Associations between elevated homocysteine, cognitive impairment, and reduced white matter volume in healthy old adults.
Elevated homocysteine has emerged as a risk factor for cognitive impairment even in healthy elderly persons. Reduced brain volume and white matter hyperintensities also occur in healthy elderly as well, but the interrelationships between these have not been well studied. We report these interrelationships in non demented, relatively healthy, community-dwelling older adults from a single East Asian population. Two hundred twenty-eight right-handed participants age 55 years and above were evaluated. Persons with medical conditions or neurological diseases other than well-controlled diabetes mellitus and hypertension were excluded. Participants underwent quantitative magnetic resonance imaging of the brain using a standardized protocol and neuropsychological evaluation. Plasma homocysteine, folate, vitamin B(12), and markers for cardiovascular risk: blood pressure, body mass index, fasting blood glucose, and lipid profile were measured. Elevated homocysteine was associated with reduced global cerebral volume, larger ventricles, reduced cerebral white matter volume, and lower cognitive performance in several domains. Elevated homocysteine was associated with reduced white matter volume (β = -20.80, t = -2.9, df = 223, p = 0.004) and lower speed of processing (β = -0.38, t = -2.1, df = 223, p = 0.03), even after controlling for age, gender, and education. However, the association between homocysteine and lower speed of processing disappeared after controlling for white matter volume. Elevated homocysteine was not associated with white matter hyperintensity volume or with hippocampal volume. Although homocysteine and folate levels were correlated, their effects on white matter volume were dissociated. In non demented, relatively healthy adults, elevated homocysteine is associated with lower cognitive scores and reduced cerebral white matter volume. These effects can be dissociated from those related to white matter hyperintensities or reduced folate level.


Akkermansia muciniphila ameliorates the age-related decline in colonic mucus thickness and attenuates immune activation in accelerated aging Ercc1 -/Δ7 mice.
The use of Akkermansia muciniphila as potential therapeutic intervention is receiving increasing attention. Health benefits attributed to this bacterium include an improvement of metabolic disorders and exerting anti-inflammatory effects. The abundance of A. muciniphila is associated with a healthy gut in early mid- and later life. However, the effects of A. muciniphila on a decline in intestinal health during the aging process are not investigated yet. We supplemented accelerated aging Ercc1 -/Δ7 mice with A. muciniphila for 10 weeks and investigated histological, transcriptional and immunological aspects of intestinal health. The thickness of the colonic mucus layer increased about 3-fold after long-term A. muciniphila supplementation and was even significantly thicker compared to mice supplemented with Lactobacillus plantarum WCFS1. Colonic gene expression profiles pointed towards a decreased expression of genes and pathways related to inflammation and immune function, and suggested a decreased presence of B cells in colon. Total B cell frequencies in spleen and mesenteric lymph nodes were not altered after A. muciniphila supplementation. Mature and immature B cell frequencies in bone marrow were increased, whereas B cell precursors were unaffected. These findings implicate that B cell migration rather than production was affected by A. muciniphila supplementation. Gene expression profiles in ileum pointed toward a decrease in metabolic- and immune-related processes and antimicrobial peptide production after A. muciniphila supplementation. Besides, A. muciniphila decreased the frequency of activated CD80+CD273- B cells in Peyer's patches. Additionally, the increased numbers of peritoneal resident macrophages and a decrease in Ly6Cint monocyte frequencies in spleen and mesenteric lymph nodes add evidence for the potentially anti-inflammatory properties of A. muciniphila. Altogether, we show that supplementation with A. muciniphila prevented the age-related decline in thickness of the colonic mucus layer and attenuated inflammation and immune-related processes at old age. This study implies that A. muciniphila supplementation can contribute to a promotion of healthy aging.


Scattering noise estimation of range-gated imaging system in turbid condition.
The range-gated imaging systems are reliable underwater imaging system with the capability to minimize backscattering effect from turbid media. The tail-gating technique has been developed to fine tune the signal to backscattering ratio and hence improve the gated image quality. However, the tail-gating technique has limited image quality enhancement in high turbidity levels. In this paper, we developed a numerical model of range-gated underwater imaging system for near target in turbid medium. The simulation results matched the experimental work favorably. Further investigation using this numerical model shows that the multiple scattering components of the backscattering noise dominate for propagation length larger than 4.2 Attenuation Length (AL). This has limited the enhancement of tail-gating technique in high turbidity conditions.


Conversion ratios of n-3 fatty acids between plasma and erythrocytes: a systematic review and meta-regression.
As biomarkers of dietary intake or disease risk factor, n-3 fatty acid (FA) can be measured in plasma phospholipids (PL), total lipids (TL) or erythrocytes. However, the numeric relationships between n-3 FA in these lipid pools are not clear. Our goal was to derive conversion ratios for plasma and erythrocyte n-3 FA. Potential studies were identified through systematic literature search in PubMed, Embase and the Cochrane Library of Systematic reviews (1950 to October 2014). In all, fifty-six studies reporting n-3 in healthy individuals were included, of which thirty-four articles reported plasma PL and erythrocytes, and twenty-two reported plasma TL and erythrocytes. Meta-regressions were performed to quantify the ratio between plasma and erythrocyte n-3 FA weight percentages, controlling for covariates including age, sex and study design. The conversion ratios from plasma PL to erythrocytes for EPA, DHA, DPA and total n-3 PUFA are 0·75, 1·16, 2·32 and 1·22; the corresponding conversion ratios from plasma TL to erythrocytes are 1·00, 2·10, 3·85 and 2·08, respectively. The conversion ratios were validated using reported values from the literature and measured data from fifty individuals. The relative error of the predicted results were within 10 % of the mean reported values except for EPA, and the individual measured data except for DPA, in plasma TL. The conversion ratios between plasma PL and erythrocytes were more stable compared with plasma TL. Such conversion ratios will be useful for nutritionists or public health professionals to assess FA profiles of different populations using data collected with different methodologies.


Safeguarding the administration of high-dose chemotherapy: a national practice survey by the American Society for Blood and Marrow Transplantation.
Overdoses of high-dose chemotherapy before hematopoietic cell transplantation are serious adverse events, but their frequency and etiology are unknown. The American Society for Blood and Marrow Transplantation (ASBMT) conducted an anonymous national survey to identify errors in safety practices during the administration of high-dose chemotherapy. The questionnaire was returned from 115 (68%) of 170 hematopoietic transplant centers in the United States. Ninety-four of the programs were university or affiliated centers, 19 were community hospitals, and 41 were founded since 1990. A total of 7650 transplants were reported for 1994: 22% of the programs performed 1-20 transplants, 60% performed 21-100 transplants, and 18% performed more than 100 transplants. Fifteen of the 115 responding centers reported a total of 18 patients inadvertently given overdoses of cisplatin (n=3), carboplatin (n=2), busulfan (n=2), cytosine arabinoside (n=2), cyclophosphamide (n=2), interleukin-2 (n=2), or other agents (n=5) between 1989 and 1994. Cumulative drug doses given as a daily dose (six cases) and nursing infusion errors (six cases) were the most common errors. The estimated chemotherapy overdose error rate was 0.06%, or 6 cases/10,000 transplants, with 95% confidence limits of 0.03-0.11%. The overdose rate among more experienced centers in operation before 1990 was lower than that among newer centers (p < 0.01). Large centers (> 100 transplants performed in 1994) experienced errors at rates lower than those in medium-sized centers (21-100 transplants, p = 0.03). Although the number of events was small in this self-reporting survey, overdoses were noted in 13% of the responding centers, especially among more recently established units. Safety practices need to emphasize multidisciplinary checkpoints at the physician, pharmacist, nursing, and institutional levels. Based on these survey results, ASBMT recommendations for further safeguards for high-dose chemotherapy administration are proposed.


Pediatric Firearm-Related Injuries in the United States.
Pediatric firearm-related deaths and injuries are a national public health crisis. In this Special Review Article, we characterize the epidemiology of firearm-related injuries in the United States and discuss public health programs, the role of pediatricians, and legislative efforts to address this health crisis. Firearm-related injuries are leading causes of unintentional injury deaths in children and adolescents. Children are more likely to be victims of unintentional injuries, the majority of which occur in the home, and adolescents are more likely to suffer from intentional injuries due to either assault or suicide attempts. Guns are present in 18% to 64% of US households, with significant variability by geographic region. Almost 40% of parents erroneously believe their children are unaware of the storage location of household guns, and 22% of parents wrongly believe that their children have never handled household guns. Public health interventions to increase firearm safety have demonstrated varying results, but the most effective programs have provided free gun safety devices to families. Pediatricians should continue working to reduce gun violence by asking patients and their families about firearm access, encouraging safe storage, and supporting firearm-related injury prevention research. Pediatricians should also play a role in educating trainees about gun violence. From a legislative perspective, universal background checks have been shown to decrease firearm homicides across all ages, and child safety laws have been shown to decrease unintentional firearm deaths and suicide deaths in youth. A collective, data-driven public health approach is crucial to halt the epidemic of pediatric firearm-related injury.


Spatially-varying surface roughness and ground-level air quality in an operational dispersion model.
Urban form controls the overall aerodynamic roughness of a city, and hence plays a significant role in how air flow interacts with the urban landscape. This paper reports improved model performance resulting from the introduction of variable surface roughness in the operational air-quality model ADMS-Urban (v3.1). We then assess to what extent pollutant concentrations can be reduced solely through local reductions in roughness. The model results suggest that reducing surface roughness in a city centre can increase ground-level pollutant concentrations, both locally in the area of reduced roughness and downwind of that area. The unexpected simulation of increased ground-level pollutant concentrations implies that this type of modelling should be used with caution for urban planning and design studies looking at ventilation of pollution. We expect the results from this study to be relevant for all atmospheric dispersion models with urban-surface parameterisations based on roughness.


Reducing agents improve enzymatic hydrolysis of cellulosic substrates in the presence of pretreatment liquid.
Enzymatic hydrolysis of pretreated lignocellulosic substrates has emerged as an interesting option to produce sugars that can be converted to liquid biofuels and other commodities using microbial biocatalysts. Lignocellulosic substrates are pretreated to make them more accessible to cellulolytic enzymes, but the pretreatment liquid partially inhibits subsequent enzymatic hydrolysis. The presence of pretreatment liquid from Norway spruce resulted in a 63% decrease in the enzymatic saccharification of Avicel compared to when the reaction was performed in a buffered aqueous solution. The addition of 15mM of a reducing agent (hydrogen sulfite, dithionite, or dithiothreitol) to reaction mixtures with the pretreatment liquid resulted in up to 54% improvement of the saccharification efficiency. When the reducing agents were added to reaction mixtures without pretreatment liquid, there was a 13-39% decrease in saccharification efficiency. In the presence of pretreatment liquid, the addition of 15mM dithionite to Avicel, α-cellulose or filter cake of pretreated spruce wood resulted in improvements between 25 and 33%. Positive effects (6-17%) of reducing agents were also observed in experiments with carboxymethyl cellulose and 2-hydroxyethyl cellulose. The approach to add reducing agents appears useful for facilitating the utilization of enzymes to convert cellulosic substrates in industrial processes.


