Rad6 overexpression induces multinucleation, centrosome amplification, abnormal mitosis, aneuploidy, and transformation.
We have isolated by differential RNA display a cDNA that is up-regulated in metastatic mammary tumor lines. This cDNA corresponds to HR6B, the yeast homologue of Rad6, a ubiquitin-conjugating enzyme, and a key player in postreplication repair and induced mutagenesis in the yeast. We show that Rad6 protein expressed in metastatic tumor lines is wild type and functional, because it is able to catalyze the transfer of ubiquitin to histone H2b and is predominantly localized in the nucleus as compared with cytoplasmic localization in normal or nonmetastatic mammary cells. This pattern of Rad6 protein expression/localization is not restricted to breast cancer cell lines, because human breast carcinomas display similar patterns of Rad6 up-regulation and nuclear localization suggesting that deregulation in expression of Rad6 may be an important step in transformation to malignant phenotype. Constitutive overexpression of exogenous human HR6B cDNA into normal-behaving MCF10A human breast epithelial cells induced cell-cell fusion that resulted in generation of multinucleated cells, centrosome amplification, multipolar mitotic spindles, aneuploidy, and ability for anchorage-independent growth. Double immunofluorescence labeling experiments demonstrated the colocalization of Rad6 protein with gamma-tubulin on centrosomes. This physical association of Rad6 with centrosomes is maintained throughout the interphase and mitotic phases of the cell cycle. The Rad6 protein exhibits notable alterations in distribution during interphase and mitotic stages of the cell cycle that are compatible with its function as a transcription factor. These findings suggest that Rad6 is an important ubiquitin-conjugating enzyme that may play a significant role in the maintenance of genomic integrity of mammalian cells and that an imbalance in the levels and activity of Rad6 could lead to chromosomal instability and transformation in vitro.


Connexin32 inhibits gastric carcinogenesis through cell cycle arrest and altered expression of p21Cip1 and p27Kip1.
Gap junctions and their structural proteins, connexins (Cxs), have been implicated in carcinogenesis. To explore the involvement of Cx32 in gastric carcinogenesis, immunochemical analysis of Cx32 and proliferation marker Ki67 using tissue-microarrayed human gastric cancer and normal tissues was performed. In addition, after Cx32 overexpression in the human gastric cancer cell line AGS, cell proliferation, cell cycle analyses, and p21(Cip1) and p27(Kip1) expression levels were examined by bromodeoxyuridine assay, flow cytometry, real-time RT-PCR, and western blotting. Immunohistochemical study noted a strong inverse correlation between Cx32 and Ki67 expression pattern as well as their location. In vitro, overexpression of Cx32 in AGS cells inhibited cell proliferation significantly. G1 arrest, up-regulation of cell cycle-regulatory proteins p21(Cip1) and p27(Kip1) was also found at both mRNA and protein levels. Taken together, Cx32 plays some roles in gastric cancer development by inhibiting gastric cancer cell proliferation through cell cycle arrest and cell cycle regulatory proteins.


The renin-angiotensin system and vascular fibrinolytic balance.
Haemostasis depends on an intricate relation among plasma coagulation and fibrinolytic factors, blood cells, vessel walls, extracellular matrix, blood viscosity, and blood flow. Hormonal systems, such as the renin-angiotensin system (RAS) act as a protector against acute fluid volume loss by inducing both vasoconstriction to maintain blood pressure and the retention of salt and water to restore normal fluid volume. Studies have confirmed that this cascade of events is brought into action by the RAS in coordination and sequentially. Inappropriate activation of the RAS may occur, however, in patients with left ventricular dysfunction and heart failure. Angiotensin-converting enzyme inhibitors are indicated to counteract the vasoconstriction and salt and water retention associated with activation of the RAS.


Prevalence and risk factors of vertebral compression fractures in female SLE patients.
Our objective was to determine the frequency of and factors associated with prevalent vertebral compression fractures in female systemic lupus erythematosus (SLE) patients attending rheumatologists in western Sweden. In this cross sectional study 150 women were included. They were examined with x-ray of thoracic and lumbar spine (Th4 to L4). A reduction of at least 20% of any vertebral height, assessed by Genant's semiquantitative method, was defined as a fracture. Bone mineral density (BMD) was measured by dual energy x-ray absorptiometry (DXA). Median patient age was 47 years (20 to 82) and disease duration 11 years (1 to 41). Only 6 (4%) women had a history of clinical compressions whereas 43 (29%) had at least one radiological fracture each. The patients with at least one fracture at any site were characterized by older age (P < 0.001), being postmenopausal (P < 0.01), higher Systemic Lupus International Collaborative Clinics Damage Index (P < 0.05), lower BMD total hip and femoral neck (P < 0.05), more peripheral fractures (P < 0.01), medication with bisphosphonates (P <0.05) and calcium and vitamin D3 (P < 0.05). There were no significant differences regarding current or cumulative glucocorticosteroid dose between the groups. In logistic regression analyses high age remained as a risk factor of at least one vertebral fracture at any site whereas low BMD in total hip was associated with vertebral fracture in the lumbar spine. Radiological compression fractures are common but seldom diagnosed in SLE patients. High age and low BMD in total hip, but not in spine, was associated with vertebral fractures.


An exploration strategy improves the diversity of de novo ligands using deep reinforcement learning: a case for the adenosine A2A receptor.
Over the last 5 years deep learning has progressed tremendously in both image recognition and natural language processing. Now it is increasingly applied to other data rich fields. In drug discovery, recurrent neural networks (RNNs) have been shown to be an effective method to generate novel chemical structures in the form of SMILES. However, ligands generated by current methods have so far provided relatively low diversity and do not fully cover the whole chemical space occupied by known ligands. Here, we propose a new method (DrugEx) to discover de novo drug-like molecules. DrugEx is an RNN model (generator) trained through reinforcement learning which was integrated with a special exploration strategy. As a case study we applied our method to design ligands against the adenosine A2A receptor. From ChEMBL data, a machine learning model (predictor) was created to predict whether generated molecules are active or not. Based on this predictor as the reward function, the generator was trained by reinforcement learning without any further data. We then compared the performance of our method with two previously published methods, REINVENT and ORGANIC. We found that candidate molecules our model designed, and predicted to be active, had a larger chemical diversity and better covered the chemical space of known ligands compared to the state-of-the-art.


Insulin therapy and GH-IGF-I axis disorders in diabetes: impact of glycaemic control and hepatic insulinization.
In Type 1 diabetes, high circulating growth hormone (GH) in conjunction with low plasma insulin-like growth factor-I (IGF-I) is indicative of a hepatic GH-resistance profile since the liver is the main source of circulating IGF-I. The reduction in specific growth hormone binding protein (GHBP), corresponding to the extracellular domain of the GH receptor, provides an indirect indication of the hepatic density of GH receptors, as does the reduction in IGFBP-3, the major IGF binding protein, which is GH-dependent. Type 1 diabetes is also associated with high levels of IGFBP-1, a binding protein down-regulated by insulin. Although most of these abnormalities have been described in situations of poor glycaemic control, hyperglycaemia does not seem to be the predominant factor in their pathogenesis. Even intensified subcutaneous insulin therapy does not normalize GH, IGF-I, GHBP and IGFBP-3 plasma levels. Some indirect evidence suggests that portal insulinopenia plays a role in the hepatic GH-resistance profile of Type 1 diabetes, i.e. discrepancies between the abnormalities reported in Type 1 and Type 2 diabetes, and the inverse relationship between residual insulin secretion in Type 1 diabetes and some of these abnormalities. Intraperitoneal insulin therapy administered to Type 1 diabetic patients by implantable pumps (without modification of glycaemic control) can improve GHBP activity, practically normalize plasma IGF-I and normalize IGFBP-3. The improvement in GH-IGF-I axis disorders obtained with intraperitoneal insulin therapy (which allows primary portal insulin absorption) provides direct evidence of the central role of portal insulin in the regulation of this system.


Specificity in Mesograzer-Induced Defences in Seagrasses.
Grazing-induced plant defences that reduce palatability to herbivores are widespread in terrestrial plants and seaweeds, but they have not yet been reported in seagrasses. We investigated the ability of two seagrass species to induce defences in response to direct grazing by three associated mesograzers. Specifically, we conducted feeding-assayed induction experiments to examine how mesograzer-specific grazing impact affects seagrass induction of defences within the context of the optimal defence theory. We found that the amphipod Gammarus insensibilis and the isopod Idotea chelipes exerted a low-intensity grazing on older blades of the seagrass Cymodocea nodosa, which reflects a weak grazing impact that may explain the lack of inducible defences. The isopod Synischia hectica exerted the strongest grazing impact on C. nodosa via high-intensity feeding on young blades with a higher fitness value. This isopod grazing induced defences in C. nodosa as indicated by a consistently lower consumption of blades previously grazed for 5, 12 and 16 days. The lower consumption was maintained when offered tissues with no plant structure (agar-reconstituted food), but showing a reduced size of the previous grazing effect. This indicates that structural traits act in combination with chemical traits to reduce seagrass palatability to the isopod. Increase in total phenolics but not in C:N ratio and total nitrogen of grazed C. nodosa suggests chemical defences rather than a modified nutritional quality as primarily induced chemical traits. We detected no induction of defences in Zostera noltei, which showed the ability to replace moderate losses of young biomass to mesograzers via compensatory growth. Our study provides the first experimental evidence of induction of defences against meso-herbivory that reduce further consumption in seagrasses. It also emphasizes the relevance of grazer identity in determining the level of grazing impact triggering resistance and compensatory responses of different seagrass species.


Deletion mapping suggests that the 1p22 melanoma susceptibility gene is a tumor suppressor localized to a 9-Mb interval.
Loss of the short arm of chromosome 1 is frequently observed in many tumor types, including melanoma. We recently localized a third melanoma susceptibility locus to chromosome band 1p22. Critical recombinants in linked families localized the gene to a 15-Mb region between D1S430 and D1S2664. To map the locus more finely we have performed studies to assess allelic loss across the region in a panel of melanomas from 1p22-linked families, sporadic melanomas, and melanoma cell lines. Eighty percent of familial melanomas exhibited loss of heterozygosity (LOH) within the region, with a smallest region of overlapping deletions (SRO) of 9 Mb between D1S207 and D1S435. This high frequency of LOH makes it very likely that the susceptibility locus is a tumor suppressor. In sporadic tumors, four SROs were defined. SRO1 and SRO2 map within the critical recombinant and familial tumor region, indicating that one or the other is likely to harbor the susceptibility gene. However, SRO3 may also be significant because it overlaps with the markers with the highest 2-point LOD score (D1S2776), part of the linkage recombinant region, and the critical region defined in mesothelioma. The candidate genes PRKCL2 and GTF2B, within SRO2, and TGFBR3, CDC7, and EVI5, in a broad region encompassing SRO3, were screened in 1p22-linked melanoma kindreds, but no coding mutations were detected. Allelic loss in melanoma cell lines was significantly less frequent than in fresh tumors, indicating that this gene may not be involved late in progression, such as in overriding cellular senescence, necessary for the propagation of melanoma cells in culture.


