To be or not to be? A caregiver's question: the lived experience of a stroke family during the first 18 months poststroke.
Disability following a stroke often requires family, commonly a spouse, to provide care enabling the stroke survivor to return home. Immediate or extended family and friends may help provide direct care or support the primary caregiver. While family members share the common stroke experience, this is lived within the context of separate lives. Research examining the individual nuances, roles and contribution of family and/or friends forming part of collective stroke networks, has largely been overlooked. This New Zealand study aimed to explore the lived experience of three stroke family members during the 18 months following a first-ever stroke. Hermeneutic phenomenology guided the study. Informed consent was obtained prior to individual interviews conducted 6 weeks, 12 months and 18 months poststroke. Three main themes emerged: (i) Being prepared, (ii) Where you stand changes the view and (iii) Relinquishing and reclaiming. Being prepared revealed how these family members anticipated the stroke and drew on personal and professional experience in facing the phenomenon. The second theme showed the influence of expectation and positionality on family members' experiences. Relinquishing and reclaiming identified loss, grief and a quest for equity in the synthesis of competing stroke survivor and caregiver desires. Familiarity with the caregiving role and experience of unreliable community services led stroke family members to question the primary caregiver's ability to resume caregiving following a family member's stroke. The primary caregiver was physically and emotionally spent, and rest home care for the survivor became the only self-preserving option. A new caring arrangement was formulated seeking equity for both stroke survivor and caregiver.


Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems.
Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump's bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.


Primary human colonic epithelial cells are transiently protected from anoikis by a Src-dependent mechanism.
Complete loss of cell anchorage triggers apoptosis in primary human colonic epithelial cells (CEC), a phenomenon known as anoikis. Besides the induction of pro-apoptotic events, activation of survival pathways was observed in detached intestinal epithelial cell lines, providing a transient apoptosis protection. However, nothing is known about molecular mechanisms protecting primary CEC from anoikis. In this study intact CEC crypts were isolated and kept in suspension, a condition which leads to the loss of cell-cell anchorage and induces anoikis. To reconstitute cell-cell contacts, cells were centrifuged to form cell aggregates. Induction of apoptosis was assessed by caspase-3 activity assay; activation of survival pathways was analyzed by Western blot. Immediately after loss of cell anchorage a rapid activation of survival proteins was observed before active caspase-3 could be detected. Src hyperactivation significantly contributed to transient protection from anoikis in CEC because its inhibition reversed the protecting effect of re-establishment of cell contacts. Basal levels of active Src in CEC from patients with inflammatory bowel disease were markedly reduced compared to control patients. These results demonstrate that loss of cell anchorage activates survival pathways in primary human CEC providing transient anoikis protection. Src is an important mediator of this mechanism and therefore constitutes a key regulatory molecule coordinating survival signals mediated by cell adhesion in primary human CEC.


Revisiting the Complexity of GLP-1 Action from Sites of Synthesis to Receptor Activation.
Glucagon-like peptide-1 (GLP-1) is produced in gut endocrine cells and in the brain, and acts through hormonal and neural pathways to regulate islet function, satiety, and gut motility, supporting development of GLP-1 receptor (GLP-1R) agonists for the treatment of diabetes and obesity. Classic notions of GLP-1 acting as a meal-stimulated hormone from the distal gut are challenged by data supporting production of GLP-1 in the endocrine pancreas, and by the importance of brain-derived GLP-1 in the control of neural activity. Moreover, attribution of direct vs indirect actions of GLP-1 is difficult, as many tissue and cellular targets of GLP-1 action do not exhibit robust or detectable GLP-1R expression. Furthermore, reliable detection of the GLP-1R is technically challenging, highly method dependent, and subject to misinterpretation. Here we revisit the actions of GLP-1, scrutinizing key concepts supporting gut vs extra-intestinal GLP-1 synthesis and secretion. We discuss new insights refining cellular localization of GLP-1R expression and integrate recent data to refine our understanding of how and where GLP-1 acts to control inflammation, cardiovascular function, islet hormone secretion, gastric emptying, appetite, and body weight. These findings update our knowledge of cell types and mechanisms linking endogenous vs pharmacological GLP-1 action to activation of the canonical GLP-1R, and the control of metabolic activity in multiple organs.


Use of cerebrospinal fluid and serum samples impregnated on FTATM Elute filter paper for the diagnosis of infections caused by Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae.
The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTATM Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo's national surveillance for routine diagnosis were selected for this study. The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory's standard DNA extraction, and processing the same samples after being dried and stored on FTATM card, and DNA extracted following the manufacturer's instructions. The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTATM cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory's standard methodology, results showed high concordance, with Kappa index ranges of 0.9877-1.00 for CSF, and 0.8004-1.00 for serum samples. The use of FTATM cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost.


Rice-eating pattern and the risk of metabolic syndrome especially waist circumference in Korean Genome and Epidemiology Study (KoGES).
Metabolic syndrome poses a serious health threat in Asian countries. Rice is a staple food in Korea, and carbohydrate intake is associated with the risk of MetS. We hypothesized that various rice-eating patterns in a carbohydrate-based diet would have different effects on the risk of MetS. Participants were 26,006 subjects enrolled in the Korean Genome and Epidemiology Study between 2004 and 2006. They were classified into four dietary patterns - white rice, rice with beans, rice with multi-grains, and mixed based on their food frequency questionnaire responses. We compared metabolic risk traits according to the rice-eating patterns. Nutrients consumption and the presence of MetS risk factors differed according to rice-eating patterns. In men odds ratio(OR) for central obesity was slightly elevated in mixed group(1.18). In women, the risk for central obesity and abnormal fasting glucose were lower in the rice with beans group (adjusted OR =0.79, 0.83 respectively) and central obesity in rice with multi-grains(adjusted OR=0.91) than the white rice group. In postmenopausal women, ORs for central obesity (0.78) and abnormal fasting glucose (0.75) in the rice with beans group and ORs for central obesity (0.83), abnormal HDL-cholesterol (0.87) and MetS(0.85) in the rice with multi-grains group was lower than those in white rice group. In premenopausal women, the risk for central obesity (OR=0.77) was reduced in the rice with beans group. The risk for MetS was lower in the rice with beans and rice with multi-grains groups compared with the white rice group, particularly in postmenopausal women.


Spatial properties of neurons in the monkey striate cortex.
Contrast sensitivity as a function of spatial frequency was determined for 138 neurons in the foveal region of primate striate cortex. The accuracy of three models in describing these functions was assessed by the method of least squares. Models based on difference-of-Gaussians (DOG) functions where shown to be superior to those based on the Gabor function or the second differential of a Gaussian. In the most general case of the DOG models, each subregion of a simple cell's receptive field was constructed from a single DOG function. All the models are compatible with the classical observation that the receptive fields of simple cells are made up of spatially discrete 'on' and 'off' regions. Although the DOG-based models have more free parameters, they can account better for the variety of shapes of spatial contrast sensitivity functions observed in cortical cells and, unlike other models, they provide a detailed description of the organization of subregions of the receptive field that is consistent with the physiological constraints imposed by earlier stages in the visual pathway. Despite the fact that the DOG-based models have spatially discrete components, the resulting amplitude spectra in the frequency domain describe complex cells just as well as simple cells. The superiority of the DOG-based models as a primary spatial filter is discussed in relation to popular models of visual processing that use the Gabor function or the second differential of a Gaussian.


Retinoblastoma: correlation of invasion of the optic nerve and choroid with prognosis and metastases.
The histological sections of 74 retinoblastoma patients were reviewed. The extent of optic nerve invasion was correlated with choroidal/scleral extension. Choroidal invasion carries 100% survival provided that the sclera, iris, and whole optic nerve are not also involved. Invasion of the optic nerve beyond the lamina cribrosa also carries 100% survival provided that the resection line is free and that invasion does not involve the sclera or iris. Plentiful rosettes were usually found in those tumours that had not extended beyond the choroid or as far as the resection line of the optic nerve and were therefore associated with a good prognosis. The absence of rosettes did not necessarily indicate a poor prognosis. The site of metastases was related to optic nerve and scleral/extrascleral extension. The various systems of staging retinoblastoma are compared, and a new system is proposed to cover the whole spectrum of the disease and to offer a reliable guide to prognosis and treatment.


Evidence for myoblast-extrinsic regulation of slow myosin heavy chain expression during muscle fiber formation in embryonic development.
Vertebrate muscles are composed of an array of diverse fast and slow fiber types with different contractile properties. Differences among fibers in fast and slow MyHC expression could be due to extrinsic factors that act on the differentiated myofibers. Alternatively, the mononucleate myoblasts that fuse to form multinucleated muscle fibers could differ intrinsically due to lineage. To distinguish between these possibilities, we determined whether the changes in proportion of slow fibers were attributable to inherent differences in myoblasts. The proportion of fibers expressing slow myosin heavy chain (MyHC) was found to change markedly with time during embryonic and fetal human limb development. During the first trimester, a maximum of 75% of fibers expressed slow MyHC. Thereafter, new fibers formed which did not express this MyHC, so that the proportion of fibers expressing slow MyHC dropped to approximately 3% of the total by midgestation. Several weeks later, a subset of the new fibers began to express slow MyHC and from week 30 of gestation through adulthood, approximately 50% of fibers were slow. However, each myoblast clone (n = 2,119) derived from muscle tissues at six stages of human development (weeks 7, 9, 16, and 22 of gestation, 2 mo after birth and adult) expressed slow MyHC upon differentiation. We conclude from these results that the control of slow MyHC expression in vivo during muscle fiber formation in embryonic development is largely extrinsic to the myoblast. By contrast, human myoblast clones from the same samples differed in their expression of embryonic and neonatal MyHCs, in agreement with studies in other species, and this difference was shown to be stably heritable. Even after 25 population doublings in tissue culture, embryonic stage myoblasts did not give rise to myoblasts capable of expressing MyHCs typical of neonatal stages, indicating that stage-specific differences are not under the control of a division dependent mechanism, or intrinsic "clock." Taken together, these results suggest that, unlike embryonic and neonatal MyHCs, the expression of slow MyHC in vivo at different developmental stages during gestation is not the result of commitment to a distinct myoblast lineage, but is largely determined by the environment.


