Estimating quantitative genetic parameters in wild populations: a comparison of pedigree and genomic approaches.
The estimation of quantitative genetic parameters in wild populations is generally limited by the accuracy and completeness of the available pedigree information. Using relatedness at genomewide markers can potentially remove this limitation and lead to less biased and more precise estimates. We estimated heritability, maternal genetic effects and genetic correlations for body size traits in an unmanaged long-term study population of Soay sheep on St Kilda using three increasingly complete and accurate estimates of relatedness: (i) Pedigree 1, using observation-derived maternal links and microsatellite-derived paternal links; (ii) Pedigree 2, using SNP-derived assignment of both maternity and paternity; and (iii) whole-genome relatedness at 37 037 autosomal SNPs. In initial analyses, heritability estimates were strikingly similar for all three methods, while standard errors were systematically lower in analyses based on Pedigree 2 and genomic relatedness. Genetic correlations were generally strong, differed little between the three estimates of relatedness and the standard errors declined only very slightly with improved relatedness information. When partitioning maternal effects into separate genetic and environmental components, maternal genetic effects found in juvenile traits increased substantially across the three relatedness estimates. Heritability declined compared to parallel models where only a maternal environment effect was fitted, suggesting that maternal genetic effects are confounded with direct genetic effects and that more accurate estimates of relatedness were better able to separate maternal genetic effects from direct genetic effects. We found that the heritability captured by SNP markers asymptoted at about half the SNPs available, suggesting that denser marker panels are not necessarily required for precise and unbiased heritability estimates. Finally, we present guidelines for the use of genomic relatedness in future quantitative genetics studies in natural populations.


Dietary Linoleic Acid: An Omega-6 Fatty Acid Essential for Liver Regeneration in Buffalo Rats.
Rodents are currently the most common animals used for hepatic surgical resection studies that investigate liver regeneration, chronic liver disease, acute liver failure, hepatic metastasis, hepatic function, and hepatic cancer. Our previous work has shown that dietary consumption of linoleic acid (LA) stimulates the growth of rodent and human tumors in vivo. Here we compared 3 diets - a 5% corn oil diet (control), a diet deficient in essential fatty acids (EFAD), and an EFAD supplemented with LA in amounts equal to those in the control diet (EFAD+LA). We hypothesized that consumption of the LA provided in the EFAD+LA diet would elevate plasma levels of LA and stimulate regeneration in rats after a 70% hepatectomy (HPX), and that regeneration would not occur in the EFAD rats. Each diet group was comprised of 30 male and 30 female Buffalo rats (BUFF/CrCrl). Rats were fed one of the 3 diets and water ad libitum. After 8 wk on the assigned diet, rats were underwent a 70% HPX. On days 4 and 21 after HPX, 30 male and 30 female rats from each diet group were anesthetized for in vivo study and then were euthanized for tissue collection. For the in vivo study, arterial and venous blood samples were collected from the liver. LA-, glucose-, and O₂ -uptake, and lactate- and CO₂ -output were significantly higher in LA-replete rats as compared with LA-deficient rats. After a 70% HPX, the remaining liver mass in control and EFAD+LA groups had doubled at day 4, reaching 60% of the original total weight, and had regenerated completely at day 21. However, no regeneration occurred in the EFAD group. At day 4 the portions of livers removed from the control and EFAD+LA groups had significantly higher content of LA, protein, cAMP, and DNA as compared with their livers on day 21. [³ H]thymidine incorporation into liver DNA was significantly higher in the 2 LA-replete groups, with male values greater than female values, as compared with LA-deficient group. These data indicate that liver regeneration after HPX is dependent on dietary LA. Understanding the mechanisms of LA-dependent liver regeneration in rats supports our current efforts to enhance successful surgical resection therapies in humans.


True arrestins and arrestin-fold proteins: a structure-based appraisal.
Arrestin-clan proteins are folded alike, a feature responsible for their recent grouping in a single clan. In human, it includes the well-characterized visual and β-arrestins, the arrestin domain-containing proteins (ARRDCs), isoforms of the retromer subunit VPS26, and DSCR3, a protein involved in Down syndrome. A new arrestin-fold-predicted protein, RGP1, described here may join the clan. Unicellular organisms like the yeast Saccharomyces cerevisiae or the amoeba Dictyostelium discoideum harbor VPS26, DSCR3, and RGP1 isoforms as well as arrestin-related trafficking adaptors or ADCs, but true arrestins are missing. Functionally, members of the arrestin clan have generally a scaffolding role in various membrane protein trafficking events. Despite their similar structure, the mechanism of cargo recognition and internalization and the nature of recruited partners differ for the different members. Based on the recent literature, true arrestins (visual and β-arrestins), ARRDCs, and yeast ARTS are the closest from a functional point of view.


BioSwitch: a tool for the detection of bistability and multi-steady state behaviour in signalling and gene regulatory networks.
Multi-steady state behaviour, and in particular multi-stability, provides biological systems with the capacity to take reliable decisions (such as cell fate determination). A problem arising frequently in systems biology is to elucidate whether a signal transduction mechanism or a gene regulatory network has the capacity for multi-steady state behaviour, and consequently for a switch-like response to stimuli. Bifurcation diagrams are a powerful instrument in non-linear analysis to study the qualitative and quantitative behaviour of equilibria including bifurcation into different equilibrium branches and bistability. However, in the context of signalling pathways, the inherent large parametric uncertainty hampers the (direct) use of standard bifurcation tools. We present BioSwitch, a toolbox to detect multi-steady state behaviour in signalling pathways and gene regulatory networks. The tool combines results from chemical reaction network theory with global optimization to efficiently detect whether a signalling pathway has the capacity to undergo a saddle node bifurcation, and in case of multi-stationarity, provides the exact coordinates of the bifurcation where to start a numerical continuation analysis with standard bifurcation tools, leading to two different branches of equilibria. Bistability detection in the G1/S transition pathway of Saccharomyces cerevisiae is included as an illustrative example. BioSwitch runs under the popular MATLAB computational environment, and is available at https://sites.google.com/view/bioswitch.


Spin echoes in the regime of weak dephasing.
This article analyzes possibilities and limits of spin echoes beyond Hahn's theory. The regime of weak dephasing is explored with the purpose of combining the enhanced signal and reduced artifacts of spin echoes with the speed and flexibility of the fast low angle shot sequence. In the regime of weak dephasing, an upper boundary of the echo time is derived analytically. This limit is verified with optimal control pulses, which are also used to heuristically examine the transition to Hahn's regime of complete dephasing. The potential of the proposed pulses is demonstrated in proof-of-concept spin echo fast low angle shot images of a volunteer's lung and head. It was found that the time between the end of the pulse and the spin echo can exceed the duration of the composite radio frequency-pulse, which stands in contrast to Hahn echoes where those measures are at most equal. The maximum echo time was found to mainly depend on the total amount of dephasing. In vivo spin echo fast low angle shot images show an increased pulmonary signal as well as reduced artifacts in areas affected by susceptibility differences. The spin dynamics in the regime of weak dephasing was investigated and the feasibility of spin echo fast low angle shot imaging was demonstrated in vivo.


A phase II study of vorinostat (MK-0683) in patients with polycythaemia vera and essential thrombocythaemia.
Inhibition of histone deacetylases may be an important target in patients with myeloproliferative neoplasms. This investigator-initiated, non-randomized, open-label phase II multi-centre study included 63 patients (19 essential thrombocythaemia, 44 polycythaemia vera) from 15 centres. The primary objective was to evaluate if vorinostat was followed by a decline in clonal myeloproliferation as defined by European Leukaemia Net. Thirty patients (48%) completed the intervention period (24 weeks of therapy). An intention-to-treat response rate of 35% was identified. Pruritus was resolved [19% to 0% (P = 0·06)] and the prevalence of splenomegaly was lowered from 50% to 27% (P = 0·03). Sixty-five per cent of the patients experienced a decrease in JAK2 V617F allele burden (P = 0·006). Thirty-three patients (52% of patients) discontinued study drug before end of intervention due to adverse events (28 patients) or lack of response (5 patients). In conclusion, vorinostat showed effectiveness by normalizing elevated leucocyte and platelet counts, resolving pruritus and significantly reducing splenomegaly. However, vorinostat was associated with significant side effects resulting in a high discontinuation rate. A lower dose of vorinostat in combination with conventional and/or novel targeted therapies may be warranted in future studies.


Analysis of the mechanical response of biomimetic materials with highly oriented microstructures through 3D printing, mechanical testing and modeling.
Many biomineralized organisms have evolved highly oriented nanostructures to perform specific functions. One key example is the abrasion-resistant rod-like microstructure found in the radular teeth of Chitons (Cryptochiton stelleri), a large mollusk. The teeth consist of a soft core and a hard shell that is abrasion resistant under extreme mechanical loads with which they are subjected during the scraping process. Such remarkable mechanical properties are achieved through a hierarchical arrangement of nanostructured magnetite rods surrounded with α-chitin. We present a combined biomimetic approach in which designs were analyzed with additive manufacturing, experiments, analytical and computational models to gain insights into the abrasion resistance and toughness of rod-like microstructures. Staggered configurations of hard hexagonal rods surrounded by thin weak interfacial material were printed, and mechanically characterized with a cube-corner indenter. Experimental results demonstrate a higher contact resistance and stiffness for the staggered alignments compared to randomly distributed fibrous materials. Moreover, we reveal an optimal rod aspect ratio that lead to an increase in the site-specific properties measured by indentation. Anisotropy has a significant effect (up to 50%) on the Young's modulus in directions parallel and perpendicular to the longitudinal axis of the rods, and 30% on hardness and fracture toughness. Optical microscopy suggests that energy is dissipated in the form of median cracks when the load is parallel to the rods and lateral cracks when the load is perpendicular to the rods. Computational models suggest that inelastic deformation of the rods at early stages of indentation can vary the resistance to penetration. As such, we found that the mechanical behavior of the system is influenced by interfacial shear strain which influences the lateral load transfer and therefore the spread of damage. This new methodology can help to elucidate the evolutionary designs of biomineralized microstructures and understand the tolerance to fracture and damage of chiton radular teeth.