Gold-catalyzed annulations of allenes with N-hydroxyanilines to form indole derivatives with benzaldehyde as a promoter.
Gold-catalyzed syntheses of 2,3-disubstituted indole derivatives from N-hydroxyanilines and allenes are described; these reactions require benzaldehyde as an additive to generate nitrones in situ. Our control experiments indicate that nitrones and water were indispensable in the reactions whereas N-hydroxyanilines alone were inactive nucleophiles. This synthetic method is compatible with allenes and N-hydroxyanilines with a reasonable range, further highlighting its synthetic utility.


A multicompartmental model of in vivo adipose tissue glycerol kinetics and capillary permeability in lean and obese humans.
Lipolysis of adipose tissue triglycerides releases glycerol. Twenty-four volunteers, of whom 6 were obese and 13 were women, received a primed-constant infusion of 2H5-glycerol for 120 min during postabsorptive steady-state conditions. Arterial, abdominal venous, and interstitial (microdialysis) samples were taken, and a four-compartment model was applied to assess subcutaneous abdominal adipose tissue glycerol kinetics. Adipose tissue blood flow was measured using 133Xe washout. Venous glycerol concentrations (median 230 micromol/l [interquartile range 210-268]) were consistently greater than those of arterial blood (69.1 micromol/l [56.5-85.5]), while glycerol isotopic enrichments (tracer-to-tracee ratio) were greater in arterial blood (8.34% [7.44-10.1]) than venous blood (2.34% [1.71-2.69], P < 0.01). Microdialysate glycerol enrichment was 1.44% (1.11-1.79), indicating incomplete permeability of glycerol between capillary blood and interstitium. Calculated interstitial glycerol concentrations were between 270 micromol/l (256-350) and 332 micromol/l (281-371) (examining different boundary conditions). The calculated capillary diffusion capacity (ps) was between 2.21 ml . 100 g tissue(-1) . min(-1) (1.31-3.13) and 3.09 ml . 100 g tissue(-1) . min(-1) (1.52-4.90) and correlated inversely with adiposity (Rs< or = -0.45, P < 0.05). Our results support previous estimates of interstitial glycerol concentration within adipose tissue and reveal capillary diffusion capacity is reduced in obesity.


The use of DiazaCon™ to limit fertility by reducing serum cholesterol in female grey squirrels, Sciurus carolinensis.
The grey squirrel, Sciurus carolinensis Gmelin, is an invasive alien species introduced into Great Britain in the late nineteenth century and into Northern Italy during the early twentieth century. Grey squirrels have displaced the native European red squirrel, Sciurus vulgaris L., throughout much of Great Britain and have a significant impact on trees and woodlands through bark-stripping activity. In Britain, eradication is no longer an option at a regional scale, but fertility control offers a non-lethal approach to reducing negative impacts. The cholesterol mimic DiazaCon™ has been successfully used to inhibit reproduction in some species. These studies aimed to evaluate whether DiazaCon™ is effective in inhibiting reproduction in grey squirrels. DiazaCon™ reduced serum cholesterol levels in female grey squirrels at a range of doses. The period of effect increased with increasing dose. Reproduction rate was not significantly different between treatment and control groups owing to a lack of breeding in controls. DiazaCon™ has potential to reduce serum cholesterol levels enough and for a sufficient period to reduce fertility in female grey squirrels. Information on baseline physiology and blood chemistry of grey squirrels is required to inform interpretation of the level of significance of the effect.


Metabolic effects of troglitazone therapy in type 2 diabetic, obese, and lean normal subjects.
To characterize metabolic effects of troglitazone in type 2 diabetic, obese, and lean subjects, and examine the effects of troglitazone 2-3 weeks after discontinuation. Nine type 2 diabetic, nine obese, and nine lean subjects underwent baseline metabolic studies including an 8-h meal-tolerance test (MTT) and a 5-h glucose clamp. Subjects then received troglitazone (600 mg/day) for 12 weeks and subsequently had repeat metabolic studies. Diabetic subjects remained off hypoglycemic agents for 2-3 weeks and then underwent a 5-h glucose clamp. In diabetic subjects, fasting plasma glucose was reduced (P<0.05) and insulin-stimulated glucose disposal (Rd) was enhanced by treatment (P<0.02). The area under the MTT 8-h plasma glucose curve declined with therapy (P<0.001), and its change was positively correlated with the improvement in Rd (r = 0.75, P<0.05). There was also a positive correlation between the change in fasting hepatic glucose output (HGO) and the change in fasting plasma glucose with treatment (r = 0.92, P<0.001). Discontinuation of therapy for 2-3 weeks did not significantly affect fasting plasma glucose or insulin-stimulated glucose Rd. In obese subjects, insulin-stimulated glucose Rd improved with therapy (P<0.001), allowing for maintenance of euglycemia by lower plasma insulin concentrations (P<0.05). In lean subjects, an increase in fasting HGO (P<0.001) and glucose clearance (P<0.01) was observed. Troglitazone lowers fasting and postprandial plasma glucose in type 2 diabetes by affecting both fasting HGO and peripheral insulin sensitivity. Its effects are evident 2-3 weeks after discontinuation. In obese subjects, its insulin sensitizing effects suggest a role for its use in the primary prevention of type 2 diabetes.


Quantitation of human immunodeficiency virus type 1 infection kinetics.
Tissue culture infections of CD4-positive human T cells by human immunodeficiency virus type 1 (HIV-1) proceed in three stages: (i) a period following the initiation of an infection during which no detectable virus is produced; (ii) a phase in which a sharp increase followed by a peak of released progeny virions can be measured; and (iii) a final period when virus production declines. In this study, we have derived equations describing the kinetics of HIV-1 accumulation in cell culture supernatants during multiple rounds of infection. Our analyses indicated that the critical parameter affecting the kinetics of HIV-1 infection is the infection rate constant k = Inn/ti, where n is the number of infectious virions produced by one cell (about 10(2)) and ti is the time required for one complete cycle of virus infection (typically 3 to 4 days). Of particular note was our finding that the infectivity of HIV-1 during cell-to-cell transmission is 10(2) to 10(3) times greater than the infectivity of cell-free virus stocks, the inocula commonly used to initiate tissue culture infections. We also demonstrated that the slow infection kinetics of an HIV-1 tat mutant is not due to a longer replication time but reflects the small number of infectious particles produced per cycle.


Effects of practitioner-delivered brief counseling and computer-generated tailored letters on cigarettes per day among smokers who do not quit--a quasi-randomized controlled trial.
It is still unclear how brief counseling for smoking cessation, combined with proactive recruitment of participants, impacts on those smokers not reaching the primary treatment goal of tobacco abstinence. Thus, within a quasi-randomized controlled trial, we examined the effects of (1) practitioner-delivered brief advice and (2) computer-generated tailored letters on cigarettes per day (CPD) among participants not succeeding in quitting. A total of 34 general practices (participation rate 87%) were randomly selected in a German region. Within these practices, 1499 daily smoking patients aged 18-70 years (participation rate 80%) agreed to participate in a smoking cessation intervention trial. Allocation to study condition was based on time of practice attendance. Latent growth analyses were performed on the subsample of 1334 (89%) smokers who did not reach 6-month prolonged abstinence within the 2-year follow-up period. CPD was assessed at baseline and at 6-, 12-, 18-, and 24-month follow-ups. Both interventions led to small but significant reductions in CPD, and they did not differ in efficacy. Treatment effects occurred within the first 6 months and could be sustained by the continuing smokers until the 24-month follow-up. Present results complement earlier findings of increased abstinence rates in the total sample. It can be concluded that, even if applied to unselected samples of smokers, from which only a minority initially intends to change, both brief counseling strategies are able to significantly decrease tobacco consumption. They hence appear to provide a means to reducing tobacco-related disease among general medical practice patients.


The 2009 Lindau Nobel Laureate meeting: Martin Chalfie, Chemistry 2008.
American Biologist Martin Chalfie shared the 2008 Nobel Prize in Chemistry with Roger Tsien and Osamu Shimomura for their discovery and development of the Green Fluorescent Protein (GFP). Martin Chalfie was born in Chicago in 1947 and grew up in Skokie Illinois. Although he had an interest in science from a young age--learning the names of the planets and reading books about dinosaurs--his journey to a career in biological science was circuitous. In high school, Chalfie enjoyed his AP Chemistry course, but his other science courses did not make much of an impression on him, and he began his undergraduate studies at Harvard uncertain of what he wanted to study. Eventually he did choose to major in Biochemistry, and during the summer between his sophomore and junior years, he joined Klaus Weber's lab and began his first real research project, studying the active site of the enzyme aspartate transcarbamylase. Unfortunately, none of the experiments he performed in Weber's lab worked, and Chalfie came to the conclusion that research was not for him. Following graduation in 1969, he was hired as a teacher Hamden Hall Country Day School in Connecticut where he taught high school chemistry, algebra, and social sciences for 2 years. After his first year of teaching, he decided to give research another try. He took a summer job in Jose Zadunaisky's lab at Yale, studying chloride transport in the frog retina. Chalfie enjoyed this experience a great deal, and having gained confidence in his own scientific abilities, he applied to graduate school at Harvard, where he joined the Physiology department in 1972 and studied norepinephrine synthesis and secretion under Bob Pearlman. His interest in working on C. elegans led him to post doc with Sydney Brenner, at the Medical Research Council Laboratory of Molecular Biology in Cambridge, England. In 1982 he was offered position at Columbia University. When Chalfie first heard about GFP at a research seminar given by Paul Brehm in 1989, his lab was studying genes involved in the development and function of touch-sensitive cells in C. elegans. He immediately became very excited about the idea of expressing the fluorescent protein in the nematode, hoping to figure out where the genes were expressed in the live organism. At the time, all methods of examining localization, such as antibody staining or in situ hybridization, required fixation of the tissue or cells, revealing the location of proteins only at fixed points in time. In September 1992, after obtaining GFP DNA from Douglas Prasher, Chalfie asked his rotation student, Ghia Euskirchen to express GFP in E. coli, unaware that several other labs were also trying to express the protein, without success. Chalfie and Euskirchen used PCR to amplify only the coding sequence of GFP, which they placed in an expression vector and expressed in E.coli. Because of her engineering background, Euskirchen knew that the microscope in the Chalfie lab was not good enough to use for this type of experiment, so she captured images of green bacteria using the microscope from her former engineering lab. This work demonstrated that GFP fluorescence requires no component other than GFP itself. In fact, the difficulty that other labs had encountered stemmed from their use of restriction enzyme digestions for subcloning, which brought along an extra sequence that prevented GFP's fluorescent expression. Following Euskirchen's successful expression in E. coli, Chalfie's technician Yuan Tu went on to express GFP in C. elegans, and Chalfie published the findings in Science in 1994. Through the study of C. elegans and GFP, Chalfie feels there is an important lesson to be learned about the importance basic research. Though there has been a recent push for clinically-relevant or patent-producing (translational) research, Chalfie warns that taking this approach alone is a mistake, given how "woefully little" we know about biology. He points out the vast expanse of the unknowns in biology, noting that important discoveries such as GFP are very frequently made through basic research using a diverse set of model organisms. Indeed, the study of GFP bioluminescence did not originally have a direct application to human health. Our understanding of it, however, has led to a wide array of clinically-relevant discoveries and developments. Chalfie believes we should not limit ourselves: "We should be a little freer and investigate things in different directions, and be a little bit awed by what we're going to find."