Single domain intracellular antibodies from diverse libraries: emphasizing dual functions of LMO2 protein interactions using a single VH domain.
Interfering intracellular antibodies are valuable for biological studies as drug surrogates and as potential macromolecular drugs per se. Their application is still limited because of the difficulty of acquisition of functional intracellular antibodies. We describe the use of the new intracellular antibody capture procedure (IAC(3)) to facilitate direct isolation of functional single domain antibody fragments using four independent target molecules (LMO2, TP53, CRAF1, and Hoxa9) from a set of diverse libraries. Initially, these have variability in only one of the three antigen-binding CDR regions of VH or VL and first round single domains are affinity matured by iterative randomization of the two other CDRs and reselection. We highlight the approach using a single domain binding to LMO2 protein. Our results show that interfering with LMO2 protein function demonstrates a role specifically in erythroid differentiation, confirm a necessary and sufficient function for LMO2 as a cancer therapy target in T-cell neoplasia and allowed for the first time production of soluble recombinant LMO2 protein by co-expression with intracellular domain antibodies. Co-crystallization of LMO2 and the anti-LMO2 VH protein was successful. These results demonstrate that this third generation IAC(3) offers a robust toolbox for various biomedical applications and consolidates functional features of the LMO2 protein complex, which includes the importance of Lmo2-Ldb1 protein interaction.


Structural basis for integration of GluD receptors within synaptic organizer complexes.
Ionotropic glutamate receptor (iGluR) family members are integrated into supramolecular complexes that modulate their location and function at excitatory synapses. However, a lack of structural information beyond isolated receptors or fragments thereof currently limits the mechanistic understanding of physiological iGluR signaling. Here, we report structural and functional analyses of the prototypical molecular bridge linking postsynaptic iGluR δ2 (GluD2) and presynaptic β-neurexin 1 (β-NRX1) via Cbln1, a C1q-like synaptic organizer. We show how Cbln1 hexamers "anchor" GluD2 amino-terminal domain dimers to monomeric β-NRX1. This arrangement promotes synaptogenesis and is essential for D: -serine-dependent GluD2 signaling in vivo, which underlies long-term depression of cerebellar parallel fiber-Purkinje cell (PF-PC) synapses and motor coordination in developing mice. These results lead to a model where protein and small-molecule ligands synergistically control synaptic iGluR function.


Increased activation of the hypoxia-inducible factor pathway in varicose veins.
Venous hypoxia has been postulated to contribute to varicose vein (VV) formation. Direct measurements of vein wall oxygen tension have previously demonstrated that the average minimum oxygen tensions were significantly lower in VVs compared with non-varicose veins (NVVs). Hypoxia-inducible factors (HIFs) are nuclear transcriptional factors that regulate the expression of several genes of oxygen homeostasis. This study aimed to investigate if hypoxia was associated with VVs by assessing the expression of HIF-1α, HIF-2α, HIF target genes, and upstream HIF regulatory enzymes in VVs and NVVs, and their regulation by hypoxia. VVs and NVVs were surgically retrieved and immediately snap-frozen or used for organ culture preparation. The relative expression of HIF-1α, HIF-2α, HIF target genes, and HIF regulatory enzymes in VVs and NVVs was analyzed with quantitative polymerase chain reaction (Q-PCR) and Western blot. VV and NVV organ ex vivo cultures were exposed to 16 hours of normoxia, hypoxia (oxygen tension 1%), or the hypoxia mimetic dimethyloxallyl glycine (DMOG) 1 mM in normoxia. The vein organ cultures were then analyzed for HIF-1α, HIF-2α, and their target gene expression with Q-PCR and Western blot. HIF-1α and HIF-2α mRNA were significantly upregulated in VVs compared with NVVs (89.8 ± 18.6 vs 10.4 ± 7.2 and 384.9 ± 209.4 vs 8.1 ± 4.2, respectively). HIF target gene mRNA expression was also significantly elevated in VVs compared with NVVs, namely glucose transporter-1 (GLUT-1; 8.7 ± 2.1 vs 1.0 ± 0.3), carbonic anhydrase-9 (CA9; 8.5 ± 2.1 vs 2.8 ± 1.2), vascular endothelial growth factor (VEGF; 7.5 ± 2.1 vs 0.9 ± 0.2), and BCL2/adenovirus E1B 19-kDa protein-interacting protein 3 (BNIP-3; 4.5 ± 0.7 vs 1.4 ± 0.3). The upregulation of HIF-1α, HIF-2α, and HIF target genes in VVs was also reflected at protein level. Of the HIF regulatory enzymes, the expression of prolyl-hydroxylase domain (PHD)-2 and PHD-3 was found to be elevated in VVs compared with NVVs. Exposure of VV and NVV organ cultures to hypoxia or DMOG was associated with increases in HIF-1α and HIF-2α protein and HIF target gene expression compared with normoxia only. The study concluded, we believe for the first time, an increased activation of the HIF pathway, with upregulation of the expression of HIF-1α and HIF-2α transcription factors, and HIF target genes, in VVs compared with NVVs. Exposure of VVs and NVVs to hypoxic conditions was associated with increased expression of HIF-1α and HIF-2α protein and HIF target genes. The data suggest that the HIF pathway may be associated with several pathophysiologic changes in the VV wall, and that hypoxia may be a feature contributing to VV pathogenesis.


Replacing commercial concentrate by Ficus thonningii improved productivity of goats in Ethiopia.
Ficus thonningii (FT) is an important multipurpose fodder tree providing economic and ecological benefits across arid and semi-arid areas in Africa. Despite its availability in many Sub-Saharan African countries, there is lack of information on its effect on animal productivity. Twenty-four male weaned highland goats of age 7 ± 1.5 months were randomly assigned to four dietary treatments in a completely randomized block design. All animals were fed wheat straw, water, and salt licks ad libitum. The control group (T1) was fed concentrate mixture at 2 % of their body weight, while in T2, T3, and T4, 25, 50, and 75 % of the weight of concentrate (DM basis), respectively, was replaced by sun dried FT leaf meal. FT leaf meal had acceptable levels crude protein (CP), neutral detergent fiber (NDF), and tannins and has resulted in increased body weight in all treatments. Goats fed T3 diet showed significantly higher (P < 0.05) values in terms of feed intake, body weight gain, some carcass attributes, and local meat quality indicators than the rest of the treatments. An increase in proportion of FT leaf meal beyond 50 %, however, resulted in decreased body weight gain, and other carcass parameters, despite increased feed intake. Therefore, F. thonningii can be used to replace commercial concentrate mixture up to 50 % to improve feed intake and productivity of Ethiopian highland goats.


Biochemical evidence for the requirement of Hoogsteen base pairing for replication by human DNA polymerase iota.
Because of the near geometric identity of Watson-Crick (W-C) GxC and AxT base pairs, a given DNA polymerase forms the four possible correct base pairs with nearly identical catalytic efficiencies. However, human DNA polymerase iota (Pol iota), a member of the Y family of DNA polymerases, exhibits a marked template specificity, being more efficient at incorporating the correct nucleotide opposite template purines than opposite pyrimidines. By using 7-deazaadenine and 7-deazaguanine as the templating residues, which disrupt Hoogsteen base pair formation, we show that, unlike the other DNA polymerases belonging to the A, B, or Y family, DNA synthesis by Pol iota is severely inhibited by these N7-modified bases. These observations provide biochemical evidence that, during normal DNA synthesis, template purines adopt a syn conformation in the Pol iota active site, enabling the formation of a Hoogsteen base pair with the incoming pyrimidine nucleotide. Additionally, mutational studies with Leu-62, which lies in close proximity to the templating residue in the Pol iota ternary complex, have indicated that both factors, steric constraints within the active site and the stability provided by the hydrogen bonds in the Hoogsteen base pair, contribute to the efficiency of correct nucleotide incorporation opposite template purines by Pol iota.


Cross-national patterns of gender differences in mathematics: a meta-analysis.
A gender gap in mathematics achievement persists in some nations but not in others. In light of the underrepresentation of women in careers in science, technology, mathematics, and engineering, increasing research attention is being devoted to understanding gender differences in mathematics achievement, attitudes, and affect. The gender stratification hypothesis maintains that such gender differences are closely related to cultural variations in opportunity structures for girls and women. We meta-analyzed 2 major international data sets, the 2003 Trends in International Mathematics and Science Study and the Programme for International Student Assessment, representing 493,495 students 14-16 years of age, to estimate the magnitude of gender differences in mathematics achievement, attitudes, and affect across 69 nations throughout the world. Consistent with the gender similarities hypothesis, all of the mean effect sizes in mathematics achievement were very small (d < 0.15); however, national effect sizes showed considerable variability (ds = -0.42 to 0.40). Despite gender similarities in achievement, boys reported more positive math attitudes and affect (ds = 0.10 to 0.33); national effect sizes ranged from d = -0.61 to 0.89. In contrast to those of previous tests of the gender stratification hypothesis, our results point to specific domains of gender equity responsible for gender gaps in math. Gender equity in school enrollment, women's share of research jobs, and women's parliamentary representation were the most powerful predictors of cross-national variability in gender gaps in math. Results are situated within the context of existing research demonstrating apparently paradoxical effects of societal gender equity and highlight the significance of increasing girls' and women's agency cross-nationally.


A case of parietal lobe atrophy.
Bilateral parietal lobe atrophy was revealed by CT scan in a 48 year-old man, with a history of learning difficulties since childhood, encopretic up to the age of eight, and left-handedness. Nevertheless, he earned his living as a skilled potter, though unable to cope with the administration of the business.


