Molecular mechanisms accounting for fibrinogen deficiency: from large deletions to intracellular retention of misfolded proteins.
Fibrinogen, the soluble precursor of fibrin, which is the main protein constituent of the blood clot, is synthesized in hepatocytes in the form of a hexamer composed of two sets of three polypeptides (Aalpha, Bbeta, and gamma). Each polypeptide is encoded by a distinct gene, FGA, FGB and FGG, all three clustered in a region of 50 kb on 4q32. Congenital afibrinogenemia is characterized by the complete absence of fibrinogen. The first causative mutation for this disease was identified in Geneva in a non-consanguineous Swiss family in 1999: the four patients were homozygous for a large deletion in the fibrinogen cluster, which eliminated almost the entire FGA genomic sequence. Mutations in the fibrinogen genes may lead to deficiency of fibrinogen by several mechanisms: acting at the DNA level, at the RNA level by affecting mRNA splicing or stability, or at the protein level by affecting protein synthesis, assembly or secretion. Recent reviews have provided helpful updates for the rapidly growing number of causative mutations identified in patients with fibrinogen deficiencies, either afibrinogenemia or hypofibrinogenemia. The aim of this review is to highlight specifically the subset of mutations that allow fibrinogen chain synthesis and hexamer assembly but impair secretion. Indeed, functional studies of these mutations have shed light on the specific sequences and structures in the fibrinogen molecule involved in the quality control of fibrinogen secretion.


Dietary Patterns Are Associated with Cardiovascular and Cancer Mortality among Swiss Adults in a Census-Linked Cohort.
Defining dietary guidelines requires a quantitative assessment of the influence of diet on the development of diseases. The aim of the study was to investigate how dietary patterns were associated with mortality in a general population sample of Switzerland. We included 15,936 participants from two population-based studies (National Research Program 1A (NRP1A) and Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA)-1977 to 1993) who fully answered a simplified 24-h dietary recall. Mortality data were available through anonymous record linkage with the Swiss National Cohort (follow-up of up to 37.9 years). Multiple correspondence analysis and hierarchical clustering were used to define data-driven qualitative dietary patterns. Mortality hazard ratios were calculated for all-cause, cancer and cardiovascular mortality using Cox regression. Two patterns were characterized by a low dietary variety ("Sausage and Vegetables", "Meat and Salad"), two by a higher variety ("Traditional", "High-fiber foods") and one by a high fish intake ("Fish"). Males with unhealthy lifestyle (smokers, low physical activity and high alcohol intake) were overrepresented in the low-variety patterns and underrepresented in the high-variety and "Fish" patterns. In multivariable-adjusted models, the "Fish" (hazard ratio = 0.82, 95% CI (0.68-0.99)) and "High-fiber foods" (0.85 (0.72-1.00)) patterns were associated with lower cancer mortality. In men, the "Fish" (0.73 (0.55-0.97)) and "Traditional" (0.76 (0.59-0.98)) patterns were associated with lower cardiovascular mortality. In summary, our results support the notion that dietary patterns affect mortality and that these patterns strongly cluster with other health determinants.


Impella Support for Acute Myocardial Infarction Complicated by Cardiogenic Shock.
Percutaneous mechanical circulatory support devices are increasingly used in acute myocardial infarction complicated by cardiogenic shock (AMI-CS), despite limited evidence for their effectiveness. The aim of this study was to evaluate outcomes associated with use of the Impella device compared with intra-aortic balloon pump (IABP) and medical treatment in patients with AMI-CS. Data of patients with AMI-CS treated with the Impella device at European tertiary care hospitals were collected retrospectively. All patients underwent early revascularization and received optimal medical treatment. Using IABP-SHOCK II (Intraaortic Balloon Pump in Cardiogenic Shock II) trial inclusion and exclusion criteria, 372 patients were identified and included in this analysis. These patients were matched to 600 patients from the IABP-SHOCK II trial. The following baseline criteria were used as matching parameters: age, sex, mechanical ventilation, ejection fraction, prior cardiopulmonary resuscitation, and lactate. Primary end point was 30-day all-cause mortality. In total, 237 patients treated with an Impella could be matched to 237 patients from the IABP-SHOCK II trial. Baseline parameters were similarly distributed after matching. There was no significant difference in 30-day all-cause mortality (48.5% versus 46.4%, P=0.64). Severe or life-threatening bleeding (8.5% versus 3.0%, P<0.01) and peripheral vascular complications (9.8% versus 3.8%, P=0.01) occurred significantly more often in the Impella group. Limiting the analysis to IABP-treated patients as a control group did not change the results. In this retrospective analysis of patients with AMI-CS, the use of an Impella device was not associated with lower 30-day mortality compared with matched patients from the IABP-SHOCK II trial treated with an IABP or medical therapy. To further evaluate this, a large randomized trial is warranted to determine the effect of the Impella device on outcome in patients with AMI-CS. URL: https://www.clinicaltrials.gov . Unique identifier: NCT03313687.


Role of survey response rates on valid inference: an application to HIV prevalence estimates.
Nationally-representative surveys suggest that females have a higher prevalence of HIV than males in most African countries. Unfortunately, these results are made on the basis of surveys with non-ignorable missing data. This study evaluates the impact that differential survey nonresponse rates between males and females can have on the point estimate of the HIV prevalence ratio of these two classifiers. We study 29 Demographic and Health Surveys (DHS) from 2001 to 2010. Instead of employing often used multiple imputation models with a Missing at Random assumption that may not hold in this setting, we assess the effect of ignoring the information contained in the missing HIV information for males and females through three proposed statistical measures. These measures can be used in settings where the interest is comparing the prevalence of a disease between two groups. The proposed measures do not utilize parametric models and can be implemented by researchers of any level. They are: (1) an upper bound on the potential bias of the usual practise of using reported HIV prevalence estimates that ignore subjects who have missing HIV outcomes. (2) Plausible range intervals to account for nonresponses, without any additional parametric modeling assumptions. (3) Prevalence ratio inflation factors to correct the point estimate of the HIV prevalence ratio, if estimates of nonresponders' HIV prevalences were known. In 86% of countries, males have higher upper bounds of HIV prevalence than females, this is consonant with males possibly having higher infection rates than females. Additionally, 74% of surveys have a plausible range that crosses 1.0, suggesting a plausible equivalence between male and female HIV prevalences. It is quite reasonable to conclude that there is so much DHS nonresponse in evaluating the HIV status question, that existing data is plausibly generated by the situation where the virus is equally distributed between the sexes.


Production of fibronectin by normal and malignant human mammary epithelial cells.
The production and retention of fibronectin by primary cultures of cells derived from the human breast has been analyzed. Two examples of each of the following cell types were examined: (a) normal epithelium from milk; (b) metastatic breast cancer cells in pleural effusions; (c) fibroblasts; (d) tissue macrophages of milk. Cell-associated fibronectin could be detected by indirect immunofluorescent staining on normal and malignant mammary epithelium and on mammary fibroblasts, but not on milk macrophages. Immune precipitation followed by gel electrophoresis of 35S-labeled cell lysates and conditioned medium confirmed that fibronectin was indeed synthesized by both types of epithelial cells and by fibroblasts, but not by macrophages, and that much of the protein was released into the medium. Quantitative analysis with radioimmune assay of the fibronectin on cells and in media showed that both normal and malignant epithelial cells synthesized levels of protein comparable to that produced by fibroblasts, but only a small fraction (less than 10%) of the material synthesized was retained by the cells. Growth on collagen-coated plastic increased the percentage of fibronectin retained by normal and malignant epithelium but did not affect retention by fibroblasts.


A descriptive profile of abused female sex workers in India.
This descriptive study presents the profiles of abused female sex workers (FSWs) in Chennai, India. Of 100 abused FSWs surveyed using a structured questionnaire, severe forms of violence by intimate partners were reported by most (98%) respondents. Of the total sample, 76% experienced violence by clients. Sexual coercion experiences of the FSWs included verbal threats (77%) and physical force (87%) by intimate partners and forced unwanted sexual acts (73%) by clients. While 39% of the women consumed alcohol before meeting a client, 26% reported that their drunkenness was a trigger for violence by clients. The findings suggest that there is an urgent need to integrate services, along with public-health interventions among FSWs to protect them from violence. Recognition of multiple identities of women in the contexts of intimate relationships versus sex work is vital in helping women to stay safe from adverse effects on health.