Desire for motherhood: exploring HIV-positive women's desires, intentions and decision-making in attaining motherhood.
With access to prevention of parent-to-child transmission (PPTCT) and antiretroviral therapy (ARV), people living with HIV/AIDS are better able to consider childbearing and parenthood. However, there is limited understanding of the reproductive healthcare needs and the impact of infection on the fertility desires of women living with HIV/AIDS. Research on the relationship between fertility and HIV/AIDS has been largely clinical, focusing on the ability of women living with HIV/AIDS (WLHA) to conceive or their pregnancy outcomes. This paper describes the findings of a qualitative study undertaken in Namakkal district, Tamil Nadu, India that aimed to explore fertility desires, intentions and fertility decision-making in WLHA and the barriers they face in fulfilling these desires. In-depth interviews were held with selected 43 currently married WLHA aged 18-35 years and 10 key informants. The women were classified according to whether or not they had living children and, within that, whether they had experienced abortion, wanted more or any children or were pregnant. The main factors distinguishing women who wanted to have a child and those who did not were their levels of anxiety about the future and available family support. Women who indicated that they did not have family support and were stigmatised by the family were reluctant to opt for a pregnancy as they were not sure of the future, including child care in event of parental death. In contrast, those women who decided to have a child did so based on family support, especially when family members offered to take care of the child in the future in the event of parental death. Awareness and access to PPTCT and ARV was another key factor guiding the final decision on child bearing. Findings highlight the need for further research on issues faced by WLHA in fulfilling their fertility desires and intentions and for programmes that both enable WLHA to exercise informed choice in meeting their fertility desires and sensitise healthcare providers about these needs.


Meanings of "acceptance" for patients with long-term pain when starting rehabilitation.
The study aimed to elucidate the meaning of acceptance in relation to the lived body and sense of self when entering a pain rehabilitation programme. Six women and three men with long-term pain were interviewed. The interviews were analysed according to interpretative phenomenological analysis. The analysis revealed three different meaning structures, first: acceptance as a process of personal empowerment, "the only way forward". Here, the individuals expressed that the body felt integrated: a trusting cooperation between self and body gave rise to hope. Second: acceptance as an equivocal project, a possible but challenging way forward. The hopeful insight was there, acknowledging that acceptance was the way to move forward, but there was also uncertainty and doubt about one's ability with a body ambiguous and confusing, difficult but important to understand. Third, in acceptance as a threat and a personal failure, "no way forward" the integration of the aching body in sense of self was impossible and pain was incomprehensible, unacceptable and unfair. Pain was the cause of feeling stuck in the body, affecting the sense of self and the person's entire life. The meaning of acceptance was related to acceptance of the persistency of pain, to how the individual related to the lived body and the need for changes in core aspects of self, and to the issue of whether to include others in the struggle of learning to move on with a meaningful life. Healthcare professionals should be aware that individuals with long-term pain conceptualize and hold different meanings of acceptance when starting rehabilitation; this should be considered and addressed in rehabilitation programmes. The meaning given to acceptance is related to the experience of the lived body and the sense of self, as well as to getting legitimization/acceptance by others; therefore these aspects need to be considered during rehabilitation. The process of achieving acceptance seems to embrace different processes which can be understood as, and facilitated by, an embodied learning process. The bodily existential challenges presented in the present study, for example to develop an integrated and cooperative relationship with the painful body, can inspire health professionals to develop interventions and communication strategies focusing on the lived body. A wide range of competencies in rehabilitation clinics seems to be needed.


Recognizing Action Units for Facial Expression Analysis.
Most automatic expression analysis systems attempt to recognize a small set of prototypic expressions, such as happiness, anger, surprise, and fear. Such prototypic expressions, however, occur rather infrequently. Human emotions and intentions are more often communicated by changes in one or a few discrete facial features. In this paper, we develop an Automatic Face Analysis (AFA) system to analyze facial expressions based on both permanent facial features (brows, eyes, mouth) and transient facial features (deepening of facial furrows) in a nearly frontal-view face image sequence. The AFA system recognizes fine-grained changes in facial expression into action units (AUs) of the Facial Action Coding System (FACS), instead of a few prototypic expressions. Multistate face and facial component models are proposed for tracking and modeling the various facial features, including lips, eyes, brows, cheeks, and furrows. During tracking, detailed parametric descriptions of the facial features are extracted. With these parameters as the inputs, a group of action units (neutral expression, six upper face AUs and 10 lower face AUs) are recognized whether they occur alone or in combinations. The system has achieved average recognition rates of 96.4 percent (95.4 percent if neutral expressions are excluded) for upper face AUs and 96.7 percent (95.6 percent with neutral expressions excluded) for lower face AUs. The generalizability of the system has been tested by using independent image databases collected and FACS-coded for ground-truth by different research teams.


The full-time clinical faculty: what goes around, comes around.
In his 1910 report entitled Medical Education in the United States and Canada, Abraham Flexner advanced an ideal model of medical education that included a university-based, full-time, salaried faculty whose time was devoted to teaching and research. This article traces the evolution of the "full-time" concept for clinical faculty and describes factors that have affected its implementation. Between 1910 and the 1930s, the full-time system for clinical faculty was implemented at a limited number of medical schools, but lack of financing made the system generally unworkable. The implementation of the "geographic" full-time concept during the 1940s to 1960s allowed faculty to be considered full-time while earning much of their income from clinical practice. Even then, there were concerns that medical schools would bring pressure on such faculty to increase their clinical activity for the purpose of supporting the institution. After the rise of private and public payers, clinical practice income came to be an explicit and increasingly important source of medical school revenue. This stimulated a significant expansion in the number of full-time clinical faculty over the next 40 years. In the 100 years following the Flexner Report, clinical faculty became "full-time" and "salaried," but not in the way Flexner imagined. Instead of deriving their salaries from the resources of the medical school, they are significantly contributing to institutional financing through their practice. Flexner's concern about the "distraction" of clinical practice interfering with faculty participation in education has come full circle, remaining a primary issue in medical education today.


Evaluation of LBM415 (NVP PDF-713), a novel peptide deformylase inhibitor, for treatment of experimental Mycoplasma pneumoniae pneumonia.
Mycoplasma pneumoniae is a major cause of community-acquired pneumonia. We evaluated the efficacy of LBM415, a novel peptide deformylase inhibitor antimicrobial agent, for the treatment of M. pneumoniae pneumonia in a mouse model. Eight-week-old BALB/c mice were intranasally inoculated once with 10(7) CFU of M. pneumoniae. Groups of mice were treated with LBM415 (50 mg/kg of body weight) or placebo subcutaneously daily for 13 days, starting 24 h after inoculation. Groups of mice were evaluated at the baseline; at days of treatment 1, 3, 6, and 13; and at 7 days after treatment. The MIC of LBM415 against M. pneumoniae was <0.005 microg/ml. LBM415-treated mice had significantly lower bronchoalveolar lavage fluid M. pneumoniae concentrations than placebo-treated mice on days 6 and 13 of treatment. Compared with placebo treatment, therapy with LBM415 significantly decreased lung histopathology scores at days 3, 6, and 13 of treatment and at 7 days after treatment. Airway obstruction was significantly lower in LBM415-treated mice than in placebo-treated mice on days 1, 3, and 6 of treatment and after 7 days of therapy, while airway hyperresponsiveness was significantly lower only on day 3 of therapy. The bronchoalveolar lavage fluid concentrations of tumor necrosis factor alpha, gamma interferon (IFN-gamma), interleukin-6 (IL-6), IL-12, KC (functional IL-8), monocyte chemotactic protein 1, macrophage inflammatory protein 1alpha, monokine induced by IFN-gamma, and IFN-inducible protein 10 were significantly reduced in LBM415-treated mice compared with the levels in placebo-treated mice. There were no differences in the bronchoalveolar lavage fluid concentrations of granulocyte-macrophage colony-stimulating factor, IL-1beta, IL-2, IL-4, IL-5, and IL-10 between the two groups of mice. LBM415 therapy had beneficial microbiologic, histologic, respiratory, and immunologic effects on acute murine M. pneumoniae pneumonia.


Angina associated with left main coronary artery compression in pulmonary hypertension.
Chest pain is a common complaint in patients with pulmonary arterial hypertension (PAH). Left main coronary artery (LMCA) compression by an enlarged pulmonary artery trunk (PAT) has been associated with angina, but appropriate diagnostic and treatment approaches remain poorly defined. We present two cases of angina caused by LMCA compression from an enlarged pulmonary artery, one of which also presented with new, severe left ventricular systolic dysfunction attributed to myocardial ischemia. Diagnosis of LMCA stenosis was made via coronary angiography followed by computed tomography-gated coronary angiography (CT-CA), which confirmed pulmonary artery enlargement as the source of extrinsic compression. Restoring LMCA patency with percutaneous intervention and/or aggressive treatment of pulmonary hypertension led to significant improvement in angina, cardiac function and quality of life. Given the negative impact on cardiac function, prompt diagnosis and treatment of extrinsic LMCA compression should be considered a priority.


