusa, a geographically isolated population. Lampedusa, located between the African coast and Sicily, consists of a young genetic isolate (<20 generations) with an exponential growth in the last generations. We analyzed the association between the FokI vitamin D receptor (VDR) gene polymorphism, previously proposed as a predictor of bone mass, with parameters of bone mass and turnover in a cohort of pre- and postmenopausal women living on Lampedusa. In 424 women (277 postmenopausal and 147 premenopausal), allelic frequencies were 49% for the F allele and 51% for the f allele. Using analysis of covariance, we found that subjects with ff genotype exhibited a significantly (P < 0.001) lower lumbar spine bone mass, by dual-energy X-ray absorptiometry, and lower values of bone ultrasonographic parameters (speed of sound and broadband ultrasound attenuation) relative to those with Ff and FF genotypes. Conversely, osteocalcin and serum cross-laps were significantly higher in ff and Ff compared to FF genotype. Our data suggest that FokI VDR polymorphism may contribute to the determination of bone mass and turnover in both pre- and postmenopausal women in this geographically isolated population.


Sensitive and Reversible Detection of Methanol and Water Vapor by In Situ Electrochemically Grown CuBTC MOFs on Interdigitated Electrodes.
The in situ electrochemical growth of Cu benzene-1,3,5-tricarboxylate (CuBTC) metal-organic frameworks, as an affinity layer, directly on custom-fabricated Cu interdigitated electrodes (IDEs) is described, acting as a transducer. Crystalline 5-7 µm thick CuBTC layers are grown on IDEs consisting of 100 electrodes with a width and a gap of both 50 µm and a height of 6-8 µm. These capacitive sensors are exposed to methanol and water vapor at 30 °C. The affinities show to be completely reversible with higher affinity toward water compared to methanol. For exposure to 1000 ppm methanol, a fast response is observed with a capacitance change of 5.57 pF at equilibrium. The capacitance increases in time followed diffusion-controlled kinetics (k = 2.9 mmol s-0.5 g-1CuBTC ). The observed capacitance change with methanol concentration follows a Langmuir adsorption isotherm, with a value for the equilibrium affinity Ke = 174.8 bar-1 . A volume fraction fMeOH = 0.038 is occupied upon exposure to 1000 ppm of methanol. The thin CuBTC affinity layer on the Cu-IDEs shows fast, reversible, and sensitive responses to methanol and water vapor, enabling quantitative detection in the range of 100-8000 ppm.


The clinical benefits of cyclosporine C2-level monitoring: a systematic review.
Monitoring of cyclosporine microemulsion (Neoral) using 2-hour postdose (C2) levels is alleged to improve clinical outcomes, but the efficacy of this strategy is uncertain. A systematic literature search was performed for trials directly comparing patients monitored with C2 levels with those monitored by trough (C0) levels. Primary outcomes assessed were renal function and acute rejection. A total of 29 studies met the inclusion criteria. Only 10 of these were randomized controlled trials. Overall quality was poor and this precluded meta-analysis. The most consistent finding in de novo renal, hepatic, and cardiac transplant recipients is a higher mean cyclosporine dose in the early postoperative period in C2 monitored patients. There is no clear evidence that this leads to impaired renal function. In the majority of studies, the monitoring strategy had no significant effect on the rate of acute rejection. In stable transplant recipients, the majority of studies show a reduction in mean cyclosporine dose with adoption of C2 monitoring. No obvious clinical benefit was derived from this reduction in dose. In de novo transplant patients, there is little evidence from prospective studies to support the theoretical benefits of C2 monitoring. Potential dose reductions in stable patients may reduce costs, but no short-term clinical benefit is seen. Quality of studies in this area is poor, and the practical limitations of C2 monitoring mean that further evidence is required before a strategy for the administration of cyclosporine based on C2 levels can be recommended.


Solid-phase combinatorial synthesis using microarrays of microcompartments with light-induced on-chip cell screening.
The process of drug discovery includes individual synthesis and characterization of drug candidates, followed by a biological screening, which is separated from synthesis in space and time. This approach suffers from low throughput and associated high costs, which in turn lead to inefficiency in the field of drug discovery. Here, we present a miniaturized platform combining combinatorial solid-phase synthesis with high-throughput cell screenings. The method is based on the formation of nanoporous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) layers patterned with hydrophilic spots separated from each other by superhydrophobic liquid-impermeable barriers. The porous polymer inside the hydrophilic spots is used as a support to conduct solid-phase synthesis. The hydrophilic spots can be then filled with droplets containing either reagents for synthesis or live cells. Upon irradiation with UV light, products of solid-phase synthesis are released from the porous polymer because of the photo-cleavable linkers used and diffuse into separate droplets. The light-induced release of the products allows the control of the release spatially, temporally, and quantitatively. To demonstrate the versatility and usability of the platform for various cell lines, we have successfully implemented peptide synthesis to create an exemplary chemical library and demonstrated high cell viability after the UV-triggered small-molecule release.