Understanding state variation in health insurance dynamics can help tailor enrollment strategies for ACA expansion.
The number and types of people who become eligible for and enroll in the Affordable Care Act's (ACA's) health insurance expansions will depend in part on the factors that cause people to become uninsured for different lengths of time. We used a small-area estimation approach to estimate differences across states in percentages of adults losing health insurance and in lengths of their uninsured spells. We found that nearly 50 percent of the nonelderly adult population in Florida, Nevada, New Mexico, and Texas--but only 18 percent in Massachusetts and 22 percent in Vermont--experienced an uninsured spell between 2009 and 2012. Compared to people who lost private coverage, those with public insurance were more likely to experience an uninsured spell, but their spells of uninsurance were shorter. We categorized states based on estimated incidence of uninsured spells and the spells' duration. States should tailor their enrollment outreach and retention efforts for the ACA's coverage expansions to address their own mix of types of coverage lost and durations of uninsured spells.


The pertussis epidemic: informing strategies for prevention of severe disease.
To assess the impact of Bordetella pertussis infections in South Australia during an epidemic and determine vulnerable populations, data from notification reports for pertussis cases occurring between July 2008 and December 2009 were reviewed to determine the distribution of disease according to specific risk factors and examine associations with hospitalizations. Although the majority (66%) of the 6230 notifications for pertussis occurred in adults aged >24 years, the highest notification and hospitalization rate occurred in infants aged <1 year. For these infants, factors associated with hospitalization included being aged <2 months [relative risk (RR) 2·3, 95% confidence interval (CI) 1·60-3·32], Indigenous ethnicity (RR 1·7, 95% CI 1·03-2·83) and receiving fewer than two doses of pertussis vaccine (RR 4·1, 95% CI 1·37-12·11). A combination of strategies aimed at improving direct protection for newborns, vaccination for the elderly, and reducing transmission from close contacts of infants are required for prevention of severe pertussis disease.


Curcumin Attenuates Iron Accumulation and Oxidative Stress in the Liver and Spleen of Chronic Iron-Overloaded Rats.
Iron overload is now recognized as a health problem in industrialized countries, as excessive iron is highly toxic for liver and spleen. The potential use of curcumin as an iron chelator has not been clearly identified experimentally in iron overload condition. Here, we evaluate the efficacy of curcumin to alleviate iron overload-induced hepatic and splenic abnormalities and to gain insight into the underlying mechanisms. Three groups of male adult rats were treated as follows: control rats, rats treated with iron in a drinking water for 2 months followed by either vehicle or curcumin treatment for 2 more months. Thereafter, we studied the effects of curcumin on iron overload-induced lipid peroxidation and anti-oxidant depletion. Treatment of iron-overloaded rats with curcumin resulted in marked decreases in iron accumulation within liver and spleen. Iron-overloaded rats had significant increases in malonyldialdehyde (MDA), a marker of lipid peroxidation and nitric oxide (NO) in liver and spleen when compared to control group. The effects of iron overload on lipid peroxidation and NO levels were significantly reduced by the intervention treatment with curcumin (P<0.05). Furthermore, the endogenous anti-oxidant activities/levels in liver and spleen were also significantly decreased in chronic iron overload and administration of curcumin restored the decrease in the hepatic and splenic antioxidant activities/levels. Our study suggests that curcumin may represent a new horizon in managing iron overload-induced toxicity as well as in pathological diseases characterized by hepatic iron accumulation such as thalassemia, sickle cell anemia, and myelodysplastic syndromes possibly via iron chelation, reduced oxidative stress derived lipid peroxidation and improving the body endogenous antioxidant defense mechanism.


MUC16 contributes to the metastasis of pancreatic ductal adenocarcinoma through focal adhesion mediated signaling mechanism.
MUC16, a heavily glycosylated type-I transmembrane mucin is overexpressed in several cancers including pancreatic ductal adenocarcinoma (PDAC). Previously, we have shown that MUC16 is significantly overexpressed in human PDAC tissues. However, the functional consequences and its role in PDAC is poorly understood. Here, we show that MUC16 knockdown decreases PDAC cell proliferation, colony formation and migration in vitro. Also, MUC16 knockdown decreases the tumor formation and metastasis in orthotopic xenograft mouse model. Mechanistically, immunoprecipitation and immunofluorescence analyses confirms MUC16 interaction with galectin-3 and mesothelin in PDAC cells. Adhesion assay displayed decreased cell attachment of MUC16 knockdown cells with recombinant galectin-1 and galectin-3 protein. Further, CRISPR/Cas9-mediated MUC16 knockout cells show decreased tumor-associated carbohydrate antigens (T and Tn) in PDAC cells. Importantly, carbohydrate antigens were decreased in the region that corresponds to MUC16 and suggests for the decreased MUC16-galectin interactions. Co-immunoprecipitation also revealed a novel interaction between MUC16 and FAK in PDAC cells. Interestingly, we observed decreased expression of mesenchymal and increased expression of epithelial markers in MUC16-silenced cells. Additionally, MUC16 loss showed a decreased FAK-mediated Akt and ERK/MAPK activation. Altogether, these findings suggest that MUC16-focal adhesion signaling may play a critical role in facilitating PDAC growth and metastasis.


Skeletal Muscle Abnormalities and Iron Deficiency in Chronic Heart FailureAn Exercise 31P Magnetic Resonance Spectroscopy Study of Calf Muscle.
Heart failure (HF) is often associated with iron deficiency (ID). Skeletal muscle abnormalities are common in HF, but the potential role of ID in this phenomenon is unclear. In addition to hemopoiesis, iron is essential for muscle bioenergetics. We examined whether energetic abnormalities in skeletal muscle in HF are affected by ID and if they are responsive to intravenous iron. Forty-four chronic HF subjects and 25 similar healthy volunteers underwent 31P magnetic resonance spectroscopy of calf muscle at rest and during exercise (plantar flexions). Results were compared between HF subjects with or without ID. In 13 ID-HF subjects, examinations were repeated 1 month after intravenous ferric carboxymaltose administration (1000 mg). As compared with controls, HF subjects displayed lower resting high-energy phosphate content, lower exercise pH, and slower postexercise PCr recovery. Compared with non-ID HF, ID-HF subjects had lower muscle strength, larger PCr depletion, and more profound intracellular acidosis with exercise, consistent with an earlier metabolic shift to anaerobic glycolysis. The exercise-induced PCr drop strongly correlated with pH change in HF group ( r=-0.71, P<0.001) but not in controls ( r=0.13, P=0.61, interaction: P<0.0001). Short-term iron administration corrected the iron deficit but had no effect on muscle bioenergetics assessed 1 month later. HF patients display skeletal muscle myopathy that is more severe in those with iron deficiency. The presence of ID is associated with greater acidosis with exercise, which may explain early muscle fatigue. Further study is warranted to identify the strategy to restore iron content in skeletal muscle.


The effect of trauma center care on pediatric injury mortality in California, 1999 to 2011.
Trauma centers (TCs) have been shown to decrease mortality in adults, but this has not been demonstrated at a population level in all children. We hypothesized that seriously injured children would have increased survival in a TC versus nontrauma center (nTC), but there would be no increased benefit from pediatric-designated versus adult TC care. This was a retrospective study of the unmasked California Office of Statewide Health and Planning Department patient discharge database (1999-2011). DRG International Classification of Diseases-9th Rev. (ICD-9) diagnostic codes indicating trauma were identified for children (0-18 years), and injury severity was calculated from ICD-9 codes using validated algorithms. To adjust for hospital case mix, we selected patients with ICD-9 codes that were capable of causing death and which appeared at both TCs and nTCs. Instrumental variable (IV) analysis using differential distance between the child's residence to a TC and to the nearest hospital was applied to further adjust for unobservable differences in TC and nTC populations. Instrumental variable regression models analyzed the association between mortality and TC versus nTC care as well as for pediatric versus adult TC designations, adjusting for demographic and clinical variables. Unadjusted mortality for the entire population of children with nontrivial trauma (n = 445,236) was 1.2%. In the final study population (n = 77,874), mortality was 5.3%, 3.8% in nTCs and 6.1% in TCs. IV regression analysis demonstrated a 0.79 percentage point (95% confidence interval, -0.80 to -0.30; p = 0.044) decrease in mortality for children cared for in TC versus nTC. No decrease in mortality was demonstrated for children cared for in pediatric versus adult TCs. Our IV TC outcome models use improved injury severity and case mix adjustment to demonstrate decreased mortality for seriously injured California children treated in TCs. These results can be used to take evidence-based steps to decrease disparities in pediatric access to, and subsequent outcomes for, trauma care. Therapeutic/care management, level III.


Rapid Viral Diagnosis of Orthopoxviruses by Electron Microscopy: Optional or a Must?
Diagnostic electron microscopy (DEM) was an essential component of viral diagnosis until the development of highly sensitive nucleic acid amplification techniques (NAT). The simple negative staining technique of DEM was applied widely to smallpox diagnosis until the world-wide eradication of the human-specific pathogen in 1980. Since then, the threat of smallpox re-emerging through laboratory escape, molecular manipulation, synthetic biology or bioterrorism has not totally disappeared and would be a major problem in an unvaccinated population. Other animal poxviruses may also emerge as human pathogens. With its rapid results (only a few minutes after arrival of the specimen), no requirement for specific reagents and its "open view", DEM remains an important component of virus diagnosis, particularly because it can easily and reliably distinguish smallpox virus or any other member of the orthopoxvirus (OPV) genus from parapoxviruses (PPV) and the far more common and less serious herpesviruses (herpes simplex and varicella zoster). Preparation, enrichment, examination, internal standards and suitable organisations are discussed to make clear its continuing value as a diagnostic technique.


c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells.
The emergence of anti-estrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease. Identifying factors that convey cell survival in this setting may guide improvements in treatment. Estrogen (E2) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods (MCF-7:5C cells), but the mechanisms underlying E2-induced stress in this setting have not been elucidated. Here, we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor (ER, ESR1) in its activation of stress responses induced by E2 in MCF-7:5C cells. E2 elevated phosphorylation of c-Src, which was blocked by 4-hydroxytamoxifen (4-OHT), suggesting that E2 activated c-Src through the ER. We found that E2 activated the sensors of the unfolded protein response (UPR), IRE1α (ERN1) and PERK kinase (EIF2AK3), the latter of which phosphorylates eukaryotic translation initiation factor-2α (eIF2α). E2 also dramatically increased reactive oxygen species production and upregulated expression of heme oxygenase HO-1 (HMOX1), an indicator of oxidative stress, along with the central energy sensor kinase AMPK (PRKAA2). Pharmacologic or RNA interference-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK, blocked E2-induced ROS production, and inhibited E2-induced apoptosis. Together, our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis. This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer.