Acute mobile phone effects on pre-attentive operation.
There is a debate whether electromagnetic field (EMF) emitted by mobile phones (MP) have an effect on cognitive functions. Since the auditory P50 component of event-related potentials (ERPs) reflects pre-attentive processing and working memory (WM) operation, the present study was designed to investigate whether the exposure to MP-EMF affects the patterns of the P50 component of ERPs elicited during a WM test. The P50 elicited during a WM task and evoked by two warning stimuli low and high frequency (500 and 3000 Hz) has been assessed in 19 normal subjects (10 women and 9 men) both without and with exposure to a 900 MHz signal, emitted by a dipole antenna placed near the subjects. Results showed that the presence of MP-EMFs induced statistically significant increase in the amplitude of P50 evoked by the low frequency stimuli, at Fp1 and O1 electrode leads as compared to themselves without MP-EMF exposure. In contrast the exposure to MP-EMFs revealed statistically significant decrease of the amplitude of P50 evoked by the high frequency stimuli, at Fp1 electrode lead as compared to themselves without MP-EMF exposure. These findings provide evidence that the MP-EMF emitted by mobile phone affect pre-attentive information processing as it is reflected in P50 evoked potential. The basis of such an effect is unclear, although several possibilities exist and call for potential directions of future research.


[Drug addiction research using intravenous self-administration procedure in mice].
A major clinical problem in treating drug abusers or addicts is the high rate of relapse to abuse even long after abstinence. In drug dependence research, drug self-administration displays excellent face validity, good construct validity, and appealing predictive validity with respect to drug consumption in humans. Because of the technical feasibility of intravenous drug self-administration, it is limited in mice relative to larger animals. Nevertheless, technical innovations made in the last 10 years have vastly improved the feasibility of long-term studies in mice. In addition, mice with targeted gene mutations, such as transgenic and knockout mice, provide a powerful tool for investigating candidate genes that may be involved in abuse of drugs in humans. Therefore, it is essential for researchers to extend the extinction-reinstatement procedure in mice. However, there are few reports that proven the relationship between genetic factor and relapsing behavior using drug self-administration procedure in mice. Based on recent documented information and our own experiences, in the present review, we described some procedural considerations for the successful establishment of drug self-administration procedure in mice, and then summarized some behavioral characteristics of genetic mouse models under the drug self-administration, and reinstatement procedure.


Eumycetoma of the foot caused by Exophiala jeanselmei in a Guinean woman.
Eumycetomas are chronic infectious entities characterized by presence of mycotic grains in (sub-)cutaneous tissues, after accidental inoculation of an exogenous filamentous fungus in the skin. The lesions evolve towards painless pseudotumor of the soft parts. We report the original case of a Guinean woman exhibiting eumycetoma of the right foot. Both laboratory tests identified a dematiaceous fungus, Exophiala jeanselmei, as the responsible infectious agent. A medical treatment with voriconazole alone was sufficient to notice a substantial clinical improvement. This finding is unusual as E. jeanselmei is uncommon in Guinea-Conakry, and as optimal treatment rather associate antifungal azoles and surgical excision.


Somatostatin receptors on human lymphocytes and leukaemia cells.
Receptors for somatostatin were identified on mitogen-activated human peripheral blood lymphocytes (PBL) and human leukaemic cells in 87.5% of lymphoblastic leukaemia and in 12.5% of non-lymphocytic leukaemia, using a somatostatin radiobinding assay. The specific binding of 125I-somatostatin of these cells increased linearly with the cell numbers and was suppressed by non-iodinated somatostatin. We investigated the distribution of fluorescent somatostatin to mitogen-activated PBL by using a fluorescence-activated cell sorter (FACS). Over 95% of the cell populations bound fluorescent somatostatin and no distinct predilection was found among certain lymphocyte subpopulations and somatostatin receptor-positive cells. Scatchard analysis showed a single class (low affinity) of binding site on mitogen-activated PBL and two classes (high and low affinity) of specific binding sites on lymphoblastic leukaemia cells.


High throughput screening of gene functions in mammalian cells using reversely transfected cell arrays: review and protocol.
Reversely transfected cell microarrays (RTCM) have been introduced as a method for parallel high throughput analysis of gene functions in mammalian cells. Hundreds to thousands of different recombinant DNA or RNA molecules can be transfected into different cell clusters at the same time on a single glass slide with this method. This allows either the simultaneous overexpression or--by using the recently developed RNA interference (RNAi) techniques--knockdown of a huge number of target genes. A growing number of sophisticated detection systems have been established to determine quantitatively the effects of the transfected molecules on the cell phenotype. Several different cell types have been successfully used for this procedure. This review summarizes the presently available knowledge on this technique and provides a laboratory protocol.


De novo splice site variant of ARID1B associated with pathogenesis of Coffin-Siris syndrome.
Coffin-Siris syndrome is an extremely rare syndrome associated with developmental and congenital anomalies. It is caused by heterozygous pathogenic variants of ARID1A, ARID1B, SMARCA4, SMARCB1, SMARCE1, and SOX11. This case study presents the whole exome sequencing of a patient with characteristic clinical features of Coffin-Siris syndrome. Analysis included Sanger sequencing of complementary DNA and bioinformatic analysis of the variant. Analysis of cDNA Sanger sequencing data revealed that the donor splice site variant led to skipping of exon 19. Further, bioinformatic analysis predicted abnormal splicing in a translational frameshift of 11 amino acids and the creation of a premature termination codon. Results found a novel de novo splice site variant c.5025+2T>C in the ARID1B and truncated 1 633 amino acid protein NP_065783.3:p. (Thr1633Valfs*11). Truncated ARID1B resulted in loss of the BAF250 domain, which is part of SWI/SNF-like ATP-dependent chromatin remodeling complex. The severe clinical manifestation presented by the proband was attributed to the disappearance of the BAF250 domain in the ARID1B protein. Our finding provides strong evidence that this pathogenic variant of exon 19 caused a frameshift mutation in the ARID1B at the terminal exon, resulting in the expression of a severe phenotype of CSS.


Contribution of prepregnancy body mass index and gestational weight gain to caesarean birth in Canada.
Overweight and obese women are known to be at increased risk of caesarean birth. This study estimates the contribution of prepregnancy body mass index (BMI) and gestational weight gain (GWG) to caesarean births in Canada. We analyzed data from women in the Canadian Maternity Experiences Survey who had a singleton term live birth in 2005-2006. Adjusted odds ratios for caesarean birth across BMI and GWG groups were derived, separately for nulliparous women and parous women with and without a prior caesarean. Population attributable fractions of caesarean births associated with above normal BMI and excess GWG were calculated. The overall caesarean birth rate was 25.7%. Among nulliparous and parous women without a previous caesarean birth, rates in obese women were 45.1% and 9.7% respectively, and rates in women who gained above their recommended GWG were 33.5% and 8.0% respectively. Caesarean birth was more strongly associated with BMI than with GWG. However, due to the high prevalence of excess GWG (48.8%), the proportion of caesareans associated with above normal BMI and excess GWG was similar [10.1% (95% CI: 9.9-10.2) and 10.9% (95% CI: 10.7-11.1) respectively]. Overall, one in five (20.2%, 95% CI: 20.0-20.4) caesarean births was associated with above normal BMI or excess GWG. Overweight and obese BMI and above recommended GWG are significantly associated with caesarean birth in singleton term pregnancies in Canada. Strategies to reduce caesarean births must include measures to prevent overweight and obese BMI prior to conception and promote recommended weight gain throughout pregnancy.


Factors associated with survival to hospital discharge for cats treated surgically for thoracic trauma.
OBJECTIVE To determine the most common types of injuries in cats surgically treated for thoracic trauma, complications associated with surgical treatment, and factors associated with mortality rate and evaluate the effectiveness of the animal trauma triage (ATT) scoring system for predicting outcome. DESIGN Retrospective case series with nested observational study. ANIMALS 23 client-owned cats surgically treated for thoracic trauma at 7 veterinary teaching hospitals between 1990 and 2014. PROCEDURES Medical records were reviewed to collect data on signalment, medical history, clinical signs and physical examination findings at initial evaluation, clinicopathologic findings, initial emergency treatments and diagnostic tests performed, type of trauma sustained, imaging findings, surgery details, postoperative complications, duration of hospitalization, and cause of death, if applicable. All variables were evaluated for associations with survival to hospital discharge. RESULTS Types of trauma that cats had sustained included dog bite or attack (n = 8 [35%]), motor vehicle accident (6 [26%]), other animal attack (2 [9%]), impalement injury or fall (2 [9%]), projectile penetrating trauma (1 [4%]), or unknown origin (4 [17%]). Intrathoracic surgery was required for 65% (15/23) of cats. The overall perioperative mortality rate was 13% (3/23). Mean ± SD ATT scores for surviving and nonsurviving cats were 6.4 ± 2.2 and 10.0 ± 1.7, respectively. Nineteen of 20 cats with no cardiopulmonary arrest survived to discharge, compared with 1 of 3 cats with cardiopulmonary arrest. Only these 2 variables were significantly associated with outcome. CONCLUSIONS AND CLINICAL RELEVANCE The perioperative mortality rate was low in this series of cats with thoracic trauma; however, those with cardiopulmonary arrest were less likely to survive to hospital discharge than other cats. Cats with a low ATT score were more likely to survive than cats with a high ATT score.


Regulation of cell specific expression of calcyclin (S100A6) in nerve cells and other tissues.
Many of the small, acidic, calcium binding S100 proteins present in the brain specifically map different anatomical regions and cell types and their overexpression is implicated in pathological changes. Similarly to other members of the S100 protein family, calcyclin (S100A6) is expressed in a cell specific manner and is found in subpopulations of neurons and astrocytes in the brain and in epithelial cells and fibroblasts. In this article we review data concerning the cell specific expression of S100 protein genes and present experimental results on the regulation of the calcyclin gene. We have performed promoter deletion studies to locate regions within the calcyclin gene promoter responsible for transcriptional regulation. The results demonstrate that the 3 kb long calcyclin gene promoter lacks a cell specific cis-acting element and drives the expression of the reporter gene also in cells that do not express endogenous calcyclin. The expression is modulated by positive and negative elements acting uniformly in the four different cell lines studied. The first intron of the calcyclin gene was found to have an inhibitory influence on expression regardless of cell type. It was also shown that calcyclin expression can be induced in calcyclin-negative cells by treatment with 5-azacytidine suggesting the involvement of gene methylation in its cell specific expression. The results are discussed in light of the data available on the regulation of other S100 genes.