Effect of magnesium supplementation on blood rheology in NOS inhibition-induced hypertension model.
This study investigated the effects of magnesium on blood rheological properties and blood pressure in nitric oxide synthase (NOS) inhibition-induced hypertension model. Hypertension was induced by oral administration of the nonselective NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME, 25 mg/kg/day) for 6 weeks and systolic blood pressure was measured by the tail-cuff method. The groups receiving magnesium supplementation were fed with rat chow containing 0.8% magnesium oxide during the experiment. At the end of experiment, blood samples were obtained from abdominal aorta, using ether anesthesia. Plasma and erythrocyte magnesium levels were determined by the atomic absorption spectrometer. RBC deformability and aggregation were determined by rotational ektacytometry. Plasma fibrinogen concentration was evaluated by ELISA. Whole blood and plasma viscosities were determined by viscometer and intracellular free Ca++ level was measured by using spectroflurometric method. Blood pressure was elevated in hypertensive groups and suppressed by magnesium therapy. Plasma viscosity and RBC aggregation were found to be higher in hypertensive rats than control animals and these parameters significantly decreased in magnesium supplemented hypertensive animals. Other measurements were not different between experimental groups. These results confirm that blood pressure, plasma viscosity and RBC aggregation increased in NOS inhibition-induced hypertension model and oral magnesium supplementation improved these parameters.


Genetic analysis of human immunodeficiency virus type 1 and 2 (HIV-1 and HIV-2) mixed infections in India reveals a recent spread of HIV-1 and HIV-2 from a single ancestor for each of these viruses.
DNA sequences encoding the surface envelope glycoproteins of human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) were amplified by PCR from uncultured peripheral blood mononuclear cells obtained from patients with serologically defined HIV-1/HIV-2 mixed infections from Bombay, India. HIV-1-specific PCR products were obtained in seven of seven randomly chosen doubly reactive cases, while HIV-2-specific sequences were detected in five of seven cases (71%). DNA sequence analysis showed that the HIV-1 gp120 coding sequences were closely related to each other (nucleotide sequence divergence of between 3.1 and 6.8%). Phylogenetic tree analysis placed the Indian strains within the C subtype of HIV-1, being most similar to sequences previously found in East and South Africa. The HIV-2 sequences were also closely related to each other, with an overall sequence divergence of between 5.6 and 10.5%. The low level of nucleotide divergence among Indian HIV-1 and HIV-2 sequences suggests a fairly recent introduction of each virus into this population from a single point of entry in each case. The HIV-2 sequences reported here represent the first analysis of Asian HIV-2 strains and confirm the serological pattern previously detected in India. These data show that a substantial spread of HIV-2, together with HIV-1, has appeared outside Africa in a population hitherto unexposed to HIV. These findings imply that further spread of HIV-2 worldwide is to be expected and have important implications for future vaccine and therapy development.


Knowledge, Attitudes and Practices of Primary Care Physicians Regarding Tobacco Dependence Treatment in Muscat Governorate, Oman: A cross-sectional study.
This study aimed to assess the knowledge, attitudes and practices (KAPs) of primary care physicians (PCPs) regarding tobacco dependence treatment (TDT) and to identify barriers to TDT practice in Muscat, Oman. This cross-sectional study was carried out from September 2019 to December 2019 at all 27 health centres in Muscat. All 313 PCPs working in these health centres were invited to participate. An adapted version of a previously validated 36-item questionnaire was used to assess KAPs regarding TDT services. A total of 255 PCPs completed the survey (response rate: 81.5%) and they demonstrated poor knowledge (40%) but positive attitudes (60%) about TDT services. Of these, the majority were female (85.9%), had never smoked (99.2%) and were general practitioners (64.3%). The mean age was 34.3 ± 6.3 years. Only 28.2% had previously received TDT training and less than half displayed accurate TDT-related knowledge, with undergraduate medical school representing the main source of such knowledge (58.8%). While 96.9% of the PCPs agreed on the importance of TDT, only 28.6% reported feeling sufficiently confident to provide such services themselves. Lack of training (83.5%) and time (71.4%) were the main barriers to TDT practice. Overall, PCPs in Muscat demonstrated poor knowledge but positive attitudes with regard to TDT; however, further knowledge and training is required in order to ensure the effective and accurate provision of TDT services. These findings provide baseline data which may be of value for further research and in overcoming barriers to TDT practice in Oman; for instance, establishing cessation services as part of the package of care in primary health centres could address the time constraints barriers identified by the PCPs.