Establishing a model of primary bovine hepatocytes with responsive growth hormone receptor expression.
The liver becomes resistant to growth hormone before parturition in dairy cows (uncoupling of the somatotropic axis). However, the mechanism of growth hormone insensitivity has not been fully described. The aim of the present study was to improve a previous model of adult bovine hepatocytes in a sandwich culture system to ensure growth hormone receptor (GHR) expression. First, we modified the protocol for hepatocyte retrieval and tested the effect of short (18 min) and long (up to 30 min) warm ischemia on hepatocyte viability. Second, we used medium additives that affect GHR expression in vivo-insulin (INS), dexamethasone (DEX), both (INS+DEX), or no hormone additives (CTRL)-to ensure the functionality of hepatocytes, as measured by lactate dehydrogenase activity and urea concentration in the medium. We also used reverse transcriptase PCR of hepatocytes to evaluate expression of albumin (ALB), hepatocyte nuclear factor 4α (HNF4A), nuclear factor-κ-B-inhibitor α (NFKBIA), cytosolic phosphoenolpyruvate carboxykinase (PCK1), and vimentin (VIM) mRNA. Moreover, we analyzed the expression of GHRtot (GHR), GHR1A, insulin-like growth factor-1 (IGF1), and IGF binding protein-2 (IGFBP2) in response to exposure to media with the different compositions. Modification of the protocol (changes in rinsing and perfusion times, buffer composition, and the volume and standardization of collagenase) led to increased cell counts and cell viability. Short warm ischemia with the modified protocol significantly increased cell count (4.7 × 107 ± 1.9 × 107 vs. 3.5 × 106 ± 1.5 × 106 vital cells/g of liver) and viability (79.1 ± 8.4 vs. 37.1 ± 8.9%). Therefore, we gathered hepatocytes from the liver after short warm ischemia with the modified protocol. For these hepatocytes, lactate dehydrogenase activity was lower in media with INS and with DEX than in media with INS+DEX or CTRL; urea concentrations were highest at d 4 for INS+DEX. As well, HNF4A and ALB were more highly expressed in hepatocytes cultured with INS and INS+DEX than in those cultured with DEX or CTRL, and the substitution of DEX suppressed VIM and NFKBIA expression but increased PCK1 expression. The expression of GHR, GHR1A, and IGF1 was suppressed by dexamethasone (DEX and INS+DEX), whereas INS distinctly increased GHR, GHR1A, and IGF1 mRNA expression. Hepatocytes in a sandwich culture showed influenceable GHR expression; this study provides a model that can be used in studies examining factors that influence the expression and signal transduction of GHR in dairy cows.


Risk of dementia after anaesthesia and surgery.
The potential relationship between anaesthesia, surgery and onset of dementia remains elusive. To determine whether the risk of dementia increases after surgery with anaesthesia, and to evaluate possible associations among age, mode of anaesthesia, type of surgery and risk of dementia. The study cohort comprised patients aged 50 years and older who were anaesthetised for the first time since 1995 between 1 January 2004 and 31 December 2007, and a control group of randomly selected patients matched for age and gender. Patients were followed until 31 December 2010 to identify the emergence of dementia. Relative to the control group, patients who underwent anaesthesia and surgery exhibited an increased risk of dementia (hazard ratio = 1.99) and a reduced mean interval to dementia diagnosis. The risk of dementia increased in patients who received intravenous or intramuscular anaesthesia, regional anaesthesia and general anaesthesia. The results of our nationwide, population-based study suggest that patients who undergo anaesthesia and surgery may be at increased risk of dementia.


Mouse Breast Carcinoma Monocytic/Macrophagic Myeloid-Derived Suppressor Cell Infiltration as a Consequence of Endothelial Dysfunction in Shb-Deficient Endothelial Cells Increases Tumor Lung Metastasis.
Metastasis reflects both the inherent properties of tumor cells and the response of the stroma to the presence of the tumor. Vascular barrier properties, either due to endothelial cell (EC) or pericyte function, play an important role in metastasis in addition to the contribution of the immune system. The Shb gene encodes the Src homology-2 domain protein B that operates downstream of tyrosine kinases in both vascular and immune cells. We have investigated E0771.lmb breast carcinoma metastasis in mice with conditional deletion of the Shb gene using the Cdh5-CreERt2 transgene, resulting in inactivation of the Shb-gene in EC and some hematopoietic cell populations. Lung metastasis from orthotopic tumors, tumor vascular and immune cell characteristics, and immune cell gene expression profiles were determined. We found no increase in vascular leakage that could explain the observed increase in metastasis upon the loss of Shb expression. Instead, Shb deficiency in EC promoted the recruitment of monocytic/macrophagic myeloid-derived suppressor cells (mMDSC), an immune cell type that confers a suppressive immune response, thus enhancing lung metastasis. An MDSC-promoting cytokine/chemokine profile was simultaneously observed in tumors grown in mice with EC-specific Shb deficiency, providing an explanation for the expanded mMDSC population. The results demonstrate an intricate interplay between tumor EC and immune cells that pivots between pro-tumoral and anti-tumoral properties, depending on relevant genetic and/or environmental factors operating in the microenvironment.


A quantitative ultrastructural study of microtubule content and secretory granule accumulation in parathyroid glands of phosphate- and colchicine-treated rats.
Microtubule involvement in secretory events of the parathyroid gland was investigated in rats treated with colchicine and/or phosphorus, agents which have been shown to modify parathyroid secretion. Quantitative ultrastructural techniques were used in an effort to assess the cytoplasmic microtubule and secretory granule content of chief cells 3 h after treatment, when hypocalcemia was well established. After cochicine administration, the chief cells appeared to have lost all assembled microtubules and accumulated greater than normal amounts of cytoplasmic secretory granules. On the other hand, phosphorus treatment was associated with increased microtubule content although the cytoplasmic content of secretory granules remained unchanged. When colchicine and phosphorus were given concomitantly, microtubules were again absent, but the secretory granule content of the cells was markedly increased. These data provide direct evidence that colchicine disrupts assembled microtubules in chief cells of rat parathyroids; the consequence of this effect appears to be a blockage of hormone release which is reflected in the accumulation of secretory granules in the cell. The fact that microtubules also show a significant increase in content when hormone release from chief cells is presumed to increase, suggests that microtubules may participate in the physiological control of parathyroid hormone secretion.


A tRNA-specific function for tRNA methyltransferase Trm10 is associated with a new tRNA quality control mechanism in Saccharomyces cerevisiae.
In Saccharomyces cerevisiae a single homolog of the tRNA methyltransferase Trm10 performs m1G9 modification on 13 different tRNAs. Here we provide evidence that the m1G9 modification catalyzed by S. cerevisiae Trm10 plays a biologically important role for one of these tRNA substrates, tRNATrp. Overexpression of tRNATrp (and not any of 38 other elongator tRNAs) rescues growth hypersensitivity of the trm10Δ strain in the presence of the antitumor drug 5-fluorouracil (5FU). Mature tRNATrp is depleted in trm10Δ cells, and its levels are further decreased upon growth in 5FU, while another Trm10 substrate (tRNAGly) is not affected under these conditions. Thus, m1G9 in S. cerevisiae is another example of a tRNA modification that is present on multiple tRNAs but is only essential for the biological function of one of those species. In addition to the effects of m1G9 on mature tRNATrp, precursor tRNATrp species accumulate in the same strains, an effect that is due to at least two distinct mechanisms. The levels of mature tRNATrp are rescued in the trm10Δmet22Δ strain, consistent with the known role of Met22 in tRNA quality control, where deletion of met22 causes inhibition of 5'-3' exonucleases that catalyze tRNA decay. However, none of the known Met22-associated exonucleases appear to be responsible for decay of hypomodified tRNATrp, based on inability of mutants of each enzyme to rescue growth of the trm10Δ strain in the presence of 5FU. Thus, the surveillance of tRNATrp appears to constitute a distinct tRNA quality control pathway in S. cerevisiae.