Effects of Org OD 14 on pituitary and peripheral beta-endorphin in castrated rats and post-menopausal women.
The aim of the first part of this study was to evaluate the effects of a new synthetic steroid (7 alpha,17 alpha)-17-hydroxy-7-methyl-19-norpregn-5(10)-en-20-yn-3-one (Org OD 14), on anterior pituitary (AP) and neurointermediate pituitary lobe (NIL) contents and on circulating levels of beta-endorphin (beta-EP) in rats. Three weeks after ovariectomy, groups of 9 rats were treated with either Org OD 14 (2 or 10 micrograms/day/rat for 14 days) or a placebo. In addition, 2 groups of ovariectomized rats were also treated with oestradiol benzoate (EB) (2 or 10 micrograms/day/rat for 14 days) to compare the effectiveness of the new steroid with that of a classical oestrogenic substance. beta-Ep concentrations were measured in plasma and in AP and NIL extracts by means of double-antibody radioimmunoassay (RIA), employing a specific anti-camel beta-EP (C-terminal fragment). Both doses of Org OD 14 induced a significant dose-related increase in plasma and pituitary lobe beta-EP concentrations as compared with the results on placebo treatment. By comparison, EB was active only at a dose of 10 micrograms/day. Despite the common stimulatory effects of EB and Org OD 14 on pituitary beta-EP, these findings suggest that the two steroids have different modes of action. The second part of the study investigated the changes in beta-EP and beta-lipotrophin (beta-LPH) plasma levels in a group of post-menopausal women treated for 6 months with Org OD 14 (2.5 mg/day) in comparison with the levels in a placebo-treated group. The clinical efficacy of Org OD 14 treatment in post-menopausal symptoms was confirmed, as well as its lack of or only transient effect on plasma lipids and lipoproteins. beta-EP and beta-LPH plasma levels were significantly higher in the Org OD 14-treated group than in the placebo group as from the second month until the end of the observation period.


Factors Associated with Ball Velocity and Low Back Pain During Kicking in Adolescent Soccer Players.
The factors associated with low back pain (LBP) and the relationship between LBP and ball velocity during kicking motion of adolescent soccer players remain largely unknown. This study aims to clarify the relationship between increasing ball velocity and LBP in adolescent soccer players. Adolescent soccer players were divided into two groups according to the presence and absence of LBP (LBP group, n=38 and NBP (no back pain) group, n=29, respectively). Real-time kick motion was measured using a three-dimensional motion analysis system and the angle of the lumbar spine, hip, and center of mass (COM) were calculated. Regression analysis was used to identify factors associated with ball velocity and LBP. In addition, Pearson's correlation coefficients were determined between the angle of the lumbar spine and hip, and ball velocity and position of COM in the extracted phase from regression analysis. The major factor associated with increasing ball velocity was the rotation angle of both hips (Adjusted R2=0.244) and vertical position of COM during kicking (Adjusted R2=0.262). Furthermore, the factors associated with LBP were the flexion angle of kick-side hip (OR=1.126) and abduction angle of both hips (kick-side OR=1.124; support-side OR=0.872). The factors for ball velocity and LBP were related to the maximum hip extension phase. In the hip extension phase of kicking, compared with the NBP group, the LBP group showed lesser extension and external rotation of the kick-side hip angle. In the hip flexion phase of kicking, the ball velocity was correlated with vertical (r=0.56)/anterior (r=0.46) position of COM in the NBP group. To compensate for this restricted hip motion, the LBP group could extend and rotate their lumbar spine, which may likely cause stress to this region.


Spatial regulation of protein A in Staphylococcus aureus.
Surface proteins of Staphylococcus aureus play vital roles in bacterial physiology and pathogenesis. Recent work suggests that surface proteins are spatially regulated by a YSIRK/GXXS signal peptide that promotes cross-wall targeting at the mid-cell, though the mechanisms remain unclear. We previously showed that protein A (SpA), a YSIRK/GXXS protein and key staphylococcal virulence factor, mis-localizes in a ltaS mutant deficient in lipoteichoic acid (LTA) production. Here, we identified that SpA contains another cross-wall targeting signal, the LysM domain, which, in addition to the YSIRK/GXXS signal peptide, significantly enhances SpA cross-wall targeting. We show that LTA synthesis, but not LtaS, is required for SpA septal anchoring and cross-wall deposition. Interestingly, LTA is predominantly found at the peripheral cell membrane and is diminished at the septum of dividing staphylococcal cells, suggesting a restriction mechanism for SpA septal localization. Finally, we show that D-alanylation of LTA abolishes SpA cross-wall deposition by disrupting SpA distribution in the peptidoglycan layer without altering SpA septal anchoring. Our study reveals that multiple factors contribute to the spatial regulation and cross-wall targeting of SpA via different mechanisms, which coordinately ensures efficient incorporation of surface proteins into the growing peptidoglycan during the cell cycle.


Combined effects of nucleotide-binding domain-like receptor protein 3 polymorphisms and environmental metals exposure on chronic kidney disease.
Chronic inflammation is the cause of chronic kidney disease (CKD). The nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome plays a vital role in the inflammation process and is associated with the regulatory effects of NLRP3 gene polymorphisms. This study evaluated the association between NLRP3 gene polymorphisms and CKD, and further explored whether the association of environmental metals with CKD varied by the NLRP3 genotypes. A total of 218 CKD patients and 427 age- and sex-matched healthy controls were recruited in this clinic-based case-control study. Patients were identified as having CKD if their estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73 m2 and stage 3-5 for at least 3 months. We examined the genotypes of fifteen common ssingle-nucleotide polymorphisms in NLRP3 genes. Concentrations of total urinary arsenic were examined by summing of urinary inorganic arsenic species. Concentrations of selenium, cadmium, and lead were measured from blood samples. Associations between NLRP3 polymorphisms, environmental metals exposure, and CKD were evaluated using multivariable logistic regression while controlling for confounders. We observed that the odds of carrying NLRP3 rs4925650 GA/AA genotypes, NLRP3 rs1539019 CA/AA genotypes, and NLRP3 rs10157379 CT/TT genotypes were significantly higher among CKD cases compared to controls, with the adjusted odds ratio (95% confidence interval) were 1.54 (1.01-2.36), 1.56 (1.04-2.33), and 1.59 (1.05-2.38), respectively. The significant multiplicative interactions were identified between high levels of blood lead and NLRP3 rs4925650 GA/AA genotypes; high levels of blood cadmium or low levels of plasma selenium and the NLRP3 haplotype (rs4925648, rs4925650, rs12048215, and rs10754555) C-A-A-C multiplicatively interacted to increase the risk of CKD. Our results imply that NLRP3 polymorphisms may play an important role in the development of environmental metals exposure related CKD.


[Essential thrombocythemia transformed to acute myeloblastic leukemia].
A 48-year-old woman was referred to Tohoku University Hospital in November 1981 because of leukocytosis pointed out in a group examination. At that time white blood cell count was 26.8 x 10(3)/microliters with no blasts, platelet count 268.0 x 10(4)/microliters and hemoglobin 11.4 g/dl. Bone marrow aspirates showed marked increase of megakaryocytes (15,900/microliters). Bone marrow chromosome analysis revealed 46, XX, -18, +mar without Ph1 chromosome, and DNA analysis showed no bcr rearrangement. She was diagnosed as having essential thrombocythemia and was treated with busulfan. On November 1986, she developed remarkable leukocytosis with leukemic blasts. White blood cells reached 153 x 10(3)/microliters with 33% blasts. Her blasts were positive for peroxidase staining, but negative for platelet peroxidase on electron microscopic study and platelet specific glycoproteins. A diagnosis of acute myeloblastic leukemia (M2) was made. The patient received various combination chemotherapy, which was ineffective, and she died due to pneumonia on June, 1989. In Japan, there has been reported only 8 cases of essential thrombocythemia transformed to acute leukemia. The clinical pictures of these 9 cases were discussed.


Transient Expression of a Major Ampullate Spidroin 1 Gene Fragment from Euprosthenops sp. in Mammalian Cells.
Spider silk possesses extraordinary and unsurpassed mechanical properties and several attempts have been made to artificially produce spider silk in order to manufacture strong and light engineering composites. In the field of oncology, recombinant spider silk has the potential to be used as a biomaterial for bone replacement after tumour surgery. In this study, a 636-base pair gene fragment, coding for a part of major ampullate spidroin 1 from the African spider, Euprosthenops sp., was cloned into the expression vector pSecTag2/Hygro A, designed for the production of protein in mammalian cells. COS-1 cells were subsequently transfected with the recombinant plasmids and transient expression of low amounts of the corresponding silk protein fragment was obtained. The expressed fragment contained repetitive sequences associated with intrinsic biomechanical properties and has potential as a starting material for designed biopolymers.


Effects of the COVID-19 Pandemic on Black Veterans' Mental Health: A Qualitative Investigation.
Although the disproportionate effects of the COVID-19 pandemic on Black Americans are well-documented, we know little about its effects on their day-to-day lives and sense of wellbeing, especially for those who have served in the military. We conducted qualitative interviews with 21 Black veterans to understand their experiences with the pandemic and administered questionnaires about mental health and pandemic impact. Questionnaires indicated mild depression, moderate anxiety and loneliness, and pandemic effects on social support and health care. Interviews revealed that heightened anxiety and stress were driven by not trusting others to take the pandemic seriously, workplace anxiety, comorbidities, and the pandemic's pronounced effects on Black communities. Participants shared insights into these effects, including lack of caution on the part of some and the role of systemic racism. Findings enhance our understanding of how Black veterans have experienced the pandemic and may have important implications for other underserved groups.


Diabetes, subclinical atherosclerosis and multiple cardiovascular risk factors in hard-to-reach asymptomatic patients.
To examine the association of cardiovascular disease risk factors with and their cumulative effect on coronary artery calcium in hard-to-reach asymptomatic patients with diabetes. A total of 2563 community-dwelling asymptomatic subjects from Central Appalachia participated in coronary artery calcium screening at a heart centre. Binary variable was used to indicate that coronary artery calcium was either present or absent. Independent variables consisted of demographic and modifiable risk factors and medical conditions. Descriptive statistics and multinomial logistic regression analyses were conducted. In total, 55.8% and 13.7% of study participants had subclinical atherosclerosis (coronary artery calcium ⩾1) and diabetes, respectively. The presence of coronary artery calcium was higher in subjects with diabetes (68.5%) than those without (53.8%). Compared to subjects without diabetes with coronary artery calcium = 0, obesity, hypertension, hypercholesterolaemia and smoking increased the odds of the presence of coronary artery calcium (coronary artery calcium score ⩾1) regardless of diabetes status; however, with larger odds ratios in subjects with diabetes. Compared to subjects without diabetes with coronary artery calcium score = 0, having 3, 4 and ⩾5 risk factors increased the odds of presence of coronary artery calcium in subjects with diabetes by 14.06 (confidence interval = 3.26-62.69), 32.30 (confidence interval = 7.41-140.82) and 47.12 (confidence interval = 10.35-214.66) times, respectively. There is a need for awareness about subclinical atherosclerosis in patients with diabetes and more research about coronary artery calcium in subpopulations of patients.