Extract of Kuding tea prevents high-fat diet-induced metabolic disorders in C57BL/6 mice via liver X receptor (LXR) β antagonism.
To investigate the effects of ilex kudingcha C. J. Tseng (kuding tea), a traditional beverage in China, on the metabolic disorders in C57BL/6 mice induced by high-fat diets. For the preventive experiment, the female C57BL/6 mice were fed with a standard diet (Chow), high-fat diet (HF), and high-fat diet mixed with 0.05% ethanol extract of kuding tea (EK) for 5 weeks. For the therapeutic experiment, the C57BL/6 mice were fed high-fat diet for 3 months, and then mice were split and EK was given with oral gavages for 2 weeks at 50 mg/day/kg. Body weight and daily food intake amounts were measured. At the end of treatment, the adipocyte images were assayed with a scanning electron microscope, and the fasting blood glucose, glucose tolerance test, serum lipid profile and lipids in the livers were analyzed. A reporter gene assay system was used to test the whether EK could act on nuclear receptor transcription factors, and the gene expression analysis was performed with a quantitative PCR assay. In the preventive treatment, EK blocked the body weight gain, reduced the size of the adipocytes, lowered serum triglyceride, cholesterol, LDL-cholesterol, fasting blood glucose levels and glucose tolerance in high-fat diet-fed C57BL/6 mice. In the therapeutic treatment, EK reduced the size of the white adipocytes, serum TG and fasting blood glucose levels in obese mice. With the reporter assay, EK inhibited LXRβ transactivity and mRNA expression of LXRβ target genes. We observed that EK has both preventive and therapeutic roles in metabolic disorders in mice induced with high-fat diets. The effects appear to be mediated through the antagonism of LXRβ transactivity. Our data indicate that kuding tea is a useful dietary therapy and a potential source for the development of novel anti-obesity and lipid lowering drugs.


Plasmid and chromosomal integration of four novel blaIMP-carrying transposons from Pseudomonas aeruginosa, Klebsiella pneumoniae and an Enterobacter sp.
To provide detailed genetic characterization of four novel blaIMP-carrying transposons from Pseudomonas aeruginosa, Klebsiella pneumoniae and an Enterobacter sp. P. aeruginosa 60512, K. pneumoniae 447, P. aeruginosa 12939 and Enterobacter sp. A1137 were subjected to genome sequencing. The complete nucleotide sequences of two plasmids (p60512-IMP from the 60512 isolate and p447-IMP from the 447 isolate) and two chromosomes (the 12939 and A1137 isolates) were determined, then a genomic comparison of p60512-IMP, p447-IMP and four novel blaIMP-carrying transposons (Tn6394, Tn6375, Tn6411 and Tn6397) with related sequences was performed. Transferability of the blaIMP gene and bacterial antimicrobial susceptibility were tested. Tn6394 and Tn6375 were located in p60512-IMP and p447-IMP, respectively, while Tn6411 and Tn6397 were integrated into the 12939 and A1137 chromosomes, respectively. Tn6394 was an ISPa17-based transposition unit that harboured the integron In992 (carrying blaIMP-1). In73 (carrying blaIMP-8), In73 and In992, together with the ISEcp1:IS1R-blaCTX-M-14-IS903D unit, the macAB-tolC region and the truncated aacC2-tmrB region, respectively, were integrated into the prototype transposons Tn1722, Tn1696 and Tn7, respectively, generating the Tn3-family unit transposons, Tn6375 and Tn6378, and the Tn7-family unit transposon Tn6411, respectively. Tn6397 was a large integrative and conjugative element carrying Tn6378. Complex events of transposition and homologous recombination have occurred during the original formation and further plasmid and chromosomal integration of these four transposons, promoting accumulation and spread of antimicrobial resistance genes.


Molecular and physiological effects of environmental UV radiation on fungal conidia.
Conidia are specialized structures produced at the end of the asexual life cycle of most filamentous fungi. They are responsible for fungal dispersal and environmental persistence. In pathogenic species, they are also involved in host recognition and infection. Conidial production, survival, dispersal, germination, pathogenicity and virulence can be strongly influenced by exposure to solar radiation, although its effects are diverse and often species dependent. UV radiation is the most harmful and mutagenic waveband of the solar spectrum. Direct exposure to solar radiation for a few hours can kill conidia of most fungal species. Conidia are killed both by solar UV-A and UV-B radiation. In addition to killing conidia, which limits the size of the fungal population and its dispersion, exposures to sublethal doses of UV radiation can reduce conidial germination speed and virulence. The focus of this review is to provide an overview of the effects of solar radiation on conidia and on the major systems involved in protection from and repair of damage induced by solar UV radiation. The efforts that have been made to obtain strains of fungi of interest such as entomopathogens more tolerant to solar radiation will also be reviewed.