Diabetic Retinopathy as a Risk Factor Associated with the Development of Hepatocellular Carcinoma in Nonalcoholic Fatty Liver Disease.
The risk factors associated with the development of hepatocellular carcinoma (HCC) in nonalcoholic fatty liver disease (NAFLD) are still unclear. The aim of the present study was to identify such risk factors in NAFLD patients who developed HCC. Between April 2000 and -December 2016, a total of 182 patients with NAFLD were enrolled in this study; of these, only 22 patients had HCC. To identify risk factors, univariate and multivariate analyses were performed. To identify risk factors other than the degree of fibrosis, propensity matched analysis adjusted by the NAFLD fibrosis score (NFS) was carried out on 44 patients. Multivariate and survival analyses were also performed in HCC patients. In 182 patients, multivariate analysis highlighted the NFS (OR 2.275; p < 0.001) and hypertension (OR 5.868; p = 0.037) as independent factors that were significantly associated with the development of HCC. After adjustment for the NFS, multivariate analysis identified diabetic retinopathy (OR 8.654; p = 0.017) as an independent factor that was significantly associated with the development of HCC. For predicting the development of HCC, the area under the receiver operating characteristic curve of diabetic retinopathy was significantly higher than that of diabetes (0.731 vs. 0.615; p < 0.001). In patients with HCC, multivariate analysis indicated that the NFS were significantly associated with diabetic retinopathy. Diabetic retinopathy as well as liver fibrosis is a risk factor that associates with the development of HCC in NAFLD patients. Therefore, NAFLD patients with diabetic retinopathy should undergo careful screening for HCC.


National Eye Institute Visual Function Scale in Type 2 Diabetes Patients.
Aim. To examine subscale and total scores of NEI-VFQ questionnaire of type 2 diabetes patients at different diabetic retinopathy (DRP) stages. Methods. A total number of 201 patients have been included. Prior to ophthalmological examination all patients participated in the NEI-VFQ questionnaire. The patients were divided into 5 groups according to the International Clinical Diabetic Retinopathy Disease Severity Scale (ICDRS). Results. The diabetes duration in general health scores (p = 0.029); the stage (p = 0.011); and clinically significant macular edema (CSME) (p = 0.019) in general vision were found to be the most efficient factors. In near vision activities the most efficient factors were near vision acuity (NVA) (p = 0.0001) and DRP stage (p = 0.020). EDTRS visual acuity was found to be the most efficient factor in vision specific role difficulties (p = 0.034) and dependency (p = 0.011) whereas Snellen visual acuity was found to be among the most effective factors in distance activities (DA) (p = 0.014) and total scores (p = 0.026). Discussion. Difference was based not on the diabetes duration, clinically significant cataract (CSCat), CSME presence, and DRP stage but on the visual acuity levels of the better seeing eye of the patients.


Identification of miRNAs that specifically target tumor suppressive KLF6-FL rather than oncogenic KLF6-SV1 isoform.
The Krüppel like factor 6 (KLF6) gene encodes multiple protein isoforms derived from alternative mRNA splicing, most of which are intimately involved in hepatocarcinogenesis and tumor progression. Recent bioinformatics analysis shows that alternative mRNA splicing of the KLF6 gene produces around 16 alternatively spliced variants with divergent or even opposing functions. Intriguingly, the full-length KLF6 (KLF6-FL) is a tumor suppressor gene frequently inactivated in liver cancer, whereas KLF6 splice variant 1 (KLF6-SV1) is an oncogenic isoform with antagonistic function against KLF6-FL. Compelling evidence indicates that miRNA, the small endogenous non-coding RNA (ncRNA), acts as a vital player in modulating a variety of cellular biological processes through targeting different mRNA regions of protein-coding genes. To identify the potential miRNAs specifically targeting KLF6-FL, we utilized bioinformatics analysis in combination with the luciferase reporter assays and screened out two miRNAs, namely miR-210 and miR-1301, specifically targeted the tumor suppressive KLF6-FL rather than the oncogenic KLF6-SV1. Our in vitro experiments demonstrated that stable expression of KLF6-FL inhibited cell proliferation, migration and angiogenesis while overexpression of miR-1301 promoted cell migration and angiogenesis. Further experiments demonstrated that miR-1301 was highly expressed in liver cancer cell lines as well as clinical specimens and we also identified the potential methylation and histone acetylation for miR-1301 gene. To sum up, our findings unveiled a novel molecular mechanism that specific miRNAs promoted tumorigenesis by targeting the tumor suppressive isoform KLF6-FL rather than its oncogenic isoform KLF6-SV1.


Pathophysiology of ANCA-associated Vasculitis.
Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis is characterized as inflammation of small-sized to medium-sized blood vessels and encompasses several clinicopathologic entities including granulomatosis with polyangiitis, microscopic polyangiitis, eosinophilic granulomatosis with polyangiitis, and renal-limited ANCA-associated vasculitis. Over the past several decades, significant progress has been made in understanding the pathophysiology of ANCA-associated vasculitis. Although neutrophils contain a multitude of granular proteins, clinically significant autoantibodies are only recognized against myeloperoxidase and proteinase 3, both of which are present in the azurophilic granules. The propensity to develop these antibodies depends on a variety of predisposing factors such as microbial infection, genetic factors, environmental agents, and therapeutic drugs among others. These factors are usually associated with production of proinflammatory cytokines with capacity to prime the neutrophils. As a result a high proportion of neutrophils in circulation may be primed resulting in exposure of cytoplasmic proteins including myeloperoxidase and proteinase 3 on the surface of the neutrophils. Primed neutrophils are activated by interaction with ANCA in circulation. Activated neutrophils attach to and transmigrate through endothelium and accumulate within the vessel wall. These neutrophils degranulate and produce reactive oxygen radicals and ultimately die, causing tissue injury. Endothelial injury results in leakage of serum proteins and coagulation factors causing fibrinoid necrosis. B cells produce ANCAs, as well as neutrophil abnormalities and imbalances in different T-cell subtypes with excess of Th17, which perpetuate the inflammatory process.


Nest desertion is not predicted by cuckoldry in the Eurasian penduline tit.
Engagement in extra-pair copulations is an example of the abundant conflicting interests between males and females over reproduction. Potential benefits for females and the risk of cuckoldry for males are expected to have important implications on the evolution of parental care. However, whether parents adjust parental care in response to parentage remains unclear. In Eurasian penduline tits Remiz pendulinus, which are small polygamous songbirds, parental care is carried out either by the male or by the female. In addition, one third of clutches is deserted by both male and female. Desertion takes place during the egg-laying phase. Using genotypes of nine microsatellite loci of 443 offspring and 211 adults, we test whether extra-pair paternity predicts parental care. We expect males to be more likely to desert cuckolded broods, whereas we expect females, if they obtain benefits from having multiple sires, to be more likely to care for broods with multiple paternity. Our results suggest that parental care is not adjusted to parentage on an ecological timescale. Furthermore, we found that male attractiveness does not predict cuckoldry, and we found no evidence for indirect benefits for females (i.e., increased growth rates or heterozygosity of extra-pair offspring). We argue that male Eurasian penduline tits may not be able to assess the risk of cuckoldry; thus, a direct association with parental care is unlikely to evolve. However, timing of desertion (i.e., when to desert during the egg-laying phase) may be influenced by the risk of cuckoldry. Future work applying extensive gene sequencing and quantitative genetics is likely to further our understanding of how selection may influence the association between parentage and parental care.


Hematology and serum chemistry of cottontail rabbits of southern Illinois.
In 1983 and 1984 blood was collected from 79 cottontail rabbits (Sylvilagus floridanus) confined to an outdoor enclosure in southern Illinois to establish reference values for hematology and serum chemistry. Packed cell volume, sodium, potassium, chloride, glucose, calcium, carbon dioxide, blood urea nitrogen, creatinine, uric acid, cholesterol, albumin, bilirubin, alkaline phosphatase, aspartate transaminase, alanine aminotransaminase, total protein, albumin/globulin ratio, and osmolality were measured. Sex and age (adult versus juvenile) of rabbit as well as season (June to September versus October to May) and method of capture (trap versus shot) variously affected most hematology and serum chemistry variables.