Diet, metabolic polymorphisms and dna adducts: the EPIC-Italy cross-sectional study.
DNA adducts in peripheral leukocytes are considered a reliable indicator of internal dose exposure to genotoxic agents and, possibly, of cancer risk. We investigated their association with diet and other individual characteristics in healthy adults. The prospective study EPIC-Italy, a section of a larger European project, enrolled 47,749 men and women, aged 35-64 years, in 5 centres: all provided individual information about dietary and life-style habits and a blood sample. In a cross-sectional study, approximately 100 volunteers were randomly selected from each of the three main geographical study areas (Northern, Central and Southern Italy). DNA adducts and four polymorphic metabolic genotypes were determined in peripheral leukocytes by using (32)P-postlabelling technique and PCR methods. Among 309 subjects (153 men), 72.8% had detectable levels of DNA adducts (mean: 8.1 +/- 0.6 per 10(9) nucleotides). Strong negative associations emerged with the reported frequency of consumption of fresh fruit and vegetables, olive oil, and the intake of antioxidants. DNA adducts were higher in subjects with GSTT1 null genotype (p = 0.05). Significant differences between study centres emerged in multivariate analyses (mean levels: 11.0, 10.0, 7.2, 6.5 and 5.2 for Florence, Naples, Turin, Varese and Ragusa, respectively). A possible opposite seasonal variation was found according to latitude: adduct levels tended to be lower in winter in Florence and the southern centres, and during summer in the two northern centres. Frequent consumption of fresh fruit and vegetables is associated with reduced levels of DNA adducts, possibly contributing to the role of diet in modulating cancer risk.


Frontier America's health system challenges and population health outcomes.
The objective of this cross-sectional descriptive study was to examine and compare the county-level characteristics including demographic factors, health system factors, and population health outcomes of frontier and nonfrontier counties in the United States. All counties in the United States were studied using the merged County Health Rankings 2011 and the Area Resource File 2009 databases. Of a total of 3,141 counties in the County Health Rankings 2011 database, 438 were identified as frontier counties using the conventional definition of fewer than 7 persons per square mile. Frontier counties were found to have a significantly higher proportion of elderly, Hispanic, and Native American residents than nonfrontier counties. Frontier counties have lower household income and lower levels of illiteracy. Frontier counties also have significantly fewer primary care physicians and higher uninsurance rates. Although frontier counties have a lower percentage of ZIP codes with healthy food and recreational facilities, the incidence of obesity is lower in frontier areas. Empirical literature on the population health outcomes and health system factors of frontier areas is limited. Frontier communities in the United States face significant challenges in terms of having populations with a higher need for primary care such as the elderly and poor. In addition, they face access barriers due to geographic remoteness. The availability of reliable data on population outcomes will enable policy makers to monitor the health status of frontier populations and to design solutions to the access issues that these populations face.


Current health-related quality of life is lower in former Division I collegiate athletes than in non-collegiate athletes.
College athletes participate in physical activity that may increase chronic stress and injury and induce overtraining. However, there is little known about how previous injuries that have occurred during college may limit current physical activity and/or decrease their subsequent health-related quality of life (HRQoL). To evaluate HRQoL in former United States National Collegiate Athletic Association Division I athletes and nonathletes with the Patient-Reported Outcomes Measurement Information System (PROMIS) and a demographics questionnaire. Cohort study (prognosis); Level of evidence, 2. The study sample was recruited through alumni databases at a large Midwestern university and consisted of 2 cohorts: (1) former Division I athletes and (2) nonathletes who participated in recreational activity, club sports, or intramurals while attending college. Participants answered a survey constructed with a web-based system. All individuals contacted were between the ages of 40 and 65 years. Study participants responded to the questions on the PROMIS scales for sleep, anxiety, depression, fatigue, pain interference, physical function, and satisfaction with participation in social roles. The PROMIS was developed to fill the void of HRQoL being evaluated by multiple instruments. An additional questionnaire was constructed to record demographic and exercise information. Initially, 1280 former Division I athletes and nonathletes were contacted; 638 surveys were returned (49.8%). Surveys eligible for analyses (71.6%) were completed by 232 former Division I athletes (mean age ± SD, 53.36 ± 7.11 years) and 225 nonathletes (mean age ± SD, 53.60 ± 6.79 years). Univariate analyses for the effect of group was significantly related to PROMIS scales for physical function, depression, fatigue, sleep disturbances, and pain interference (P < .05). The overall scores were significantly worse for the former Division I athletes than for the nonathletes on 5 of the 7 scales. In addition, the former Division I athletes reported significantly more limitations in daily activities and more major and chronic injuries than did the nonathlete controls. According to these data, former Division I athletes have decreased HRQoL compared with nonathletes. Sports encourage physical activity, which help promote a healthy lifestyle. Moderate activity and exercise should be encouraged. However, the demands of Division I athletics may result in injuries that linger into adulthood and possibly make participants incapable of staying active as they age, thereby lowering their HRQoL.


Algae as nutritional and functional food sources: revisiting our understanding.
Global demand for macroalgal and microalgal foods is growing, and algae are increasingly being consumed for functional benefits beyond the traditional considerations of nutrition and health. There is substantial evidence for the health benefits of algal-derived food products, but there remain considerable challenges in quantifying these benefits, as well as possible adverse effects. First, there is a limited understanding of nutritional composition across algal species, geographical regions, and seasons, all of which can substantially affect their dietary value. The second issue is quantifying which fractions of algal foods are bioavailable to humans, and which factors influence how food constituents are released, ranging from food preparation through genetic differentiation in the gut microbiome. Third is understanding how algal nutritional and functional constituents interact in human metabolism. Superimposed considerations are the effects of harvesting, storage, and food processing techniques that can dramatically influence the potential nutritive value of algal-derived foods. We highlight this rapidly advancing area of algal science with a particular focus on the key research required to assess better the health benefits of an alga or algal product. There are rich opportunities for phycologists in this emerging field, requiring exciting new experimental and collaborative approaches.


Development of a longitudinal injury prevention curriculum for undergraduate medical students.
Although injury is a major cause of death and disability, concepts of injury prevention have not been present in the curricula of most medical schools. There have been recent calls in the literature, including a 2005 report from the Association of American Medical Colleges, for the addition of injury prevention concepts into medical school education. This report describes the process of development of a longitudinal injury prevention curriculum in one medical school. The curriculum committee felt that adding the material in a longitudinal fashion would better correlate the material to the rest of the curriculum as well as provide efficiency in the already crowded undergraduate programme of study. The report describes the content of the longitudinal curriculum developed as well as initial evaluation of the material.


Compensation of Signal Spillover in Suspension and Imaging Mass Cytometry.
The advent of mass cytometry increased the number of parameters measured at the single-cell level while decreasing the extent of crosstalk between channels relative to dye-based flow cytometry. Although reduced, spillover still exists in mass cytometry data, and minimizing its effect requires considerable expert knowledge and substantial experimental effort. Here, we describe a novel bead-based compensation workflow and R-based software that estimates and corrects for interference between channels. We performed an in-depth characterization of the spillover properties in mass cytometry, including limitations defined by the linear range of the mass cytometer and the reproducibility of the spillover over time and across machines. We demonstrated the utility of our method in suspension and imaging mass cytometry. To conclude, our approach greatly simplifies the development of new antibody panels, increases flexibility for antibody-metal pairing, opens the way to using less pure isotopes, and improves overall data quality, thereby reducing the risk of reporting cell phenotype artifacts.


The therapeutic potential and mechanisms of action of quercetin in relation to lipopolysaccharide-induced sepsis in vitro and in vivo.
Sepsis caused by Gram-negative bacterial infection is characterized by extensive inflammatory cytokine production, which leads to multiple organ failure and a high lethality rate. Therefore, compounds that are able to alleviate profound inflammatory responses may have therapeutic potential in relation to sepsis. Quercetin, one of the flavonoids found widely in the human diet, has been reported to have many health benefits, but the mechanisms underlying its biological effects remain obscure. In the present study, our aim was to investigate the molecular mechanisms by which quercetin inhibits lipopolysaccharide (LPS)-induced pro-inflammatory cytokine production and to evaluate the capacity of quercetin to attenuate the mortality rate in a mice model of lethal sepsis. Our results show that quercetin significantly attenuates LPS-induced production of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 macrophages. The LPS-stimulated phosphorylations of the inhibitors of κB kinase (IKKs), Akt, and c-Jun N-terminal kinase (JNK) are also inhibited by quercetin. Quercetin causes a significant reduction in the phosphorylation and degradation of inhibitor of κBα (IκBα) and in the nuclear level of nuclear factor-κB (NF-κB), the latter being associated with decreased NF-κB binding activity. Most importantly, acute administration of quercetin reduces the lethality rate and circulating levels of TNF-α and IL-1β in C57BL/6J mice with endotoxemia induced by LPS, whereas chronic dietary supplementation with quercetin shows no inhibitory effect on serum TNF-α and IL-1β levels. These findings provide clues that quercetin may be a promising agent for the prevention of systemic inflammatory diseases such as sepsis.


The experience of living with knee osteoarthritis: exploring illness and treatment beliefs through thematic analysis.
Knee osteoarthritis (OA) is a major cause of disability in older adults. However, there is limited research on the daily experience of living with knee OA. We aimed to offer insight into the beliefs of patients with knee OA about their illness and treatment. Twenty-four semi-structured interviews were conducted with 17 women and 7 men with physician-diagnosed knee OA, aged between 48 and 84 years (mean = 62, SD = 7). The audio-taped interviews lasted from 30 min to 1 h, and were transcribed verbatim. The data were subjected to thematic analysis. The transcripts were independently coded by two researchers to increase reliability of coding. Six themes were developed and two of these are examined in further detail: (i) Illness representation and (ii) Beliefs about the medical and surgical control of pain. Illness representation comprised beliefs about people's understanding of OA and their pain experience, as well as expectations about the course of illness. The second theme presented experiences of limited pain relief and concerns about the use of drugs and surgery. Exploring illness representations and beliefs about medical and surgical control of pain may provide the basis for initiating psychological interventions for people with knee OA. People with knee OA place pain at the core of their living, and hold beliefs about knee OA being an incurable disease of a progressive nature, linked to specific causal factors. People with OA have concerns and worries about use of medication to control pain, and are ambivalent towards relying on medical or surgical interventions. Exploring and addressing patient illness beliefs and treatment expectations may help improve concordance with and outcomes from intervention.