Sedentary behavior after stroke: A new target for therapeutic intervention.
Over the last 10 years, evidence has emerged that too much sedentary time (e.g. time spent sitting down) has adverse effects on health, including an increased risk of cardiovascular disease incidence and mortality. A considerable amount of media attention has been given to the topic. The current UK activity guidelines recommend that all adults should minimize the amount of time spent being sedentary for extended periods. How best to minimize sedentary behavior is a focus of ongoing research. Understanding the impact of sedentary behaviors on the health of people with stroke is vital as they are some of the most sedentary individuals in society. Implementing strategies to encourage regular, short breaks in sedentary behaviors has potential to improve health outcomes after stroke. Intervention work already conducted with adults and older adults suggests that sedentary behaviors can be changed. A research priority is to explore the determinants of sedentary behavior in people with stroke and to develop tailored interventions.


Influence of Bovine Serum Lipids and Fetal Bovine Serum on the Expression of Cell Surface Markers in Cultured Bovine Preadipocytes.
To establish the influence of fetal bovine serum (FBS) and bovine serum lipids (BSL) on cell differentiation marker expression, bovine adipose-derived stem cells from subcutaneous tissue were incubated for 14 days in 4 types of differentiation media containing 10% FBS and 10 µL/mL BSL (TRT-1), no FBS and 10 µL/mL of BSL (TRT-2), 10% FBS and no BSL (TRT-3), or no supplements (TRT-4). Cells were subjected to Nile red staining, immunocytochemistry (CD73, CD90, CD105, DLK1, FabP4), and quantitative real-time PCR (CD73, CD90, CD105, FabP4). The number of cells presenting FabP4 and the percentage of mature adipocytes with large lipid droplets were increased in TRT-2, accompanied by a robust increase in FabP4 mRNA abundance and a decrease in DLK1-positive cells. In preadipocytes, CD73 was present around the nucleus and translocated towards cell membranes during differentiation. Although the percentage of CD73-positive cells was not different among treatments, its mRNA abundance, immunocytochemical staining intensity, and translocation towards cell membranes were decreased when the medium contained no FBS (TRT-2 and TRT-4). All cells showed a diffuse distribution of CD90 and CD105 and remained positive for these markers irrespective of the treatment. However, the CD90 and CD105 mRNA abundance was decreased in TRT-2 and TRT-4; i.e., in media containing no FBS. The presence of FBS increased the absolute number of cell nuclei as assessed by DAPI fluorescence. Our results suggest that bovine subcutaneous preadipocytes display typical stem cell markers. The differentiation into mature adipocytes is promoted by BSL, whereas FBS endorses cell proliferation.


Factors associated with life satisfaction in Parkinson's disease.
To identify factors associated with life satisfaction (LS) in people with Parkinson's disease (PD), including a specific focus on those with late-stage PD. The study included 251 persons with PD (median age 70 years; PD duration 8 years). Analyses involved the total sample and a subsample with late-stage PD, that is Hoehn and Yahr stages IV and V (n=62). LS was assessed with item 1 of the Life Satisfaction Questionnaire (LiSat-11). Simple logistic regression analyses were performed for both the total sample and for the subsample with late-stage PD. For the total sample, a multivariable logistic regression analysis was also performed. In the total sample, 12 of the 20 independent variables were significantly associated with LS: need of help with ADL; walking difficulties; number of non-motor symptoms (NMS); fatigue; depressive symptoms; general self-efficacy; motor symptoms; pain; PD severity; freezing episodes; gender (woman); and fluctuations. When controlling for age and gender in the multivariable logistic regression model, depressive symptoms were negatively associated with high LS and general self-efficacy was positively associated with high LS. In late-stage PD, simple logistic regression analyses (controlling for age and gender) identified the following factors as associated with LS: number of NMS, general self-efficacy, walking difficulties and fatigue. This study provides new knowledge on factors associated with LS in a PD sample, including those with late-stage PD. As the ultimate goal for PD care should be improvement in LS, the results have direct clinical implication.


Genetic analysis of the dTDP-rhamnose biosynthesis region of the Escherichia coli VW187 (O7:K1) rfb gene cluster: identification of functional homologs of rfbB and rfbA in the rff cluster and correct location of the rffE gene.
The O-repeating unit of the Escherichia coli O7-specific lipopolysaccharide is made of galactose, mannose, rhamnose, 4-acetamido-4,6-dideoxyglucose, and N-acetyglucosamine. We have recently characterized the genes involved in the biosynthesis of the sugar precursor GDP-mannose occurring in the E. coli O7:K1 strain VW187 (C. L. Marolda and M. A. Valvano, J. Bacteriol. 175:148-158, 1993). In the present study, we identified and sequenced the rfbBDAC genes encoding the enzymes for the biosynthesis of another precursor, dTDP-rhamnose. These genes are localized on the upstream end of the rfbEcO7 region, and they are strongly conserved compared with similar genes found in various enteric and nonenteric bacteria. Upstream of rfbB we identified a DNA segment containing the rfb promoter and a highly conserved untranslated leader sequence also present in the promoter regions of other surface polysaccharide gene clusters. Also, we have determined that rfbB and rfbA have homologs, rffG (o355) and rffH (o292), respectively, located on the rff cluster, which is involved in the synthesis of enterobacterial common antigen. We provide biochemical evidence that rffG and rffH encode dTDP-glucose dehydratase and glucose-1-phosphate thymidylyltransferase activities, respectively, and we also show that rffG complemented the rfbB defect in the O7+ cosmid pJHCV32. We also demonstrate that rffG is distinct from rffE and map the rffE gene to the second gene of the rff cluster.


DOCK8 is essential for T-cell survival and the maintenance of CD8+ T-cell memory.
Deficiency in the guanine nucleotide exchange factor dedicator of cytokinesis 8 (DOCK8) causes a human immunodeficiency syndrome associated with recurrent sinopulmonary and viral infections. We have recently identified a DOCK8-deficient mouse strain, carrying an ethylnitrosourea-induced splice-site mutation that shows a failure to mature a humoral immune response due to the loss of germinal centre B cells. In this study, we turned to T-cell immunity to investigate further the human immunodeficiency syndrome and its association with decreased peripheral CD4(+) and CD8(+) T cells. Characterisation of the DOCK8-deficient mouse revealed T-cell lymphopenia, with increased T-cell turnover and decreased survival. Egress of mature CD4(+) thymocytes was reduced with increased migration of these cells to the chemokine CXCL12. However, despite the two-fold reduction in peripheral naïve T cells, the DOCK8-deficient mice generated a normal primary CD8(+) immune response and were able to survive acute influenza virus infection. The limiting effect of DOCK8 was in the normal survival of CD8(+) memory T cells after infection. These findings help to explain why DOCK8-deficient patients are susceptible to recurrent infections and provide new insights into how T-cell memory is sustained.


Multimodal prevention of first psychotic episode through N-acetyl-l-cysteine and integrated preventive psychological intervention in individuals clinically at high risk for psychosis: Protocol of a randomized, placebo-controlled, parallel-group trial.
Meta-analyses indicate positive effects of both antipsychotic and cognitive-behavioural interventions in subjects clinically at high risk (CHR) for psychosis in terms of a delay or prevention of psychotic disorders. However, these effects have been limited regarding social functioning and the relative efficacy of both types of interventions remains unclear. Furthermore, neuroprotective substances seem to be a promising alternative agent in psychosis-prevention as they are associated with few and weak side-effects. In this multi-centre randomized controlled trial (RCT), we investigate the effects of two interventions on transition to psychosis and social functioning: (a) an integrated preventive psychological intervention (IPPI) including stress-/symptom-management and social-cognitive remediation; (b) N-acetyl-l-cysteine (NAC) as a pharmacological intervention with glutamatergic, neuroprotective and anti-inflammatory capabilities. This is a double-blind, placebo-controlled RCT with regard to NAC and a single-blind RCT with regard to IPPI using a 2 × 2-factorial design to investigate the individual and combined preventive effects of both interventions. To this aim, a total of 200 CHR subjects will be randomized stratified by site to one of four conditions: (a) IPPI and NAC; (b) IPPI and Placebo; (c) NAC and psychological stress management; (d) Placebo and psychological stress management. Interventions are delivered over 26 weeks with a follow-up period of 12 months. This paper reports on the rationale and protocol of an indicated prevention trial to detect the most effective and tolerable interventions with regard to transition to psychosis as well as improvements in social functioning, and to evaluate the synergistic effects of these interventions.


Assessment of non-clinical attributes in paramedicine using multiple mini-interviews.
Non-clinical attributes are increasingly emphasised as an important factor in paramedic practice. However, the assessment of these attributes often lacks the evidence base to support it. Exploring the relationship between non-clinical attributes and clinical skills is also of theoretical and practical importance. To first seek evidence of reliability and validity for the assessment of non-clinical attributes using the multiple mini-interview (MMI) in paramedic contexts and second, to explore the association between non-clinical attributes and clinical skills in paramedicine. Entry to practice level paramedic candidates completed a 10-station MMI to assess non-clinical attributes on day 1 and a 10-station simulation-based assessment (SBA) of clinical skills on day 2. Both were assessed using different global rating scales. Our primary outcomes included MMI inter-station reliability (calculated using generalisability theory) and Pearson's correlation between non-clinical attributes and clinically focused skills. 30 trainees completed the MMI and 26 of the 30 completed the SBA. Inter-station reliability for the MMI reached 0.77. Pearson's correlations (disattenuated correlations in parentheses) between the overall MMI score and mean SBA global rating scores reached r=0.31 (r=0.48) and ranged by dimension from r=-0.11 (-0.17) (procedural skills) to r=0.54(r=0.83) (communication). The MMI demonstrated evidence of reliability and validity for the assessment of non-clinical attributes in paramedic contexts. Non-clinical attributes and paramedic clinical skills are likely distinct but related constructs, each contributing to the construct of paramedic practice. Programmes of assessment should include both to ensure the construct of paramedic practice is adequately represented.