Effect of opacifiers and UV absorbers on pigmented maxillofacial silicone elastomer, part 1: color stability after artificial aging.
Much dissatisfaction with the color instability and reduced lifetime of extraoral maxillofacial prostheses due to degradation has been reported. The purpose of this study was to compare the effect of a UV mineral-based light protecting agent (LP) on the color stability of pigmented maxillofacial silicone elastomer MDX-4210/Type A after artificial aging to 2 widely used opacifiers. Forty-five groups were established (n=225 total). Three different types of opacifiers (LP, titanium white dry pigment [TW], or silicone intrinsic white [SW]) were added to silicone MDX-4210/type A at 3 concentrations (5%, 10%, or 15%) and subsequently combined with each of 5 colors (no pigments [control], red, blue, yellow, or mixed pigments). Artists' oil pigment was used with LP and TW, while intrinsic silicone pigment was used to color SW. Before and after an energy exposure of 450 kJ/m(2), CIE L*a*b* values were measured with a spectrophotometer. The CIELAB 50:50% perceptibility (ΔE*=1.1) and acceptability threshold (ΔE*=3.0) were used to interpret color changes (ΔE*). Color differences after aging were subjected to 3-way ANOVA. Means were compared by the Fisher PLSD intervals at α=.05. The ΔE* values of all groups were below the acceptability threshold of ΔE*=3.0, except for the control group of SW at 10%, which showed the greatest color change (ΔE*=3.1). When mixed pigment groups were considered, at 5% concentration, LP showed the smallest color change, followed by SW and TW (P<.05); at 10%, no significant differences among the 3 opacifiers were noted (P>.05); at 15%, LP showed the smallest color change, followed by TW and SW (P<.05). All 3 opacifiers at all concentrations protected pigmented silicone MDX4-4210/Type A from color degradation. The LP group showed the smallest color changes.


Extracorporeal membrane oxygenation in the management of post-pneumonectomy air leak and adult respiratory distress syndrome of the non-operated lung.
Post-pneumonectomy air leak and severe respiratory failure of the non-operated lung is considered to be a life-threatening complication of lung surgery. We present the case report of a 68-year-old man who underwent a right pneumonectomy for spinocellular carcinoma. Refractory respiratory failure occurred following bronchial stump air leakage and adult respiratory distress syndrome (ARDS) of the non-operated lung. Established veno-venous extracorporeal membrane oxygenation (VV ECMO) was utilized to maintain tissue oxygenation while re-do surgery was performed. The leaking bronchial stump was closed with an azygos vein patch and, subsequently, weaning off ECMO was accomplished 7 days later. The patient fully recovered and he is limited only by mild exertional dyspnea at 24 months follow-up after the initial surgery.


Wheelchair cushions for spinal cord-injured individuals.
Pressure sore prevention is a major objective in the rehabilitation of individuals with spinal cord injury. Wheelchair cushions are frequently prescribed to relieve pressure and reduce the risk of pressure sores in this population. In this study, 251 subjects with paraplegia and quadriplegia were evaluated to decide which wheelchair cushions were suitable. Criteria for the comparative evaluation of cushions included not only magnitude and distribution of pressure but also factors such as wheelchair compatibility, ease of transfer, activities, and independence. Although the Roho cushion was prescribed most frequently, it was not recommended for all subjects. This study provides additional evidence that no single cushion is optimal for all people with spinal cord injury. Rather, objective measurements and clinical judgments are essential elements of a complete evaluation.


Postpartum Depression Among Somali Women in Norway.
Postpartum depression (PPD) has been described as the most common complication experienced postpartum, affecting about 10-15 % of all new mothers. Factors like a history of mental illness, and experienced recent adverse life events has been associated with an increased risk for developing PPD. Immigrant women in Western countries have been found to have a marked higher prevalence of PPD compared to the general population. In Norway the prevalence of PPD in the general population has been found to be around 8-10 %, and among Pakistani immigrants a rate of 7.6 % was found. Somali people in Norway are the second largest immigrant group in Norway with a non-Western background. No study on PPD and associated factors among Somali women has been found in the literature. The aim of the study was to assess PPD and associated factors among Somali women in greater Oslo region, Norway. A cross-sectional survey was conducted; recruiting new mothers through all maternity wards in the Oslo region. Data was collected with interview-administrated questionnaires. PPD was assessed using Edinburgh Postnatal Depression Scale (EPDS), defining those scoring ≥10 to have a possible PPD. Of the 80 eligible women identified, 39 (49 %) consented to participate, and completed the study. Of the 39 respondents 3 (7.7 %) were assessed to have a possible PPD. Most important associated factors found were history of mental illness, having experienced technical assistance during delivery, self-rated health and experienced economical problems last 12 months. A low prevalence of PPD was found, and both the prevalence and its associated factors should be interpreted with caution. The associated factors do not have enough power to give any strength to the associations. However, some of the results can be used in develop new hypotheses with regard to PPD among Somali women as immigrants in a Western society.


High-cocoa polyphenol-rich chocolate improves blood pressure in patients with diabetes and hypertension.
The aim was to examine the effects of high-cocoa polyphenol-rich chocolate on lipid profiles, weight, blood pressure, glycemic control, and inflammation in individuals with Type 2 diabetes and hypertension. Sixty individuals [32 in dark chocolate group (DCG) and 28 in white chocolate group (WCG)] with Type 2 diabetes on stable medication were enrolled in a randomized, placebo-controlled double-blind study. Subjects were randomized to consume 25 g DCG or WCG for 8 weeks. Changes in weight, blood pressure, glycemic control, lipid profile, and high sensitive C-reactive protein (hsCRP) were measured at the beginning and end of the intervention. This clinical trial was registered at the Iranian registry of clinical trials. In DCC group, compared with baseline, serum levels of Apo A-1 (P = 0.045) was increased and fasting blood sugar (FBS) (P = 0.027), hemoglobin A1c (HbA1c) (P = 0.025), Apo B (P = 0.012) and Log of hsCRP (P = 0.043) levels were decreased at the end of study. No changes were seen within the WCG in studied parameters. High polyphenol chocolate consumption compared to white chocolate resulted in significant decrease in of systolic (-5.93 ± 6.25 vs. -1.07 ± 7.97 mmHg, P = 0.004) and diastolic blood pressure (-6.4 ± 6.25 vs. 0.17 ± 7.9 mmHg, P = 0.002), FBS (-7.84 ± 19.15 vs. 4.00 ± 20.58 mg/dl, P = 0.019) over the course of 8 weeks of daily chocolate consumption neither weight nor body mass index and TG levels altered from baseline. High polyphenol chocolate is effective in improving TG levels in hypertensive patients with diabetes and decreasing blood pressure and FBS without affecting weight, inflammatory markers, insulin resistance or glycemic control.


New Insights Into Drug Repurposing for COVID-19 Using Deep Learning.
The coronavirus disease 2019 (COVID-19) has continued to spread worldwide since late 2019. To expedite the process of providing treatment to those who have contracted the disease and to ensure the accessibility of effective drugs, numerous strategies have been implemented to find potential anti-COVID-19 drugs in a short span of time. Motivated by this critical global challenge, in this review, we detail approaches that have been used for drug repurposing for COVID-19 and suggest improvements to the existing deep learning (DL) approach to identify and repurpose drugs to treat this complex disease. By optimizing hyperparameter settings, deploying suitable activation functions, and designing optimization algorithms, the improved DL approach will be able to perform feature extraction from quality big data, turning the traditional DL approach, referred to as a "black box," which generalizes and learns the transmitted data, into a "glass box" that will have the interpretability of its rationale while maintaining a high level of prediction accuracy. When adopted for drug repurposing for COVID-19, this improved approach will create a new generation of DL approaches that can establish a cause and effect relationship as to why the repurposed drugs are suitable for treating COVID-19. Its ability can also be extended to repurpose drugs for other complex diseases, develop appropriate treatment strategies for new diseases, and provide precision medical treatment to patients, thus paving the way to discover new drugs that can potentially be effective for treating COVID-19.


Missed diagnosis of subarachnoid hemorrhage in the emergency department.
Subarachnoid hemorrhage (SAH) can be devastating, yet its initial presentation may be limited to common symptoms and subtle signs, potentially leading to misdiagnosis. Little is known about population rates of misdiagnosis of SAH, or hospital factors that may contribute to it. We estimated the population-based rate of missed SAH among emergency department (ED) patients and examined its relationship with hospital characteristics. We studied persons admitted with a nontraumatic SAH to all Ontario hospitals over 3 years (April 2002 to March 2005). SAH was defined as missed if the patient had an ED visit related to the SAH (based on a prespecified definition) in the 14 days before admission. We examined the association between hospital teaching status and missed SAH and explored whether annual ED volume of SAH or CT availability explained this association. Of 1507 patients diagnosed with SAH, 5.4% (95% CI, 4.3 to 6.6) had a missed diagnosis. The risk was significantly higher among patients triaged as low acuity (odds ratio 2.65; 95% CI, 1.46 to 4.80), as well as in nonteaching hospitals (adjusted odds ratio 2.12; 95% CI, 1.02, 4.44). Neither ED SAH volume nor on-site CT availability explained the effect of teaching status. About 1 in 20 SAH patients are missed during an ED visit. Lower acuity patients are at higher risk of misdiagnosis, suggesting the need for heightened suspicion among patients with minimal clinical findings. The risk is also greater in nonteaching hospitals, but this is not explained by the annual volume of SAHs seen in the ED or access to CT.