Performance on WISC-III and WAIS-R NI vocabulary subtests assessed with event-related brain potentials: an innovative method of assessment.
Assessment of intellectual functions is essential for rehabilitative interventions with traumatic brain injury and stroke patients. However, many of these patients are difficult or impossible to assess because of aphasia and physical disabilities that prevent their performance on traditionally administered tests of mental functioning. We present normative evidence obtained from healthy individuals for the efficacy of a new method of assessing such patients using standardized and validated tests formatted for computer presentation and using direct measures of brain activity (event-related brain potentials, ERP) as the measurable response instead of verbal or motor behavior.


A prospective study of dietary calcium, dairy products and prostate cancer risk (Finland).
High dietary intakes of calcium and dairy products have been hypothesized to enhance prostate cancer risk, but available prospective data regarding these associations are inconsistent. We examined dietary intakes of calcium and dairy products in relation to risk of prostate cancer in the Alpha-Tocopherol, Beta-Carotene (ATBC) Cancer Prevention Study, a cohort of 29,133 male smokers aged 50-69 years at study entry. Dietary intake was assessed at baseline using a validated 276-item food use questionnaire. Cox proportional hazards regression was used to adjust for known or suspected risk factors for prostate cancer. During 17 years of follow-up, we ascertained 1,267 incident cases of prostate cancer. High versus low intake of dietary calcium was associated with a marked increase in prostate cancer risk. The multivariate relative risk (RR) of prostate cancer for > or =2,000 mg/day compared to <1,000 mg/day of calcium intake was 1.63 (95% confidence interval (CI), 1.27-2.10; p trend < 0.0001). Total dairy intake was also positively associated with risk of prostate cancer. The multivariate RR of prostate cancer comparing extreme quintiles of intake was 1.26 (95% CI, 1.04-1.51; p trend = 0.03). However, no association with total dairy intake remained after we adjusted for calcium (p trend = 0.17). Findings were similar by stage and grade of prostate cancer. The results from this large prospective study suggest that intake of calcium or some related component contained in dairy foods is associated with increased prostate cancer risk.


Obstructive sleep apneas in relation to severity of cervical spinal cord injury.
Thirty-three subjects (28 men, five women) with complete or incomplete cervical cord injury representing a wide range of neurological impairment were investigated with regard to the prevalence of Obstructive Sleep Apnea (OSA). The relation between OSA and neurological function, respiratory capacity, body mass index and symptoms associated with OSA were studied. Overnight sleep recordings employed combined oximetry and respiratory movement monitoring. Pulmonary function tests included static and dynamic spirometry, maximal static inspiratory and expiratory pressures at the mouth. The subjects answered a questionnaire concerning sleep quality and tiredness. The prevalence of OSA was 15% (5/33) in this nonobese cervical cord injury study population. Nine percent of the subjects (3/33) fulfilled the criteria for obstructive sleep apnea syndrome, but daytime sleepiness or fatigue were also common in subjects without OSA. There was an inverse correlation between oxygen desaturation index and American Spinal Injury Association (ASIA) motor score in the subjects with complete injury, while there was no such correlation in the whole study group. There were significant correlations between maximal inspiratory and expiratory pressures and vital capacity and between ASIA motor score and vital capacity.


Using CPAP in COVID-19 patients outside of the intensive care setting: a comparison of survival and outcomes between dialysis and non-dialysis dependent patients.
SARS-CoV-2 (COVID-19) is a novel coronavirus associated with high mortality rates. The use of Continuous Positive Airway Pressure (CPAP) has been recognised as a management option for severe COVID-19 (NHS, Specialty guides for patient management during the coronavirus pandemic Guidance for the role and use of non-invasive respiratory support in adult patients with coronavirus (confirmed or suspected), https://www.nice.org.uk/guidance/ng159 ). We offered ward-based CPAP to COVID-19, dialysis patients not suitable for escalation to ICU. The aim of the study was to evaluate the use of CPAP for COVID-19 dialysis patients compared to non-dialysis COVID-19 patients outside of the intensive care setting. We further aimed to investigate factors associated with improved outcomes. Data was collected from a single centre (Royal Preston Hospital, UK), from March to June 2020. Treatment outcomes were compared for dialysis and non-dialysis dependent patients who received CPAP with limitations on their escalation and resuscitation status. Kaplan-Meier survival curves and Cox regression models were used to compare outcomes. The primary study outcome was 30 day mortality. Confounders including length of admission, systemic anticoagulation and ultrafiltration volumes on dialysis were also analysed. Over the study period, 40 dialysis patients tested positive for COVID-19, with 30 requiring hospital admission. 93% (n = 28) required supplementary oxygen and 12% (n = 9) required CPAP on the ward. These patients were compared to a serial selection of 14 non-dialysis patients treated with CPAP during the same period. Results showed a significant difference in 30 day survival rates between the two groups: 88.9% in the dialysis group vs. 21.4% in the non-dialysis group. Statistical modelling showed that anticoagulation was also an important factor and correlated with better outcomes. This is to the best of our knowledge, the largest series of COVID-19 dialysis patients treated with CPAP in a ward-based setting. In general, dialysis dependent patients have multiple co-morbidities including cardiovascular disease and diabetes mellitus making them vulnerable to COVID-19 and not always suitable for treatment in ICU. We showed a significantly lower 30 day mortality rate with the use of CPAP in the dialysis group (11.1%) compared to the non-dialysis group (78.6%). Despite a small sample size, we believe this study provides impetus for further work clarifying the role of CPAP in treating COVID-19 dialysis dependent patients.


Norwegian e-Infrastructure for Life Sciences (NeLS).
The Norwegian e-Infrastructure for Life Sciences (NeLS) has been developed by ELIXIR Norway to provide its users with a system enabling data storage, sharing, and analysis in a project-oriented fashion. The system is available through easy-to-use web interfaces, including the Galaxy workbench for data analysis and workflow execution. Users confident with a command-line interface and programming may also access it through Secure Shell (SSH) and application programming interfaces (APIs). NeLS has been in production since 2015, with training and support provided by the help desk of ELIXIR Norway. Through collaboration with NorSeq, the national consortium for high-throughput sequencing, an integrated service is offered so that sequencing data generated in a research project is provided to the involved researchers through NeLS. Sensitive data, such as individual genomic sequencing data, are handled using the TSD (Services for Sensitive Data) platform provided by Sigma2 and the University of Oslo. NeLS integrates national e-infrastructure storage and computing resources, and is also integrated with the SEEK platform in order to store large data files produced by experiments described in SEEK. In this article, we outline the architecture of NeLS and discuss possible directions for further development.


Are participant characteristics from ISCOLE study sites comparable to the rest of their country?
The International Study of Childhood Obesity, Lifestyle and the Environment (ISCOLE) provides robust, multi-national information on physical activity, diet and weight status in 9-11-year-old children around the world. The purpose of this analysis was to examine the similarities and differences between participant characteristics from ISCOLE sites and data from nationally representative surveys from ISCOLE countries (Australia, Brazil, Canada, China, Colombia, Finland, Kenya, India, Portugal, South Africa, the United Kingdom and the United States). Distributions of characteristics were assessed within each ISCOLE country-level database, and compared with published data from national or regional surveys, where available. Variables of comparison were identified a priori and included body mass index (BMI), physical activity (accelerometer-determined steps per day) and screen time (child-report). Of 12 countries, data on weight status (BMI) were available in 8 countries, data on measured physical activity (steps per day) were available in 5 countries and data on self-reported screen time were available in 9 countries. The five ISCOLE countries that were part of the Health Behaviour in School-aged Children Survey (that is, Canada, Finland, Portugal, the United Kingdom (England) and the United States) also provided comparable data on self-reported physical activity. Available country-specific data often used different measurement tools or cut-points, making direct comparisons difficult. Where possible, ISCOLE data were re-analyzed to match country-level data, but this step limited between-country comparisons. From the analyses performed, the ISCOLE data do not seem to be systematically biased; however, owing to limitations in data availability, data from ISCOLE should be used with appropriate caution when planning country-level population health interventions. This work highlights the need for harmonized measurement tools around the world while accounting for culturally specific characteristics, and the need for collaboration across study centers and research groups.


Thrombosis in giant basilar tip aneurysms during coil embolization.
Two patients with giant basilar tip aneurysms underwent coil embolization, one with both platinum fiber and platinum Guglielmi detachable coils and the other with Guglielmi detachable coils only. In both cases, spontaneous intraaneurysmal thrombosis occurred outside the coil mass, presumably a result of disruption of the intraaneurysmal flow pattern.


Minicore myopathy in children: a clinical and histopathological study of 19 cases.
Minicore myopathy is a congenital myopathy characterized by multifocal areas of degeneration in muscle fibres. Genetic heterogeneity expected on the basis of clinical variability awaits further resolution. We reviewed 19 cases in order to further delineate the phenotype. Marked hypotonia was the predominant presenting feature, with evidence of antenatal onset in 30% of cases. Weakness was most pronounced axially and proximally, often more severely affecting the shoulder girdle. Mild facial involvement was frequent. Varying degrees of scoliosis were obvious in all patients older than 10 years. In addition, two patients who were also the most severely affected had complete external ophthalmoplegia. One patient showed marked distal involvement. Respiratory failure developed in half of all patients after 10 years of age and correlated strongly with the degree of scoliosis. Cardiac involvement occurred mainly secondary to respiratory impairment. The course appeared static in most cases. Loss of independent walking was observed only in one case at the age of 10 years. On ultrasound scan, differential involvement within the quadriceps was documented in several patients. Variability in fibre size, type 1 predominance and atrophy with occasional type 2 hypertrophy were prominent but nonspecific histological changes. Apart from typical minicores, a marked increase in internal nuclei was the most prominent histological feature. With the exception of one family in which two generations were affected, inheritance appeared autosomal-recessive or sporadic in all cases.