Long-chain polyunsaturated fatty acids in childhood developmental and psychiatric disorders.
Both omega-3 and omega-6 long-chain PUFA (LC-PUFA) are crucial to brain development and function, but omega-3 LC-PUFA in particular are often lacking in modern diets in developed countries. Increasing evidence, reviewed here, indicates that LC-PUFA deficiencies or imbalances are associated with childhood developmental and psychiatric disorders including ADHD, dyslexia, dyspraxia, and autistic spectrum disorders. These conditions show a high clinical overlap and run in the same families, as well as showing associations with various adult psychiatric disorders in which FA abnormalities are already implicated, such as depression, other mood disorders, and schizophrenia. Preliminary evidence from controlled trials also suggests that dietary supplementation with LC-PUFA might help in the management of these kinds of childhood behavioral and learning difficulties. Treatment with omega-3 FA appears most promising, but the few small studies published to date have involved different populations, study designs, treatments, and outcome measures. Large-scale studies are now needed to confirm the benefits reported. Further research is also required to assess the durability of such treatment effects, to determine optimal treatment compositions and dosages, and to develop reliable ways of identifying those individuals most likely to benefit from this kind of treatment. Childhood developmental and psychiatric disorders clearly reflect multifactorial influences, but the study of LC-PUFA and their metabolism could offer important new approaches to their early identification and management. Heterogeneity and comorbidity are such, however, that a focus on specific traits or symptoms may prove more fruitful than an exclusive reliance on current diagnostic categories.


Dispersion and alignment of nanorods in cylindrical block copolymer thin films.
Although significant progress has been made in controlling the dispersion of spherical nanoparticles in block copolymer thin films, our ability to disperse and control the assembly of anisotropic nanoparticles into well-defined structures is lacking in comparison. Here we use a combination of experiments and field theoretic simulations to examine the assembly of gold nanorods (AuNRs) in a block copolymer. Experimentally, poly(2-vinylpyridine)-grafted AuNRs (P2VP-AuNRs) are incorporated into poly(styrene)-b-poly(2-vinylpyridine) (PS-b-P2VP) thin films with a vertical cylinder morphology. At sufficiently low concentrations, the AuNRs disperse in the block copolymer thin film. For these dispersed AuNR systems, atomic force microscopy combined with sequential ultraviolet ozone etching indicates that the P2VP-AuNRs segregate to the base of the P2VP cylinders. Furthermore, top-down transmission electron microscopy imaging shows that the P2VP-AuNRs mainly lie parallel to the substrate. Our field theoretic simulations indicate that the NRs are strongly attracted to the cylinder base where they can relieve the local stretching of the minority block of the copolymer. These simulations also indicate conditions that will drive AuNRs to adopt a vertical orientation, namely by increasing nanorod length and/or reducing the wetting of the short block towards the substrate.


Regulation of GNRH production by estrogen and bone morphogenetic proteins in GT1-7 hypothalamic cells.
Recent studies have shown that bone morphogenetic proteins (BMPs) are important regulators in the pituitary-gonadal endocrine axis. We here investigated the effects of BMPs on GNRH production controlled by estrogen using murine GT1-7 hypothalamic neuron cells. GT1-7 cells expressed estrogen receptor alpha (ERalpha; ESR1 as listed in MGI Database), ERbeta (ESR2 as listed in MGI Database), BMP receptors, SMADs, and a binding protein follistatin. Treatment with BMP2 and BMP4 had no effect on Gnrh mRNA expression; however, BMP6 and BMP7 significantly increased Gnrh mRNA expression as well as GnRH production by GT1-7 cells. Notably, the reduction of Gnrh expression caused by estradiol (E(2)) was restored by cotreatment with BMP2 and BMP4, whereas it was not affected by BMP6 or BMP7. E(2) activated extracellular signal-regulated kinase (ERK) 1/2 and stress-activated protein kinase/c-Jun NH(2)-terminal kinase (SAPK/JNK) signaling but did not activate p38-mitogen-activated protein kinase (MAPK) signaling in GT1-7 cells. Inhibition of ERK1/ERK2 reversed the inhibitory effect of estrogen on Gnrh expression, whereas SAPK/JNK inhibition did not affect the E(2) actions. Expression levels of Eralpha and Erbeta were reduced by BMP2 and BMP4, but were increased by BMP6 and BMP7. Treatment with an ER antagonist inhibited the E(2) effects on Gnrh suppression including reduction of E(2)-induced ERK phosphorylation, suggesting the involvement of genomic ER actions in Gnrh suppression. BMP2 and BMP4 also suppressed estrogen-induced phosphorylation of ERK1/ERK2 and SAPK/JNK signaling, suggesting that BMP2 and BMP4 downregulate estrogen effects by attenuating ER-MAPK signaling. Considering that BMP6 and BMP7 increased the expression of alpha1E-subunit of R-type calcium channel (Cacna1e), which is critical for GNRH secretion, it is possible that BMP6 and BMP7 directly stimulate GNRH release by GT1-7 cells. Collectively, a newly uncovered interaction of BMPs and ER may be involved in controlling hypothalamic GNRH production and secretion via an autocrine/paracrine mechanism.


Role of β/δ101Gln in regulating the effect of temperature and allosteric effectors on oxygen affinity in woolly mammoth hemoglobin.
The oxygen affinity of woolly mammoth hemoglobin (rHb WM) is less affected by temperature change than that of Asian elephant hemoglobin (rHb AE) or human normal adult hemoglobin (Hb A). We report here a biochemical-biophysical study of Hb A, rHb AE, rHb WM, and three rHb WM mutants with amino acid substitutions at β/δ101 (β/δ101Gln→Glu, Lys, or Asp) plus a double and a triple mutant, designed to clarify the role of the β/δ101 residue. The β/δ101Gln residue is important for responding to allosteric effectors, such as phosphate, inositol hexaphosphate (IHP), and chloride. The rHb WM mutants studied generally have higher affinity for oxygen under various conditions of pH, temperature, and salt concentration, and in the presence or absence of organic phosphate, than do rHb WM, rHb AE, and Hb A. Titrations for the O2 affinity of these mutant rHbs as a function of chloride concentration indicate a lower heterotopic effect of this anion due to the replacement of β/δ101Gln in rHb WM. The alkaline Bohr effect of rHb WM and its mutants is reduced by 20-50% compared to that of Hb A and is independent of changes in temperature, in contrast to what has been observed in the hemoglobins of most mammalian species, including human. The results of our study on the temperature dependence of the O2 affinity of rHb WM and its mutant rHbs illustrate the important role of β/δ101Gln in regulating the functional properties of these hemoglobins.


The PhenoGen informatics website: tools for analyses of complex traits.
With the advent of "omics" (e.g. genomics, transcriptomics, proteomics and phenomics), studies can produce enormous amounts of data. Managing this diverse data and integrating with other biological data are major challenges for the bioinformatics community. Comprehensive new tools are needed to store, integrate and analyze the data efficiently. The PhenoGen Informatics website http://phenogen.uchsc.edu is a comprehensive toolbox for storing, analyzing and integrating microarray data and related genotype and phenotype data. The site is particularly suited for combining QTL and microarray data to search for "candidate" genes contributing to complex traits. In addition, the site allows, if desired by the investigators, sharing of the data. Investigators can conduct "in-silico" microarray experiments using their own and/or "shared" data. The PhenoGen website provides access to tools that can be used for high-throughput data storage, analyses and interpretation of the results. Some of the advantages of the architecture of the website are that, in the future, the present set of tools can be adapted for the analyses of any type of high-throughput "omics" data, and that access to new tools, available in the public domain or developed at PhenoGen, can be easily provided.


Interprofessional psychiatric teams: is multidimensionality evident in treatment conferences?
Interprofessional teamwork is practised when the care needs of patients are complex. Little is known about the extent to which team competence really determines patient interventions. The aim of the study was to examine the degree of multidimensionality in patient discussions in psychiatry, and to how different professions contribute. Psychiatric teams were observed during 30 team meetings. A content analysis was used to examine the amount of attention given to medical, psychological and social aspects. The results indicated difficulties in achieving multidimensionality in patient discussions during team meetings. The descriptive element of the discussion was dominated by the social aspect, to which all professions contributed. The analytical element was dominated by the psychological aspect, also to which all the professions contributed. In suggesting interventions, medical interventions were emphasized, principally by the physicians. Decisions on interventions concerned equally medical, social and psychological aspects. An interprofessional composition of teams offers no guarantee that interventions will be of a multidimensional nature. The results are discussed in relation to previous research and practical implications.


Depression and anxiety during the year before death from cancer.
Previous studies of depression and anxiety during the year before death have reported different findings. We therefore aimed to study depression and anxiety in patients who had died from cancer and had previously attended cancer clinics. We analysed routine data on 4869 deceased patients who had completed the Hospital Anxiety and Depression Scale (HADS) as part of their routine cancer care. The HADS data were linked with demographic, cancer and mortality data from national registries. We used data from all HADS completed in the last year of life to investigate the relationships between mean depression (HADS-D) and anxiety (HADS-A) scores and the percentages of high scores (≥11 on each subscale) and time to death (Analysis 1). This analysis used multivariable linear regression with cubic splines and robust standard errors to allow for multiple HADS from the same patients. We also investigated within-patient changes in scores (Analysis 2) in a subset of patients who had completed more than one HADS. In Analysis 1, modelled mean HADS-D scores increased by around 2.5 and the percentage of high HADS-D scores increased from 13% at six months before death to 30% at one month before death. Changes in HADS-A were smaller and occurred later. In Analysis 2, similar patterns were observed in individual patients' HADS scores. Depression should be looked for and treated in patients with cancer and a prognosis of six months or less, in order to maximise the quality of patients' remaining life.


Sensory neural pathways revisited to unravel the temporal dynamics of the Simon effect: A model-based cognitive neuroscience approach.
The Simon task is one of the most prominent interference tasks and has been extensively studied in experimental psychology and cognitive neuroscience. Despite years of research, the underlying mechanism driving the phenomenon and its temporal dynamics are still disputed. Within the framework of the review, we adopt a model-based cognitive neuroscience approach. We first go over key findings in the literature of the Simon task, discuss competing qualitative cognitive theories and the difficulty of testing them empirically. We then introduce sequential sampling models, a particular class of mathematical cognitive process models. Finally, we argue that the brain architecture accountable for the processing of spatial ('where') and non-spatial ('what') information, could constrain these models. We conclude that there is a clear need to bridge neural and behavioral measures, and that mathematical cognitive models may facilitate the construction of this bridge and work towards revealing the underlying mechanisms of the Simon effect.