Changes in 2-hydroxyestrone and 16alpha-hydroxyestrone metabolism with flaxseed consumption: modification by COMT and CYP1B1 genotype.
Consumption of the phytoestrogen lignans, structurally similar to estrogen, has been associated with alterations in gene expression and estrogen metabolism. Furthermore, lignan consumption, subsequent changes in metabolizing enzyme expression, and genetic variability in these enzymes may alter estrogen metabolism and modify disease risk. Therefore, we investigated the effect of flaxseed on hydroxyestrone metabolite excretion by catechol-O-methyltransferase (COMT) and cytochrome P450 1B1 (CYP1B1) genotype. We conducted an intervention among 132 healthy, postmenopausal women, ages 46 to 75 years. Participants consumed 10 g ground flaxseed daily for 7 consecutive days. Blood and urine samples were collected at baseline and after the 7-day intervention. COMT Val(158)Met and CYP1B1 Leu(432)Val genotypes were determined using PCR-RFLP methods. Urinary 2-hydroxyestrone (2OHE1) and 16alpha-hydroxyestrone (16OHE1) were quantified by ELISA assay. The effect of genotype on intervention-related changes in estrogen metabolites was assessed with the Kruskal-Wallis test. Compared with baseline levels, postintervention levels of urinary 2OHE1 (ng/mg creatinine; mean +/- SD, 16.1 +/- 10.6 versus 9.3 +/- 6.9, postintervention and baseline, respectively; P < 0.01) and 2OHE1/16OHE1 ratios (mean +/- SD, 2.73 +/- 1.47 versus 1.54 +/- 0.75, postintervention and baseline, respectively; P < 0.01) were significantly higher. The change in 2OHE1/16OHE1 increased with increasing numbers of variant alleles for COMT (mean change: Val/Val, 0.90; Val/Met, 1.15; and Met/Met, 1.50; P = 0.17, Kruskal-Wallis) and especially CYP1B1 (mean change: Leu/Leu, 0.89; Leu/Val, 1.32; and Val/Val, 1.51; P = 0.04, Kruskal-Wallis). Our findings suggest that variation in hormone-related genes may modify the effect of dietary lignan exposures on estrogen metabolism.


Dendrimer pre-treatment enhances the skin permeation of chlorhexidine digluconate: Characterisation by in vitro percutaneous absorption studies and Time-of-Flight Secondary Ion Mass Spectrometry.
Skin penetration and localisation of chlorhexidine digluconate (CHG) within the skin have been investigated in order to better understand and optimise the delivery using a nano polymeric delivery system of this topically-applied antimicrobial drug. Franz-type diffusion cell studies using in vitro porcine skin and tape stripping procedures were coupled with Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) to visualise the skin during various treatments with CHG and polyamidoamine dendrimers (PAMAM). Pre-treatment of the skin with PAMAM dendrimers significantly increased the amount and depth of permeation of CHG into the skin in vitro. The effect observed was not concentration dependant in the range 0.5-10mM PAMAM. This could be important in terms of the efficiency of treatment of bacterial infection in the skin. It appears that the mechanism of enhancement is due to the PAMAM dendrimer disrupting skin barrier lipid conformation or by occluding the skin surface. Franz-type diffusion cell experiments are complimented by the detailed visualisation offered by the semi-quantitative ToF-SIMS method which provides excellent benefits in terms of sensitivity and fragment ion specificity. This allows a more accurate depth profile of chlorhexidine permeation within the skin to be obtained and potentially affords the opportunity to map the co-localisation of permeants with skin structures, thus providing a greater ability to characterise skin absorption and to understand the mechanism of permeation, providing opportunities for new and more effective therapies.


Management of cutaneous hemangiomas: a retrospective analysis of 1109 cases and comparison of conventional dose prednisolone with high-dose methylprednisolone therapy.
The effectiveness of the different pharmacological agents and different doses of systemic corticosteroids was analyzed. A total of 1109 patients (median age 8 months; F/M: 2.3) with hemangioma, followed up in our unit for 23 years, were evaluated retrospectively. Forty-five of them received systemic corticosteroids. Two different pharmacological agents, prednisolone (in 26 patients) and methyl prednisolone (in 19 patients), had been used in three different regimens. Groups were compared according to the final results and rebound regrowth. Response was considered good or excellent in 16 patients (36%). Then were no differences in response to therapy among the three regimens. No difference was found in response to therapy between prednisolone and methylprednisolone and the two different doses of the methylprednisolone. Rebound regrowth was significantly higher in methyl prednisolone than in the prednisolone group (p = .045). In multivariate analysis the dimension of the lesion (p = .0065) and age at initiation of treatment (p = .0041) were the most important factors affecting the response. In conclusion, the systemic corticosteroids are effective in 36% of patients, independent of dosage and pharmacological agents and duration of the therapy. The dimension of the lesion and age at initiation of treatment are the most important factors affecting the response to treatment.


Cancer risk in diabetic patients treated with metformin: a systematic review and meta-analysis.
A growing body of evidence has suggested that metformin potentially reduces the risk of cancer. Our objective was to enhance the precision of estimates of the effect of metformin on the risk of any-site and site-specific cancers in patients with diabetes. We performed a search of MEDLINE, EMBASE, ISI Web of Science, Cochrane Library, and ClinicalTrials.gov for pertinent articles published as of October 12, 2011, and included them in a systematic review and meta-analysis. We calculated pooled risk ratios (RRs) for overall cancer mortality and cancer incidence. Of the 21,195 diabetic patients reported in 6 studies (4 cohort studies, 2 RCTs), 991 (4.5%) cases of death from cancer were reported. A total of 11,117 (5.3%) cases of incident cancer at any site were reported among 210,892 patients in 10 studies (2 RCTs, 6 cohort studies, 2 case-control studies). The risks of cancer among metformin users were significantly lower than those among non-metformin users: the pooled RRs (95% confidence interval) were 0.66 (0.49-0.88) for cancer mortality, 0.67 (0.53-0.85) for all-cancer incidence, 0.68 (0.53-0.88) for colorectal cancer (n = 6), 0.20 (0.07-0.59) for hepatocellular cancer (n = 4), 0.67 (0.45-0.99) for lung cancer (n = 3). The use of metformin in diabetic patients was associated with significantly lower risks of cancer mortality and incidence. However, this analysis is mainly based on observational studies and our findings underscore the more need for long-term RCTs to confirm this potential benefit for individuals with diabetes.


A study of the effect of hemorrhage on the cardiorespiratory actions of halothane, isoflurane and sevoflurane in the dog.
To compare the cardiorespiratory changes induced by equipotent concentrations of halothane (HAL), isoflurane (ISO) and sevoflurane (SEVO) before and after hemorrhage. Prospective, randomized clinical trial. Twenty-four healthy adult dogs weighing 15.4 +/- 3.4 kg (mean +/- SD). Animals were randomly allocated to one of three groups (n = 8 per group). In each group, anesthesia was maintained with 1.5 minimum alveolar concentration of HAL (1.3%), ISO (1.9%) and SEVO (3.5%) in oxygen. Controlled ventilation was performed to maintain eucapnia. Cardiorespiratory variables were evaluated at baseline (between 60 and 90 minutes after induction), immediately after and 30 minutes after the withdrawal of 32 mL kg(-1) of blood (40% of the estimated blood volume) over a 30-minute period. During baseline conditions, ISO and SEVO resulted in higher cardiac index (CI) than HAL. Heart rates were higher with SEVO at baseline, while mean arterial pressure (MAP) and mean pulmonary arterial pressure did not differ between groups. Although heart rate values were higher for ISO and SEVO after hemorrhage, only ISO resulted in a higher CI when compared with HAL. In ISO-anesthetized dogs, MAP was higher immediately after hemorrhage, and this was related to better maintenance of CI and to an increase in systemic vascular resistance index from baseline. Although the hemodynamic responses of ISO and SEVO are similar in normovolaemic dogs, ISO results in better maintenance of circulatory function during the early period following a massive blood loss. Inhaled anesthetics should be used judiciously in animals presented with blood loss. However, if an inhalational agent is to be used under these circumstances, ISO may provide better hemodynamic stability than SEVO or HAL.