Apoptosis gene profiling reveals spatio-temporal regulated expression of the p53/Mdm2 pathway during lens development.
Evidence is emerging for apoptosis gene expression in the lens during development. Therefore, here we used a filter array to assess expression of 243 apoptosis-related genes in the developing postnatal mouse lens using (33)P labelled cDNA synthesized from p7 and p14 mouse lenses. We demonstrated that 161 apoptosis-related genes were expressed at levels significantly above background and 20 genes were potentially significantly differentially expressed (P<0.05) by at least 2-fold between p7 and p14. We used RT-PCR to confirm expression of these genes in newborn, p7, p14 and 4 wk mouse lens cDNA samples. Expression of 19/20 of the genes examined was confirmed, while 5 genes (Huntingtin, Mdm2, Dffa, galectin-3 and Mcl-1) were confirmed as differentially regulated between p7 and p14. RT-PCR was also used to examine the expression of the chick homologues of the most-highly expressed and/or potentially differentially regulated genes in chick embryo lenses at E6-E16. The majority of genes expressed in the postnatal mouse lens were also expressed in the chick embryo lens. Western blotting confirmed developmentally regulated expression of Axl and Mcl-1 during mouse lens development and of Mdm2, Mdm4/X and p53 during mouse and chick lens development. Western blotting also revealed the presence of p53 and Mdm4/X splice variants and/or proteolytic cleavage products in the developing lens. Since Mdm2 is a regulator of the tumour suppressor gene p53, we chose to thoroughly investigate the spatio-temporal expression patterns of p53, Mdm2 and the functionally related Mdm4/X in mouse lens development at E12.5-E16.5 using immunocytochemistry. We also examined Mdm2 expression patterns during chick lens development at E6-E16 and Mdm4/X and p53 at E14. Expression of Mdm2, Mdm4/X and p53 was spatio-temporally regulated in various compartments of the developing lens in both mouse and chick, including lens epithelial and lens fibre cells, indicating potential roles for these factors in regulation of lens epithelial cell proliferation and/or lens fibre cell differentiation This study provides a thorough initial analysis of apoptosis gene expression in the postnatal mouse lens and provides a resource for further investigation of the roles in lens development of the apoptosis genes identified. Furthermore, building on the array studies, we present the first spatio-temporal analysis of expression of p53 pathway molecules (p53, Mdm2 and Mdm4/X) in both developing mouse and chick lenses, suggesting a potential role for the p53/Mdm2 pathway in lens development, which merits further functional analysis.


Does adenoid hypertrophy affect disease severity in children with allergic rhinitis?
Our study aims to evaluate the presence of adenoid hypertrophy (AH) in children with allergic rhinitis (AR) and the association of AH disease severity and clinical laboratory finding from retrospective, cross-sectional, and nonrandomized trial. The study included 566 children being treated and followed up for allergic rhinitis. Skin prick test for the same allergens was performed for all patients. Adenoid tissue was analyzed by an ENT specialist and the diagnosis was confirmed based on the patient history, endoscopic physical examination and radiology. Adenoid hypertrophy was detected in 118 (21.2 %) of the children with AR. Children with and without AH did not differ statistically and significantly by gender, age, presence of atopy in the family, exposure to smoke (p > 0.05). Comparison of the groups for AR duration demonstrated significantly higher frequency of persistent rhinitis in patients with AH (p < 0.05). Of the AR patients with AH, 90 (76.3 %) had moderate-severe rhinitis and 274 (62.6 %) AR patients without AH had moderate-severe rhinitis (p = 0.005). Itchy nose was more frequent in AR patients without AH, and nasal congestion was more common in AR patients with AH (p = 0.017 and p = 0.001, respectively). The presence of asthma was more common among AR patients without AH (p = 0.037). Intergroup comparisons for presence of atopic dermatitis, the percentage of eosinophil, serum IgE levels, the number of positive sensitivity, polysensitization, sensitivity to house dust mite, cockroach, pollens and dander yielded no significant difference (p > 0.05). On the other hand, sensitivity to Alternaria alternata was significantly more frequent in AR patients with AH (p = 0.032). The presence of AH increased the severity of the disease and prolongs disease duration. There was a negative relationship between AH and asthma in children with AR. AH is more common among children with mold sensitivity. AH should be considered and investigated particularly in non-asthmatic children with pronounced nasal congestion and A. alternata sensitivity.


Maternal adrenalectomy alters the immune and endocrine functions of fetal alcohol-exposed male offspring.
Exposure to ethanol in utero compromises the offspring's developing immune and endocrine systems. Persistent functional changes, particularly in T-cell-dependent aspects of immunity and in hypothalamic-pituitary-adrenal activity, are commonly seen. The present study examined the degree to which fetal alcohol exposure (FAE) during development suppressed the lymphocyte proliferative response to Concanavalin-A (Con A). We also examined the effect of maternal adrenalectomy on the expression of glucocorticoid-regulated genes and the response to Con A in FAE offspring. Con A-stimulated lymphocyte proliferation was stably suppressed (between 28-46%) in FAE males compared to isocalorically pair-fed offspring at 7, 21, 40, and 60 days of age. In contrast, lymphocyte proliferation in the immature or peripubertal FAE female was totally unaffected. In 60-day- old male rats, maternal adrenalectomy reversed the FAE-induced suppression of Con A-stimulated proliferation, but had no effect on lymphocyte proliferation. FAE increased anterior pituitary POMC (the precursor of ACTH) mRNA levels dramatically in males, and this increase was also reversed by maternal adrenalectomy. In both sexes, anterior pituitary glucocorticoid receptor mRNA levels were unaffected by prenatal alcohol exposure alone, but were significantly decreased in male and increased in female offspring of adrenalectomized dams ingesting alcohol. Furthermore, in male, but not female, offspring, hypothalamic levels of glucocorticoid receptor and CRF mRNA were increased significantly by FAE alone or in combination with maternal adrenalectomy. In female, but not male, offspring, maternal adrenalectomy with concomitant alcohol exposure increased anterior pituitary POMC mRNA levels compared to that in sham/pair-fed offspring. In summary, FAE induced a gender-specific impairment of Con A-stimulated lymphocyte proliferation. This deficit is present both before and after puberty, demonstrating its stability into adulthood. Furthermore, in males, maternal adrenalectomy reversed these FAE-induced deficits in T-cell function as well as the effect of FAE on anterior pituitary POMC expression. This supports the hypothesis that maternal adrenal hormones participate in the immunosuppressive "imprinting" of the FAE fetus and are, therefore, causally implicated in the sexually dimorphic T-cell dysfunction found in FAE offspring.


Authors' response: Letter to the Editor concerning OCRA as preferred method in ISO standards on biomechanical risk factors.
We thank Drs. Colombini and Occhipinti for their personal reply to our Discussion Paper (1, 2). We share the overall goal of preventing workplace injuries and welcome a discussion of the ISO process on workplace ergonomics standards; this was the primary aim of the Discussion Paper. We hope that other members of the relevant ISO working groups will also participate in the discussion. However, Drs. Colombini and Occipinti misinterpret our paper. Our aim was not to "addresses the scientific basis of ISO standards on biomechanical risk factors and more specifically the OCRA methodology". The purpose was to point out that "while the ISO process has value, it has also clear limitations when it comes to developing occupational health and safety standards that should be based on scientific principles". It is true that our paper discussed the OCRA method, but only as an example, in a single paragraph. We noted that the OCRA method was promoted as the preferred method by the ISO working group even though there were other risk assessment methods which, at the time (and currently), were at least as scientifically valid (3). The discovery that, while on the ISO working group, Drs. Colombini and Occipinti elevated the risk assessment method that they developed (OCRA) over the other methods, demonstrates one of several limitations of the ISO process, namely, the lack of attention to conflict of interest. Finally, we would like to draw attention to the note by Drs. Colombini and Occhipinti that "the ISO standards in question were actually developed by the working group, as mandated by ISO, over the period 2000‒2004". This long-elapsed time, without an update to the standard, should be a concern for all scientists given the large quantity of quality scientific literature published since then (eg, 3‒6). Fourteen years is well beyond what is recommended in the ISO guidelines. References 1. Colombini D, Occhipinti E. Scientific basis of the OCRA method for risk assessment of biomechanical overload of the upper limb, as preferred method in ISO standards on biomechanical risk factors. Scand J Work Environ Health ‒ online first. https://doi.org.10.5271/sjweh.3746 2. Armstrong T J, Burdorf I A, Descatha A, Farioli A, Graf M, Horie S, Marras W S, Potvin J R, Rempel D, Spatari G, Takala E P, Verbeek J, Violante FS. Scientific basis of ISO standards on biomechanical risk factors. Scand J Work Environ Health ‒ online first. https://doi.org/10.5271/sjweh.3718 3. Takala EP, Pehkonen I, Forsman M, Hansson GA, Mathiassen SE, Neumann WP, Sjøgaard G, Veiersted KB, Westgaard RH, Winkel J. Systematic evaluation of observational methods assessing biomechanical exposures at work. Scand J Work Environ Health. 2010;36:3-24. https://doi.org/10.5271/sjweh.2876 4. Paulsen R, Gallu T, Gilkey D, Reiser R, Murgia L, Rosecrance J. The inter-rater reliability of Strain Index and OCRA Checklist task assessments in cheese processing. Applied Ergonomics. 2015; 51,199-204. https://doi.org/10.1016/j.apergo.2015.04.019 5. Kapellusch JM, Gerr FE, Malloy EJ, Garg A, Harris-Adamson C, Bao SS, Burt SE, Dale AM, Eisen EA, Evanoff BA, Hegmann KT, Silverstein BA, Theise MS, Rempel DM. Exposure-response relationships for the ACGIH threshold limit value for hand-activity level: results from a pooled data study of carpal tunnel syndrome. Scand J Work Environ Health. 2014;40:610-20. https://doi.org/10.5271/sjweh.3456 6. Violante FS, Farioli A, Graziosi F, Marinelli F, Curti S, Armstrong TJ, Mattioli S, Bonfiglioli R. Carpal tunnel syndrome and manual work: the OCTOPUS cohort, results of a ten-year longitudinal study. Scand J Work Environ Health. 2016;42:280-90. https://doi.org/10.5271/sjweh.3566.