Comparative genomic characterization of three Streptococcus parauberis strains in fish pathogen, as assessed by wide-genome analyses.
Streptococcus parauberis, which is the main causative agent of streptococcosis among olive flounder (Paralichthys olivaceus) in northeast Asia, can be distinctly divided into two groups (type I and type II) by an agglutination test. Here, the whole genome sequences of two Japanese strains (KRS-02083 and KRS-02109) were determined and compared with the previously determined genome of a Korean strain (KCTC 11537). The genomes of S. parauberis are intermediate in size and have lower GC contents than those of other streptococci. We annotated 2,236 and 2,048 genes in KRS-02083 and KRS-02109, respectively. Our results revealed that the three S. parauberis strains contain different genomic insertions and deletions. In particular, the genomes of Korean and Japanese strains encode different factors for sugar utilization; the former encodes the phosphotransferase system (PTS) for sorbose, whereas the latter encodes proteins for lactose hydrolysis, respectively. And the KRS-02109 strain, specifically, was the type II strain found to be able to resist phage infection through the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system and which might contribute valuably to serologically distribution. Thus, our genome-wide association study shows that polymorphisms can affect pathogen responses, providing insight into biological/biochemical pathways and phylogenetic diversity.


Multiresistant Salmonella ohio infections at the University Hospital of the West Indies.
During 1982-83 there was a substantial increase in the number of S. ohio infections at the University Hospital of the West Indies, which coincided with the appearance of strains resistant to chloramphenicol, cotrimoxazole, ampicillin, neomycin and carbenicillin. Multiresistant strains of S. ohio accounted for 19.3% of all salmonella isolates during this period and all of 40 strains tested were able to transfer resistance determinants to E. coli K12 J 53-2. S. ohio was cultured from stool (60), blood (5), wounds and abscesses (4) and postmortem material (2). Eighty-six per cent of S. ohio infections occurred in children of 3 years old or less. There was a high incidence of gastroenteritis in malnourished children, a 14% incidence of localizing infections and a 7% incidence of septicaemia. Two infants with severe gastroenteritis and bronchopneumonia died. There were a number of unusual infections including two cases of septicaemia in children receiving chloramphenicol for Haemophilus influenzae meningitis, a scrotal abscess secondary to extravasation of urine and infected scabies in a child with marasmic kwashiorkor.


Nature based solutions for contaminated land remediation and brownfield redevelopment in cities: A review.
Urban industrialization has caused severe land contamination at hundreds of thousands of sites in cities all around the world, posing a serious health risk to millions of people. Many contaminated brownfield sites are being left abandoned due to the high cost of remediation. Traditional physical and chemical remediation technologies also require high energy and resource input, and can result in loss of land functionality and cause secondary pollution. Nature-based solutions (NBS) including phytoremediation and conversion of brownfield sites to public greenspaces, holds much promise in maximizing a sustainable urban renaissance. NBS is an umbrella concept that can be used to capture nature based, cost effective and eco-friendly treatment technologies, as well as redevelopment strategies that are socially inclusive, economically viable, and with good public acceptance. The NBS concept is novel and in urgent need of new research to better understand the pros and cons, and to enhance its practicality. This review article summarizes NBS's main features, key technology choices, case studies, limitations, and future trends for urban contaminated land remediation and brownfield redevelopment.


Diffuse X-ray scattering from correlated motions in a protein crystal.
Protein dynamics are integral to biological function, yet few techniques are sensitive to collective atomic motions. A long-standing goal of X-ray crystallography has been to combine structural information from Bragg diffraction with dynamic information contained in the diffuse scattering background. However, the origin of macromolecular diffuse scattering has been poorly understood, limiting its applicability. We present a finely sampled diffuse scattering map from triclinic lysozyme with unprecedented accuracy and detail, clearly resolving both the inter- and intramolecular correlations. These correlations are studied theoretically using both all-atom molecular dynamics and simple vibrational models. Although lattice dynamics reproduce most of the diffuse pattern, protein internal dynamics, which include hinge-bending motions, are needed to explain the short-ranged correlations revealed by Patterson analysis. These insights lay the groundwork for animating crystal structures with biochemically relevant motions.


Postoperative Transcatheter Interventions in Children Undergoing Congenital Heart Surgery.
Background Postoperative transcatheter interventions (TCIs) are performed after congenital heart surgery to treat residual or recurrent anatomic lesions. We used the Society of Thoracic Surgeons Congenital Heart Surgery Database to evaluate rates of postoperative TCIs, center variability, and to determine whether center approaches to postoperative TCI might be associated with outcomes. Methods and Results Patients <18 years undergoing an index operation (2010-2016) were included. We determined predischarge postoperative TCI rates and used multivariable modeling, adjusting for patient factors and case complexity, to evaluate the association between center risk-adjusted postoperative TCI rates and risk-adjusted outcomes (operative mortality, post-TCI mortality, and failure-to-rescue). Postoperative TCI was performed after 2615/105 742 (2.5%) index operations and after 1443/25 416 (5.7%) highest complexity operations (STAT [Society of Thoracic Surgeons-European Association for Cardio-Thoracic Surgery Mortality Score] Mortality Category 4 and 5). Median (interquartile range) age of patients undergoing TCI was 2.7 (0.2-8.0) months with 43% performed in neonates. There was a wide center variability across the 107 included centers with risk-adjusted rates of postoperative TCI ranging from 0.0% to 8.0% overall and 0.0% to 20.7% for STAT 4 and 5 cases. Postoperative TCI was associated with higher risk-adjusted odds of operative mortality (odds ratio, 4.06; 95% CI, 3.60-4.58). Centers with higher postoperative TCI rates had higher overall operative mortality ( R2=0.23; P=0.02) but did not have higher post-TCI mortality ( P=0.10). There was no correlation between center TCI rates and failure-to-rescue ( P=0.19). Conclusions Patients undergoing postoperative TCI represent a high-risk cohort. Wide center variability suggests the potential for improving outcomes, but further study is necessary to better understand optimal approaches.


Immunomodulatory activity of enzymatically synthesized glycogen and its digested metabolite in a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages.
Previously, we developed enzymatically synthesized glycogen (ESG) from starch, and showed its immunomodulatory and dietary fiber-like activities. In this study, we investigated the metabolism of ESG and its immunomodulatory activity using differentiated Caco-2 cells as a model of the intestinal barrier. In a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages, mRNA expression of IL-6, IL-8, IL-1β and BAFF cytokines was up-regulated in Caco-2 cells and IL-8 production in basolateral medium was induced after 24 h apical treatment with 5 mg ml(-1) of ESG. The mRNA level of iNOS was also up-regulated in RAW264.7 macrophages. After characterization of the binding of anti-glycogen monoclonal antibodies (IV58B6 and ESG1A9) to ESG and its digested metabolite resistant glycogen (RG), an enzyme-linked immunosorbent assay (ELISA) system was developed to quantify ESG and RG. Using this system, we investigated the metabolism of ESG in differentiated Caco-2 cells. When ESG (7000 kDa, 5 mg ml(-1)) was added to the apical side of Caco-2 monolayers, ESG disappeared and RG (about 3000 kDa, 3.5 mg ml(-1)) appeared in the apical solution during a 24 h incubation. Neither ESG nor RG was detected in the basolateral solution. In addition, both ESG and RG were bound to TLR2 in Caco-2 cells. In conclusion, we suggest that ESG is metabolized to a RG-like structure in the intestine, and this metabolite activates the immune system via stimulation of the intestinal epithelium, although neither ESG nor its metabolite could permeate the intestinal cells under our experimental conditions. These results provide evidence for the beneficial function of ESG as a food ingredient.


Antagonizing the Androgen Receptor with a Biomimetic Acyltransferase.
The Androgen Receptor (AR) remains the leading target of advanced prostate cancer therapies. Thiosalicylamide analogs have previously been shown to act in cells as acyltransfer catalysts that are capable of transferring cellular acetate, presumably from acetyl-CoA, to HIV NCp7. Here we explore if the cellular acetyl-transfer activity of thiosalicylamides can be redirected to other cellular targets guided by ligands for AR. We constructed conjugates of thiosalicylamides and the AR-binding small molecule tolfenamic acid, which binds the BF-3 site of AR, proximal to the coactivator "FXXLF" binding surface. The thiosalicylamide-tolfenamic acid conjugate, YZ03, but not the separate thiosalicylamide plus tolfenamic acid, significantly enhanced acetylation of endogenous AR in CWR22Rv1 cells. Further analysis confirms that Lys720, a residue critical to FXXLF coactivator peptide binding, is a site of acyl-YZ03 acetylation. Under acyl-transfer conditions, YZ03 significantly enhances the ability of BF-3 site binding ligands to inhibit AR-coactivator peptide association. These data suggest that biomimetic acyltransferases can enhance protein-protein interaction inhibitors through covalent modification of critical interfacial residues.


Two key genomic regions harbour QTLs for salinity tolerance in ICCV 2 × JG 11 derived chickpea (Cicer arietinum L.) recombinant inbred lines.
Although chickpea (Cicer arietinum L.), an important food legume crop, is sensitive to salinity, considerable variation for salinity tolerance exists in the germplasm. To improve any existing cultivar, it is important to understand the genetic and physiological mechanisms underlying this tolerance. In the present study, 188 recombinant inbred lines (RILs) derived from the cross ICCV 2 × JG 11 were used to assess yield and related traits in a soil with 0 mM NaCl (control) and 80 mM NaCl (salinity) over two consecutive years. Salinity significantly (P < 0.05) affected almost all traits across years and yield reduction was in large part related to a reduction in seed number but also a reduction in above ground biomass. A genetic map was constructed using 56 polymorphic markers (28 simple sequence repeats; SSRs and 28 single nucleotide polymorphisms; SNPs). The QTL analysis revealed two key genomic regions on CaLG05 (28.6 cM) and on CaLG07 (19.4 cM), that harboured QTLs for six and five different salinity tolerance associated traits, respectively, and imparting either higher plant vigour (on CaLG05) or higher reproductive success (on CaLG07). Two major QTLs for yield in the salinity treatment (explaining 12 and 17% of the phenotypic variation) were identified within the two key genomic regions. Comparison with already published chickpea genetic maps showed that these regions conferred salinity tolerance across two other populations and the markers can be deployed for enhancing salinity tolerance in chickpea. Based on the gene ontology annotation, forty eight putative candidate genes responsive to salinity stress were found on CaLG05 (31 genes) and CaLG07 (17 genes) in a distance of 11.1 Mb and 8.2 Mb on chickpea reference genome. Most of the genes were known to be involved in achieving osmoregulation under stress conditions. Identification of putative candidate genes further strengthens the idea of using CaLG05 and CaLG07 genomic regions for marker assisted breeding (MAB). Further fine mapping of these key genomic regions may lead to novel gene identification for salinity stress tolerance in chickpea.