Anxiety and female sexual functioning: an empirical study.
Previous research regarding anxiety and female sexual functioning has yielded conflicting conclusions. This study examined the effect of state/trait anxiety and anxiety sensitivity on sexual responding and the propensity toward sexual inhibition/excitation in women without an anxiety disorder (n = 100, M age = 28.8 years) compared with women with an anxiety disorder (panic disorder or generalized anxiety disorder, n = 30, M age = 30.2 years). Participants completed self-report measures of state and trait anxiety, anxiety sensitivity, sexual functioning, and sexual inhibition/excitation. Women with an anxiety disorder reported worse sexual functioning compared with those without an anxiety disorder (except for desire, lubrication, and pain) and a greater propensity toward sexual inhibition, because of the threat of performance failure and its consequences. Dispositional anxiety and related worries significantly predicted various types of sexual dysfunctions. Findings suggested the importance of considering the relation between anxiety and sexual functioning to design optimal prevention and therapeutic interventions for women with anxiety disorders.


Phylogenetic analysis and molecular characteristics of seven variant Chinese field isolates of PRRSV.
Porcine reproductive and respiratory syndrome (PRRS) has now been widely recognized as an economically important disease. The objective of this study was to compare the molecular and biological characteristics of porcine reproductive and respiratory syndrome virus (PRRSV) field isolates in China to those of the modified live virus (MLV) PRRS vaccine and its parent strain (ATCC VR2332). Five genes (GP2, GP3, GP4, GP5 and NSP2) of seven isolates of PRRSV from China, designated LS-4, HM-1, HQ-5, HQ-6, GC-2, GCH-3 and ST-7/2008, were sequenced and analyzed. Phylogenetic analyses based on the nucleotide sequence of the ORF2-5 and NSP2 showed that the seven Chinese isolates belonged to the same genetic subgroup and were related to the North American PRRSV genotype. Comparative analysis with the relevant sequences of another Chinese isolate (BJ-4) and North American (VR2332 and MLV) viruses revealed that these isolates have 80.8-92.9% homology with VR-2332, and 81.3-98.8% identity with MLV and 80.7-92.9% with BJ-4. All Nsp2 nonstructural protein of these seven isolates exhibited variations (a 29 amino acids deletion) in comparison with other North American PRRSV isolates. Therefore, these isolates were novel strain with unique amino acid composition. However, they all share more than 97% identity with other highly pathogenic Chinese PRRSV strains. Additionally, there are extensive amino acid (aa) mutations in the GP5 protein and the Nsp2 protein when compared with the previous isolates. These results might be useful to study the genetic diversity of PRRSV in China and to track the infection sources as well as for vaccines development.


Embedded Psychiatric Services in a Multidisciplinary Amyotrophic Lateral Sclerosis Clinic: An Assessment of Patient Needs and Perceptions.
Amyotrophic lateral sclerosis (ALS) is often associated with a range of difficult neuropsychiatric symptoms and conditions, including depression, apathy, pseudobulbar affect, and frontotemporal dementia (FTD). Despite the potential role for psychiatrists in the treatment of ALS, they are not typically involved in the ALS clinical team. The investigators describe a quality improvement intervention providing embedded psychiatric services within a multidisciplinary clinic (MDC). A psychiatrist working within an ALS MDC evaluated patients (N=116) over a 1-year period. The clinic assessed the prevalence of neuropsychiatric symptoms and conditions in patients with ALS (depression, anxiety, pseudobulbar affect, and cognitive impairment, including FTD) using standardized screening methods. Fifty-five patients and 47 family members completed surveys about perceptions of their need for psychiatric care, their experience of meeting with a psychiatrist, and their desire for future access to psychiatric care. Prevalence rates for neuropsychiatric symptoms were 14.9% for depression, 11.3% for anxiety, 19% for cognitive impairment (including FTD, 8.6%), and 36.2% for pseudobulbar affect; 62.0% of patients were being prescribed at least one psychotropic medication. Both patients and family members reported that meeting with a psychiatrist was helpful, that the treatment provided was helpful, and that they would prefer continued availability of psychiatric services in the future. The presence of cognitive impairment and use of antidepressants increased the likelihood of patients reporting a benefit from psychiatric care. Patients with ALS report a benefit from increased access to psychiatric services. The inclusion of a psychiatrist within the ALS MDC model should be considered to improve quality of care for this patient population.


Correlation of High Body Mass Index With More Advanced Localized Prostate Cancer at Radical Prostatectomy Is Not Reflected in PSA Level and PSA Density but Is Seen in PSA Mass.
Prostate cancer screening algorithms and preoperative nomograms do not include patients' body mass index (BMI). We evaluated outcomes at radical prostatectomy (RP) adjusted to BMI. Serum prostate-specific antigen (PSA) levels, PSA mass, PSA density (PSAD), and RP findings were analyzed with respect to BMI in 4,926 men who underwent RP between 2005 and 2014. In total, 1,001 (20.3%) men were normal weight, 2,547 (51.7%) were overweight, and 1,378 (28%) were obese. Median PSA levels (ng/mL) were normal weight, 5.0; overweight, 5.1; and obese, 5.2 (P = .094). Median PSA mass increased with increasing BMI: 15.9 vs 17.4 vs 19.4 μg (P < .001). Median PSAD was not significantly different: 0.11 vs 0.11 vs 0.11 ng/mL/g (P = .084). Median prostate weight increased with increasing BMI: 44 vs 45 vs 49 g (P < .001). Median prostatectomy tumor volume increased with increasing BMI: 3.9 vs 4.7 vs 5.9 cm(3) (P < .001). Overweight and obese patients had a higher Gleason score and more locally advanced cancer (P < .001). Frequency of positive surgical margins increased with higher BMIs (P < .001). Frequency of lymph node metastasis did not differ significantly (P = .088). While BMI correlates with tumor volume, Gleason score, and extent of disease at RP, there is no routinely measured clinical parameter reflecting this. Only PSA mass highlights this correlation. Thus, BMI and potentially PSA mass should be taken into account in predictive algorithms pertaining to prostate cancer and its surgical treatment.


Effect of surface modification on physical and cellular properties of PCL thin film.
Lack of suitable surface functional groups is one of the main limitations related to the cell attachment of Polycaprolactone (PCL). The aim of this research was to surface modify the PCL film using gelatin coating, via a simple physical entrapment process. In this regard, after preparation of PCL films using casting, they were immersed in each gelatin solutions. Consequently, chemical crosslinking using glutaraldehyde was performed to improve the stability of the PCL-gelatin film. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), Scanning electron microscope (SEM), contact angle measurement, strip tensile test, Dimethylthiazol-diphenyltetrazolium bromide (MTT) assay and Cell seeding were used to evaluate the quality of the coating layer, the thickness of PCL-gelatin film, the surface wettability, their mechanical properties, Cell viability and Cell attachment and proliferation respectively. Results showed that the amount of entrapped gelatin enhanced with increasing acetone in the gelatin solution. Surface modification led to a two-fold increment of mechanical strength, about 50% increase in elastic modulus, 54% in elongation and up to 11% increment in cell viability. Moreover, wettability and cell attachment of PCL film significantly enhanced, after gelatin modification. In conclusion, the simple and cost effective modification of PCL using gelatin entrapment could provide significant mechanical and biological properties making it a promising approach for development of three-dimensional scaffolds for bone tissue engineering.


Association Between Neck Circumference and Coronary Heart Disease: A Meta-analysis.
Aims: Neck circumference (NC) was found to be related to the risk factors for coronary heart disease (CHD). However, the effects of NC on CHD are still controversial. To evaluate the relationship between NC and CHD, a meta-analysis of observational studies was performed. Method: Eligible studies on the association between NC and CHD were searched in Medline, Embase, Ovid, and Web of Science databases published in English from January 1980 to December 2016. Moreover, studies published in Chinese in Wanfang and China Hospital Knowledge databases were also searched. Random effects models in the metafor package in statistical analysis software R 3.3.3 were used for the meta-analysis. Heterogeneity was analyzed with Q statistics. Results: Eight studies were selected for the meta-analysis. A larger NC was associated with a higher prevalence of CHD (OR = 1.18, 95% CI 1.04-1.34, p = 0.0108). The eight studies were further divided into three subgroups according to the criteria for diagnosing CHD. In the subgroup of coronary angiography, NC was also found to be associated with the prevalence of CHD with low heterogeneity (OR = 1.17, 95% CI 1.07-1.28, p = 0.0007, I 2 = 17.02%). However, in the subgroup of computed tomography or past history, no association between NC and CHD was found. In addition, subgroup analyses were also conducted according to the regions of the study. No association between NC and CHD was identified in either Chinese studies or Brazil studies (OR = 1.20, 95% CI 0.96-1.49; OR = 1.31, 95% CI 0.82-2.09, respectively). Conclusion: Larger NC is associated with increased risk of CHD, especially when coronary angiography was taken to diagnose CHD.


Reducing the reliance on nitrogen fertilizer for wheat production.
All crops require nitrogen (N) for the production of a photosynthetically active canopy, whose functionality will strongly influence yield. Cereal crops also require N for storage proteins in the grain, an important quality attribute. Optimal efficiency is achieved by the controlled remobilization of canopy-N to the developing grain during crop maturation. Whilst N will always be required for crop production, targeting efficient capture and use will optimise consumption of this valuable macronutrient. Efficient management of N through agronomic practice and use of appropriate germplasm are essential for sustainability of agricultural production. Both the economic demands of agriculture and the need to avoid negative environmental impacts of N-pollutants, such as nitrate in water courses or release of N-containing greenhouse gases, are important drivers to seek the most efficient use of this critical agronomic input. New cultivars optimised for traits relating to N-use efficiency rather than yield alone will be required. Targets for genetic improvement involve maximising capture, partitioning and remobilization in the canopy and to the grain, and yield per se. Whilst there is existing genetic diversity amongst modern cultivars, substantial improvements may require exploitation of a wider germplasm pool, utilizing land races and ancestral germplasm.