Crosstalk between Tumor Cells and Macrophages in Stroma Renders Tumor Cells as the Primary Source of MCP-1/CCL2 in Lewis Lung Carcinoma.
The chemokine MCP-1/CCL2 is produced by a variety of tumors and plays an important role in cancer progression. We and others previously demonstrated that the primary source of MCP-1 in several mouse tumors, including 4T1 breast cancer, M5076 sarcoma, and B16 melanoma, was stromal cells. In the present study, we identified that tumor cells were the primary source of MCP-1 in Lewis lung carcinoma (LLC), because MCP-1 mRNA was highly expressed in tumors grown in both wild type (WT) and MCP-1(-/-) mice with elevated serum MCP-1 levels. Since LLC cells isolated from tumors expressed low levels of MCP-1 in vitro, it appeared that the tumor-stromal cell interaction in a tumor microenvironment increased MCP-1 expression in LLC cells. In fact, co-culture of LLC cells with normal mouse peritoneal macrophages or normal lung cells containing macrophages increased MCP-1 expression by LLC cells. Macrophages from TNFα(-/-) mice failed to activate LLC cells and anti-TNFα neutralizing antibody abolished the effect of WT macrophages on LLC cells. When LLC cells were transplanted into TNFα(-/-) mice, the levels of MCP-1 mRNA in tumors and serum MCP-1 levels were markedly lower as compared to WT mice, and importantly, tumors grew more slowly. Taken together, our results indicate that TNFα released by tumor cell-activated macrophages is critical for increased MCP-1 production by tumors cells. Thus, disruption of tumor-stromal cell interaction may inhibit tumor progression by reducing the production of tumor-promoting proinflammatory mediators, such as MCP-1.


Elevated prothrombin results in clots with an altered fiber structure: a possible mechanism of the increased thrombotic risk.
Individuals with elevated prothrombin levels are at increased risk of venous thrombosis. To understand the mechanism behind this observation, we studied the effect of prothrombin concentration on thrombin generation and fibrin clot structure. The pattern of thrombin generation was directly related to the prothrombin level at all concentrations tested. From 0% to 300% of normal plasma levels of prothrombin, increasing the prothrombin concentration increased the initial rate, peak, and total amount of thrombin generated. Importantly, fibrin clot structure was also affected by the prothrombin concentration. Fibrin clots made from prothrombin concentrations less than 10% of plasma levels were weak and poorly formed. Fibrin clots made at 10% to 100% of plasma levels of prothrombin had similar fiber structures (mass-to-length ratio; mu). However, the fiber mass-to-length ratio decreased with increasing prothrombin levels more than 100% of plasma levels, in a dose-dependent manner. These results suggest that increased levels of prothrombin alter thrombin generation and clot structure. Specifically, elevated prothrombin levels produce clots with reduced fibrin mass-to-length ratios compared with normal clots. We hypothesize that this alteration in fibrin clot structure is an important determinant of the risk of thrombosis.


Characterizing the genetic basis of transcriptome diversity through RNA-sequencing of 922 individuals.
Understanding the consequences of regulatory variation in the human genome remains a major challenge, with important implications for understanding gene regulation and interpreting the many disease-risk variants that fall outside of protein-coding regions. Here, we provide a direct window into the regulatory consequences of genetic variation by sequencing RNA from 922 genotyped individuals. We present a comprehensive description of the distribution of regulatory variation--by the specific expression phenotypes altered, the properties of affected genes, and the genomic characteristics of regulatory variants. We detect variants influencing expression of over ten thousand genes, and through the enhanced resolution offered by RNA-sequencing, for the first time we identify thousands of variants associated with specific phenotypes including splicing and allelic expression. Evaluating the effects of both long-range intra-chromosomal and trans (cross-chromosomal) regulation, we observe modularity in the regulatory network, with three-dimensional chromosomal configuration playing a particular role in regulatory modules within each chromosome. We also observe a significant depletion of regulatory variants affecting central and critical genes, along with a trend of reduced effect sizes as variant frequency increases, providing evidence that purifying selection and buffering have limited the deleterious impact of regulatory variation on the cell. Further, generalizing beyond observed variants, we have analyzed the genomic properties of variants associated with expression and splicing and developed a Bayesian model to predict regulatory consequences of genetic variants, applicable to the interpretation of individual genomes and disease studies. Together, these results represent a critical step toward characterizing the complete landscape of human regulatory variation.


CD34+CD38- leukemic stem cell frequency to predict outcome in acute myeloid leukemia.
Current risk algorithms are primarily based on pre-treatment factors and imperfectly predict outcome in acute myeloid leukemia (AML). We introduce and validate a post-treatment approach of leukemic stem cell (LSC) assessment for prediction of outcome. LSC containing CD34+CD38- fractions were measured using flow cytometry in an add-on study of the HOVON102/SAKK trial. Predefined cut-off levels were prospectively evaluated to assess CD34+CD38-LSC levels at diagnosis (n = 594), and, to identify LSClow/LSChigh (n = 302) and MRDlow/MRDhigh patients (n = 305) in bone marrow in morphological complete remission (CR). In 242 CR patients combined MRD and LSC results were available. At diagnosis the CD34+CD38- LSC frequency independently predicts overall survival (OS). After achieving CR, combining LSC and MRD showed reduced survival in MRDhigh/LSChigh patients (hazard ratio [HR] 3.62 for OS and 5.89 for cumulative incidence of relapse [CIR]) compared to MRDlow/LSChigh, MRDhigh/LSClow, and especially MRDlow/LSClow patients. Moreover, in the NPM1mutant positive sub-group, prognostic value of golden standard NPM1-MRD by qPCR can be improved by addition of flow cytometric approaches. This is the first prospective study demonstrating that LSC strongly improves prognostic impact of MRD detection, identifying a patient subgroup with an almost 100% treatment failure probability, warranting consideration of LSC measurement incorporation in future AML risk schemes.


An Investigation of Influential Users in the Promotion and Marketing of Heated Tobacco Products on Instagram: A Social Network Analysis.
While an increasing body of the literature has documented the exposure to emerging tobacco products including heated tobacco products (HTPs) on social media, few studies have investigated the various stakeholders involved in the generation of promotional tobacco content. This study constructed a social network of Instagram users who posted IQOS content, a leading HTP brand, between 1 January and 5 April 2021 and identified users who positioned near the center of the network. We identified 4526 unique Instagram users who had created 19,951 IQOS-related posts during the study period. Nearly half of the users (42.1%) were business accounts authorized by Instagram, among which 59.0% belonged to Personal Goods and General Merchandise Stores and 18.1% belonged to Creators and Celebrities. For users with higher in-degree, out-degree, betweenness, and closeness centrality in the network, the majority of them were accounts directly associated with IQOS (e.g., containing "iqos" in username) or related to tobacco business as self-identified in the bio. Our findings further refine the social media marketing presence of tobacco products and suggest that the current self-regulatory efforts led by social media platforms are far from enough.


Depletion of sirtuin 1 (SIRT1) leads to epigenetic modifications of telomerase (TERT) gene in hepatocellular carcinoma cells.
Sirtuin 1 (SIRT1) is a nicotinamide adenine dinucleotide (NAD)-dependent deacetylase that is implicated in plethora of biological processes, including metabolism, aging, stress response, and tumorigenesis. Telomerase (TERT) is essential for telomere maintenance. Activation of TERT is considered a crucial step in tumorigenesis, and therefore it is a potential therapeutic target against cancer. We have recently found that SIRT1 expression is highly elevated in hepatocellular carcinoma, and the depletion of SIRT1 leads to substantial reduction in TERT mRNA and protein expression. However, the underlying molecular mechanism of SIRT1-dependent TERT expression remains uncharacterized. Here, we elucidated if SIRT1 regulates TERT expression via transcriptional, epigenetic and post-transcriptional mechanisms. We report that depletion of SIRT1 does not lead to significant change in transcriptional activity and CpG methylation patterns of the TERT promoter, nor does it affect mRNA stability or 3'-UTR regulation of TERT. Intriguingly, depletion of SIRT1 is associated with substantial induction of acetylated histone H3-K9 and reduction of trimethyl H3-K9 at the TERT gene, which are known to be associated with gene activation. Our data revealed that SIRT1 regulates histone acetylation and methylation at the TERT promoter. We postulated that SIRT1 may regulate TERT expression via long-range interaction, or via yet unidentified histone modifications.


Collection devices for human papillomavirus.
Human papillomavirus (HPV) testing has relied to date on samples collected by experienced health professionals. Self-administered testing devices could allow HPV testing to occur in large-scale epidemiologic studies of primary care screening populations. The purpose of this study is to determine whether a self-collection device for cervicovaginal HPV infection could be developed. A prospective randomized trial of a consecutive sampling of 93 women, 18 years or older, receiving routine cervical cancer screening and colposcopy in the urban gynecologic clinics in Philadelphia, Pennsylvania, were randomized into 2 arms. Women in arm 1 used a self-administered tampon before the physician-directed swabs of the cervix; in arm 2, women underwent the physician-directed swab testing before using the self-administered tampon. The concordance of HPV DNA positivity between sampling methods detected by a Hybrid Capture HPV tube test for both low- and high-risk types of HPV was the main outcome measure. The concordance rate (ie, women whose cultures were classified as negative on both tests or positive on both tests) for arms 1 and 2 were similar: 78.3% and 80.9%, respectively. The tampon was equivalent to the physician-directed swab in HPV detection and suggests its feasibility in long-term primary care studies of screening populations.


Meiotic errors followed by two parallel postzygotic trisomy rescue events are a frequent cause of constitutional segmental mosaicism.
Structural copy number variation (CNV) is a frequent cause of human variation and disease. Evidence is mounting that somatic acquired CNVs are prevalent, with mosaicisms of large segmental CNVs in blood found in up to one percent of both the healthy and patient populations. It is generally accepted that such constitutional mosaicisms are derived from postzygotic somatic mutations. However, few studies have tested this assumption. Here we determined the origin of CNVs which coexist with a normal cell line in nine individuals. We show that in 2/9 the CNV originated during meiosis. The existence of two cell lines with 46 chromosomes thus resulted from two parallel trisomy rescue events during postzygotic mitoses.