Limitations of Adenoviral Vector-Mediated Delivery of Gold Nanoparticles to Tumors for Hyperthermia Induction.
Novel combinatorial treatment strategies are desired to achieve tumor eradication. In this regard, nanotechnology and gene therapy hold the potential to expand the available tumor treatment options. In particular, gold nanoparticles (AuNPs) have been utilized for hyperthermic tumor cell ablation. Similarly, adenoviral (Ad) vectors have been utilized for targeting, imaging, and cancer gene therapy. Thus, to combine AuNP-mediated hyperthermia with Ad vector-based gene therapy, we have previously coupled AuNPs to Ad vectors. Herein we tested the capability of these AuNP-coupled Ad vectors for hyperthermic tumor cell ablation. Towards this end, we compared absorption characteristics of different sized AuNPs and determined that in our system 20 nm diameter AuNPs are suitable for laser induced hyperthermic tumor cell killing. In addition, we observed that AuNPs outside and inside the cell contribute differentially towards hyperthermia induction. Unfortunately, due to the limitation of delivery of required amounts of AuNPs to cells, we observed that AuNP-coupled Ad vectors are unable to kill tumor cells via hyperthermia. However, with future technological advances, it may become possible to realize the potential of the multifunctional AuNP-coupled Ad vector system for simultaneous targeting, imaging, and combined hyperthermia and gene therapy of tumors.


A connectome of a learning and memory center in the adult Drosophila brain.
Understanding memory formation, storage and retrieval requires knowledge of the underlying neuronal circuits. In Drosophila, the mushroom body (MB) is the major site of associative learning. We reconstructed the morphologies and synaptic connections of all 983 neurons within the three functional units, or compartments, that compose the adult MB's α lobe, using a dataset of isotropic 8 nm voxels collected by focused ion-beam milling scanning electron microscopy. We found that Kenyon cells (KCs), whose sparse activity encodes sensory information, each make multiple en passant synapses to MB output neurons (MBONs) in each compartment. Some MBONs have inputs from all KCs, while others differentially sample sensory modalities. Only 6% of KC>MBON synapses receive a direct synapse from a dopaminergic neuron (DAN). We identified two unanticipated classes of synapses, KC>DAN and DAN>MBON. DAN activation produces a slow depolarization of the MBON in these DAN>MBON synapses and can weaken memory recall.


[Simple and rapid screening for methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) and their metabolites in urine using direct analysis in real time (DART)-TOFMS].
An ionization technique, direct analysis in real time (DART) has recently been developed for the ambient ionization of a variety samples. The DART coupled with time-of-flight mass spectrometry (TOFMS) would be useful as a simple and rapid screening for the targeted compounds in various samples, because it provides the molecular information of these compounds without time-consuming extraction. In this study, we investigated rapid screening methods of illicit drugs and their metabolites, such as methamphetamine (MA), 3,4-methylenedioxymethamphetamine (MDMA), amphetamine (AP) and 3,4-methylenedioxyamphetamine (MDA) in human urine using DART-TOFMS. As serious matrix effects caused by urea in urine samples and ionizations of the targeted compounds were greatly suppressed in the DART-TOFMS analyses, simple pretreatment methods to remove the urea from the samples were investigated. When a pipette tip-type solid-phase extraction with a dichloromethane and isopropanol mixed solution as an eluent was used for the pretreatment, the limits of detection (LODs) of 4 compounds added to control urine samples were 0.25 µg/ml. On the other hand, the LODs of these compounds were 0.5 µg/ml by a liquid-liquid extraction using a dichloromethane and hexane mixed solution. In both extractions, the recoveries of 4 compounds from urine samples were over 70% and these extraction methods showed good linearity in the range of 0.5-5 µg/ml by GC-MS analyses. In conclusion, our proposed method using DART-TOFMS could simultaneously detect MA, MDMA and their metabolites in urine at 0.5 µg/ml without time-consuming pretreatment steps. Therefore it would be useful for screening drugs in urine with the molecular information.


Development and Characterization of Gefitinib Loaded Polymeric Nanoparticles by Ionic Gelation Method.
There is a rapidly growing interest in the development of nanoparticle drug delivery mainly for anticancer drugs as it promises to solve several problems associated with anticancer drugs such as poor water solubility, low therapeutic index, nonspecific distribution and higher systemic toxicity, etc. The objective of the study was to investigate the effect of various critical variables like, concentration of chitosan, concentration of sodium tripolyphosphate (STPP) and volume of STPP on various characteristics of gefitinib loaded nanoparticles. Thirteen formulations of the polymeric nanoparticles were prepared using various concentrations of chitosan (0.1-1% w/v), STPP (0.2-1.5% w/v) and different volumes of STPP (8-20 ml) by ionic gelation method. Mannitol (5% w/v) was used as cryoprotectant. The prepared nanoparticle formulations were characterized for various parameters like particle size, zeta potential, process yield, encapsulation efficiency, drug content, and in vitro drug release. The nanoparticle formulation NF-1 containing 0.1% w/v of chitosan and 10 ml volume of 0.2% w/v STPP showed best results in terms of particle size (123.8nm), polydispersity index (0.247), zeta potential (+30.4 mV), process yield (68.09%), drug content (74.32%), encapsulation efficiency (70.52%) and released (56.2 %) drug over a period of 24 h. The in vitro drug release analysis showed sustained release of gefitinib from nanoparticles and followed Korsmeyer-Peppas model. The nanoparticle formulation with desired characteristics can be prepared at low concentration of chitosan and STPP along with low volume of STPP. The formulated nanoparticles may prove to be the best option for the treatment of cancer.


The Largest Paracentric Inversion, the Highest Rate of Recombinant Spermatozoa. Case Report: 46,XY, inv(2)(q21.2q37.3) and Literature Review.
Carriers of inversions involving euchromatic regions are at risk of having unbalanced offspring due to meiotic crossover. In carriers, recombination can occur during gametogenesis and cause genetically unbalanced sperm and subsequently unbalanced embryos. Here we present segregation analysis results of an infertile male with 46,XY,inv(2) (q21.2q37.3) using fluorescent in situ hybridization (FISH) on sperm cells. This is the largest paracentric inversion (PAI) reported so far in a meiotic segregation analysis study. Sperm FISH revealed 28.0% recombinant spermatozoa rate for chromo-some 2, which was the highest rate in PAI carriers in the literature. Our results indicate a clear correlation between the size of the inverted segment and the frequency of the recombinant spermatozoa. The results of the FISH analysis with the information of unbalanced spermatozoa rate can provide accurate counseling on the genetic risk of infertility.


Characterization of the dynamics in the protonic conductor CsH₂PO₄ by ¹⁷O solid-state NMR spectroscopy and first-principles calculations: correlating phosphate and protonic motion.
(17)O NMR spectroscopy combined with first-principles calculations was employed to understand the local structure and dynamics of the phosphate ions and protons in the paraelectric phase of the proton conductor CsH2PO4. For the room-temperature structure, the results confirm that one proton (H1) is localized in an asymmetric H-bond (between O1 donor and O2 acceptor oxygen atoms), whereas the H2 proton undergoes rapid exchange between two sites in a hydrogen bond with a symmetric double potential well at a rate ≥10(7) Hz. Variable-temperature (17)O NMR spectra recorded from 22 to 214 °C were interpreted by considering different models for the rotation of the phosphate anions. At least two distinct rate constants for rotations about four pseudo C3 axes of the phosphate ion were required in order to achieve good agreement with the experimental data. An activation energy of 0.21 ± 0.06 eV was observed for rotation about the P-O1 axis, with a higher activation energy of 0.50 ± 0.07 eV being obtained for rotation about the P-O2, P-O3(d), and P-O3(a) axes, with the superscripts denoting, respectively, dynamic donor and acceptor oxygen atoms of the H-bond. The higher activation energy of the second process is most likely associated with the cost of breaking an O1-H1 bond. The activation energy of this process is slightly lower than that obtained from the (1)H exchange process (0.70 ± 0.07 eV) (Kim, G.; Blanc, F.; Hu, Y.-Y.; Grey, C. P. J. Phys. Chem. C 2013, 117, 6504-6515) associated with the translational motion of the protons. The relationship between proton jumps and phosphate rotation was analyzed in detail by considering uncorrelated motion, motion of individual PO4 ions and the four connected/H-bonded protons, and concerted motions of adjacent phosphate units, mediated by proton hops. We conclude that, while phosphate rotations aid proton motion, not all phosphate rotations result in proton jumps.