Causal Structure Learning in Continuous Systems.
Real causal systems are complicated. Despite this, causal learning research has traditionally emphasized how causal relations can be induced on the basis of idealized events, i.e., those that have been mapped to binary variables and abstracted from time. For example, participants may be asked to assess the efficacy of a headache-relief pill on the basis of multiple patients who take the pill (or not) and find their headache relieved (or not). In contrast, the current study examines learning via interactions with continuous dynamic systems, systems that include continuous variables that interact over time (and that can be continuously observed in real time by the learner). To explore such systems, we develop a new framework that represents a causal system as a network of stationary Gauss-Markov ("Ornstein-Uhlenbeck") processes and show how such OU networks can express complex dynamic phenomena, such as feedback loops and oscillations. To assess adult's abilities to learn such systems, we conducted an experiment in which participants were asked to identify the causal relationships of a number of OU networks, potentially carrying out multiple, temporally-extended interventions. We compared their judgments to a normative model for learning OU networks as well as a range of alternative and heuristic learning models from the literature. We found that, although participants exhibited substantial learning of such systems, they committed certain systematic errors. These successes and failures were best accounted for by a model that describes people as focusing on pairs of variables, rather than evaluating the evidence with respect to the full space of possible structural models. We argue that our approach provides both a principled framework for exploring the space of dynamic learning environments as well as new algorithmic insights into how people interact successfully with a continuous causal world.


Sprinting to the finish: Toward a theory of Human Capital Resource Complementarity.
In traditional work contexts, factors such as individuals' general competencies are used to predict indices of their performance such as yearly performance appraisals. Whereas traditional approaches to predicting individuals' performance focus on differences between individuals, a considerable proportion of variability in performance is attributable to within-person sources. However, we submit that within-person variability in performance may also be attributable to the fact that people work in different contexts. Moreover, individual performance is often the result of unrecognized team contributions. Accordingly, we advance a Human Capital Resource Complementarity (HCRC) theory to explain the alignment of human capital resources with dynamic situational features, and to illustrate the influence of team collective competencies on the performance of individual members. We then empirically test HCRC theory-derived hypotheses using a sample of 169 cyclists from 22 teams across 18 stages of the centennial Tour de France. Our results suggest that individuals' specific competencies interact with situational characteristics to predict their performance variability over time, beyond that accounted for by their general competencies. Moreover, these effects are accentuated to the extent that teammates' competencies aligned with individual competencies in a given situation. Implications for future theory building, research, and practice are discussed. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Chemical modulation of mutant mGlu1 receptors derived from deleterious GRM1 mutations found in schizophrenics.
Schizophrenia is a complex and highly heterogeneous psychiatric disorder whose precise etiology remains elusive. While genome-wide association studies (GWAS) have identified risk genes, they have failed to determine if rare coding single nucleotide polymorphisms (nsSNPs) contribute in schizophrenia. Recently, two independent studies identified 12 rare, deleterious nsSNPS in the GRM1 gene, which encodes the metabotropic glutamate receptor subtype 1 (mGlu1), in schizophrenic patients. Here, we generated stable cell lines expressing the mGlu1 mutant receptors and assessed their pharmacology. Using both the endogenous agonist glutamate and the synthetic agonist DHPG, we found that several of the mutant mGlu1 receptors displayed a loss of function that was not due to a loss in plasma membrane expression. Due to a lack of mGlu1 positive allosteric modulators (PAM) tool compounds active at human mGlu1, we optimized a known mGlu4 PAM/mGlu1 NAM chemotype into a series of potent and selective mGlu1 PAMs by virtue of a double "molecular switch". Employing mGlu1 PAMs from multiple chemotypes, we demonstrate that the mutant receptors can be potentiated by small molecules and in some cases efficacy restored to that comparable to wild type mGlu1 receptors, suggesting deficits in patients with schizophrenia due to these mutations may be amenable to intervention with an mGlu1 PAM. However, in wild type animals, mGlu1 negative allosteric modulators (NAMs) are efficacious in classic models predictive of antipsychotic activity, whereas we show that mGlu1 PAMs have no effect to slight potentiation in these models. These data further highlight the heterogeneity of schizophrenia and the critical role of patient selection strategies in psychiatric clinical trials to match genotype with therapeutic mechanism.