Self-Recognition of an Inducible Host lncRNA by RIG-I Feedback Restricts Innate Immune Response.
The innate RNA sensor RIG-I is critical in the initiation of antiviral type I interferons (IFNs) production upon recognition of "non-self" viral RNAs. Here, we identify a host-derived, IFN-inducible long noncoding RNA, lnc-Lsm3b, that can compete with viral RNAs in the binding of RIG-I monomers and feedback inactivate the RIG-I innate function at late stage of innate response. Mechanistically, binding of lnc-Lsm3b restricts RIG-I protein's conformational shift and prevents downstream signaling, thereby terminating type I IFNs production. Multivalent structural motifs and long-stem structure are critical features of lnc-Lsm3b for RIG-I binding and inhibition. These data reveal a non-canonical self-recognition mode in the regulation of immune response and demonstrate an important role of an inducible "self" lncRNA acting as a potent molecular decoy actively saturating RIG-I binding sites to restrict the duration of "non-self" RNA-induced innate immune response and maintaining immune homeostasis, with potential utility in inflammatory disease management.


Inhibition of mitophagy drives macrophage activation and antibacterial defense during sepsis.
Mitochondria have emerged as key actors of innate and adaptive immunity. Mitophagy has a pivotal role in cell homeostasis, but its contribution to macrophage functions and host defense remains to be delineated. Here, we showed that lipopolysaccharide (LPS) in combination with IFN-γ inhibited PINK1-dependent mitophagy in macrophages through a STAT1-dependent activation of the inflammatory caspases 1 and 11. In addition, we demonstrated that the inhibition of mitophagy triggered classical macrophage activation in a mitochondrial ROS-dependent manner. In a murine model of polymicrobial infection (cecal ligature and puncture), adoptive transfer of Pink1-deficient bone marrow or pharmacological inhibition of mitophagy promoted macrophage activation, which favored bactericidal clearance and led to a better survival rate. Reciprocally, mitochondrial uncouplers that promote mitophagy reversed LPS/IFN-γ-mediated activation of macrophages and led to immunoparalysis with impaired bacterial clearance and lowered survival. In critically ill patients, we showed that mitophagy was inhibited in blood monocytes of patients with sepsis as compared with nonseptic patients. Overall, this work demonstrates that the inhibition of mitophagy is a physiological mechanism that contributes to the activation of myeloid cells and improves the outcome of sepsis.


Reduced mammary tumor progression in a transgenic mouse model fed an isoflavone-poor soy protein concentrate.
Dietary exposure to soy has been associated with reduced breast cancer incidence. Soy isoflavones and protein components, such as protease inhibitors and the lunasin peptide, have been indicated as potential agents reducing carcinogenesis. In this study, the effect of soy-based diets was evaluated in a transgenic mouse model of breast carcinoma, overexpressing the neu oncogene. Neu female mice were fed for 20 wk a soy- and isoflavone-free diet (IFD), 4RF21 laboratory mouse diet, soy-based, thus isoflavone-rich (STD), or AIN-76-based semisynthetic diets with a soy protein isolate (SPI) or an isoflavone-poor soy protein concentrate (IPSP) as protein source. Mice were then sacrificed and tumors removed. Mammary tumor weights were not different in SPI versus IFD and STD fed mice. In contrast, mice fed IPSP showed reduced tumor progression versus IFD and STD groups (p < 0.05). Moreover, IPSP fed mice showed lower bromo-2'-deoxyuridine (BrdU) incorporation into breast tumor cells compared to STD and SPI fed animals (p < 0.02). Lung metastases were detected in 80% of IFD fed mice, in 70% of mice fed STD and SPI, and only in 50% of the IPSP fed animals. These results indicate that a diet containing an isoflavone-poor soy protein concentrate may inhibit breast tumor progression and metastasis development.


Regulation of inosine-5'-monophosphate dehydrogenase type II gene expression in human T cells. Role for a novel 5' palindromic octamer sequence.
Expression of the gene encoding human inosine- 5'-monophosphate dehydrogenase (IMPDH) type II, an enzyme catalyzing the rate-limiting step in the generation of guanine nucleotides, is increased more than 10-fold in activated peripheral blood T lymphocytes and is required for T cell activation. We have examined the 5'-regulatory sequences that are important for the transcriptional regulation of this gene in T cells. DNase I mapping of genomic DNA identified a hypersensitive element near the transcription initiation site. Fine mapping by in vivo footprinting demonstrated five transcription factor binding sites that are occupied in both resting and activated peripheral blood T lymphocytes; these are tandem CRE motifs, a Sp1 site, an overlapping Egr-1/Sp1 site, and a novel palindromic octamer sequence (POS). The tandem CRE and POS sites are of major functional importance as judged by mutational and electrophoretic mobility shift analyses. These data provide evidence that expression of the human IMPDH type II gene is predominantly regulated by the nuclear factors ATF-2 and an as yet unidentified POS-binding protein. Additional major protein-DNA interactions do not occur within the promoter region after T lymphocyte activation, indicating a requirement for additional protein-protein interactions and/or post-translational modifications of pre-bound transcription factors to account for the observed increase in IMPDH type II gene expression.


Numerical simulation of proton distribution with electric double layer in extended nanospaces.
Understanding the properties of liquid confined in extended nanospaces (10-1000 nm) is crucial for nanofluidics. Because of the confinement and surface effects, water may have specific structures and reveals unique physicochemical properties. Recently, our group has developed a super resolution laser-induced fluorescence (LIF) technique to visualize proton distribution with the electrical double layer (EDL) in a fused-silica extended nanochannel (Kazoe, Y.; Mawatari, K.; Sugii, Y.; Kitamori, T. Anal. Chem.2011, 83, 8152). In this study, based on the coupling of the Poisson-Boltzmann theory and site-dissociation model, the effect of specific water properties in an extended nanochannel on formation of EDL was investigated by comparison of numerical results with our previous experimental results. The numerical results of the proton distribution with a lower dielectric constant of approximately 17 were shown to be in good agreement with our experimental results, which confirms our previous observation showing a lower water permittivity in an extended nanochannel. In addition, the higher silanol deprotonation rate in extended nanochannels was also demonstrated, which is supported by our previous results of NMR and streaming current measurements. The present results will be beneficial for a further understanding of interfacial chemistry, fluid physics, and electrokinetics in extended nanochannels.


The missing pieces for better future predictions in subarctic ecosystems: A Torneträsk case study.
Arctic and subarctic ecosystems are experiencing substantial changes in hydrology, vegetation, permafrost conditions, and carbon cycling, in response to climatic change and other anthropogenic drivers, and these changes are likely to continue over this century. The total magnitude of these changes results from multiple interactions among these drivers. Field measurements can address the overall responses to different changing drivers, but are less capable of quantifying the interactions among them. Currently, a comprehensive assessment of the drivers of ecosystem changes, and the magnitude of their direct and indirect impacts on subarctic ecosystems, is missing. The Torneträsk area, in the Swedish subarctic, has an unrivalled history of environmental observation over 100 years, and is one of the most studied sites in the Arctic. In this study, we summarize and rank the drivers of ecosystem change in the Torneträsk area, and propose research priorities identified, by expert assessment, to improve predictions of ecosystem changes. The research priorities identified include understanding impacts on ecosystems brought on by altered frequency and intensity of winter warming events, evapotranspiration rates, rainfall, duration of snow cover and lake-ice, changed soil moisture, and droughts. This case study can help us understand the ongoing ecosystem changes occurring in the Torneträsk area, and contribute to improve predictions of future ecosystem changes at a larger scale. This understanding will provide the basis for the future mitigation and adaptation plans needed in a changing climate.


A correlation between the relative predisposition of MHC class II alleles to type 1 diabetes and the structure of their proteins.
In human type 1 diabetes (T1D) and in its murine model, the major histocompatibility complex (MHC) class II molecules, human leukocyte antigens (HLA)-DQ and -DR and their murine orthologues, IA and IE, are the major genetic determinants. In this report, we have ranked HLA class II molecule-associated T1D risk in a two-sided gradient from very high to very low. Very low risk corresponded to dominant protection from T1D. We predicted the protein structure of DQ by using the published crystal structures of different allotypes of the murine orthologue of DQ, IA. We discovered marked similarities both within, and cross species between T1D protective class II molecules. Likewise, the T1D predisposing molecules showed conserved similarities that contrasted with the shared patterns observed between the protective molecules. We also found striking inter-isotypic conservation between protective DQ, IA allotypes and protective DR4 subtypes. The data provide evidence for a joint action of the class II peptide-binding pockets P1, P4 and P9 in disease susceptibility and resistance with a main role for P9 in DQ/IA and for P1 and P4 in DR/IE. Overall, these results suggest shared epitope(s) in the target autoantigen(s), and common pathways in human and murine T1D.


Characterizing associations and SNP-environment interactions for GWAS-identified prostate cancer risk markers--results from BPC3.
Genome-wide association studies (GWAS) have identified multiple single nucleotide polymorphisms (SNPs) associated with prostate cancer risk. However, whether these associations can be consistently replicated, vary with disease aggressiveness (tumor stage and grade) and/or interact with non-genetic potential risk factors or other SNPs is unknown. We therefore genotyped 39 SNPs from regions identified by several prostate cancer GWAS in 10,501 prostate cancer cases and 10,831 controls from the NCI Breast and Prostate Cancer Cohort Consortium (BPC3). We replicated 36 out of 39 SNPs (P-values ranging from 0.01 to 10⁻²⁸). Two SNPs located near KLK3 associated with PSA levels showed differential association with Gleason grade (rs2735839, P = 0.0001 and rs266849, P = 0.0004; case-only test), where the alleles associated with decreasing PSA levels were inversely associated with low-grade (as defined by Gleason grade < 8) tumors but positively associated with high-grade tumors. No other SNP showed differential associations according to disease stage or grade. We observed no effect modification by SNP for association with age at diagnosis, family history of prostate cancer, diabetes, BMI, height, smoking or alcohol intake. Moreover, we found no evidence of pair-wise SNP-SNP interactions. While these SNPs represent new independent risk factors for prostate cancer, we saw little evidence for effect modification by other SNPs or by the environmental factors examined.