Biomedical Application of Electroactive Polymers in Electrochemical Sensors: A Review.
Conducting polymers are of interest due to their unique behavior on exposure to electric fields, which has led to their use in flexible electronics, sensors, and biomaterials. The unique electroactive properties of conducting polymers allow them to be used to prepare biosensors that enable real time, point of care (POC) testing. Potential advantages of these devices include their low cost and low detection limit, ultimately resulting in increased access to treatment. This article presents a review of the characteristics of conducting polymer-based biosensors and the recent advances in their application in the recognition of disease biomarkers.


Clonality of the parathyroid nodules with uremic parathyroid hyperplasia.
Clonal assessment suggests that most parathyroid adenomas and a subset of uremic parathyroid hyperplasia are monoclonal. A weakness that remains in the prior clonal studies is assessing the clonal status of the tissue fragments containing multiple nodules rather than a single nodule emerging in the uremic parathyroid hyperplasia. We applied the X chromosome-linked phosphoglycerate kinase (PGK) gene inactivation assay method for clonality to study individual nodules. Materials were obtained from 31 cases with parathyroid adenoma and 16 with uremic parathyroid hyperplasia. 17 cases were heterozygous in the PGK-1 locus. We were able to assess the clonality of 10 parathyroid adenomas and 7 hyperplastic glands. Monoclonality was demonstrable in 9 of the 10 parathyroid adenomas and in 4 of the 7 hyperplastic glands. Further analysis of 11 individual nodules microdissected from 3 monoclonal and 1 polyclonal hyperplastic glands revealed that 6 nodules were monoclonal and 5 were polyclonal. Nodules arising in a hyperplastic gland could be of monoclonal or polyclonal origin. Polyclonal and monoclonal nodules coexisted within single glands. Our findings indicate a progression from generalized hyperplasia to a monoclonal tumor in uremic parathyroid hyperplasia. Comparing clonality with the parathyroid hormone (PTH) immunoreactivity and histological features, we found that monoclonal nodules showed a homogeneous immunoreactivity against PTH antibody, whereas most of the polyclonal nodules showed a heterogeneous staining. Classic morphological criteria alone was inadequate to distinguish a monoclonal from a polyclonal nodule.


Widespread Chromosomal Losses and Mitochondrial DNA Alterations as Genetic Drivers in Hürthle Cell Carcinoma.
Hürthle cell carcinoma of the thyroid (HCC) is a form of thyroid cancer recalcitrant to radioiodine therapy that exhibits an accumulation of mitochondria. We performed whole-exome sequencing on a cohort of primary, recurrent, and metastatic tumors, and identified recurrent mutations in DAXX, TP53, NRAS, NF1, CDKN1A, ARHGAP35, and the TERT promoter. Parallel analysis of mtDNA revealed recurrent homoplasmic mutations in subunits of complex I of the electron transport chain. Analysis of DNA copy-number alterations uncovered widespread loss of chromosomes culminating in near-haploid chromosomal content in a large fraction of HCC, which was maintained during metastatic spread. This work uncovers a distinct molecular origin of HCC compared with other thyroid malignancies.


[Contact allergy to fragrances].
The incidence of allergy induced by fragrances, leading mostly to the development of contact dermatitis and urticaria chronica, has been growing in a large number of countries. In general, allergy is of non-occupational nature, however, it can also have traits of occupational exposure. The fragrance mix is used in screening for allergy to aromas, but it frequently produces false positive or false negative results of the test. The aim of the study was to assess whether the fragrance mix is suitable for detecting allergy to aromas in persons with suspected occupational allergy and also to analyze types of allergy to separate fragrances. In a group of 1937 patients diagnosed in the Nofer Institute of Occupational Medicine in Lódź, the incidence of allergy to fragrance mix was assessed. They underwent dermatological examinations and tests with use of the European standard kit (Chemotechnique Diagnostics, Malmö, Sweden) in the years 2000-2005. In addition, in the group of patients with positive tests, the type of allergy to individual aromas was analyzed using fragrance series (Chemotechnique Diagnostics, Malmö, Sweden). The positive reaction to fragrance mix was found in 99 (5.1%) patients (women--5.2% and men--4.9%). In 57.6% of patients, allergy to balsam of Peru was also observed. Allergy to at least one aroma was diagnosed in 82 (72%) persons tested with an expanded fragrance series. In the remaining 23 (28%) patients, patch tests proved to be false positive. In the latter group, 17 patients showed negative results of the test to balsam of Peru. In the study group, 65% of patients reacted to 1, 2 or 3 and 36% to more aromas. The following allergens sensitized most frequently: cynnamic alcohol, hydroxycitronellal, cinnamic aldehyde, isoeugenol, eugenol, Ylang-Ylang oil, oakmoss absolute, and jasmine. Allergic contact dermatitis was diagnosed in 69.7%, urticaria chronica in 5.1% of patients, atopic dermatitis in 4%, whereas in 21.2% no skin lesions were observed. The results of the study confirmed observations of other authors that the usefulness of fragrance mix in the diagnosis of allergy to aromas is limited (a high number of false positive results of skin tests). We noticed that a combined reaction to fragrance mix and balsam of Peru is helpful in diagnosis in persons whom a complete allergic diagnosis cannot be performed. We also found less frequent hypersensitivity to oakmoss absolute and more frequent to cinnamic aldehyde and alcohol, hydroxycitronellal, eugenol, isoeugenol in Lódź than in studies reported by other authors.


The Gut-Brain Axis: Influence of Microbiota on Mood and Mental Health.
The gut-brain axis is a bidirectional communication network that links the enteric and central nervous systems. This network is not only anatomical, but it extends to include endocrine, humoral, metabolic, and immune routes of communication as well. The autonomic nervous system, hypothalamic-pituitary-adrenal (HPA) axis, and nerves within the gastrointestinal tract, all link the gut and the brain, allowing the brain to influence intestinal activities, including activity of functional immune effector cells; and the gut to influence mood, cognition, and mental health.


Hemodynamic and neurohumoral evidence of multifaceted pathophysiology in human hypertension.
As hypertension advances, secondary pathophysiologic changes are induced in multiple organs. Consequently, we investigated the pathophysiology of the earliest forms of hypertension--e.g., borderline hypertension. Borderline hypertension is associated with abnormal autonomic control of the circulation; sympathetic drive to the heart, blood vessels, and kidney is increased, cardiac parasympathetic inhibition is decreased, and plasma norepinephrine is increased. The hemodynamic picture is one of increased cardiac output not met by adequate vasodilation. The condition of "hyperkinetic" borderline hypertension is a precursor of more severe hypertension. In due course, a transition from high cardiac output to high vascular resistance occurs, while the enhanced sympathetic tone recedes toward normal values. The mechanism of hemodynamic transition is easily understood: cardiac output decreases due to structural changes and receptor downregulation, whereas ensuing vascular hypertrophy increases vascular resistance. The apparent regression of plasma norepinephrine values is explained in the framework of our hypothesis of the "blood pressure-seeking properties of the central nervous system." Large body mass and overweight are a consistent feature of borderline hypertension. A recent study in Tecumseh, Michigan shows that weight, plasma norepinephrine, a hyperkinetic state, and plasma insulin values are correlated in the general population. The explanation of this interrelationship will greatly advance our understanding of hypertension. From the pathophysiological viewpoint, the paradoxical outcome of clinical trials involving older antihypertensive medication is not surprising. The complexity of pathophysiologic interrelationships and the fact that risk factors for atherosclerosis are increased in hypertension suggest that reduction of blood pressure cannot be expected to ameliorate all consequences of hypertension.(ABSTRACT TRUNCATED AT 250 WORDS)


Intervention Activities Associated with the Implementation of a Comprehensive School Tobacco Policy at Danish Vocational Schools: A Repeated Cross-Sectional Study.
School tobacco policies are often poorly implemented, which may explain their limited effectiveness. Further, constructs to measure implementation outcomes of school tobacco policies are missing. The Smoke-Free Vocational Schools intervention was designed to stimulate the implementation of a comprehensive school tobacco policy into routine practice. This study (1) developed implementation fidelity outcomes measures for the school tobacco policy and (2) examined associations between intervention activities and implementation fidelity at two time points. We applied a repeated cross-sectional survey study design across seven schools: the first time point was >5 months after the policy was established and the second time point > 14 months after policy establishment. The dependent/outcome variables were four binary fidelity domains as well as a total score across domains. A total of six intervention activities were measured among either students (e.g., new school-break facilities) or staff/managers (e.g., a joint workshop before policy implementation). Associations were analyzed separately for students and staff/managers using generalized linear mixed models, adjusted for confounders. A total of n = 2674 students and n = 871 staff/managers participated. The total implementation fidelity scores increased over time among both students and staff/managers. Three intervention activities were consistently associated with the total implementation fidelity score, including: new school-break facilities (BT1 = 0.08, 95% CI = 0.03; 0.12; BT2 = 0.07, 95% CI = 0.04-0.50), the joint workshop before policy implementation (BT1 = 0.13, 95% CI = 0.02; 0.25; BT2 = 0.13, 95% CI = 0.004; 0.24), and internalization of fixed procedures for enforcement (BT1 = 0.19, 95% CI = 0.13-0.26; BT2 = 0.16, 95% CI = 0.13-0.26). These findings can be applied by schools and other actors in practice. The developed implementation fidelity outcomes measures can be applied in future research on school tobacco policies.