Enhancing effect of serum ultrafiltrate on the activity of cephalosporins against gram-negative bacilli.
A few studies have suggested that the inhibitory effect of serum on activity of broad-spectrum cephalosporins is less than that predicted by the degree of protein binding. Microdilution MICs of ceftriaxone, cefoperazone, moxalactam, and ceftizoxime were therefore determined against ATCC and clinical strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus in Mueller-Hinton broth containing either human albumin (as 0, 2.5, or 5% solution) or heat-inactivated human serum (as 0, 25, 50, or 95% solution). Arithmetic linear dilutions were used to improve accuracy. For standard bacterial strains, MICs in the presence of 5% albumin were higher than in broth alone by multiples of 10.9 to 21 for ceftriaxone, 5.5 to 16.4 for cefoperazone, 1.9 to 3.7 for moxalactam, and 1.1 to 1.4 for ceftizoxime, as expected by their protein binding. MICs in the presence of 95% serum were similar to those in 5% albumin for all four drugs against S. aureus and P. aeruginosa but were 2.2- to 4.8-fold lower (P less than 0.001) against E. coli and K. pneumoniae. Similar findings were observed at lower protein concentrations and with clinical isolates, except that for some strains of P. aeruginosa MICs were lower in serum than in albumin. Individual sera from five subjects gave comparable results. The addition of serum ultrafiltrate to albumin-containing solutions reduced MICs of ceftriaxone and cefoperazone 1.6- to 7.4-fold against E. coli and K. pneumoniae (P less than 0.01) but did not alter the MICs for S. aureus. Serum may contain an ultrafiltrable component(s) that enhances the activity of third-generation cephalosporins against many gram-negative bacilli.


Inferring nonneutral evolution from human-chimp-mouse orthologous gene trios.
Even though human and chimpanzee gene sequences are nearly 99% identical, sequence comparisons can nevertheless be highly informative in identifying biologically important changes that have occurred since our ancestral lineages diverged. We analyzed alignments of 7645 chimpanzee gene sequences to their human and mouse orthologs. These three-species sequence alignments allowed us to identify genes undergoing natural selection along the human and chimp lineage by fitting models that include parameters specifying rates of synonymous and nonsynonymous nucleotide substitution. This evolutionary approach revealed an informative set of genes with significantly different patterns of substitution on the human lineage compared with the chimpanzee and mouse lineages. Partitions of genes into inferred biological classes identified accelerated evolution in several functional classes, including olfaction and nuclear transport. In addition to suggesting adaptive physiological differences between chimps and humans, human-accelerated genes are significantly more likely to underlie major known Mendelian disorders.


The Influence of Polymorphisms of Interleukin-17A and -17F Genes on Susceptibility and Activity of Rheumatoid Arthritis.
The roles of interleukin (IL)-17A and IL-17F in the pathogenesis of rheumatoid arthritis (RA) have been previously studied. However, the relationships between polymorphisms (IL-17A G197A, the IL-17F 7488A/G, and the IL-17F 7383A/G) of these genes with RA have not been clarified yet. To investigate the impacts of these polymorphisms on the severity and susceptibility of RA in a Turkish population. One hundred sixty-one patients with RA and 88 healthy sex-, age-, and ethnicity-matched controls were enrolled in this study. The erythrocyte sedimentation rate, C-reactive protein (CRP), and disease activity scores 28 (DAS28) of all participants were recorded. The IL-17A G197A, the IL-17F 7488A/G, and 7383A/G polymorphisms were determined using the polymerase chain reaction-restriction fragment length polymorphism method. We found no significant difference regarding genotypes or allelic frequency distributions of the IL-17A G197A, the IL-17F 7383A/G, and 7488A/G polymorphisms between patients and healthy controls (p>0.05). There were slight, but not significant, differences in terms of CRP levels associated with the distribution of the genotypes of the IL-17F 7488A/G, and regarding DAS28 levels according to the genotype distribution of the IL-17A G197A polymorphism (p=0.062, 0.087, 0.052, respectively). These findings suggest that future larger scale studies with increased power should be performed to determine if the IL-17F 7488A/G and the IL-17A G197A polymorphisms are associated with the disease activity in patients with RA.