C-reactive protein gene and Toll-like receptor 4 gene polymorphisms can relate to the development of psoriatic arthritis.
We aimed to determine in psoriatic arthritis (PsA) patients the Toll-like receptor (TLR) 4 and C-reactive gene (CRP) polymorphisms and allele frequency and to investigate the relationship between clinical parameters and gene polymorphisms. We enrolled in this study 31 PsA and 41 healthy control subjects. PsA diagnosis was according to CASPAR criteria. Bath ankylosing spondylitis diseases activity index, Maastricht ankylosing spondylitis enthesitis score, and Bath ankylosing spondylitis functional index were measured. C, A, and T alleles of CRP and A and G alleles of TLR 4 were determined using the analysis of melting curves after real-time PCR. CRP A, C, and T allele frequency in controls was 26.8, 73.2, and 36.6%, respectively. In the PsA patient group, A, C, and T allele frequency was 9.7, 87.1, and 12.9%, respectively. Between control and PsA groups, there was a significant difference in A, C, and T allele frequency (P = 0.008, 0.038, and 0.001, respectively). The frequency of CRP gene polymorphisms (CA, AA, CT, TA, and TT alleles) in the control group was 56.1% and in the PsA group was 22.6%. There was a significant difference between the two groups (P = 0.004). The absence of a CRP gene polymorphism was a risk factor for PsA (odds ratio 4.3, 95% CI; 1.5-12.4, P = 0.005). TLR gene haploid frequency was investigated, and all control subjects had the wild-type AA allele. PsA patient GA allele frequency was 6.5%. There was no significant difference between the two groups (P = 0.182). GA mutant allele frequency was related to PsA (odds ratio 7.03, 95% CI; 0.32-151.9, P = 0.214). We have shown that CRP gene polymorphisms are higher in control subjects than PsA patients, and TLR 4 gene polymorphisms were found to be related to PsA.


Serum leptin and bone metabolism in HIV patients treated with highly active antiretroviral therapy.
The role of leptin in bone metabolism has not yet been fully elucidated and results remain controversial. We investigated whether changes in serum leptin correlated to bone mineral density (BMD) occur in human immunodeficiency virus (HIV) patients on highly active antiretroviral therapy (HAART). The study population was 117 HIV patients (67 men, 50 women) on HAART and 50 healthy controls, all with normal body mass index (BMI). Based on whole body BMD as measured by dual energy x-ray absorptiometry (DEXA), patients were classified as having a low (< -1) T-score (L) or a normal (> -1) T-score (N); DEXA scans were also used to determine total body fat (TFM) and percent fat (F%); radioimmunologic assays were used to measure leptin, osteocalcin (OC), bone alkaline phosphatase (BAP), 1,25 (OH)2 D in serum, and pyridinium cross-links (PYD & DPD) in urine. Of the 117 HIV patients, 54 (46.1%) were classified as L and 63 (53.9%) as N; BMD in both sexes was lower (P <0.01) among the L patients than among either the N patients or the controls; 25/32 L men and 19/22 women were osteopenic, the remaining were osteoporotic. The mean TFM, F%, OC, BAP and PYD & DPD values were higher and the mean 1,25 (OH)2 D values were lower in the L than in the N patients; leptin was higher among the L men (P <0.002) and the L women (P <0.03) than in the N patients. In both sexes. leptin positively correlated with TFM, F%, BAP and PYD & DPD; however, leptin, TFM and F% correlated negatively with BMD. A negative correlation was found between 1,25 (OH)2 D and PYD & DPD in women. At follow-up assessment of 56 HIV patients continuing HAART, leptin and BAP increased and 1,25 (OH)2 D decreased, but not significantly; BMD significantly decreased in women and PYD & DPD increased in men (P <0.02). An inverse relationship was found between leptin and BMD in HIV patients with osteopenia/osteoporosis treated with HAART. While the role of leptin in bone metabolism in a setting of HIV is still unclear, an inhibitory effect of leptin associated with a negative action by HAART may be hypothesized.


Biology and external morphology of the immature stages of the butterfly Callicore pygas eucale, with comments on the taxonomy of the genus Callicore (Nymphalidae: Biblidinae).
The biology and the external morphology of the immature stages of Callicore pygas eucale (Fruhstorfer, 1916) (Lepidoptera: Nymphalidae: Biblidinae) are described. Immatures were collected on Allophylus edulis (Radlkofer) (Sapindales: Sapindaceae) in Curitiba, Paraná, Brazil, and reared in the laboratory. Morphological descriptions and illustrations are given based on observations through electronic, stereoscopic, and optic microscopes, the latter two attached to a camera lucida. Results are compared and discussed with the immature stages of other species of the subtribe Callicorina. Immature stages data provide further evidence that Callicore is paraphyletic and that generic limits within the Callicorina need revision.


Rehabilitative Soft Exoskeleton for Rodents.
Robotic exoskeletons provide programmable, consistent and controllable active therapeutic assistance to patients with neurological disorders. Here we introduce a prototype and preliminary experimental evaluation of a rehabilitative gait exoskeleton that enables compliant yet effective manipulation of the fragile limbs of rats. To assist the displacements of the lower limbs without impeding natural gait movements, we designed and fabricated soft pneumatic actuators (SPAs). The exoskeleton integrates two customizable SPAs that are attached to a limb. This configuration enables a 1 N force load, a range of motion exceeding 80 mm in the major axis, and speed of actuation reaching two gait cycles/s. Preliminary experiments in rats with spinal cord injury validated the basic features of the exoskeleton. We propose strategies to improve the performance of the robot and discuss the potential of SPAs for the design of other wearable interfaces.


Differentiated human airway organoids to assess infectivity of emerging influenza virus.
Novel reassortant avian influenza H7N9 virus and pandemic 2009 H1N1 (H1N1pdm) virus cause human infections, while avian H7N2 and swine H1N1 virus mainly infect birds and pigs, respectively. There is no robust in vitro model for assessing the infectivity of emerging viruses in humans. Based on a recently established method, we generated long-term expanding 3D human airway organoids which accommodate four types of airway epithelial cells: ciliated, goblet, club, and basal cells. We report differentiation conditions which increase ciliated cell numbers to a nearly physiological level with synchronously beating cilia readily discernible in every organoid. In addition, the differentiation conditions induce elevated levels of serine proteases, which are essential for productive infection of human influenza viruses and low-pathogenic avian influenza viruses. We also established improved 2D monolayer culture conditions for the differentiated airway organoids. To demonstrate the ability of differentiated airway organoids to identify human-infective virus, 3D and 2D differentiated airway organoids are applied to evaluate two pairs of viruses with known distinct infectivity in humans, H7N9/Ah versus H7N2 and H1N1pdm versus an H1N1 strain isolated from swine (H1N1sw). The human-infective H7N9/Ah virus replicated more robustly than the poorly human-infective H7N2 virus; the highly human-infective H1N1pdm virus replicated to a higher titer than the counterpart H1N1sw. Collectively, we developed differentiated human airway organoids which can morphologically and functionally simulate human airway epithelium. These differentiated airway organoids can be applied for rapid assessment of the infectivity of emerging respiratory viruses to human.


Reliability and validity of the modified Conconi test on concept II rowing ergometers.
The purpose of this study was to assess the reliability and validity of the modified Conconi test on Concept II rowing ergometers. Twenty-eight oarsmen conducted 3 performance tests on separate days. Reliability was assessed using the break point in heart rate (HR) linearity called the Conconi test (CT) and Conconi retest (CRT) for the noninvasive measurement of anaerobic threshold (AT). Blood lactate measurement was considered the gold standard for the assessment of the AT, and the validity of the CT was assessed by blood samples taken during an incremental load test (ILT) on ergometers. According to the results, the mean power output (PO) scores for the CT, CRT, and ILT were 234.2 +/- 40.3 W, 232.5 +/- 39.7 W, and 229.7 +/- 39.6 W, respectively. The mean HR values at the AT for the CT, CRT, and ILT were 165.4 +/- 11.2 b.min, 160.4 +/- 10.8 b.min, and 158.3 +/- 8.8 b.min, respectively. Interclass correlation coefficient (ICC) analysis indicated a significant correlation between the 3 tests with one another. Also, Bland and Altman plots showed that there was an association between noninvasive tests and the ILT PO scores and HRs (95% confidence interval [CI]). In conclusion, this study showed that the modified CT is a reliable and valid method for determining the AT of elite men rowers.


Nestin is not essential for development of the CNS but required for dispersion of acetylcholine receptor clusters at the area of neuromuscular junctions.
Nestin is expressed in many different progenitors during development including those of the CNS, heart, skeletal muscle, and kidney. The adult expression is mainly restricted to the subependymal zone and dentate gyrus of the brain, the neuromuscular junction, and renal podocytes. In addition, this intermediate filament protein has served as a marker of neural stem/progenitor cells for close to 20 years. Therefore it is surprising that its function in development and adult physiology is still poorly understood. Here we report that nestin deficiency is compatible with normal development of the CNS. The mutant mice, however, show impaired motor coordination. Furthermore, we found that the number of acetylcholine receptor clusters, the nerve length, and the endplate bandwidth are significantly increased in neuromuscular junction area of nestin-deficient mice. This is similar to the phenotype described for deficiency of cyclin-dependent kinase 5 (Cdk5), a candidate downstream affecter of nestin. Moreover, we demonstrate that nestin deficiency can rescue maintenance of acetylcholine receptor clusters in the absence of agrin, similar to Cdk5/agrin double knock-outs, suggesting that the observed nestin deficiency phenotype is the consequence of aberrant Cdk5 activity.