Increasing compliance with the World Health Organization Surgical Safety Checklist-A regional health system's experience.
In 2009, NorthShore University HealthSystem adapted the World Health Organization Surgical Safety Checklist (SSC) at each of its 4 hospitals. Despite evidence that SSC reduces intraoperative mistakes and increase patient safety, compliance was found to be low with the paper form. In November 2013, NorthShore integrated the SSC into the electronic health record (EHR). The aim was to increase communication between operating room (OR) personnel and to encourage best practices during the natural workflow of surgeons, anesthesiologists, and nurses. The purpose of this study was to examine the impact of an electronic SSC on compliance and patient safety. An anonymous OR observer selected cases at random and evaluated the compliance rate before the rollout of the electronic SSC. In June 2014, an electronic audit was performed to assess the compliance rate. Random OR observations were also performed throughout the summer in 2014. Perioperative risk events, such as consent issues, incorrect counts, wrong site, and wrong procedure were compared before and after the electronic SSC rollout. A perception survey was also administered to NorthShore OR personnel. Compliance increased from 48% (n = 167) to 92% (n = 1,037; P < .001) after the SSC was integrated into the electronic health record. Surgeons (91% vs 97%; P < .001), anesthesiologists (89% vs 100%; P < .001), and nurses (55% vs 93%; P < .001) demonstrated an increase in compliance. A comparison between risk events in the pre- and post-rollout period showed a 32% decrease (P < .01). Hospital-wide indicators including length of stay and 30-day readmissions were lower. In a survey to assess the OR personnel's perceptions of the new checklist, 76% of surgeons, 86% of anesthesiologists, and 88% of nurses believed the electronic SSC will have a positive impact on patient safety. The World Health Organization SSC is a validated tool to increase patient safety and reduce intraoperative complications. The electronic SSC has demonstrated an increased compliance rate, a reduced number of risk events, and most OR personnel believe it will have a positive impact on patient safety.


The impact of economic factors on the realities of outpatient multi-drug treatment of chronic diseases in rural areas.
The reform of the health care system in Poland introduced in 1999 brought not ony a lot of disappointments arising from the failure in performing its basic assumptions, but also from the deterioration in access to medical care by patients coming from the poorest class of the Polish population. The subjects were 209 people at the age of 52-80 years including 149 women and 60 men - inhabitants of rural areas of Lublin province. These were outpatients who attended the Endocrine Clinic in Poniatowa. The studies were conducted with the use of the standardized survey questionnaire. Of all 209 respondents requiring long-term multidrug therapies for chronic diseases, 128 (61.2% of those surveyed) reported episodic or permanent noncompliance to treatment regimens, 90 people (43%) had applied medications in a way at variance with medical recommendations. Among the reasons, economic factors were most often mentioned. Ony 21 respondents (10%) expressed the opinion that they had always possessed sufficient means to buy necessary drugs. Current regulations concerning the refunding of costs of medicines may make effective outpatient treatment of chronic diseases impossible. Almost 61.2% of the polled treated themselves contrary to medical recommendations. In most cases, improper courses of treatment were caused by financial problems.


Hepatocellular carcinoma: ten years experience among veterans in Puerto Rico.
The aim of our study is to determine the characteristics of hepatocellular carcinoma (HCC) as well as risk factors, demographics, survival rates and the use of diagnostic and therapeutic modalities among veteran patients in Puerto Rico. A retrospective study of 114 patients with Hispanic background and biopsy-proven HCC diagnosed at the VA Caribbean Healthcare System from 1992 to 2002 was performed. Demographics data, Child-Turcotte-Pugh (CTP) score, presence of cirrhosis, viral serology, alcohol and/or other liver diseases history, diagnostic modalities, lesion size, therapy, and overall survival were examined. The mean age was 66.6 years old. 82% had known underlying cirrhosis. 60% had alcoholic liver disease (ALD), 33% positive serology for hepatitis C (HCV) and 21% both. 5.3% had chronic hepatitis B virus (HBV) infection. Additional causes were not present. CTP classification was: A (42%), B (44%) and C (14%). Abdominal CT scan demonstrated most of the lesions, while ultrasound only 57%. Alfa-fetoprotein was diagnostic in 32%. Mean survival was 10.3 months, better for those with CTP score A. Only 42% of the patients received any kind of therapy. ALD is the principal underlying liver disease in our HCC patients, closely followed by chronic HCV infection. Less than half of our patients received treatment mainly due to advanced disease for which the over survival was less than a year. HCC continues to be a dreadful disease with poor prognosis for which aggressive screening should be considered for all patients with cirrhosis and advanced liver disease regardless of the cause.


Protein-based signatures of functional evolution in Plasmodium falciparum.
It has been known for over a decade that Plasmodium falciparum proteins are enriched in non-globular domains of unknown function. The potential for these regions of protein sequence to undergo high levels of genetic drift provides a fundamental challenge to attempts to identify the molecular basis of adaptive change in malaria parasites. Evolutionary comparisons were undertaken using a set of forty P. falciparum metabolic enzyme genes, both within the hominid malaria clade (P. reichenowi) and across the genus (P. chabaudi). All genes contained coding elements highly conserved across the genus, but there were also a large number of regions of weakly or non-aligning coding sequence. These displayed remarkable levels of non-synonymous fixed differences within the hominid malaria clade indicating near complete release from purifying selection (dN/dS ratio at residues non-aligning across genus: 0.64, dN/dS ratio at residues identical across genus: 0.03). Regions of low conservation also possessed high levels of hydrophilicity, a marker of non-globularity. The propensity for such regions to act as potent sources of non-synonymous genetic drift within extant P. falciparum isolates was confirmed at chromosomal regions containing genes known to mediate drug resistance in field isolates, where 150 of 153 amino acid variants were located in poorly conserved regions. In contrast, all 22 amino acid variants associated with drug resistance were restricted to highly conserved regions. Additional mutations associated with laboratory-selected drug resistance, such as those in PfATPase4 selected by spiroindolone, were similarly restricted while mutations in another calcium ATPase (PfSERCA, a gene proposed to mediate artemisinin resistance) that reach significant frequencies in field isolates were located exclusively in poorly conserved regions consistent with genetic drift. Coding sequences of malaria parasites contain prospectively definable domains subject to neutral or nearly neutral evolution on a scale that appears unrivalled in biology. This distinct evolutionary landscape has potential to confound analytical methods developed for other genera. Against this tide of genetic drift, polymorphisms mediating functional change stand out to such an extent that evolutionary context provides a useful signal for identifying the molecular basis of drug resistance in malaria parasites, a finding that is of relevance to both genome-wide and candidate gene studies in this genus.


A kindred with cerebellar ataxia and thermoanalgesia.
To characterise the clinical, neurophysiological, neuropathological and genetic features of a family with cerebellar autosomal dominant ataxia. Patients were submitted to clinical, neuroradiological and neurophysiological examinations. Molecular studies were undertaken to exclude SCAs 1-3, 6-8, 12 and 17. Studies were performed to rule out linkage to SCA4 on chromosome 16, and for all still uncharacterised SCA loci. Neuropathological examination of the proband was performed with immunocytochemistry. These patients presented a late onset cerebellar ataxia with thermoanalgesia and deep sensory loss. Unlike in SCA4, reflexes were preserved. MRI revealed cerebellar, medullar and spinal cord atrophy. Neurophysiological studies showed absence or marked reduction of the sensory nerve action potentials and somatosensory evoked potentials in lower and upper limbs but preservation of the soleus H reflex. No triplet repeat expansion mutations in the studied SCA genes were identified. Our studies ruled out linkage of the disease to the SCA4 locus on chromosome 16 and the remaining reported SCA loci. The neuropathological study of the proband revealed severe loss of Purkinje cells and dentate neurons. The inferior olive and lower cranial nerve nuclei also showed extensive cell loss. Posterior columns and spinocerebellar tracts were demyelinated. Ubiquitin immunoreactive intranuclear inclusions were absent. This kind of cerebellar ataxia, associated with thermoanalgesia as well as deep sensory loss with retained reflexes, does not associate to any known SCA loci. Therefore, we identify and describe a new form of late onset dominant spinocerebellar ataxia.


Simulated drought regimes reveal community resilience and hydrological thresholds for altered decomposition.
Future climate scenarios forecast a 10-50% decline in rainfall in Eastern Amazonia. Altered precipitation patterns may change important ecosystem functions like decomposition through either changes in physical and chemical processes or shifts in the activity and/or composition of species. We experimentally manipulated hydroperiods (length of wet:dry cycles) in a tank bromeliad ecosystem to examine impacts on leaf litter decomposition. Gross loss of litter mass over 112 days was greatest in continuously submersed litter, lowest in continuously dry litter, and intermediate over a range of hydroperiods ranging from eight cycles of 7 wet:7 dry days to one cycle of 56 wet:56 dry days. The resilience of litter mass loss to hydroperiod length is due to a shift from biologically assisted decomposition (mostly microbial) at short wet:dry hydroperiods to physicochemical release of dissolved organic matter at longer wet:dry hydroperiods. Biologically assisted decomposition was maximized at wet:dry hydroperiods falling within the range of ambient conditions (12-22 consecutive dry days) but then declined under prolonged wet:dry hydroperiods (28 and 56 dry days. Fungal:bacterial ratios showed a similar pattern as biologically assisted decomposition to hydroperiod length. Our results suggest that microbial communities confer functional resilience to altered hydroperiod in tank bromeliad ecosystems. We predict a substantial decrease in biological activity relevant to decomposition under climate scenarios that increase consecutive dry days by 1.6- to 3.2-fold in our study area, whereas decreased frequency of dry periods will tend to increase the physicochemical component of decomposition.


Application of Max-SAT-based ATPG to optimal cancer therapy design.
Cancer and other gene related diseases are usually caused by a failure in the signaling pathway between genes and cells. These failures can occur in different areas of the gene regulatory network, but can be abstracted as faults in the regulatory function. For effective cancer treatment, it is imperative to identify faults and select appropriate drugs to treat the faults. In this paper, we present an extensible Max-SAT based automatic test pattern generation (ATPG) algorithm for cancer therapy. This ATPG algorithm is based on Boolean Satisfiability (SAT) and utilizes the stuck-at fault model for representing signaling faults. A weighted partial Max-SAT formulation is used to enable efficient selection of the most effective drug. Several usage cases are presented for fault identification and drug selection. These cases include the identification of testable faults, optimal drug selection for single/multiple known faults, and optimal drug selection for overall fault coverage. Experimental results on growth factor (GF) signaling pathways demonstrate that our algorithm is flexible, and can yield an exact solution for each feature in much less than 1 second.