Mitochondrial abnormalities in patients with primary open-angle glaucoma.
Primary open-angle glaucoma (POAG) is the second most common cause of blindness. It has been linked to mutations in the myocilin (MYOC) and optineurin (OPTN) genes, although mutations have been found in <5% of patients. The pathologic mechanism(s) of POAG remain unknown but may include retinal ganglion cell apoptosis, which causes progressive damage to axons at the optic nerve head. In 27 patients with definite POAG, the MYOC and OPTN genes were sequenced, the entire mitochondrial (mt)DNA coding region was sequenced, relative mtDNA content was investigated, and mitochondrial respiratory function was assessed. Only three benign polymorphisms were identified in MYOC and OPTN in patients with POAG and in control subjects. Conversely, 27 different novel nonsynonymous mtDNA changes were found, only in patients with POAG (not control subjects), 22 of which (found in 14 patients) were potentially pathogenic. Unlike Leber hereditary optic neuropathy, most mtDNA sequence alterations in patients with POAG were transversions-sequence changes that alter the purine/pyrimidine orientation and imply oxidative stress. mtDNA content was relatively increased in 17 patients with POAG compared with age-matched control subjects, also implying a possible response to oxidative stress. Mean mitochondrial respiratory activity was decreased by 21% in patients with glaucoma compared with control subjects (P<0.001). These results reveal a spectrum of mitochondrial abnormalities in patients with POAG, implicating oxidative stress and implying that mitochondria dysfunction may be a risk factor for POAG. This concept may open up new experimental and therapeutic opportunities.


Provocative questions in osteosarcoma basic and translational biology: A report from the Children's Oncology Group.
Patients who are diagnosed with osteosarcoma (OS) today receive the same therapy that patients have received over the last 4 decades. Extensive efforts to identify more effective or less toxic regimens have proved disappointing. As we enter a postgenomic era in which we now recognize OS not as a cancer of mutations but as one defined by p53 loss, chromosomal complexity, copy number alteration, and profound heterogeneity, emerging threads of discovery leave many hopeful that an improving understanding of biology will drive discoveries that improve clinical care. Under the organization of the Bone Tumor Biology Committee of the Children's Oncology Group, a team of clinicians and scientists sought to define the state of the science and to identify questions that, if answered, have the greatest potential to drive fundamental clinical advances. Having discussed these questions in a series of meetings, each led by invited experts, we distilled these conversations into a series of seven Provocative Questions. These include questions about the molecular events that trigger oncogenesis, the genomic and epigenomic drivers of disease, the biology of lung metastasis, research models that best predict clinical outcomes, and processes for translating findings into clinical trials. Here, we briefly present each Provocative Question, review the current scientific evidence, note the immediate opportunities, and speculate on the impact that answered questions might have on the field. We do so with an intent to provide a framework around which investigators can build programs and collaborations to tackle the hardest problems and to establish research priorities for those developing policies and providing funding.


Achieving high-quality care: a view from NICE.
The National Institute for Health and Care Excellence (NICE) was established in 1999 to provide evidence-based guidance. The task of producing guidance by reviewing primary research data and using an advisory committee to develop evidence-based recommendations, is not straightforward. Guidance production is, however, less challenging than the task of putting evidence-based recommendations into practice.NICE is very sensitive to this challenge as, since 1999, over 1500 pieces of NICE guidance have been published. A number of pieces of guidance relate to heart disease, including pharmaceutical agents, new medical technologies and clinical guidelines. Examples include guidelines on acute heart failure and atrial fibrillation, and advice on technologies including edoxaban and implantable cardioverter defibrillators.The research evidence is clear that a change in practice rarely comes about as a result of simply disseminating guidance on best practice. Simple dissemination is particularly ineffective if the guidance has not been produced by a well-respected, credible organisation. It is also clear from the literature that implementation is more successful when more than one approach is taken, and when there is alignment between efforts at organisational, local and national levels.At an organisational level, there should be support from the Board for quality improvement, with ongoing measurement of progress. Resources should be provided for targeted change programmes, particularly where new guidance suggests improvements are required. A systematic process for putting change in place should include identifying barriers to change, agreeing interventions to overcome the barriers and drive forward improvement and planning for implementation and evaluation.


Effect of low molecular weight heparin (dalteparin) and fondaparinux (Arixtra) on human osteoblasts in vitro.
The prolonged administration of heparin for prevention and treatment of venous thromboembolism has been associated with a risk of heparin-induced osteoporosis. Fondaparinux is a new antithrombotic drug that specifically inhibits factor Xa. Because of the known interactions of other antithrombotic agents with bone remodelling, the effects of fondaparinux on human osteoblasts were analysed in vitro. Primary human osteoblast cell cultures were incubated with either the low molecular weight heparin dalteparin at concentrations of 30, 300 and 900 microg/ml or with fondaparinux at concentrations of 25, 50, 100, 150, 200 and 250 microg/ml. Cellular proliferation rate and protein synthesis were measured. Expression of genes encoding osteocalcin, collagen type I and alkaline phosphatase was examined by reverse transcriptase-polymerase chain reaction. Incubation with dalteparin led to a significant, dose-dependent inhibition of osteoblast proliferation, inhibition of protein synthesis, and inhibited expression of phenotype markers (osteocalcin and alkaline phosphatase genes) after 3 and 7 days. No inhibitory effects were observed in the fondaparinux-treated cells. Fondaparinux did not inhibit osteoblast proliferation in vitro and may reduce the risk of heparin-induced osteoporosis associated with long-term heparin administration.