Environmental threats, mitigation strategies and high-mountain areas.
High-mountain areas suffer from increasing environmental threats. The causes are often global in dimension but lead to specific impacts under conditions of steep and/or high-altitude terrain with strong effects from snow and ice. This paper presents a global perspective, focussing primarily on observed and projected changes in climate and then goes on to discuss key messages of greatest relevance to the highest belts of mountain regions. The paper finishes with a brief discussion of mitigation strategies.


The effect of insecticide-treated bed nets on mortality of Gambian children.
Insecticide treatment of bed nets ("mosquito nets") may be a cheap and acceptable method of reducing the morbidity and mortality caused by malaria. In a rural area of The Gambia, bed nets in villages participating in a primary health-care (PHC) scheme were treated with permethrin at the beginning of the malaria transmission season. Additionally, children aged 6 months to 5 years were randomised to receive weekly either chemoprophylaxis with maloprim or a placebo throughout the malaria transmission season. We measured mortality in children in PHC villages before and after the interventions described, and compared this with mortality in villages where no interventions occurred (non-PHC villages). About 92% of children in PHC villages slept under insecticide-treated bed nets. In the year before intervention, mortality in children aged 1-4 years was lower in non-PHC villages. After intervention, the overall mortality and mortality attributable to malaria of children aged 1-4 in the intervention villages was 37% and 30%, respectively, of that in the non-PHC villages. Among children who slept under treated nets, we found no evidence of an additional benefit of chemoprophylaxis in preventing deaths. Insecticide-treated bed nets are simple to introduce and can reduce mortality from malaria.


Vision and driving self-restriction in older adults.
To assess driving self-restriction (vision related and nonvision related) in relation to vision test performance of older adults. Cross-sectional study. Population-based cohort of community-dwelling older adults. Six hundred twenty-nine current drivers aged 55 and older had driving behavior, health, and physical function assessed and vision function tested in 1993-95. Self-report of driving restriction as vision or non-vision related and performance on a comprehensive battery of vision tests (visual acuity; contrast sensitivity; effects of illumination level, contrast, and glare on acuity; visual fields with and without attentional load; color vision; temporal sensitivity; and the effect of dim light on walking ability). Demographic, health, and functional characteristics differed significantly between restrictors and nonrestrictors but not between vision- and nonvision-related restrictors. Controlling for potential confounding, only vision-related driving self-restriction was significantly associated with reduced performance on nonstandard measures of acuity. Poor depth perception was significantly associated with restriction for both vision- and nonvision-related reasons. Poor performance on attentional visual field tests, analyzed individually and in combination with standard field tests, was not associated with driving self-restriction. Older adults with early changes in spatial vision function and depth perception appear to recognize their limitations and restrict their driving even if they do not acknowledge the visual impairment as the cause for restriction. Poor visual attention, a risk factor for crashes, may not be recognized. Additional studies of driving self-restriction in relation to risk factors for crashes in older adults may help refine this strategy of reducing driving-related injury and death.


Self-association behaviour of protein:surfactant systems in alcohol/water mixtures.
The effect of the addition of short-chain monohydric alcohols (ethanol and propan-2-ol) to the protein:surfactant system lysozyme:sodium dodecyl sulfate (Lz:SDS) in aqueous solution was investigated using a conductometric technique. A second protein:surfactant system, bovine serum albumin:SDS (BSA:SDS) was also investigated so that the effect of a different protein conformation and composition could be compared. The critical aggregation concentration (CAC) of the protein forming the complex and the critical micelle concentration (CMC *) of SDS in the presence of protein, at different alcohol concentrations, were determined. It was found in both cases that the addition of alcohol does not produce a significant change in the CAC, whereas the CMC * displays variation with alcohol concentration that shows an inversion in the ranges 0.05-0.06 ethanol mole fraction and 0.02-0.03 propan-2-ol mole fraction. This suggests that, in contrast with the CAC behaviour, the major factor that drives SDS micellization in the presence of protein is the variation in water structure. Results also suggest that it occurs in the same way for both proteins, where electrostatic interactions are the main force in the formation of the complex. Conversely, hydrophobic interactions play the dominant role at the micellization stage, and only the extent of the interaction between protein:surfactant aggregates and surfactant species seems to depend on protein nature.


Consequences of T2 relaxation during half-pulse slice selection for ultrashort TE imaging.
A "half-pulse" slice selection approach is used in the ultrashort echo time pulse sequence and is required to give minimal transverse relaxation in a two-dimensional acquisition. This method splits the normal excitation radiofrequency pulse in half and acquires a pair of images, each using one of these half-pulses. These half-pulses are used without a refocusing gradient since summing the pair of images yields images with accurate slice selection. When the radiofrequency pulse duration is similar to the sample T(2), characteristics such as the effective echo time and choice of radiofrequency pulse require careful evaluation as some of the approximations in conventional slice selection do not apply. We derive a theory that includes relaxation during excitation using Pauly's excitation k-space formalism. Further, this theory is tested on phantoms with a range of values of T(2) demonstrating the effect on the slice profile. We conclude that relaxation during excitation is significant and should be included in our estimate of the T(2) weighting of the sequence. In general, the T(2) weighting should be measured from the time of the centroid of the excitation pulse.


Increasing attendance for psychotherapy: implementation intentions and the self-regulation of attendance-related negative affect.
The present study evaluated an implementation intention intervention that aimed to increase attendance at scheduled, initial appointments for psychotherapy by helping clients to manage negative feelings about attendance. Participants received a postal questionnaire that measured their views about attending psychotherapy. One half of the sample was randomly assigned to an implementation intention induction that was embedded in the questionnaire. Intention-to-treat analysis (N=390) indicated that participants who formed implementation intentions were more likely to attend compared to controls (75% vs. 63%), and this effect was even stronger among participants who returned the questionnaire (83% vs. 57%). Whereas anticipated affective costs (e.g., shame) had a negative impact on attendance for most participants, this effect was attenuated when participants formed implementation intentions and perceived that attendance would be beneficial. Thus, implementation intention formation can help clients to deal effectively with negative feelings that might otherwise prevent them attending their first psychotherapy appointment.


Two infant vision screening programmes: prediction and prevention of strabismus and amblyopia from photo- and videorefractive screening.
Two infant vision screening programmes on total populations in the Cambridge Health District have been designed to identify manifest strabismus and strabismogenic and amblyogenic refractive errors at 7-9 months of age. The first, completed, programme used the isotropic photorefractor with cycloplegia together with a standard orthoptic examination. The second, current, programme uses the VRP-1 isotropic videorefractor to identify infants with accommodative lags which are followed up by refraction under cycloplegia. Both programmes show good agreement between infants identified at screening and retinoscopic refractions at follow-up, showing that photo- and videorefraction (with or without cycloplegia) can be effective methods for screening for ametropia in infants and young children. In each programme 5-6% of infants showed abnormal levels of hyperopia (> or = 3.5 D in any meridian), less than 1% showed anisometropia > or = 1.5 D; very few infants (0.25%) showed -3D myopia or greater. Less than 1% showed manifest strabismus. Hyperopic and anisometropic children entered a randomised controlled trial of partial refractive correction. All children identified at screening, alongside appropriate control groups, are extensively followed up to age 4 years. The first programme has found that children who were hyperopic in infancy were 13 times more likely to become strabismic, and 6 times more likely to show measurable acuity deficits by 4 years, compared with controls. Wearing a partial spectacle correction reduced these risk ratios to 4:1 and 2.5:1 respectively. The impaired acuity can be attributed, in part, to meridional amblyopia resulting from persisting astigmatism. Both hyperopic and myopic infants showed refractive changes in the direction of emmetropia between 9 months and 4 years. Wearing a partial spectacle correction did not affect this process of emmetropisation, but does provide the possibility of reducing the incidence of common pre-school vision problems.


[Study on the effect of monoclonal antibodies on Giardia lamblia in vitro].
Monoclonal antibodies (McAbs) were generated by fusion of SP2/0 myeloma cells with spleen cells from BALB/c mice immunized with Giardia lamblia trophozoites. To define the effect of the McAb on Giardia lamblia, trophozoites were subcultured axenically after contact with McAb for 1 h at 37 degrees C. The McAbs 1G5 and 1E2 (both IgG1) were able to inhibit the growth of trophozoite at the rate of 63.4% and 35%, respectively. The inhibitory effect was decreased along with the decreasing quantity of McAb (from dilution 1:45 to 1:225) added to the TYI-S-33 medium. The actions of McAbs could be eliminated by absorption with trophozoites and not be influenced by complement in fresh guinea-pig serum, thus the actions of McAbs appeared to be specific and complement independent.


Enteral nutritional support by gastrostomy tube in children with cancer.
We examined the use of gastrostomy tubes in malnourished children with cancer as part of our ongoing efforts to improve their supportive care. Patients were examined on the basis of percentage of weight loss and percentage of desirable body weight. Twenty-five patients underwent gastrostomy tube placement followed by aggressive enteral nutritional support. Gastrostomy tubes were placed at a mean of 3.5 months (range, 0.3 to 8 months) after diagnosis; mean weight loss had been 10.1% (range, to 21%) of desirable body weight. There were no immediate postoperative complications. Gastrostomy tube feedings were well tolerated by all patients. All children gained or maintained weight, and 60% of the severely malnourished children returned to a desirable body weight after an average of 4.9 months (range, 1 to 13 months). Weight gain averaged 12.9% (range, to 45.4%) of desirable body weight. The most common complications were 38 episodes of inflammation at the gastrostomy tube site during periods of severe neutropenia, which were treated successfully with topically or orally administered antibiotics, and 13 episodes of cellulitis, which required intravenously administered antibiotics. The infection rate was 1.58 episodes per 1000 days of use compared with a rate of 5.0 per 1000 days previously reported with total parenteral nutrition. The monthly costs of gastrostomy tube nutrition support were 9% of those associated with use of total parenteral nutrition. Gastrostomy tube use in children with cancer is a safe, effective, and cost-effective method of reversing malnutrition. Further investigation with larger numbers of patients is warranted.