Sensory processing disorders and social participation.
Participation in social aspects of daily life is crucial to children's development. Although disability status is recognized to affect children's ability to participate in social activities, little is understood about the impact of sensory processing disorders (SPD) on children's social participation. We examined the social participation patterns of 2 groups of children (ages 6-9): (1) children with SPD and (2) their typically developing peers. All children participated in a structured interview to report their social participation patterns, including activity patterns and social networks. We used parent and teacher questionnaires to triangulate the data gathered from the children. Results revealed that the 2 groups of children demonstrated generally similar patterns of activity preferences and use of free time but had significant differences in areas related to intensity and enjoyment of involvement and in their social networks. Implications for future research and interventions are discussed.


Microsurgical management of hypoglossal schwannomas over 3 decades: a modified grading scale to guide surgical approach.
Schwannomas originating from the hypoglossal nerve are extremely rare. Microsurgical resection with the goal for cure has traditionally been associated with a high risk of postoperative deficits. To summarize our clinical experience using tailored cranial base approaches for these formidable lesions. The clinical records of 13 patients were retrospectively reviewed. In addition, all reported patients in the literature were reviewed. The extreme lateral infrajugular transcondylar-transtubercular exposure approach was used in all of our patients. Based on our experience and literature analysis, we propose the following modified grading scale to facilitate surgical planning: type A, intradural tumors; type B, dumbbell-shaped tumors; type C, extracranial tumors; and type D, peripheral tumors. All 13 patients underwent total, near-total, or subtotal tumor resection. Eight patients were men, 5 were women (mean age, 41.7 years). Sural nerve graft reconstruction for the hypoglossal nerve was performed in 4 patients. Three of the 4 patients in whom nerve reconstruction was performed regained satisfactory movement of their tongue. In the review of the literature, the mean patient age was 45.8 years. Patients presented with tongue atrophy (91.6%), headache (60.9%), and dysphagia (31.8%). The tumors were categorized as type A in 31.7% of these patients, type B in 38.6%, type C in 6.2%, and type D in 23.4%. The extreme lateral infrajugular transcondylar-transtubercular exposure approach, which is a modification of the extreme lateral suboccipital approach, provides sufficient exposure for most intracranial dumbbell-shaped hypoglossal schwannomas. Hypoglossal nerve reconstruction using a sural nerve graft improves tongue atrophy and movement for patients with resected nerves.


Evolutionary Analyses of Base-Pairing Interactions in DNA and RNA Secondary Structures.
Pairs of nucleotides within functional nucleic acid secondary structures often display evidence of coevolution that is consistent with the maintenance of base-pairing. Here, we introduce a sequence evolution model, MESSI (Modeling the Evolution of Secondary Structure Interactions), that infers coevolution associated with base-paired sites in DNA or RNA sequence alignments. MESSI can estimate coevolution while accounting for an unknown secondary structure. MESSI can also use graphics processing unit parallelism to increase computational speed. We used MESSI to infer coevolution associated with GC, AU (AT in DNA), GU (GT in DNA) pairs in noncoding RNA alignments, and in single-stranded RNA and DNA virus alignments. Estimates of GU pair coevolution were found to be higher at base-paired sites in single-stranded RNA viruses and noncoding RNAs than estimates of GT pair coevolution in single-stranded DNA viruses. A potential biophysical explanation is that GT pairs do not stabilize DNA secondary structures to the same extent that GU pairs do in RNA. Additionally, MESSI estimates the degrees of coevolution at individual base-paired sites in an alignment. These estimates were computed for a SHAPE-MaP-determined HIV-1 NL4-3 RNA secondary structure. We found that estimates of coevolution were more strongly correlated with experimentally determined SHAPE-MaP pairing scores than three nonevolutionary measures of base-pairing covariation. To assist researchers in prioritizing substructures with potential functionality, MESSI automatically ranks substructures by degrees of coevolution at base-paired sites within them. Such a ranking was created for an HIV-1 subtype B alignment, revealing an excess of top-ranking substructures that have been previously identified as having structure-related functional importance, among several uncharacterized top-ranking substructures.


VO2 Reserve vs. Heart Rate Reserve During Moderate Intensity Treadmill Exercise.
VO2 and heart rate (HR) are widely used when determining appropriate training intensities for clinical, healthy, and athletic populations. It has been shown that if the % reserve (%R) is used, rather than % of max, HR and VO2 can be used interchangeably to accurately prescribe exercise intensities. Thus, heart rate reserve (HRR) can be prescribed if VO2 reserve (VO2R) is known. Therefore, the purpose of this study was to compare VO2 R and HRR during moderate intensity exercise (50%R). Physically active college students performed a maximal treadmill test to exhaustion. During which VO2 and HR were monitored to determine max values. Upon completion of the maximal test, calculations were made to determine the % grade expected to yield approximately 50% of the subjects VO2R. Subjects then returned to complete the submaximal test (50%R) at least two days later. The %VO2R and %HRR were calculated and compared to the predicted value as well as to each other. Statistical analysis revealed that VO2 at 50%R was significantly greater than the actual VO2 achieved, p < .001. Conversely, the mean predicted HR at 50%R was significantly less than the actual HR achieved, p < .001. In conclusion, this study indicated that VO2 could be more accurately predicted than HR during moderate intensity exercise. The weak correlation between VO2R and HRR indicates that caution should be used when relying on a HR to determine VO2.


Association between protein signals and type 2 diabetes incidence.
Understanding early determinants of type 2 diabetes is essential for refining disease prevention strategies. Proteomic technology may provide a useful approach to identify novel protein patterns potentially related to pathophysiological changes that lead up to diabetes. In this study, we sought to identify protein signals that are associated with diabetes incidence in a middle-aged population. Serum samples from 519 participants in a nested case-control selection (167 cases and 352 age-, sex- and BMI-matched normoglycemic control subjects, median follow-up 14.0 years) within the Whitehall-II cohort were analyzed by linear matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Nine protein peaks were found to be associated with incident diabetes. Rate ratios for high peak intensity ranged between 0.4 (95% CI, 0.2-0.8) and 4.0 (95% CI, 1.7-9.2) and were robust to adjustment for main potential confounders, including obesity, lipids and C-reactive protein. The proteins associated with these peaks may reflect diabetes pathogenesis. Our study exemplifies the utility of an approach that combines proteomic and epidemiological data.


Ketogenic Diet for Refractory Childhood Epilepsy: Beyond Seizures Control, the Experience of a Portuguese Pediatric Centre.
Ketogenic diet is a low carbohydrate diet, which can be used as a treatment for refractory childhood epilepsy. The first aim of this study was to evaluate its efficacy, in patients receiving ketogenic diet for at least three months, on epilepsy control, behaviour and awareness. The secondary aims were to evaluate the variation in the number of antiepileptic drugs, reasons for discontinuing the diet and adverse effects. Retrospective analysis of clinical records of patients who underwent ketogenic diet for refractory epilepsy, from October 2007 to January 2018, in a tertiary pediatric hospital. In the twenty-nine eligible patients, the mean age of initiation was 7.9 years-old (+/- 5.6). Of those, 18 had a ≥ 50% reduction of seizure activity, 19 a marked behaviour improvement and 18 improved awareness. The median number of antiepileptic drugs remained equal for the 15 patients who completed 18 months of treatment (three drugs). The main reason for discontinuing ketogenic diet was a familiar decision. The main adverse effects were hypercholesterolemia (n = 23) and hypertriglyceridemia (n = 21). Results were comparable to those of other cohorts, namely age of initiation, proportion of patients completing ketogenic diet, most frequent reasons for stopping and significant improvement of alertness and behavior. Beyond seizure control, patients experienced a marked improvement in behavior and awareness. It is necessary to develop strategies to increase the adherence of families to the diet.


Na(+)/H (+) exchanger isoform 1 induced osteopontin expression in cardiomyocytes involves NFAT3/Gata4.
Osteopontin (OPN), a multifunctional glycophosphoprotein, has been reported to contribute to the development and progression of cardiac remodeling and hypertrophy. Cardiac-specific OPN knockout mice were protected against hypertrophy and fibrosis mediated by Ang II. Recently, transgenic mice expressing the active form of the Na(+)/H(+) exchanger isoform 1 (NHE1) developed spontaneous hypertrophy in association with elevated levels of OPN. The mechanism by which active NHE1 induces OPN expression and contributes to the hypertrophic response remains unclear. To validate whether expression of the active form of NHE1 induces OPN, cardiomyocytes were stimulated with Ang II, a known inducer of both OPN and NHE1. Ang II induced hypertrophy and increased OPN protein expression (151.6 ± 28.19 %, P < 0.01) and NHE1 activity in H9c2 cardiomyoblasts. Ang II-induced hypertrophy and OPN protein expression were regressed in the presence of an NHE1 inhibitor, EMD 87580, or a calcineurin inhibitor, FK506. In addition, our results indicated that activation of NHE1-induced NFAT3 translocation into the nucleus and a significant activation of the transcription factor Gata4 (NHE1: 149 ± 28 % of control, P < 0.05). NHE1-induced activation of Gata4 was inhibited by FK506. In summary, our results suggest that activation of NHE1 induces hypertrophy through the activation of NFAT3/Gata4 and OPN expression.


Responding to Richard: Celebrity and (mis)representation of science.
Drawing on 48 in-depth interviews conducted with biologists and physicists at universities in the United Kingdom, this study examines scientists' perceptions of the role celebrity scientists play in socially contentious public debates. We examine Richard Dawkins' involvement in public debates related to the relationship between science and religion as a case to analyze scientists' perceptions of the role celebrity scientists play in the public sphere and the implications of celebrity science for the practice of science communication. Findings show that Dawkins' proponents view the celebrity scientist as a provocateur who asserts the cultural authority of science in the public sphere. Critics, who include both religious and nonreligious scientists, argue that Dawkins misrepresents science and scientists and reject his approach to public engagement. Scientists emphasize promotion of science over the scientist, diplomacy over derision, and dialogue over ideological extremism.