Hemangioblastic foci in human first trimester placenta: Distribution and gestational profile.
The human placenta is a site of both hematopoiesis and vasculogenesis. There are reports of hemangioblastic foci (HAF) in the first trimester placenta, but little published information about their spatiotemporal incidence. We have used semi-thin sections and whole mount staining techniques on archival early pregnancy hysterectomy material as well as freshly-collected termination tissue. We report a description of the distribution of HAF, their gestational profile, and some characteristics of the constituent cells. We show crypt-shaped HAF are present in villi at different levels from 4 to 11 weeks and in the chorionic plate from 4 to 9 weeks. In the villous placenta, the foci often approach closely at one end to the trophoblast basement membrane. Morphologically they show remarkable similarity to those found in the yolk sac at similar stages. In some crypts, all cells are CD34+, but CD34 and nestin progressively segregate into the endothelial lineage. Brachyury is present in less differentiated cells. The erythroid lineage is dominant, as shown by the widespread expression of CD235a/glycophorin and characteristic erythroid morphologies, indicating various degrees of differentiation. However, CD41 is also present in non-endothelial cells. Initially a discontinuous UEA-1/CD31-positive endothelium forms at the periphery of the foci. These cells appear to become integrated into the developing vasculogenic/angiogenic vessel network. We also demonstrate that, independent of HAF, vasculogenesis occurs near the tips of growing villi during the first trimester. We suggest HAF interface with the developing vascular network, producing communication channels that allow erythrocytes to enter the placental-embryonic circulation. We speculate that the erythroid cells act as oxygen reservoirs during the period before flow of maternal blood through the intervillous space of the placenta, allowing a slow feed of oxygen-rich cells to the developing embryo.


Risk of incident cardiovascular disease among users of smokeless tobacco in the Atherosclerosis Risk in Communities (ARIC) study.
Use of smokeless tobacco in the United States has been relatively constant in recent years, as tobacco companies continue aggressive marketing campaigns. The health effects of smokeless tobacco use need further documentation. Thus, the authors examined whether current use of smokeless tobacco was associated with increased incidence of cardiovascular disease (CVD) in 14,498 men and women aged 45-64 years at baseline (1987-1989) in the Atherosclerosis Risk in Communities (ARIC) Study. There were 2,572 incident CVD events (myocardial infarction, coronary revascularization, coronary death, or stroke) during a median of 16.7 years of follow-up (maximum = 19.1 years). Current use of smokeless tobacco at baseline was associated with 1.27-fold greater CVD incidence (95% confidence interval: 1.06, 1.52) than was nonuse, independently of demographic, socioeconomic, and lifestyle and other tobacco-related variables. Past use of smokeless tobacco was not associated with CVD incidence. In conclusion, current use of smokeless tobacco was associated with increased risk of CVD incidence in ARIC cigarette nonsmokers. Current users of smokeless tobacco should be informed of its harm and advised to quit the practice. Current cigarette smokers should also be given sufficient information on safe, therapeutic methods of quitting which do not include switching to smokeless tobacco.


Methods in high precision refractometry of optical glasses.
Five refractive methods and an interferometric method for measuring refractive indices of optical glasses to a precision of several units in the sixth or seventh decimal place are examined. Tables and graphs are presented as aids in evaluating the various methods. Reduction to standard conditions of temperature, pressure, and humidity is also discussed. The minimum deviation method provides greater precision than the autocollimating method for the same precision in angle measurement. The V block, immersion, and critical angle methods, being comparative methods, provide lower precision. A proposed interferometer method shows capability for seventh place accuracy.


Asylums: the historical perspective before, during, and after.
Extensive institutionalisation of people with mental disorders has a brief history lasting just 150 years. Yet asylums feature prominently in modern perceptions of psychiatry's development, on a mental map drawn in sharp contrasts between humanity and barbarity, knowledge and ignorance, and good and bad practice. This Review seeks to nuance the standard narrative of asylums by considering the voices and views of those who were in them at different historical timepoints. The arguments are to caution against dismissing inpatient care outright as simply fundamentally misconceived, and to show the ideological background to many structures and changes, which might superficially seem merely clinical and instrumental. The aim of this Review is to bring out what current mental health systems can learn from an evidence-based analysis of asylums' actual successes and failures in the past.