The effect of mobility on sexual risk behaviour and HIV infection: a cross-sectional study of men who have sex with men in southern India.
Mobility is an important factor contributing to the spread of HIV among key population at risk for HIV; however, research linking this relationship among men who have sex men (MSM) is scarce in India. This study examines the association between mobility and sexual risk behaviour and HIV infection among MSM in southern India. Data are drawn from a cross-sectional biobehavioural survey of 1608 self-identified MSM from four districts of Andhra Pradesh in India, recruited through a probability-based sampling in 2009-2010. Logistic regression models were used to estimate odds ratios and 95% CIs for sexual risk behaviours (unprotected sex with any male partner) and HIV infection based on the mobility status (travelled and had sex in the past year) after adjusting for sociodemographics and risk behaviours. Of the 1608 MSM, one-fourth (26%) were mobile. Of these, three-fourths had travelled across districts but within the state (56%), and one-fifth (20%) across states. As compared to non-mobile MSM, a higher proportion of MSM who were mobile across districts (adjusted (OR=1.42, 95% CI 1.04 to 1.95) or states (adjusted OR=3.20, 95% CI 1.65 to 6.17) reported having unprotected sex with any male sexual partner. Further, mobility across districts (adjusted OR=1.43, 95% CI 1.01 to 2.03) or states (adjusted OR=2.45, 95% CI 1.46 to 4.10) was significantly associated with HIV infection. Mobile MSM have a higher likelihood of contracting HIV. Interventions extending the ways to reach out to MSM with greater mobility may augment ongoing efforts to reduce the spread of HIV/AIDS in India.


Metachronous Contralateral Testicular Cancer in the Cisplatin Era: A Population-Based Cohort Study.
It is hypothesized that cisplatin-based chemotherapy (CBCT) reduces the occurrence of metachronous contralateral (second) germ cell testicular cancer (TC). However, studies including treatment details are lacking. The aim of this study was to assess the second TC risk, emphasizing the impact of previous TC treatment. Based on the Cancer Registry of Norway, 5,620 men were diagnosed with first TC between 1980 and 2009. Treatment data regarding TC were retrieved from medical records. Cumulative incidences of second TC were estimated, and standardized incidence ratios were calculated. The effect of treatment intensity was investigated using Cox proportional hazard regression. Median follow-up was 18.0 years, during which 218 men were diagnosed with a second TC after median 6.2 years. Overall, the 20-year crude cumulative incidence was 4.0% (95% CI, 3.5 to 4.6), with lower incidence after chemotherapy (CT) (3.2%; 95% CI, 2.5 to 4.0) than after surgery only (5.4%; 95% CI, 4.2 to 6.8). The second TC incidence was also lower for those age ≥ 30 years (2.8%; 95% CI, 2.3 to 3.4) at first TC diagnosis than those age < 30 years (6.0%; 95% CI, 5.0 to 7.1). Overall, the second TC risk was 13-fold higher compared with the risk of developing TC in the general male population (standardized incidence ratio, 13.1; 95% CI, 11.5 to 15.0). With surgery only as reference, treatment with CT significantly reduced the second TC risk (hazard ratio [HR], 0.55). For each additional CBCT cycle administered, the second TC risk decreased significantly after three, four, and more than four cycles (HRs, 0.53, 0.41, and 0.21, respectively). Age at first TC diagnosis and treatment intensity influenced the second TC risk, with significantly reduced risks after more than two CBCT cycles.


Comprehensive genomic characterization of head and neck squamous cell carcinomas.
The Cancer Genome Atlas profiled 279 head and neck squamous cell carcinomas (HNSCCs) to provide a comprehensive landscape of somatic genomic alterations. Here we show that human-papillomavirus-associated tumours are dominated by helical domain mutations of the oncogene PIK3CA, novel alterations involving loss of TRAF3, and amplification of the cell cycle gene E2F1. Smoking-related HNSCCs demonstrate near universal loss-of-function TP53 mutations and CDKN2A inactivation with frequent copy number alterations including amplification of 3q26/28 and 11q13/22. A subgroup of oral cavity tumours with favourable clinical outcomes displayed infrequent copy number alterations in conjunction with activating mutations of HRAS or PIK3CA, coupled with inactivating mutations of CASP8, NOTCH1 and TP53. Other distinct subgroups contained loss-of-function alterations of the chromatin modifier NSD1, WNT pathway genes AJUBA and FAT1, and activation of oxidative stress factor NFE2L2, mainly in laryngeal tumours. Therapeutic candidate alterations were identified in most HNSCCs.


Partner support and maternal depression in the context of the Iowa floods.
A systematic investigation of the role of prenatal partner support in perinatal maternal depression was conducted. Separate facets of partner support were examined (i.e., received support and support adequacy) and a multidimensional model of support was applied to investigate the effects of distinct types of support (i.e., informational, physical comfort, emotional/esteem, and tangible support). Both main and stress-buffering models of partner support were tested in the context of prenatal maternal stress resulting from exposure to a natural disaster. Questionnaire data were analyzed from 145 partnered women using growth curve analytic techniques. Results indicate that received support interacts with maternal flood stress during pregnancy to weaken the association between stress and trajectories of maternal depression from pregnancy to 30 months postpartum. Support adequacy did not interact with stress, but was associated with levels of depressive symptoms controlling for maternal stress and received support. Results demonstrate the distinct roles of various facets and types of support for a more refined explanatory model of prenatal partner support and perinatal maternal depression. Results inform both main effect and stress buffering models of partner support as they apply to the etiology of perinatal maternal depression, and highlight the importance of promoting partner support during pregnancy that matches support preferences.


Multiwavelength erbium-doped fiber laser based on graphene oxide.
A multiwavelength erbium-doped fiber (EDF) laser based on graphene oxide (GO) has been proposed, to the best of our knowledge, for the first time, to generate an output of stable wavelengths. The structure mainly comprises a few layers of GO between two single-mode fibers incorporated into a capillary device and a Lyot comb filter. GO can show a good nonlinear optical effect, which is beneficial to suppress the mode competition caused by the EDF and stabilize the multiwavelength output. With assistance from the GO device, 11 stable simultaneous lasing signals with a power nonuniformity of about 1.5 dB are obtained. Wavelength spacing is about 0.42 nm and the linewidth of each wavelength is less than 0.07 nm.


Huperzine a in the treatment of Alzheimer's disease and vascular dementia: a meta-analysis.
The objective of our study was to perform an updated meta-analysis of placebo-controlled RCTs of Huperzine A (Hup A) on patients with Alzheimer's disease (AD) and vascular dementia (VD), in order to provide the basis and reference for clinical rational drug use. The primary outcome measures assessed were minimental state examination (MMSE) and activities of daily living scale (ADL). Eight AD trials with 733 participants and two VD trials with 92 participants that met our inclusion criteria were identified. The results showed that Hup A could significantly improve the MMSE and ADL score of AD and VD patients, and longer durations would result in better efficacy for the patients with AD. It seemed that there was significant improvement of cognitive function measured by memory quotient (MQ) in patients with AD. Most adverse effects in AD were generally of mild to moderate severity and transient. Compared to the patients with AD, Hup A may offer fewer side effects for participants with VD in this study. Therefore, Hup A is a well-tolerated drug that could significantly improve cognitive performance in patients with AD or VD, but we need to use it with caution in the clinical treatment.


Effect of Cryopreservation and Post-Cryopreservation Somatic Embryogenesis on the Epigenetic Fidelity of Cocoa (Theobroma cacao L.).
While cocoa plants regenerated from cryopreserved somatic embryos can demonstrate high levels of phenotypic variability, little is known about the sources of the observed variability. Previous studies have shown that the encapsulation-dehydration cryopreservation methodology imposes no significant extra mutational load since embryos carrying high levels of genetic variability are selected against during protracted culture. Also, the use of secondary rather than primary somatic embryos has been shown to further reduce the incidence of genetic somaclonal variation. Here, the effect of in vitro conservation, cryopreservation and post-cryopreservation generation of somatic embryos on the appearance of epigenetic somaclonal variation were comparatively assessed. To achieve this we compared the epigenetic profiles, generated using Methylation Sensitive Amplified Polymorphisms, of leaves collected from the ortet tree and from cocoa somatic embryos derived from three in vitro conditions: somatic embryos, somatic embryos cryopreserved in liquid nitrogen and somatic embryos generated from cryoproserved somatic embryos. Somatic embryos accumulated epigenetic changes but these were less extensive than in those regenerated after storage in LN. Furthermore, the passage of cryopreserved embryos through another embryogenic stage led to further increase in variation. Interestingly, this detected variability appears to be in some measure reversible. The outcome of this study indicates that the cryopreservation induced phenotypic variability could be, at least partially, due to DNA methylation changes. Phenotypic variability observed in cryostored cocoa somatic-embryos is epigenetic in nature. This variability is partially reversible, not stochastic in nature but a directed response to the in-vitro culture and cryopreservation.


The alarmin IL-33 promotes regulatory T-cell function in the intestine.
FOXP3(+) regulatory T cells (Treg cells) are abundant in the intestine, where they prevent dysregulated inflammatory responses to self and environmental stimuli. It is now appreciated that Treg cells acquire tissue-specific adaptations that facilitate their survival and function; however, key host factors controlling the Treg response in the intestine are poorly understood. The interleukin (IL)-1 family member IL-33 is constitutively expressed in epithelial cells at barrier sites, where it functions as an endogenous danger signal, or alarmin, in response to tissue damage. Recent studies in humans have described high levels of IL-33 in inflamed lesions of inflammatory bowel disease patients, suggesting a role for this cytokine in disease pathogenesis. In the intestine, both protective and pathological roles for IL-33 have been described in murine models of acute colitis, but its contribution to chronic inflammation remains ill defined. Here we show in mice that the IL-33 receptor ST2 is preferentially expressed on colonic Treg cells, where it promotes Treg function and adaptation to the inflammatory environment. IL-33 signalling in T cells stimulates Treg responses in several ways. First, it enhances transforming growth factor (TGF)-β1-mediated differentiation of Treg cells and, second, it provides a necessary signal for Treg-cell accumulation and maintenance in inflamed tissues. Strikingly, IL-23, a key pro-inflammatory cytokine in the pathogenesis of inflammatory bowel disease, restrained Treg responses through inhibition of IL-33 responsiveness. These results demonstrate a hitherto unrecognized link between an endogenous mediator of tissue damage and a major anti-inflammatory pathway, and suggest that the balance between IL-33 and IL-23 may be a key controller of intestinal immune responses.