The morphology and genetic characterization of Iheringascaris goai n. sp. (Nematoda: Raphidascarididae) from the intestine of the silver whiting and spotted catfish off the central west coast of India.
In this study a new species of nematode, Iheringascaris goai n. sp., is reported from two fish hosts, including silver whiting, Sillago sihama, and spotted catfish, Arius maculatus, caught off the Central West Coast of India at Goa. The new species can be differentiated morphologically from I. inquies, the most closely related species collected from cohabiting marine fish. The distinguishing characteristics are distinct cuticular striations, a unilateral excretory system, the presence of dentigerous ridges on the inner margin of the lips and the ratio of oesophagus to body length. In males, the ratio of spicules to body length is higher and the number of pre-anal papillae is less in comparison to those in I. inquies. In addition, the tail curves ventrad in males, while in females, the vulva is post-equatorial. The sequence alignment of 18S rDNA and cytochrome c oxidase subunit I with sequences of known species selected from the same superfamily shows a significant difference. The morphological and molecular differences reported here can, therefore, be used to assign the specimen to a new species.


Interferon-Gamma (IFNg) +874A/T Polymorphism Does Not Significantly Affect the Severity of Periodontitis.
Interferon-gamma (IFNg) is an immune-regulatory cytokine with a role in host responses to periodontitis. Genetic factors have been reported to modify the corresponding protein expression. The objective of this study was to evaluate the association and role of IFNg polymorphisms, such as IFNg +874 A/T, and the susceptibility to periodontitis. A total of 100 unrelated subjects were included in the present study. Genomic deoxyribonucleic acid (DNA) was obtained from peripheral blood of 43 patients with mild periodontitis and 57 patients with severe periodontitis. The determined clinical parameters of periodontitis included probing depth, clinical attachment loss, and papilla bleeding index. The oral hygiene indicators were also assessed. The level of IFNg was determined from the gingival crevicular fluid by enzyme-linked immunosorbent assay technique. The IFNg +874 A/T polymorphisms were analyzed from peripheral blood by the method of restriction fragment length polymorphism-polymerase chain reaction. Statistical analysis of the results was conducted using chi-squared testing for categorical data. Independent t-tests and Mann-Whitney U tests were used for numeric data. Kruskal-Wallis testing was used to compare genotypes concerning for IFNg +874 A/T polymorphism. A p-value < 0.05 was assumed for statistical significance. Analysis of the IFNg +874 A/T polymorphism showed no significant differences with the level of IFNg. No significant differences were observed either in IFNg +874 A/T polymorphism between the subjects with mild periodontitis and those with severe periodontitis (p > 0.05). The subjects with severe periodontitis showed marginally but not significantly higher levels of IFNg compared with subjects with mild periodontitis (p > 0.05). The polymorphism of IFNg +874 A/T was not associated with the level of IFNg nor with the risk of periodontitis in this study.


Clinical outcomes of dialysis patients after implantation of DES: meta analysis and systematic review of literature.
Studies on percutaneous transluminal cardiac angioplasty (PTCA) in patients with end stage renal disease (ESRD) on hemodialysis have suggested high rates of procedural complications and restenosis. Bare metal stent percutaneous coronary intervention (PCI) has significantly reduced restenosis and subsequent target lesion revascularization in these patients, although not to the level of non dialysis controls. The introduction of drug-eluting stents (DES) has dramatically reduced restenosis rates compared with bare metal stents (BMS) in patients with various clinical and angiographic characteristics, however their impact on patients with ESRD on dialysis is unclear due to consistent exclusion of this population from major trials. The purpose of this study was therefore to compare the outcomes of PCI with DES and BMS when used for ESRD patients on dialysis, by meta analytical techniques. Comparative studies published between January 2002 and January 2009 of DES vs. BMS in ESRD patients on dialysis were identified using an electronic search and reviewed using a random effects model. The primary endpoints of our study were the hard endpoints of mortality, myocardial infarction (MI) and target lesion revascularization (TLR). A secondary endpoint of our analysis was late luminal loss. In hospital mortality and MI were also assessed. Heterogeneity was assessed using Cochrane Q and I(2) statistics. Five reports comprising 641 patients (279 DES, and 362 BMS) were included in the analysis. All the studies were non-randomized comparisons between DES and BMS. The length of follow-up was in the range between 9 and 12 months. In hospital clinical outcomes were similar between the two groups. At follow-up there was a trend towards lower TLR (OR 0.50, CI 0.27-0.93, P=0.011 I(2)=48%) and decreased late luminal loss (WMD -0.34, CI -0.58 -0.10 P=0.09, I(2)=58%) in patients undergoing PCI with implantation of DES. There was no difference in the rates of all cause mortality (OR 0.66, CI 0.40-1.08, P=0.070 I(2)= 0%), and MI (OR 1.35, CI 0.52-3.52, P=0.53, I(2)%) between the two groups. In ESRD patients on dialysis undergoing PCI, DES are safe and reduce repeat revascularizations. The limited number of patients and limited quality of primary studies included, however need careful interpretation of our results. Further well designed, large randomized controlled trials are required to establish the strategy of management in ESRD patients undergoing PCI.


Using Laguerre expansion within point-process models of heartbeat dynamics: a comparative study.
Point-process models have been recognized as a distinguished tool for the instantaneous assessment of heartbeat dynamics. Although not thoroughly linked to the physiology, nonlinear models also yield a more accurate quantification of cardiovascular control dynamics. Here, we propose a Laguerre expansion of the linear and nonlinear Wiener-Volterra kernels in order to account for the nonlinear and non-gaussian information contained in the ECG-derived heartbeat series while using a reduced number of parameters. Within an Inverse-Gaussian probability model, up to quadratic nonlinearities were considered to continuously estimate the dynamic spectrum and bispectrum. Results performed on 10 subjects undergoing a stand-up protocol show that this novel methodology improves on the algorithmic performances and, at the same time, more accurately characterizes sympatho-vagal changes to posture.


A novel electroluminescent device based on a reduced graphene oxide wrapped phosphor (ZnS:Cu,Al) and hexagonal-boron nitride for high-performance luminescence.
Reduced graphene oxide (rGO) has recently emerged as a very promising family of exotic carbon material with augmented performance in electronic and optoelectronic devices. Herein, we report an efficient and novel inorganic electroluminescent device geometry, where a new phosphor composite, reduced graphene oxide wrapped ZnS:Cu,Al, acts as an active emitting layer and an exfoliated hexagonal boron nitride (h-BN) as a dielectric layer. The roles of rGO in the active layer as a conductive support and local electric field enhancing agent are attributed to its wrinkles being unraveled compared with other carbon exotic nano-forms such as carbon nanotubes, graphite, charcoal and activated carbon, which significantly improves the brightness of the device (∼50 cd m-2 for 0.50 wt% rGO/ZnS:Cu,Al at 10 kHz and 110 V with an external quantum efficiency of ∼6.3% ± 0.1% and current efficiency of ∼0.81 ± 0.09 cd A-1). This new and facile strategy to construct the luminescent devices could be a paradigm shift towards cost effective, highly stable in air (for several days) and energy efficient next generation display devices.


Computer-assisted surgery is not more accurate or precise than conventional arthroscopic ACL reconstruction: a prospective randomized clinical trial.
Accurate and precise tunnel placement is critical to the success of anterior cruciate ligament (ACL) reconstruction. A new development, computer-assisted surgery, aids in placement of the ACL bone tunnels during surgery. Our hypothesis was that computer-assisted ACL reconstruction would allow more accurate and precise tunnel placement compared with conventional surgery. In a prospective, double-blind, randomized clinical study, 100 patients eligible for ACL reconstruction with a transtibial technique were stratified by surgeon and randomized to either conventional or computer-assisted surgery. Measurement of femoral and tibial tunnel placement with use of three-dimensional computed tomography (CT) was used as the primary outcome to compare conventional ACL surgery with computer-assisted surgery. The placement of the femoral tunnel did not differ between groups (mean, 39.7% of the proximal-distal distance on the intracondylar axis [Blumensaat line] in the conventional group compared with 39.0% in the computer-assisted surgery group; p = 0.70). The anterior-posterior positioning of the tibial tunnel on the tibial plateau also did not differ significantly (38.9% in the conventional group compared with 38.2% in the computer-assisted surgery group; p = 0.58). There was no significant difference in the precision of either the femoral or the tibial tunnel placement between the two groups. There was no significant difference in either the accuracy or the precision of tunnel placement between conventional and computer-assisted ACL reconstruction.


The human L-type calcium channel Cav1.3 regulates insulin release and polymorphisms in CACNA1D associate with type 2 diabetes.
Voltage-gated calcium channels of the L-type have been shown to be essential for rodent pancreatic beta cell function, but data about their presence and regulation in humans are incomplete. We therefore sought to elucidate which L-type channel isoform is functionally important and its association with inherited diabetes-related phenotypes. Beta cells of human islets from cadaver donors were enriched using FACS to study the expression of the genes encoding voltage-gated calcium channel (Cav)1.2 and Cav1.3 by absolute quantitative PCR in whole human and rat islets, as well as in clonal cells. Single-cell exocytosis was monitored as increases in cell capacitance after treatment with small interfering (si)RNA against CACNA1D (which encodes Cav1.3). Three single nucleotide polymorphisms (SNPs) were genotyped in 8,987 non-diabetic and 2,830 type 2 diabetic individuals from Finland and Sweden and analysed for associations with type 2 diabetes and insulin phenotypes. In FACS-enriched human beta cells, CACNA1D mRNA expression exceeded that of CACNA1C (which encodes Cav1.2) by approximately 60-fold and was decreased in islets from type 2 diabetes patients. The latter coincided with diminished secretion of insulin in vitro. CACNA1D siRNA reduced glucose-stimulated insulin release in INS-1 832/13 cells and exocytosis in human beta cells. Phenotype/genotype associations of three SNPs in the CACNA1D gene revealed an association between the C allele of the SNP rs312480 and reduced mRNA expression, as well as decreased insulin secretion in vivo, whereas both rs312486/G and rs9841978/G were associated with type 2 diabetes. We conclude that the L-type calcium channel Cav1.3 is important in human glucose-induced insulin secretion, and common variants in CACNA1D might contribute to type 2 diabetes.