Recreational exposure to low concentrations of microcystins during an algal bloom in a small lake.
We measured microcystins in blood from people at risk for swallowing water or inhaling spray while swimming, water skiing, jet skiing, or boating during an algal bloom. We monitored water samples from a small lake as a Microcystis aeruginosa bloom developed. We recruited 97 people planning recreational activities in that lake and seven others who volunteered to recreate in a nearby bloom-free lake. We conducted our field study within a week of finding a 10-microg/L microcystin concentration. We analyzed water, air, and human blood samples for water quality, potential human pathogens, algal taxonomy, and microcystin concentrations. We interviewed study participants for demographic and current health symptom information. Water samples were assayed for potential respiratory viruses (adenoviruses and enteroviruses), but none were detected. We did find low concentrations of Escherichia coli, indicating fecal contamination. We found low levels of microcystins (2 microg/L to 5 microg/L) in the water and (<0.1 ng/m(3)) in the aerosol samples. Blood levels of microcystins for all participants were below the limit of detection (0.147 microg/L). Given this low exposure level, study participants reported no symptom increases following recreational exposure to microcystins. This is the first study to report that water-based recreational activities can expose people to very low concentrations of aerosol-borne microcystins; we recently conducted another field study to assess exposures to higher concentrations of these algal toxins.


Capillary electrophoresis characterization of molecularly imprinted polymer particles in fast binding with 17β-estradiol.
Molecularly imprinted polymer (MIP) submicron particles were synthesized, using either ethylene glycol dimethacrylate or trimethylolpropane trimethacrylate as a cross-linker, specifically for recognition of 17β-estradiol (E2). HPLC with fluorescence detection (HPLC-FD) results showed that 90(±5)% of E2 bound onto these particles after 2 min of incubation, and 96(±3)% after long equilibrium. The binding capacity was 8(±3) μmol/g for MIP particles prepared using ethylene glycol dimethacrylate, and 33-43(±8) μmol/g for using trimethylolpropane trimethacrylate. CE separation of MIP and non-imprinted polymer particles was successful when 50 mM borate buffer (pH 8.5) containing 0.005% w/v EOTrol™ LN in reverse polarity (-30 kV) was used. The electrophoretic mobilities of MIP and non-imprinted polymer particles, together with dynamic light scattering measurement of particle sizes, allowed for an estimation of their surface charges. Automated injection of E2 and particles in mixture set a lower limit of 20(±1) s on incubation time for the study of fast binding kinetics. The presence of E2 and bisphenol A (BPA) together tested the selectivity of MIP particles, when the two compounds competed for available binding cavities or sites. Addition of E2 after BPA confirmed E2 occupation of the specific binding cavities, via displacement of BPA.


Translational control and metastatic progression: enhanced activity of the mRNA cap-binding protein eIF-4E selectively enhances translation of metastasis-related mRNAs.
To form metastases, tumors must break from the primary tumor site, invade surrounding tissues, enter and survive within the circulation and ultimately colonize a distal tissue. Each of these steps requires the cooperative function of numerous proteins--proteins that facilitate angiogenesis (e.g., VEGF), cell survival (e.g., Bcl-2), invasion (e.g., MMPs), and autocrine growth stimulation (e.g., c-myc, cyclin D1). Although expression of these proteins is regulated at many levels by disparate stimuli, translation of these key malignancy-related proteins is regulated primarily by the activity of the mRNA cap-binding protein eIF-4E, the rate-limiting member of the eIF-4F translation initiation complex. By binding the cap structure at the 5' terminus of cellular mRNAs, eIF-4E recruits mRNAs to the eIF-4F complex, which then scans from the 5' cap through the untranslated region (5'UTR), unwinding secondary structure to reveal the translation initiation codon and to enable ribosome loading. Messenger RNAs with short unstructured 5' UTRs are more easily translated than mRNAs harboring lengthy, highly structured 5' UTRs, as these prohibit efficient scanning and start codon recognition. As such, the translation of these mRNAs, which typically encode proteins involved in angiogenesis (e.g., VEGF), tumor growth (cyclin D1) and survival (Bcl-2), is suppressed except when eIF-4E is engaged with the eIF-4F complex--a common event in many human and experimental cancers. This review focuses on the hypothesis that enhanced eIF-4E function contributes to metastatic progression by selectively upregulating the translation of key malignancy-related proteins that together conspire to drive the metastatic process.