Short report: do intestinal nematodes increase the risk for multibacillary leprosy?
Intestinal helminths are known to subvert the host's immune response towards a Th2 response, which in turn may lead to both eosinophilia and high immunoglobulin E titers often associated with these parasites. Mycobacterium leprae infection may lead to different clinical and pathological forms. Multibacillary forms are associated with Th2 cytokines, whereas paucibacillary forms are associated with Th1 cytokines. We report a significantly higher frequency of intestinal helminthic infections in patients with the lepromatous form, a multibacillary form of leprosy (odds ratio, 2.99; 95% confidence interval, 1.82-4.95; P = 0.006) when compared with patients with paucibacillary leprosy or to a control group without leprosy. A direct correlation was also found between mycobacterial index and the frequency of intestinal helminths. Our results suggest that the presence of intestinal helminths may facilitate the establishment of M. leprae infection or the progression to more severe forms of leprosy.


Electron densities by the maximum entropy method (MEM) for various types of prior densities: a case study on three amino acids and a tripeptide.
Dynamic model densities according to Mondal et al. [(2012), Acta Cryst. A68, 568-581] are presented for independent atom models (IAM), IAMs after high-order refinements (IAM-HO), invariom (INV) models and multipole (MP) models of α-glycine, DL-serine, L-alanine and Ala-Tyr-Ala at T ≃ 20 K. Each dynamic model density is used as prior in the calculation of electron density according to the maximum entropy method (MEM). We show that at the bond-critical points (BCPs) of covalent C-C and C-N bonds the IAM-HO and INV priors produce reliable MEM density maps, including reliable values for the density and its Laplacian. The agreement between these MEM density maps and dynamic MP density maps is less good for polar C-O bonds, which is explained by the large spread of values of topological descriptors of C-O bonds in static MP densities. The density and Laplacian at BCPs of hydrogen bonds have similar values in MEM density maps obtained with all four kinds of prior densities. This feature is related to the smaller spatial variation of the densities in these regions, as expressed by small magnitudes of the Laplacians and the densities. It is concluded that the use of the IAM-HO prior instead of the IAM prior leads to improved MEM density maps. This observation shows interesting parallels to MP refinements, where the use of the IAM-HO as an initial model is the accepted procedure for solving MP parameters. A deconvolution of thermal motion and static density that is better than the deconvolution of the IAM appears to be necessary in order to arrive at the best MP models as well as at the best MEM densities.


Cardiorenal actions of neutral endopeptidase inhibition in experimental congestive heart failure.
The present studies were designed to determine the action of neutral endopeptidase inhibition (NEP-I), an inhibitor of the degradation of atrial natriuretic factor (ANF), in congestive heart failure (CHF). Studies were conducted in two groups of anesthetized dogs with CHF induced by 8 days of rapid right ventricular pacing. Group 1 (n = 5) received a specific NEP-I (SQ 28,603) at two doses administered sequentially -30 mg/kg followed by a 60 mg/kg i.v. bolus. Group 2 (n = 5) received intravenous infusion of exogenous ANF (100 ng/kg/min) to achieve increases in plasma ANF concentration as observed in group 1. NEP-I resulted in a diuresis and natriuresis (p less than 0.05) with increases in the fractional excretion of sodium and fractional excretion of lithium, the latter a marker for proximal tubule sodium delivery. Such tubular actions occurred in the absence of increases in glomerular filtration rate or renal blood flow but were associated with significant increases in urinary ANF and urinary cyclic GMP. Plasma ANF increased after the 30 mg/kg NEP-I dose. In contrast, in group 2 with exogenous ANF and despite a marked increase in plasma ANF, no natriuresis was observed. Arterial pressure did not change in either group. These studies demonstrate for the first time in CHF that NEP-I may potentiate the natriuretic action of endogenous ANF by a mechanism that is independent of systemic or renal hemodynamics and does not parallel increases in plasma ANF. These studies support an important therapeutic role for NEP-I in CHF.


Avoiding COVID-19 complications with diabetic patients could be achieved by multi-dose Bacillus Calmette-Guérin vaccine: a case study of beta cells regeneration.
Diabetes mellitus (DM) is one of the major risk factors for COVID-19 complications as it is one of the chronic immune-compromising conditions especially if patients have uncontrolled diabetes, poor HbA1c and/or irregular blood glucose levels. Diabetic patients' mortality rates with COVID-19 are higher than those of cardiovascular or cancer patients. Recently, Bacillus Calmette-Guérin (BCG) vaccine has shown successful results in reversing diabetes in both rats and clinical trials based on different mechanisms from aerobic glycolysis to beta cells regeneration. BCG is a multi-face vaccine that has been used extensively in protection from tuberculosis (TB) and leprosy and has been repositioned for treatment of bladder cancer, diabetes and multiple sclerosis. Recently, COVID-19 epidemiological studies confirmed that universal BCG vaccination reduced morbidity and mortality in certain geographical areas. Countries without universal policies of BCG vaccination (Italy, Nederland, USA) have been more severely affected compared to countries with universal and long-standing BCG policies that have shown low numbers of reported COVID-19 cases. Some countries have started clinical trials that included a single dose BCG vaccine as prophylaxis from COVID-19 or an attempt to minimize its side effects. This proposed research aims to use BCG vaccine as a double-edged weapon countering both COVID-19 and diabetes, not only as protection but also as therapeutic vaccination. The work includes a case study of regenerated pancreatic beta cells based on improved C-peptide and PCPRI laboratory findings after BCG vaccination for a 9 year old patient. The patient was re-vaccinated based on a negative tuberculin test and no scar at the site of injection of the 1st BCG vaccination at birth. The authors suggest and invite the scientific community to take into consideration the concept of direct BCG re-vaccination (after 4 weeks) because of the reported gene expressions and exaggerated innate immunity consequently. As the diabetic MODY-5 patient (mutation of HNF1B, Val2Leu) was on low dose Riomet® while eliminating insulin gradually, a simple analytical method for metformin assay was recommended to ensure its concentration before use as it is not approved yet by the Egyptian QC labs.


Activation of HIF1α Rescues the Hypoxic Response and Reverses Metabolic Dysfunction in the Diabetic Heart.
Type 2 diabetes (T2D) impairs hypoxia-inducible factor (HIF)1α activation, a master transcription factor that drives cellular adaptation to hypoxia. Reduced activation of HIF1α contributes to the impaired post-ischemic remodeling observed following myocardial infarction in T2D. Molidustat is an HIF stabilizer currently undergoing clinical trials for the treatment of renal anemia associated with chronic kidney disease; however, it may provide a route to pharmacologically activate HIF1α in the T2D heart. In human cardiomyocytes, molidustat stabilized HIF1α and downstream HIF target genes, promoting anaerobic glucose metabolism. In hypoxia, insulin resistance blunted HIF1α activation and downstream signaling, but this was reversed by molidustat. In T2D rats, oral treatment with molidustat rescued the cardiac metabolic dysfunction caused by T2D, promoting glucose metabolism and mitochondrial function, while suppressing fatty acid oxidation and lipid accumulation. This resulted in beneficial effects on post-ischemic cardiac function, with the impaired contractile recovery in T2D heart reversed by molidustat treatment. In conclusion, pharmacological HIF1α stabilization can overcome the blunted hypoxic response induced by insulin resistance. In vivo this corrected the abnormal metabolic phenotype and impaired post-ischemic recovery of the diabetic heart. Therefore, molidustat may be an effective compound to further explore the clinical translatability of HIF1α activation in the diabetic heart.


The mount st. Helens volcanic eruption of 18 may 1980: large short-term surface temperature effects.
The surface temperature effects of the 18 May 1980 eruption of Mount St. Helens Volcano were examinedfor 1 day immediately after the eruption; 24-hour temperature differences and Model Output Statistics errors as well as the detailed temporal evolution of surface temperature at selected stations were used. During the daytime hours immediately after the eruption, the temperature was suppressed by the volcanic plume by as much as 8 degrees C. That night, low-level volcanic dust produced temperature enhancements of up to 8 degrees C. These effects quickly diminished the next day as the volcanic dust cloud dissipated and moved toward the east. The net local effect of the eruption appears to be warming, in contrast to cooling which might be expected over climatic time scales.


The effects of quinine and chloroquine antimalarial treatments in the first trimester of pregnancy.
Quinine (n = 246) was used to treat uncomplicated Plasmodium falciparum and chloroquine (n = 130) was used to treat P. vivax, in a total of 376 episodes of malaria in the first trimester of pregnancy, in 300 Karen women (Thailand, 1995-2000). Parasites were still present on day 6 or 7 in 4.7% (11/234) of episodes treated with quinine. The overall 28 day parasite reappearance rate following quinine was 28.7% (60/209) for primary treatments and 44% (11/25) for re-treatments. Quinine treatment resulted in a high rate of gametocyte carriage: person-gametocyte-weeks = 42.5 (95% CI 27.8-62.1) per 1000 woman-weeks. For P. vivax, the reappearance rate for all episodes by day 28 was 4.5% (5/111). Significantly more women complained of tinnitus following quinine treatment compared to on admission: 64.5% (78/121) vs 31.6% (59/187), P < 0.001. Using survival analysis, the community rate of spontaneous abortion in women who never had malaria in pregnancy, 17.8% (16.5-19.0), did not differ significantly from rates in women treated with quinine: 22.9% (95% CI 15.5-30.3), or chloroquine: 18.3% (95% CI 9.3-27.3), P = 0.42. Pregnancies exposed to quinine or chloroquine and carried to term did not have increased rates of congenital abnormality, stillbirth or low birthweight. These results suggest that therapeutic doses of quinine and chloroquine are safe to use in the first trimester of pregnancy.


Improved control of multiple-antibiotic-resistance-related microbial risk in swine manure wastes by autothermal thermophilic aerobic digestion.
In this study, we microbiologically evaluated antibiotic resistance and pathogenicity in livestock (swine) manure as well as its biologically stabilized products. One of new livestock manure stabilization techniques is ATAD (Autothermal Thermophilic Aerobic Digestion). Because of its high operation temperature (60-65 degrees C), it has been speculated to have effective microbial risk control in livestock manure. This hypothesis was tested by evaluating microbial risk in ATAD-treated swine manure. Antibiotic resistance, multiple antibiotic resistance (MAR), and pathogenicity were microbiologically examined for swine manure as well as its conventionally stabilized (anaerobically fermented) and ATAD-stabilized products. In the swine manure and its conventionally stabilized product, antibiotic resistant (tetracycline-, kanamycine-, ampicillin-, and rifampicin-resistant) bacteria and the pathogen indicator bacteria were detected. Furthermore, approximately 2-5% of the Staphylococcus and Salmonella colonies from their selective culture media were found to exhibit a MAR-phenotypes, suggesting a serious level of microbe induced health risk. In contrast, after the swine manure was stabilized with a pilot-scale ATAD treatment for 3 days at 60-65 degrees C, antibiotic resistant bacteria, pathogen indicator bacteria, and MAR-exhibiting pathogens were all undetected. These findings support the improved control of microbial risk in livestock wastes by ATAD treatment.