Infrared reflectance-absorbance spectroscopy of thin films formed by forced dewetting of solid-fluid interfaces.
An infrared reflectance-absorbance spectroscopy method for characterizing the ultrathin fluid film retained on a surface upon forced dewetting from a fluid has been developed for investigation of interfacial molecular structure at reflective substrates. This report details the optical considerations and constraints necessary to acquire IR spectral data from nanometer-thick films retained upon forced dewetting of a solid substrate from a fluid into a vapor-saturated environment. The feasibility of this method is demonstrated through successful spectral acquisition from Ag surfaces modified with 11-mercaptoundecanol forcibly dewet from water. The IR spectral results clearly illustrate that information is acquired only from the interfacial region with no contribution from the bulk liquid. Residual layer thicknesses calculated from IR absorbance values are substantiated by ellipsometry. The spectra make clear that the molecular structure of the residual layer is distinctly different from that of the bulk liquid, confirming that this method is viable for interfacial structure elucidation of thin fluid films at a variety of solid substrates.


Assembly of a reference transcriptome for the gymnosome pteropod Clione limacina and profiling responses to short-term CO2 exposure.
The gymnosome (unshelled) pteropod Clione limacina is a pelagic predatory mollusc found in polar and sub-polar regions. It has been studied for its distinctive swimming behavior and as an obligate predator on the closely related thecosome (shelled) pteropods. As concern about ocean acidification increases, it becomes useful to compare the physiological responses of closely-related calcifying and non-calcifying species to acidification. The goals of this study were thus to generate a reference transcriptome for Clione limacina, to expose individuals to CO2 for a period of 3days, and to explore differential patterns of gene expression. Our Trinity assembly contained 300,994 transcripts of which ~26% could be annotated. In total, only 41 transcripts were differentially expressed following the CO2 treatment, consistent with a limited physiological response of this species to short-term CO2 exposure. The differentially expressed genes identified in our study were largely distinct from those identified in previous studies of thecosome pteropods, although some similar transcripts were identified, suggesting that comparison of these transcriptomes and responses may provide insight into differences in responses to ocean acidification among phylogenetically and functionally distinct molluscan lineages.


Lycium Barbarum Polysaccharides and Wolfberry Juice Prevent DEHP-Induced Hepatotoxicity via PXR-Regulated Detoxification Pathway.
Environmental di(2-Ethylhexyl) phthalate (DEHP) is widely used in various industries as a plasticizer, and has been reported to induce reproductive and developmental toxicities in organisms. The purpose of this study was to evaluate the detoxification capacity of Lycium barbarum polysaccharides (LBP) and wolfberry juice (WJ) against DEHP-induced hepatotoxicity. Two groups of rats were purchased to study two different intervention method experiments: LBP (50, 100, 200 mg/kg·bw) intervention before DEHP (2000 mg/kg·bw) exposure, and LBP (200 mg/kg·bw) or WJ (8 mL/kg·bw) intervention after DEHP (3000 mg/kg·bw) exposure. The rats were exposed to DEHP once, while the intervention lasted for seven days. At the end of the intervention, enzyme-linked immunosorbent assay (ELISA) was used to measure the related index. The LBP intervention before DEHP exposure experiment (the first experimental method) found that LBP group rats showed a strong capacity toward DEHP detoxification, evidenced by the significant upregulation of activities and concentrations of the partner retinoid, X receptor alpha (RXRα), and downstream regulators Cytochrome P4502E1 (CYP2E1), Cytochrome P4503A1 (CYP3A1), Glutathione S-Transferase Pi (GSTpi), and UDP-glucuronosyltransferase 1 (UGT1) in a dose-dependent manner. The LBP and WJ intervention after DEHP exposure experiment (the second intervention experiment) found that WJ could downregulate pregnane X receptor (PXR), and upregulate downstream regulators, CYP2E1, CYP3A1, and Glutathione S-Transferase (GST) with the extension of intervention time, to alleviate the toxicity of DEHP. However, the intervention effect of WJ was more obvious than that of LBP. These results suggested that LBP and WJ might be effective detoxification agents against DEHP-induced toxic effects, by activating PXR and PXR-related detoxifying enzymes.


A Combined Metabolomics and Fluxomics Analysis Identifies Steps Limiting Oil Synthesis in Maize Embryos.
Enhancing fatty acid synthesis (FAS) in maize (Zea mays) has tremendous potential nutritional and economic benefits due to the rapidly growing demand for vegetable oil. In maize kernels, the endosperm and the embryo are the main sites for synthesis and accumulation of starch and oil, respectively. So far, breeding efforts to achieve elevated oil content in maize have resulted in smaller endosperms and therefore lower yield. Directly changing their carbon metabolism may be the key to increasing oil content in maize kernels without affecting yield. To test this hypothesis, the intracellular metabolite levels were compared in maize embryos from two different maize lines, ALEXHO S K SYNTHETIC (Alex) and LH59, which accumulate 48% and 34% of oil, respectively. Comparative metabolomics highlighted the metabolites and pathways that were active in the embryos and important for oil production. The contribution of each pathway to FAS in terms of carbon, reductant, and energy provision was assessed by measuring the carbon flow through the metabolic network (13C-metabolic flux analysis) in developing Alex embryos to build a map of carbon flow through the central metabolism. This approach combined mathematical modeling with biochemical quantification to identify metabolic bottlenecks in FAS in maize embryos. This study describes a combination of innovative tools that will pave the way for controlling seed composition in important food crops.


Awakening consumer stewardship of health benefits: prevalence and differentiation of new health plan models.
Despite widespread publicity of consumer-directed health plans, little is known about their prevalence and the extent to which their designs adequately reflect and support consumerism. We examined three types of consumer-directed health plans: health reimbursement accounts (HRAs), premium-tiered, and point-of-care tiered benefit plans. We sought to measure the extent to which these plans had diffused, as well as to provide a critical look at the ways in which these plans support consumerism. Consumerism in this context refers to efforts to enable informed consumer choice and consumers' involvement in managing their health. We also wished to determine whether mainstream health plans-health maintenance organization (HMO), point of service (POS), and preferred provider organization (PPO) models-were being influenced by consumerism. Our study uses national survey data collected by Mercer Human Resource Consulting from 680 national and regional commercial health benefit plans on HMO, PPO, POS, and consumer-directed products. We defined consumer-directed products as health benefit plans that provided (1) consumer incentives to select more economical health care options, including self-care and no care, and (2) information and support to inform such selections. We asked health plans that offered consumer-directed products about 2003 enrollment, basic design features, and the availability of decision support. We also asked mainstream health plans about their activities that supported consumerism (e.g., proactive outreach to inform or influence enrollee behavior, such as self-management or preventive care, reminders sent to patients with identified medical conditions.) We analyzed survey responses for all four product lines in order to identify those plans that offer health reimbursement accounts (HRAs), premium-tiered, or point-of-care tiered models as well as efforts of mainstream health plans to engage informed consumer decision making. The majority of enrollees in consumer-directed health plans are in tiered models (primarily point-of-care tiered networks) rather than HRAs. Tiers are predominantly determined based on both cost and quality criteria. Enrollment in HRAs has grown substantially, in part because of the entry of mainstream managed care plans into the consumer-directed market. Health reimbursement accounts, tiered networks, and traditional managed care plans vary in their capacity to support consumers in managing their health risks and selection of provider and treatment options, with HRAs providing the most and mainstream plans the least. While enrollment in consumer-directed health plans continues to grow steadily, it remains a tiny fraction of all employer-sponsored coverage. Decision support in these plans, a critical link to help consumers make more informed choices, is also still limited. This lack may be of concern in light of the fact that only a minority of such plans report that they monitor claims to protect against underuse. Tiered benefit models appear to be more readily accepted by the market than HRAs. If they are to succeed in optimizing consumers' utility from health benefit spending, careful attention needs to be paid to how well these models inform consumers about the consequences of their selections.


Time Investment for Program Implementation to Manage Neuropsychiatric Symptoms: An Observational Longitudinal Study in In-Home and Residential Care Settings.
There are no studies on how the same psychosocial dementia care program is adapted to both in-home and residential care settings. To evaluate the time investment required by professionals to implement a psychosocial dementia care program to manage neuropsychiatric symptoms. A prospective observational study design was used. The program consisted of 1) a one-day training course, 2) three interdisciplinary discussion meetings in five months, and 3) a web-based tool for the continued assessment of neuropsychiatric symptoms. Care professionals implemented the intervention in in-home (19 in-home care management agencies and 14 multiple in-home service providers) and residential care settings (19 group homes and eight nursing homes) in Japan from October 2019 to February 2020. The level of neuropsychiatric symptoms for the participants was evaluated using the Neuropsychiatric Inventory (NPI: 0-144). The time investment was reported by participating professionals. A total of 125 persons with dementia were included at baseline. Neuropsychiatric symptoms were significantly decreased at the final follow-up in all types of providers (Cohen's drm = 0.44-0.61). The mean (SD) time required for the five-month implementation was 417.9 (219.8) minutes. There was a mean (SD) decrease of 8.6 (14.0) points in the total NPI score among the 103 persons with completed interventions. The time investment was significantly lower in in-home care management agencies than in group homes, and lower in follow-ups than at baseline assessment. The program implementation may incur a substantial time investment regardless of setting. An additional benefit scheme to reward the time investment would be helpful to encourage implementation until the follow-ups.


[Development of delayed-type hypersensitivity to hog cholera].
Leukocyte migration inhibition test showed that pigs with classical swine fever develop delayed type hypersensitivity. The highest migration inhibition (65-85%) was observed in animals with the acute form, on day 3 after infection, or directly before death, or in animals immunized with reactogenic strain with clinical signs of disease. In some pigs with acute form the migrating capacity of leukocytes was restored on days 7-8 postinfection. Leukocyte migration inhibition factor is detected in the sera of pigs starting from day 3 after infection with the virulent strain. The degree of delayed type hypersensitivity correlated with the outcome of classical swine fever.


A thyroxine binding globulin (TBG)-like protein in the sera of developing and adult rats.
We report evidence based on equilibrium binding, electrophoretic, autoradiographic studies, that the rat possesses a major high affinity thyroid hormone binding protein, with an electrophoretic mobility and binding properties similar to those of the human thyroxine binding globulin (TBG). We show that in the sera of postnatal developing animals, the thyroxine and the triiodothyronine binding activities increase up to 10 times over adult or foetal levels, due to a high transient post-natal surge of the rat TBG. In the adult serum, the TBG persists in decreased amounts: it then yields the predominant role as thyroxine carrier to the thyroid binding prealbumin, but retains the major role as binder of triiodothyronine i.e. of the biologically active thyroid hormone.