A DNA template recognition protein: partial purification from mouse liver and stimulation of DNA polymerase alpha.
A protein that specifically enhances up to 13-fold the rate of copying of poly(dT) template by DNA polymerase alpha was partially purified from chromatin of regenerating mouse liver cells. This stimulatory protein, designated herein factor D, also increases 2-3-fold the activity of polymerase alpha with heat-denatured DNA and with primed, circular single-stranded phi X174 DNA. However, factor D has no detectable effect on the copying by polymerase alpha of poly(dG), poly(dA), and poly(dC) templates. Activity of mouse DNA polymerase beta is not affected by factor D with all the tested templates. In contrast to polymerase alpha, factor D is resistant to inactivation by N-ethylmaleimide and calcium ions, but it is readily heat-inactivated at 46 degrees C and is inactivated by trypsin digestion. Partially purified factor D is not associated with detectable activities of DNA polymerase, DNA primase, deoxyribonucleotidyl terminal transferase, and endo- or exodeoxyribonuclease.


Chromosomal rearrangements and the first indication of an ♀X1 X1 X2 X2 /♂X1 X2 Y sex chromosome system in Rineloricaria fishes (Teleostei: Siluriformes).
Rineloricaria is the most diverse genus within the freshwater fish subfamily Loricariinae, and it is widely distributed in the Neotropical region. Despite limited cytogenetic data, records from southern and south-eastern Brazil suggest a high rate of chromosomal rearrangements in this genus, mirrored in remarkable inter- and intraspecific karyotype variability. In the present work, we investigated the karyotype features of Rineloricaria teffeana, an endemic representative from northern Brazil, using both conventional and molecular cytogenetic techniques. We revealed different diploid chromosome numbers (2n) between sexes (33♂/34♀), which suggests the presence of an ♀X1 X1 X2 X2 /♂X1 X2 Y multiple sex chromosome system. The male-limited Y chromosome was the largest and the only biarmed element in the karyotype, implying Y-autosome fusion as the most probable mechanism behind its origination. C-banding revealed low amounts of constitutive heterochromatin, mostly confined to the (peri)centromeric regions of most chromosomes (including the X2 and the Y) but also occupying the distal regions of a few chromosomal pairs. The chromosomal localization of the 18S ribosomal DNA (rDNA) clusters revealed a single site on chromosome pair 4, which was adjacent to the 5S rDNA cluster. Additional 5S rDNA loci were present on the autosome pair 8, X1 chromosome, and in the presumed fusion point on the Y chromosome. The probe for telomeric repeat motif (TTAGGG)n revealed signals of variable intensities at the ends of all chromosomes except for the Y chromosome, where no detectable signals were evidenced. Male-to-female comparative genomic hybridization revealed no sex-specific or sex-biased repetitive DNA accumulations, suggesting a presumably low level of neo-Y chromosome differentiation. We provide evidence that rDNA sites might have played a role in the formation of this putative multiple sex chromosome system and that chromosome fusions originate through different mechanisms among different Rineloricaria species.


Estimating the strength of selective sweeps from deep population diversity data.
Selective sweeps are typically associated with a local reduction of genetic diversity around the adaptive site. However, selective sweeps can also quickly carry neutral mutations to observable population frequencies if they arise early in a sweep and hitchhike with the adaptive allele. We show that the interplay between mutation and exponential amplification through hitchhiking results in a characteristic frequency spectrum of the resulting novel haplotype variation that depends only on the ratio of the mutation rate and the selection coefficient of the sweep. On the basis of this result, we develop an estimator for the selection coefficient driving a sweep. Since this estimator utilizes the novel variation arising from mutations during a sweep, it does not rely on preexisting variation and can also be applied to loci that lack recombination. Compared with standard approaches that infer selection coefficients from the size of dips in genetic diversity around the adaptive site, our estimator requires much shorter sequences but sampled at high population depth to capture low-frequency variants; given such data, it consistently outperforms standard approaches. We investigate analytically and numerically how the accuracy of our estimator is affected by the decay of the sweep pattern over time as a consequence of random genetic drift and discuss potential effects of recombination, soft sweeps, and demography. As an example for its use, we apply our estimator to deep sequencing data from human immunodeficiency virus populations.


The monoclonal antibody E587 recognizes growing (new and regenerating) retinal axons in the goldfish retinotectal pathway.
E587 is a new monoclonal antibody against a 200 kDa cell-surface glycoprotein in the fish retinotectal pathway. The E587 antigen probably belongs to the class of cell adhesion molecules, and more specifically, to the family of L1-like molecules. The immunopurified protein is recognized by the antibody against the HNK1/L2 sugar epitope (associated with most cell adhesion molecules) and by a polyclonal antiserum against chick G4, which is related to the cell adhesion molecule L1 in mouse. Moreover the NH2-terminal sequence of E587 shows similarity with L1 and Ng-CAM. The E587 immunostaining pattern in the fish retinotectal pathway suggests that the E587 antigen is a growth-associated molecule on fish retinal axons. In fish embryos, all retinal axons are labeled. In adult fish, however, only the young axons from newly added ganglion cells carry E587 staining. After optic nerve transection (ONS) and retinal axonal regeneration, all axons reexpress the E587 antigen into their terminal processes in the tectal retinorecipient layers. The reexpression of the E587 antigen is temporally regulated, and E587 immunoreactivity declines by 7 months and disappears at 12 months after ONS. We hypothesize that the E587 antigen may mediate axon-axon associations. In its restricted appearance on young axons in normal adult fish, it may contribute to the selective fasciculation of the newest axons with young axons and thus participate in the creation of the age-related fiber organization in the fish optic nerve.


Assessing oral cancer early detection: clarifying dentists' practices.
This study explores new methods for assessing in greater detail what dentists do when they perform oral cancer early detection examinations. It clarifies practice behaviors and opens opportunities to identify factors that facilitate thorough early detection examinations by clinicians and to assess the relative effectiveness of different examination procedures. A 38-item survey instrument was e-mailed to dentists in a western US, multistate dental practice group. Questionnaires were received by 241 dentists, and 102 responded. An Oral Cancer Knowledge scale (0 to 14) was generated from correct responses on oral cancer general knowledge. An Oral Cancer Examination Thoroughness scale was calculated from the two dimensions of reported usage and frequency of procedures in oral cancer examinations. Nearly all responding dentists were in general practice (90%), with a median year of graduation from dental school of 1994. The Oral Cancer Knowledge scores ranged from 5 to 14 with a mean of 10.4. The mean Thoroughness of Examination score was 11.34 (range 0 to 20). The two scales were not statistically correlated (r = -0.015, P = 0.883). Statistically, recency of continuing education was significantly associated with knowledge (P = 0.0284) and appears to be marginally associated with thoroughness (P = 0.075). This study documents considerable variability in dentists' knowledge and thoroughness of examinations. The scales provide tools for future studies for improving understanding of early detection of oral cancer in clinical practice.


Inflammation Promotes a Conversion of Astrocytes into Neural Progenitor Cells via NF-κB Activation.
Brain inflammation, a common feature in neurodegenerative diseases, is a complex series of events, which can be detrimental and even lead to neuronal death. Nonetheless, several studies suggest that inflammatory signals are also positively influencing neural cell proliferation, survival, migration, and differentiation. Recently, correlative studies suggested that astrocytes are able to dedifferentiate upon injury and may thereby re-acquire neural stem cell (NSC) potential. However, the mechanism underlying this dedifferentiation process upon injury remains unclear. Here, we report that during the early response of reactive gliosis, inflammation induces a conversion of mature astrocytes into neural progenitors. A TNF treatment induces the decrease of specific astrocyte markers, such as glial fibrillary acidic protein (GFAP) or genes related to glycogen metabolism, while a subset of these cells re-expresses immaturity markers, such as CD44, Musashi-1, and Oct4. Thus, TNF treatment results in the appearance of cells that exhibit a neural progenitor phenotype and are able to proliferate and differentiate into neurons and/or astrocytes. This dedifferentiation process is maintained as long as TNF is present in the culture medium. In addition, we highlight a role for Oct4 in this process, since the TNF-induced dedifferentiation can be prevented by inhibiting Oct4 expression. Our results show that activation of the NF-κB pathway through TNF plays an important role in the dedifferentiation of astrocytes via the re-expression of Oct4. These findings indicate that the first step of reactive gliosis is in fact a dedifferentiation process of resident astrocytes mediated by the NF-κB pathway.


Oral and inhaled corticoid treatment for wheezing in the first year of life.
To evaluate the prevalence of corticoid utilization for the treatment of wheezing in infants less than 12 months old and to analyze factors associated with this practice. This was a cross-sectional study that administered the validated questionnaire from the International Study on the Prevalence of Wheezing in Infants to 1,261 infants aged 12 to 15 months in Belo Horizonte, Brazil. Proportions and 95% confidence intervals were calculated and the chi-square test was used to detect associations between variables. Six hundred and fifty-six (52%) infants, 53% male and 48.2% white, exhibited wheezing during the first year of life. Mean age at first episode was 5.11±2.89 months. There was a high rate of morbidity, with many emergency visits (71%) and hospitalizations (27.8%). Also common were a family history of asthma and atopic disease (32.2 to 71%) and exposure to passive smoking (41.5%) and to mould (47.3%). The prevalence rates for corticoid use, whether via oral route (48.7%) or inhaled (51.3%), were elevated and were higher in the group that suffered three or more episodes. Children suffering greater morbidity were more likely to be prescribed a corticoid (p < 0.05). The high frequency of corticoid use highlights the need to establish specific criteria for the treatment of wheezing in the first years of life in order to avoid extrapolation of asthma treatments to other conditions that are transitory and self-limiting and in which using corticoids could involve more risk than benefit.