Angular tuning bias of vibrissa-responsive cells in the paralemniscal pathway.
One of the most salient features of primary vibrissal afferents is their sensitivity to the direction in which the vibrissae move. Directional sensitivity is also well conserved in brainstem, thalamic, and cortical neurons of the lemniscal pathway, indicating that this property plays a key role in the organization of the vibrissal system. Here, we show that directional tuning is also a fundamental feature of second-order interpolaris neurons that give rise to the paralemniscal pathway. Quantitative assessment of responses to vibrissa deflection revealed an anisotropic organization of receptive fields with regard to topography, response magnitude, and the degree of angular tuning. Responses evoked by all vibrissae within the receptive field of each cell exhibited a high consistency of direction preference, but a striking difference in angular tuning preference was found among cells that reside in the rostral and caudal divisions of the interpolaris nucleus. Although in caudal interpolaris vectors of angular preference pointed in all directions, in rostral interpolaris virtually all vectors pointed upward, revealing a strong preference for this direction. Control experiments showed that the upward bias did not rely on a preferential innervation of rostral cells by upwardly tuned primary vibrissa afferents, nor did it rely on a direction-selective recruitment of feedforward inhibition. We thus propose that the upward preference bias of rostral cells, which project to the posterior group of the thalamus, emerges from use-dependent synaptic processes that relate to the kinematics of whisking.


A Novel Combination Treatment Targeting BCL-XL and MCL1 for KRAS/BRAF-mutated and BCL2L1-amplified Colorectal Cancers.
Colorectal cancer is the third most commonly diagnosed cancer in the world, and exhibits heterogeneous characteristics in terms of genomic alterations, expression signature, and drug responsiveness. Although there have been considerable efforts to classify this disease based on high-throughput sequencing techniques, targeted treatments for specific subgroups have been limited. KRAS and BRAF mutations are prevalent genetic alterations in colorectal cancers, and patients with mutations in either of these genes have a worse prognosis and are resistant to anti-EGFR treatments. In this study, we have found that a subgroup of colorectal cancers, defined by having either KRAS or BRAF (KRAS/BRAF) mutations and BCL2L1 (encoding BCL-XL) amplification, can be effectively targeted by simultaneous inhibition of BCL-XL (with ABT-263) and MCL1 (with YM-155). This combination treatment of ABT-263 and YM-155 was shown to have a synergistic effect in vitro as well as in in vivo patient-derived xenograft models. Our data suggest that combined inhibition of BCL-XL and MCL1 provides a promising treatment strategy for this genomically defined colorectal cancer subgroup. Mol Cancer Ther; 16(10); 2178-90. ©2017 AACR.


Cloning and characterization of the scute (sc) gene of Drosophila subobscura.
The scute (sc) gene, a member of the achaete-scute complex of Drosophila melanogaster, has dual functions: sisterless (sis-b) function required for sex determination and dosage compensation and scute function, which is involved in neurogenesis. The sc homologue of D. subobscura was cloned. It lacks introns and encodes a single 1.7-kb transcript slightly larger than that of D. melanogaster (1.6 kb). The sc protein of D. subobscura is slightly larger than that of D. melanogaster (382 vs. 345 amino acids). Sequence comparisons between both species show the Sc protein to have a highly conserved bHLH domain. Outside this domain, amino acid replacements are not randomly distributed. Two additional conserved domains, of 20 and 36 amino acids, are present near the C-terminal end. They may represent domains confering specificity upon the Sc protein with respect to other proteins of the achaete-scute complex. In its 3' untranslated region, Sc RNA contains uridine stretches, putative Sxl protein DNA-binding sites. The D. subobscura Sc protein can cooperate with other D. melanogaster bHLH proteins because D. subobscura sc supplies sis-b function when introduced into D. melanogaster transgenic flies mutant for sc.


Efficacy of home-based visuomotor feedback training in stroke patients with chronic hemispatial neglect.
Hemispatial neglect is a severe cognitive condition frequently observed after a stroke, associated with unawareness of one side of space, disability and poor long-term outcome. Visuomotor feedback training (VFT) is a neglect rehabilitation technique that involves a simple, inexpensive and feasible training of grasping-to-lift rods at the centre. We compared the immediate and long-term effects of VFT vs. a control training when delivered in a home-based setting. Twenty participants were randomly allocated to an intervention (who received VFT) or a control group (n = 10 each). Training was delivered for two sessions by an experimenter and then patients self-administered it for 10 sessions over two weeks. Outcome measures included the Behavioural Inattention Test (BIT), line bisection, Balloons Test, Landmark task, room description task, subjective straight-ahead pointing task and the Stroke Impact Scale. The measures were obtained before, immediately after the training sessions and after four-months post-training. Significantly greater short and long-term improvements were obtained after VFT when compared to control training in line bisection, BIT and spatial bias in cancellation. VFT also produced improvements on activities of daily living. We conclude that VFT is a feasible, effective, home-based rehabilitation method for neglect patients that warrants further investigation with well-designed randomised controlled trials on a large sample of patients.


The practice of adult genetics: a 7-year experience from a single center.
The purpose of our study is to familiarize the reader with genetic disorders commonly seen in adults and identify challenges and barriers that limit provision of services. We conducted a retrospective chart analysis of patients seen in the adult Genetics clinics from January 2004 to December 2010 in a metropolitan medical center consisting of an academic private clinic and a county hospital clinic. During the study period, a total of 1,552 patients (n = 1,108 private clinic patients; n = 444 county clinic patients) were evaluated and managed. Of these, 790 and 280 were new patient visits at the private clinic and county clinic, respectively. Approximately 35% (374/1,070) of new patients were seen for cancer-related indications, while neurological indications accounted for approximately 14% (153/1,070) in both clinics. Cardiology-related indications accounted for approximately 13% (145/1,070) of patients, followed closely by chromosomal and syndromic indications for which almost 9% (96/1,070) of new patients were seen. Approximately 8% (90/1,070) of new patients were seen for musculoskeletal indications. We saw increased clinic growth during the study period and found that the most common indications for referral are: (1) Personal/family history of cancer (2) neurological (3) cardiovascular (CV) (4) chromosomal/syndromic and (5) musculoskeletal. A number of challenges were identified, including coordination of services, feasibility of testing, and an overall higher complexity of care with increased clinic scheduling time requirements. Through this review, we demonstrate the demand for adult genetics services and propose some guidelines to address the challenges of management in the adult genetics patient population.


An evaluation of age-related differences in quality of life preferences in patients with non-Hodgkin's lymphoma.
Health related quality of life is an important outcome measure. With aging, patients may experience changes in physical, socioeconomic and psychological functioning. This pilot study examined whether age influences the level of importance that patients with non-Hodgkin's lymphoma assign to questions addressing aspects of traditional quality of life domains. A questionnaire assessing six domains (physical, appearance, toxicity, social, financial, psychological) with 29 items was given to 76 outpatients with non-Hodgkin's lymphoma. Each question asked how important the content of the item was to the individual. Mean item scores were compared between patients aged < 65 and > 65 years. Reliability ranged from 0.57 (social domain) to 0.83 (physical domain). Test-retest reliability for the entire questionnaire was 0.63. Although there was a suggestion that older patients scored the items relating to faith, appearance to others, intimacy and toxicity trade-offs differently than younger patients, when accounting for multiple comparisons in this study, no apparent differences were seen in any of the items between age groups. It appears that in this group of patients with lymphoma, age does not obviously influence the preferences of patients for items contained in quality of life assessment.


Continuous modulations of femtosecond laser-induced periodic surface structures and scanned line-widths on silicon by polarization changes.
Large-area, uniform laser-induced periodic surface structures (LIPSS) are of wide potential industry applications. The continuity and processing precision of LIPSS are mainly determined by the scanning intervals of adjacent scanning lines. Therefore, continuous modulations of LIPSS and scanned line-widths within one laser scanning pass are of great significance. This study proposes that by varying the laser (800 nm, 50 fs, 1 kHz) polarization direction, LIPSS and the scanned line-widths on a silicon (111) surface can be continuously modulated with high precision. It shows that the scanned line-width reaches the maximum when the polarization direction is perpendicular to the scanning direction. As an application example, the experiments show large-area, uniform LIPSS can be fabricated by controlling the scanning intervals based on the one-pass scanned line-widths. The simulation shows that the initially formed LIPSS structures induce directional surface plasmon polaritons (SPP) scattering along the laser polarization direction, which strengthens the subsequently anisotropic LIPSS fabrication. The simulation results are in good agreement with the experiments, which both support the conclusions of continuous modulations of the LIPSS and scanned line-widths.


Prediction of complete gene structures in human genomic DNA.
We introduce a general probabilistic model of the gene structure of human genomic sequences which incorporates descriptions of the basic transcriptional, translational and splicing signals, as well as length distributions and compositional features of exons, introns and intergenic regions. Distinct sets of model parameters are derived to account for the many substantial differences in gene density and structure observed in distinct C + G compositional regions of the human genome. In addition, new models of the donor and acceptor splice signals are described which capture potentially important dependencies between signal positions. The model is applied to the problem of gene identification in a computer program, GENSCAN, which identifies complete exon/intron structures of genes in genomic DNA. Novel features of the program include the capacity to predict multiple genes in a sequence, to deal with partial as well as complete genes, and to predict consistent sets of genes occurring on either or both DNA strands. GENSCAN is shown to have substantially higher accuracy than existing methods when tested on standardized sets of human and vertebrate genes, with 75 to 80% of exons identified exactly. The program is also capable of indicating fairly accurately the reliability of each predicted exon. Consistently high levels of accuracy are observed for sequences of differing C + G content and for distinct groups of vertebrates.