Thrombopoietic cytokines in patients with iron deficiency anemia with or without thrombocytosis.
Iron deficiency anemia is a cause of reactive thrombocytosis. A moderate increase in platelet numbers is common but sometimes counts may exceed 1,000 x 10(9)/l. The mechanisms causing reactive thrombocytosis are unclear. In this study, we evaluated 15 women with iron deficiency anemia and thrombocytosis (platelets >450 x 10(9)/l) and 16 women with iron deficiency anemia with normal platelet counts. Serum samples were taken before oral iron replacement therapy, after 1 and 3 months and at the end of replacement therapy. Thrombopoietin, erythropoietin (EPO), leukemia inhibitory factor, interleukin-6 and interleukin-11 levels were assayed. There was no change in the levels of thrombopoietic cytokines except for EPO. The correlation between high EPO levels and high platelet counts may suggest that EPO increases platelet counts, but the same EPO level changes can also be demonstrated in women with iron deficiency anemia but normal initial platelet counts. The fact that the levels of other cytokines remained unchanged during treatment suggests that either these cytokines have no effect on reactive thrombocytosis or the change in platelet counts in our patients is in a narrow range and is thus not affected by the cytokine levels.


DYRK1A regulates B cell acute lymphoblastic leukemia through phosphorylation of FOXO1 and STAT3.
DYRK1A is a serine/threonine kinase encoded on human chromosome 21 (HSA21) that has been implicated in several pathologies of Down syndrome (DS), including cognitive deficits and Alzheimer's disease. Although children with DS are predisposed to developing leukemia, especially B cell acute lymphoblastic leukemia (B-ALL), the HSA21 genes that contribute to malignancies remain largely undefined. Here, we report that DYRK1A is overexpressed and required for B-ALL. Genetic and pharmacologic inhibition of DYRK1A decreased leukemic cell expansion and suppressed B-ALL development in vitro and in vivo. Furthermore, we found that FOXO1 and STAT3, transcription factors that are indispensable for B cell development, are critical substrates of DYRK1A. Loss of DYRK1A-mediated FOXO1 and STAT3 signaling disrupted DNA damage and ROS regulation, respectively, leading to preferential cell death in leukemic B cells. Thus, we reveal a DYRK1A/FOXO1/STAT3 axis that facilitates the development and maintenance of B-ALL.


The Sexual Harassment-Suicide Connection in the U.S. Military: Contextual Effects of Hostile Work Environment and Trusted Unit Leaders.
Sexual harassment has been associated with suicidal behaviors, and with the rise in suicides in the U.S. military, sexual harassment's role in suicide has been of growing interest. Lacking are studies that examine group- or unit-level variables in the relationship of sexual harassment to suicidal behaviors (thoughts, plans, and attempts). In this study, survey data from soldiers (12,567 soldiers in 180 company-sized units) who completed the Unit Risk Inventory administered during calendar year 2010 were analyzed using hierarchical linear modeling. At the individual level, sexual harassment was associated with a fivefold increase for risk of suicide. Reporting that leaders could be trusted was associated with a decreased suicide risk by about one-third. There was no statistically significant interaction between sexual harassment and trusted leaders in predicting the suicidal behaviors. At the group level, units or companies having higher levels of sexual harassment also had soldiers three times more at risk for suicide. A cross-leveling effect was also observed: Among units having higher levels of sexual harassment, the negative correlation (buffering effect of unit leaders on suicidal behaviors) was diminished. Implications of findings for preventing sexual harassment and suicide risk are discussed.


Cerebral chromomycosis.
A 52-year-old man was admitted to the hospital with headache, confusion, and early papilledema. He was receiving prednisone and azathioprine for interstitial pulmonary fibrosis. A right temporoparietal cerebral abscess was diagnosed by computed tomographic scan, brain scan, and cerebral angiography. Fungal cultures of pus obtained at operation grew a pigmented fungus, Cladosporium trichoides. Three additional abscesses were found in the right cerebellar hemisphere at autopsy after the patient died of an unrelated cause. Chromomycosis occurring at sites other than skin is extremely rare. No lesions were detected outside the CNS in our patient. C trichoides has not been previously recognized as an agent causing disease in compromised hosts.


Mathematical modeling and simulation of thyroid homeostasis: Implications for the Allan-Herndon-Dudley syndrome.
A mathematical model of the pituitary-thyroid feedback loop is extended to deepen the understanding of the Allan-Herndon-Dudley syndrome (AHDS). The AHDS is characterized by unusual thyroid hormone concentrations and a mutation in the SLC16A2 gene encoding for the monocarboxylate transporter 8 (MCT8). This mutation leads to a loss of thyroid hormone transport activity. One hypothesis to explain the unusual hormone concentrations of AHDS patients is that due to the loss of thyroid hormone transport activity, thyroxine (T 4) is partially retained in thyroid cells. This hypothesis is investigated by extending a mathematical model of the pituitary-thyroid feedback loop to include a model of the net effects of membrane transporters such that the thyroid hormone transport activity can be considered. A nonlinear modeling approach based on the Michaelis-Menten kinetics and its linear approximation are employed to consider the membrane transporters. The unknown parameters are estimated through a constrained parameter optimization. In dynamic simulations, damaged membrane transporters result in a retention of T 4 in thyroid cells and ultimately in the unusual hormone concentrations of AHDS patients. The Michaelis-Menten modeling approach and its linear approximation lead to similar results. The results support the hypothesis that a partial retention of T 4 in thyroid cells represents one mechanism responsible for the unusual hormone concentrations of AHDS patients. Moreover, our results suggest that the retention of T 4 in thyroid cells could be the main reason for the unusual hormone concentrations of AHDS patients.


Phosphorylation of chlamydomonas reinhardi chloroplast membrane proteins in vivo and in vitro.
Phosphorylation of thylakoid membrane proteins in the chloroplast of wild-type and mutant strains of Chlamydomonas reinhardi has been studied in vivo and in vitro. Intact cells or purified membranes were labeled with [32P]orthophosphate or [gamma-32P]ATP, respectively, and the presence of phosphorylated polypeptides was detected by autoradiography after membrane fractionation by SDS PAGE. The 32P was esterified to serine and threonine residues. At least six polypeptides were phosphorylated in vitro and in vivo, and corresponded to components of the photosystem II complex contributing to the formation of the light-harvesting-chlorophyll (LHC) a,b-protein complex, the DCMU binding site (32-35 kdaltons), and the reaction center (26 kdaltons). In agreement with previous reports (Alfonzo, et al., 1979, Plant Physiol., 65:730-734; and Bennett, 1979, FEBS (Fed. Eur. Biochem. Soc.) Lett., 103:342-344), the membrane-bound protein kinase was markedly stimulated by light in vitro via a mechanism requiring photosystem II activity. Phosphorylation of thylakoid membrane polypeptides in vivo was, however, completely independent of illumination. Similar amounts of phosphate were incorporated into the photosynthetic membranes of cells incubated in the dark, in white light with or without 3-(3,4-dichlorophenyl-1,1-dimethyl urea (DCMU), or in red or far-red light. Different turnovers of the phosphate were observed in the light and dark, and a phosphoprotein phosphatase involved in this turnover process was also associated with the membrane. Comparison of the amount of esterified phosphate per protein in vivo and the maximum incorporation in isolated membranes revealed that only a small fraction of the available sites could be phosphorylated in vitro. In contrast to the DCMU binding site, the LHC and 26-kdalton polypeptide were not phosphorylated in vivo when the reaction center II polypeptides of 44-54 kdaltons were missing. The finding that all the phosphoproteins appear to be components of the photosystem II complex and are only partially dephosphorylated in vivo suggests strongly that protein phosphorylation might play an important role in the maintenance of the organizational integrity of this complex. The observation that the LHC is not phosphorylated in the absence of the reaction center lends support to this idea.


[Concordance among Mini-Examen Cognoscitivo and Mini-Mental State Examination in cognitive impairment screening].
To compare the concordance between two cognitive impairment (CI) screening tests: the 30-point Folstein's Mini Mental State Examination (MMSE) and their validated and slightly modified spanish translation, the 35-point Lobo's Mini-Examen Cognoscitivo (MEC). Analytical cross-sectional multicenter study. Primary care. Randomized sample of subjects aged 65 years or more attended in 54 primary care centers in Catalonia. Inclusion of 3,167 subjects. Institutionalized patients were excluded. After a training period, primary care doctors and nurses administered MMSE and MEC simultaneously to their own patients. Standardization of variables. Age 74 (6.1) years-old, 1,611 (50.9%) women, 954 (30.1%) illiterate or without primary education. Prevalence of CI according to MMSE was 16.52% (n = 449) and according to MEC 5,49% (n = 165) (P<.001). Although intraclass correlation coefficient was 0,864 (95% CI, 0.855-0.873), the kappa index at score 24 for both tests was 0.468, but it increased up to 0.788 taking 20 and 23 scores for MMSE and MEC, respectively. At multivariate analysis, low educational level, and age more than 80 years-old predict a bad concordance among both tests. In spite of good correlation between MMSE and MEC, both detect different CI prevalences with the cut-off point at 23/24. Their agreement is only moderated in practice, because we interpret tests in a dichotomic way (CI versus no-CI). In our experience, we cannot use them indistinctly with the cut-off point at 23/24, especially in subjects with low educational level or aged 80 or more years.


Challenges and Opportunities of Nanotechnology as Delivery Platform for Tocotrienols in Cancer Therapy.
Plant-derived phytonutrients have emerged as health enhancers. Tocotrienols from the vitamin E family gained high attention in recent years due to their multi-targeted biological properties, including lipid-lowering, neuroprotection, anti-inflammatory, antioxidant, and anticancer effects. Despite well-defined mechanism of action as an anti-cancer agent, their clinical use is hampered by poor pharmacokinetic profile and low oral bioavailability. Delivery systems based on nanotechnology were proven to be advantageous in elevating the delivery of tocotrienols to tumor sites for enhanced efficacy. To date, preclinical development of nanocarriers for tocotrienols include niosomes, lipid nanoemulsions, nanostructured lipid carriers (NLCs) and polymeric nanoparticles. Active targeting was explored via the use of transferrin as targeting ligand in niosomes. In vitro, nanocarriers were shown to enhance the anti-proliferative efficacy and cellular uptake of tocotrienols in cancer cells. In vivo, improved bioavailability of tocotrienols were reported with NLCs while marked tumor regression was observed with transferrin-targeted niosomes. In this review, the advantages and limitations of each nanocarriers were critically analyzed. Furthermore, a number of key challenges were identified including scale-up production, biological barriers, and toxicity profiles. To overcome these challenges, three research opportunities were highlighted based on rapid advancements in the field of nanomedicine. This review aims to provide a wholesome perspective for tocotrienol nanoformulations in cancer therapy directed toward effective clinical translation.


Failure of regulation results in an amplified oxidation burst by neutrophils in children with primary nephrotic syndrome.
The mechanism responsible for proteinuria in non-genetic idiopathic nephrotic syndrome (iNS) is unknown. Animal models suggest an effect of free radicals on podocytes, and indirect evidence in humans confirm this implication. We determined the oxidative burst by blood CD15+ polymorphonucleates (PMN) utilizing the 5-(and-6)-carboxy-2',7'-dichlorofluorescin diacetate (DCF-DA) fluorescence assay in 38 children with iNS. Results were compared with PMN from normal subjects and patients with renal pathologies considered traditionally to be models of oxidative stress [six anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis, seven post-infectious glomerulonephritis]. Radicals of oxygen (ROS) production was finally determined in a patient with immunodeficiency, polyendocrinopathy, enteropathy X-linked (IPEX) and in seven iNS children after treatment with Rituximab. Results demonstrated a 10-fold increase of ROS production by resting PMN in iNS compared to normal PMN. When PMN were separated from other cells, ROS increased significantly in all conditions while a near-normal production was restored by adding autologous cells and/or supernatants in controls, vasculitis and post-infectious glomerulonephritis but not in iNS. Results indicated that the oxidative burst was regulated by soluble factors and that this regulatory circuit was altered in iNS. PMN obtained from a child with IPEX produced 100 times more ROS during exacerbation of clinical symptoms and restored to a near normal-level in remission. Rituximab decreased ROS production by 60%. In conclusion, our study shows that oxidant production is increased in iNS for an imbalance between PMN and other blood cells. Regulatory T cells (Tregs) and CD20 are probably involved in this regulation. Overall, our observations reinforce the concept that oxidants deriving from PMN are implicated in iNS.


The use of a brief depression screen in older emergency department patients.
To determine the sensitivity and specificity of a brief two-question depression screen for the detection of depression in older emergency department (ED) patients, and to determine the prevalence of depression in this population. This was a prospective, observational study. Participants included a convenience sampling of ED patients 70 years and older presenting to an urban teaching hospital over a 17-month period. Exclusions were refusal to participate, inability to communicate, and critical illness. Subjects were screened for depression with the previously validated Short Form Geriatric Depression Scale (SFGDS). Standardized scores on the SFGDS were used to determine the prevalence of depression. Patients were then given a previously published two-question depression screen, and results were compared with the SFGDS as the criterion standard. Sensitivity, specificity, and prevalence data are reported as proportions with 95% confidence intervals (95% CIs). Two hundred sixty-seven of 327 eligible patients were enrolled. Forty-four (17%; 95% CI = 12% to 21%) scored positive for depression on the SFGDS. The sensitivity of the brief two-question depression screen was 84% (37/44; 95% CI = 70% to 93%), with a specificity of 61% (136/223; 95% CI = 55% to 67%) using a cutoff score of at least one of two positive responses. Depression is fairly prevalent in older ED patients. The brief two-question depression screen, using a cutoff score of at least one positive response, is promising for ED use. However, given lower specificity, patients scoring positive for depression should be followed up with a more specific tool such as the self-administered SFGDS prior to referral for further evaluation and treatment.


Distribution and abundance of metabotropic glutamate receptor subtype 2 in rat brain revealed by [3H]LY354740 binding in vitro and quantitative radioautography: correlation with the sites of synthesis, expression, and agonist stimulation of [35S]GTPgammas binding.
Until recently, there was a lack of selective radioligands for the subtypes of metabotropic glutamate (mGlu) receptors. [(3)H]LY354740 ((+)-2-aminobicyclo[3,1,0]hexane-2,6-dicarboxylic acid), a selective agonist for group II receptors (mGlu2 and -3, which are negatively coupled to cAMP production), has now been used to map their brain distribution and abundance by in vitro binding and quantitative radioautography. The selective cation dependence of its binding allowed the discrimination between mGlu2 and mGlu3 receptor labeling. Thus, in the presence of Ca(2+) and Mg(2+) ions, the agonist bound selectively to mGlu2 receptors as evidenced by: 1) the correlative distribution and abundance of binding sites (highest in the lacunosum moleculare of the hippocampus and lowest in white matter) with mGlu2 receptor mRNA and protein revealed by in situ hybridization histochemistry and immunohistochemistry, respectively; 2) its selective pharmacology; and 3) the distribution of LY354740-stimulated [(35)S]GTPgammaS binding (25-97% above basal, according to the brain region), revealing G protein-coupled receptor coupling to G(i) proteins. Nonspecific binding (in the presence of 10 muM DCG-IV, a group II-selective, mGlu2-preferring, receptor agonist) was <10% of total. In adjacent sections, the distribution of binding sites for [(3)H]DCG-IV was very similar. This extensive study paves the way for investigations of the regional expression and regulation of mGlu2 receptors in human CNS diseases, such as Alzheimer's disease, which may reveal their functional roles and identify potential therapeutic drug targets. Indeed, it has recently been demonstrated (Higgins et al. [2004] Neuropharmacology 46:907-917) that pharmacological manipulation of mGlu2 receptors influences cognitive performance in the rodent.


BAG6/BAT3 modulates autophagy by affecting EP300/p300 intracellular localization.
We recently reported that BAG6/BAT3 (BCL2-associated athanogene 6) is essential for basal and starvation-induced autophagy in E18.5 bag6(-/-) mouse embryos and in mouse embryonic fibroblasts (MEFs) through the modulation of the EP300/p300-dependent acetylation of TRP53 and autophagy-related (ATG) proteins. We observed that BAG6 increases TRP53 acetylation during starvation and pro-autophagic TRP53-target gene expression. BAG6 also decreases the EP300 dependent-acetylation of ATG5, ATG7, and LC3-I, posttranslational modifications that inhibit autophagy. In addition, in the absence of BAG6 or when using a mutant of BAG6 exclusively located in the cytoplasm, autophagy is inhibited, ATG7 is hyperacetylated, TRP53 acetylation is abrogated, and EP300 accumulates in the cytoplasm indicating that BAG6 is involved in the regulation of the nuclear localization of EP300. We also reported that the interaction between BAG6 and EP300 occurs in the cytoplasm rather than the nucleus. Moreover, during starvation, EP300 is transported to the nucleus in a BAG6-dependent manner. We concluded that BAG6 regulates autophagy by controlling the localization of EP300 and its accessibility to nuclear (TRP53) and cytoplasmic (ATGs) substrates.


The prognosis of inflammatory bowel disease.
The complications of ulcerative colitis generally develop during the first two years of disease. The mortality is higher than expected and the highest likelihood of colectomy also occurs early in the disease. Mortality in Crohn's disease is greater than expected, especially in males. For both conditions, the overall mortality has decreased steadily, and currently is less than 5%. Ulcerative colitis is curable with proctocolectomy and ileostomy. In Crohn's disease, intestinal resection and reanastomosis is followed by recurrence in the majority of patients. The recurrence rate after proctocolectomy and ileostomy for Crohn's disease of the colon also is considerable, ranging from 20% to 35%. In ulcerative colitis, the more colon involved, the more frequent and more serious are the complications. In Crohn's disease, the anatomic pattern of disease tends to predict the type and extent of complications. Both ulcerative colitis and Crohn's disease appear to follow a more severe course in children and adolescents with "inflammatory bowel disease." Patients with either ulcerative colitis or Crohn's disease are at increased risk for the later development of cancer. In ulcerative colitis, the excess risk is limited to colorectal cancer. Patients with Crohn's disease have increased cancer rates for both the small and large bowel. Finally, most patients with these diseases are able to maintain normal occupations and enjoy reasonably stable social and economic situations. The successful adaptation of patients with inflammatory bowel disease is influenced by a hopeful, optimistic personality and by an encouraging, supportive physician.


The impact of supplementation in winter-run chinook salmon on effective population size.
Supplementation of young raised at a protected site, such as a hatchery, may influence the effective population size of an endangered species. A supplementation program for the endangered winter-run chinook salmon from the Sacramento River, California, has been releasing fish since 1991. A breeding protocol, instituted in 1992, seeks to maximize the effective population size from the captive spawners by equaling their contributions to the released progeny. As a result, the releases in 1994 and 1995 appear not to have decreased the overall effective population size and may have increased it somewhat. However, mistaken use of non-winter-run chinook spawners resulted in artificial crosses between runs with a potential reduction in effective population size, and imprinting of the released fish on Battle Creek, the site of the hatchery, resulted in limiting the contribution of the released fish to the target mainstem population. Rapid genetic analysis of captured spawners and a new rearing facility on the Sacramento River should alleviate these problems and their negative effect on the effective population size in future years.


Production of atypical category exemplars in patients with schizophrenia.
Previous studies have revealed semantic memory impairments in patients with schizophrenia, and suggested that certain of these impairments were related to thought disorganization. One explanation offered for this is a broadening of the boundaries of semantic categories in schizophrenia. We selected 16 semantic categories, and required a sample of 41 schizophrenia patients and 43 healthy control subjects to produce one exemplar from each category. The typicality of the subjects' responses was rated. The exemplars produced by the patients were on average less typical than those produced by the healthy controls. No significant association between typicality of the response and thought disorganization was revealed in the patient sample. Affective flattening, alogia, and anhedonia were significantly and inversely associated with the typicality score, that is, higher ratings of these symptoms were associated with more typical responses. Our results suggest that a broadening of semantic category boundaries is observed in patients with schizophrenia, but is unrelated to thought disorganization. This semantic abnormality is not a feature of the patients with high ratings of certain negative symptoms.


Influence of continuous magnetic field on the separation of ephedrine enantiomers by molecularly imprinted polymers.
A set of polymers was imprinted with (-)-ephedrine using UV initiation, under the influence of a constant external magnetic field with intensities ranging from 0 to 1.55 T. Synthesised materials were characterised by X-ray crystallography, infrared spectroscopy, swelling and surface area. Recognition properties were assessed by the ability to discriminate between (+) and (-)-ephedrine and by Scatchard analyses on chromatographic mode. It was shown that polymer morphology and recognition properties are affected by the magnetic field. This resulted in considerable improvements in the chromatographic resolution of ephedrine enantiomers by materials synthesised under the influence of magnetic field. Apparently the magnetic field improved the ordering of the polymer structure and facilitated the formation of more uniform imprinting sites.