Simplified antibiotic regimens for the management of clinically diagnosed severe infections in newborns and young infants in first-level facilities in Karachi, Pakistan: study design for an outpatient randomized controlled equivalence trial.
Infection in young infants is a major cause of morbidity and mortality in low-middle income countries, with high neonatal mortality rates. Timely case management is lifesaving, but the current standard of hospitalization for parenteral antibiotic therapy is not always feasible. Alternative, simpler antibiotic regimens that could be used in outpatient settings have the potential to save thousands of lives. This trial aims to determine whether 2 simplified antibiotic regimens are equivalent to the reference therapy with 7 days of once-daily (OD) intramuscular (IM) procaine penicillin and gentamicin for outpatient management of young infants with clinically presumed systemic bacterial infection treated in primary health-care clinics in 5 communities in Karachi, Pakistan. The reference regimen is close to the current recommendation of the hospital-based intravenous ampicillin and gentamicin therapy for neonatal sepsis. The 2 comparison arms are (1) IM gentamicin OD and oral amoxicillin twice daily for 7 days; and (2) IM penicillin and gentamicin OD for 2 days, followed by oral amoxicillin twice daily for 5 days; 2250 "evaluable" infants will be enrolled. The primary outcome of this trial is treatment failure (death, deterioration or lack of improvement) within 7 days of enrollment. Results are expected by early 2014. This trial will determine whether simplified antibiotic regimens with fewer injections in combination with high-dose amoxicillin are equivalent to 7 days of IM procaine penicillin and gentamicin in young infants with clinical severe infection. Results will have program and policy implications in countries with limited access to hospital care and high burden of neonatal deaths.


Genetic Variants Underlying Risk of Intracranial Aneurysms: Insights from a GWAS in Portugal.
Subarachnoid hemorrhage (SAH) is a life-threatening event that most frequently leads to severe disability and death. Its most frequent cause is the rupture of a saccular intracranial aneurysm (IA), which is a blood vessel dilation caused by disease or weakening of the vessel wall. Although the genetic contribution to IA is well established, to date no single gene has been unequivocally identified as responsible for IA formation or rupture. We aimed to identify IA susceptibility genes in the Portuguese population through a pool-based multistage genome-wide association study. Replicate pools were allelotyped in triplicate in a discovery dataset (100 IA cases and 92 gender-matched controls) using the Affymetrix Human SNP Array 6.0. Top SNPs (absolute value of the relative allele score difference between cases and controls |RASdiff|≥13.0%) were selected for technical validation by individual genotyping in the discovery dataset. From the 101 SNPs successfully genotyped, 99 SNPs were nominally associated with IA. Replication of technically validated SNPs was conducted in an independent replication dataset (100 Portuguese IA cases and 407 controls). rs4667622 (between UBR3 and MYO3B), rs6599001 (between SCN11A and WDR48), rs3932338 (214 kilobases downstream of PRDM9), and rs10943471 (96 kilobases upstream of HTR1B) were associated with IA (unadjusted allelic chi-square tests) in the datasets tested (discovery: 6.84E-04≤P≤1.92E-02, replication: 2.66E-04≤P≤2.28E-02, and combined datasets: 6.05E-05≤P≤5.50E-04). Additionally, we confirmed the known association with IA of rs1333040 at the 9p21.3 genomic region, thus validating our dataset. These novel findings in the Portuguese population warrant further replication in additional independent studies, and provide additional candidates to more comprehensively understand IA etiopathogenesis.


Converting a light-driven proton pump into a light-gated proton channel.
There are two types of membrane-embedded ion transport machineries in nature. The ion pumps generate electrochemical potential by energy-coupled active ion transportation, while the ion channels produce action potential by stimulus-dependent passive ion transportation. About 80% of the amino acid residues of the light-driven proton pump archaerhodopsin-3 (AR3) and the light-gated cation channel channelrhodopsin (ChR) differ although they share the close similarity in architecture. Therefore, the question arises: How can these proteins function differently? The absorption maxima of ion pumps are red-shifted about 30-100 nm compared with ChRs, implying a structural difference in the retinal binding cavity. To modify the cavity, a blue-shifted AR3 named AR3-T was produced by replacing three residues located around the retinal (i.e., M128A, G132V, and A225T). AR3-T showed an inward H(+) flux across the membrane, raising the possibility that it works as an inward H(+) pump or an H(+) channel. Electrophysiological experiments showed that the reverse membrane potential was nearly zero, indicating light-gated ion channeling activity of AR3-T. Spectroscopic characterization of AR3-T revealed similar photochemical properties to some of ChRs, including an all-trans retinal configuration, a strong hydrogen bond between the protonated retinal Schiff base and its counterion, and a slow photocycle. From these results, we concluded that the functional determinant in the H(+) transporters is localized at the center of the membrane-spanning domain, but not in the cytoplasmic and extracellular domains.


Leg tourniquets modify the on-kinetics of oxygen uptake during cycle exercise.
Obstructive cardiovascular disease decelerates the on-kinetics of the body's oxygen uptake rate (VO2), either by reducing the O2 delivery or by degrading the exercise capacity of skeletal muscle. This study sought to determine whether obstructed blood flow decelerates the on-kinetics of VO2 by reducing O2 delivery. Breath-by-breath measurements of VO2 were studied in eight healthy youth (age, 16.5 +/- 0.6 years) during two-legged square-wave exercise on the cycle ergometer. The square protocol was 5 minutes of unloaded exercise followed by an immediate increase to 5 minutes of loaded exercise at 78 +/- 5 W. In the experimental trials, thigh cuffs were inflated 30 seconds before the onset of loaded exercise. Control trials replicated the square-wave exercise with deflated thigh cuffs. The on-kinetics of VO2 were biphasic in the control trial and triphasic in the experimental trial. Experimental VO2 was significantly higher than control values in the final 90 seconds of exercise (P<.05). The difference between experimental and control VO2 was attributable to inflated thigh cuffs. At no time was experimental VO2 significantly lower than control VO2. The clinical implication of the results is that skeletal myopathy, not reduced blood flow, is essential for decelerating the VO2 on-kinetics of patients with obstructive vascular disease. The warranted conclusion is that square-wave exercise with inflated thigh cuffs stimulates a third phase of VO2 on-kinetics.


No Alterations of Brain Structural Asymmetry in Major Depressive Disorder: An ENIGMA Consortium Analysis.
Asymmetry is a subtle but pervasive aspect of the human brain, and it may be altered in several psychiatric conditions. MRI studies have shown subtle differences of brain anatomy between people with major depressive disorder and healthy control subjects, but few studies have specifically examined brain anatomical asymmetry in relation to this disorder, and results from those studies have remained inconclusive. At the functional level, some electroencephalography studies have indicated left fronto-cortical hypoactivity and right parietal hypoactivity in depressive disorders, so aspects of lateralized anatomy may also be affected. The authors used pooled individual-level data from data sets collected around the world to investigate differences in laterality in measures of cortical thickness, cortical surface area, and subcortical volume between individuals with major depression and healthy control subjects. The authors investigated differences in the laterality of thickness and surface area measures of 34 cerebral cortical regions in 2,256 individuals with major depression and 3,504 control subjects from 31 separate data sets, and they investigated volume asymmetries of eight subcortical structures in 2,540 individuals with major depression and 4,230 control subjects from 32 data sets. T1-weighted MRI data were processed with a single protocol using FreeSurfer and the Desikan-Killiany atlas. The large sample size provided 80% power to detect effects of the order of Cohen's d=0.1. The largest effect size (Cohen's d) of major depression diagnosis was 0.085 for the thickness asymmetry of the superior temporal cortex, which was not significant after adjustment for multiple testing. Asymmetry measures were not significantly associated with medication use, acute compared with remitted status, first episode compared with recurrent status, or age at onset. Altered brain macro-anatomical asymmetry may be of little relevance to major depression etiology in most cases.


[Urodynamic test with self reduction of the cystocele in the diagnosis of lower urinary tract obstruction].
The association of cystocele and urodynamic lower urinary tract obstruction is frequent, occasionally not being possible to rule out intrinsic obstruction of bladder neck and urethra. With the aim to confirm the obstructive character of the cystocele we performed at test consisting in manual reduction of the cystocele by the patient herself, to check if by this simple manoeuvre the urodynamic parameters of obstruction disappear or diminish. 24 patients consulting for sensation of vaginal bulge, with a mean age of 66 years, participated in the study. The initial diagnosis of lower urinary tract obstruction and cystocele was obtained after video-urodynamic tests. The urodynamic test with self reduction of the cystocele was based on cystomanometry and voiding pressure-flow tests. The parameters of the study included presence, amplitude and bladder capacity during detrusor involuntary contractions in the cystomanometry; in the voiding pressure/flow test the parameters of the study were the URA as a urethra resistance parameter, the type of obstruction with the Chess classification, and Wmax and W80-W20 as parameters of detrusor contractility. A higher frequency of severe (58.3%) than moderate (41.6%) cystocele was demonstrated. No significant relation with age was demonstrated. The URA significantly diminished (p<0.01) with self reduction: from a median value of 30.5 before to 15.5 H2O cm after reduction. Although it was observed in all grades of cystocele, this reduction was greater in the severe ones. The most frequent type of obstruction was the constrictive (62.5%), over compressive (4.2%), mixed (12.5%) and unobstructed (4.2%). The constrictive obstruction significantly disappeared or diminished after reduction (p < 0.05) to a 45.8% of the cases, the compressive to 0%, the mixed to 4.2% and the nonobstructive to 50%. No significant relationship between these data and grade of cystocele was demonstrated. On the other hand, no significant differences were demonstrated with cystocele self reduction in the other urodynamic parameters (detrusor hyperactivity and contractility, Wmax and W80-W20), neither in their relationship with the grade of cystocele. This test could be very useful in the diagnosis of lower urinary tract obstruction for its validity and significance, apart from being an easy to perform and reproducible test.


The leading-edge vortex on a rotating wing changes markedly beyond a certain central body size.
Stable attachment of a leading-edge vortex (LEV) plays a key role in generating the high lift on rotating wings with a central body. The central body size can affect the LEV structure broadly in two ways. First, an overall change in the size changes the Reynolds number, which is known to have an influence on the LEV structure. Second, it may affect the Coriolis acceleration acting across the wing, depending on the wing-offset from the axis of rotation. To investigate this, the effects of Reynolds number and the wing-offset are independently studied for a rotating wing. The three-dimensional LEV structure is mapped using a scanning particle image velocimetry technique. The rapid acquisition of images and their correlation are carefully validated. The results presented in this paper show that the LEV structure changes mainly with the Reynolds number. The LEV-split is found to be only minimally affected by changing the central body radius in the range of small offsets, which interestingly includes the range for most insects. However, beyond this small offset range, the LEV-split is found to change dramatically.