Small-angle neutron scattering solution structures of NADPH-dependent sulfite reductase.
Sulfite reductase (SiR), a dodecameric complex of flavoprotein reductase subunits (SiRFP) and hemoprotein oxidase subunits (SiRHP), reduces sulfur for biomass incorporation. Electron transfer within SiR requires intra- and inter-subunit interactions that are mediated by the relative position of each protein, governed by flexible domain movements. Using small-angle neutron scattering, we report the first solution structures of SiR heterodimers containing a single copy of each subunit. These structures show how the subunits bind and how both subunit binding and oxidation state impact SiRFP's conformation. Neutron contrast matching experiments on selectively deuterated heterodimers allow us to define the contribution of each subunit to the solution scattering. SiRHP binding induces a change in the position of SiRFP's flavodoxin-like domain relative to its ferredoxin-NADP+ reductase domain while compacting SiRHP's N-terminus. Reduction of SiRFP leads to a more open structure relative to its oxidized state, re-positioning SiRFP's N-terminal flavodoxin-like domain towards the SiRHP binding position. These structures show, for the first time, how both SiRHP binding to, and reduction of, SiRFP positions SiRFP for electron transfer between the subunits.


Global patterns and climatic controls of forest structural complexity.
The complexity of forest structures plays a crucial role in regulating forest ecosystem functions and strongly influences biodiversity. Yet, knowledge of the global patterns and determinants of forest structural complexity remains scarce. Using a stand structural complexity index based on terrestrial laser scanning, we quantify the structural complexity of boreal, temperate, subtropical and tropical primary forests. We find that the global variation of forest structural complexity is largely explained by annual precipitation and precipitation seasonality (R² = 0.89). Using the structural complexity of primary forests as benchmark, we model the potential structural complexity across biomes and present a global map of the potential structural complexity of the earth´s forest ecoregions. Our analyses reveal distinct latitudinal patterns of forest structure and show that hotspots of high structural complexity coincide with hotspots of plant diversity. Considering the mechanistic underpinnings of forest structural complexity, our results suggest spatially contrasting changes of forest structure with climate change within and across biomes.


Prognostic value of p16(INK4a) as a marker of clinical evolution in patients with cervical intraepithelial neoplasia grade 3 (CIN 3) treated by cervical conization.
The aim of this study was to assess the prognostic value of p16(INK4a) as a marker of post-conization relapse in patients treated for cervical intraepithelial neoplasia grade 3 (CIN 3). A retrospective study of 76 women with CIN 3 diagnoses, treated at the Hospital of Santa Casa de Misericórdia of São Paulo (Brazil) between January 2003 and September 2004, was performed. The study samples were obtained from cervical conization procedures, where paraffin blocks containing areas with the greatest amount of neoplastic tissue were selected. Immunohistochemical techniques were used on individual paraffin blocks for each case to detect p16(INK4a) protein expression. The p16(INK4a) cell counts were performed in 10 different high-amplification fields (400x) by light microscopy and total cell count expressed as number of cells per mm(2) . Patients involved in this study were followed up at the colposcopy outpatient unit for at least 48 months after cervical conization. The correlation of p16(INK4a) values with post-conization evolution in the patients (disease relapse or disease free) was determined. A significantly higher count of cells expressing p16(INK4a) was found in those patients with disease relapse during follow-up (p < 0.001). The variables age, number of gestations, and births correlated positively with number of cells expressing p16(INK4a) cells (p < 0.001; p = 0.001; 0.009, respectively). No correlation was found for the variables menopause, hormonal contraception, or smoking (p = 0.369, 0.425 and 0.853, respectively). p16(INK4a) can be considered a biomarker of cervical intraepithelial neoplasia grade 3 cases presenting high risk of relapse or evolution to invasive carcinoma.


Anatomy and physiology of supernumerary cercal afferents in crickets: implications for pattern formation.
The afferent projection of supernumerary sensory neurons was examined in the cricket cercal sensory system. When a right cercus was exchanged for a left cercus the growth of supernumerary cerci was induced. The supernumerary cerci which formed tended to appear medial and/or lateral in a simple left leads to right exchange and were always right handed. Distal to the region where transplant and supernumerary meet, receptors on the supernumerary structure appear in a recognizable pattern typical of control cerci, and individual receptors homologous in position to control receptors could be identified. The supernumerary neurons associated with these identified receptors projected to the same area of neuropil as their homologs and thus uniquely identified neurons by all of the usual criteria were produced by the regeneration process. When the response properties of one postsynaptic neuron were examined it was shown that supernumerary afferents innervated it in a normal manner. The supernumerary neurons provide tests of various hypotheses for the assembly of ordered afferent projections. First, their axon trajectories are often atypical, but the terminal arbors grow into their proper areas in spite of this. Thus axon trajectory can be uncoupled from target area and is therefore unlikely to be a decisive factor in growth of the arbor. Second, supernumerary neurons are born later in development than their control homologs but their aroborizations are completely normal, adding to the existing evidence against a timing mechanism in the assembly of this afferent projection. Third, supernumerary neurons share target regions with their homologs, indicating a lack of competitive interactions. The most economical hypothesis to explain the precise mapping between the cercus and CNS holds that one step in the differentiation of insect sensory neurons is position dependent and that this step determines the choice of a target region within the CNS.


E test as an alternative to conventional MIC determination for surveillance of drug resistant Streptococcus pneumoniae.
A commercial E test was compared with the standard agar dilution method for the determination of minimum inhibitory concentration (MIC) of penicillin, erythromycin, chloramphenicol and cefotaxime for 36 strains of Streptococcus pneumoniae from patients with invasive diseases. Additional strains were tested for MIC values for penicillin (6), erythromycin (14) and cefotaxime (13) for a better statistical evaluation. Besides, 5 reference standards with predetermined MIC values obtained from WHO pneumococcal reference center at Copenhagen, Denmark were tested for penicillin and erythromycin, for quality assessment using both agar dilution as well as E test methods. An overall agreement within +/- 2 dilutions was noted for 97 per cent of the strains tested for all the antimicrobials. A high degree of correlation was noted for erythromycin (r = 1), penicillin (r = 0.99), chloramphenicol (r = 0.95) and cefotaxime (r = 0.9). In MIC determination of a single antimicrobial for diagnostic purpose, E test was found to be more cost effective than conventional agar dilution method. E test was simple to perform, easy to interpret and a valid method for MIC determination of antimicrobials for S. pneumoniae in our center.


Low complication rate of colostomy closures.
Colostomy closure, a procedure often relegated to the less experienced member of the surgical team, is not a benign procedure. Early reports in the literature have clearly indicated that colostomy closure is attended by an exorbitantly high complication rate. The complication rate of 12% at the Medical Center of Central Georgia from 1972 to 1977 was very much lower than those reported in the literature in the past decade. We believe this relatively low rate of complication is basically due to: (1) proper mechanical and chemical preparation of the colon preoperatively, (2) delayed primary closure of wounds, and (3) meticulous attention to detail in surgical technic.


Hearing loss impacts neural alpha oscillations under adverse listening conditions.
Degradations in external, acoustic stimulation have long been suspected to increase the load on working memory (WM). One neural signature of WM load is enhanced power of alpha oscillations (6-12 Hz). However, it is unknown to what extent common internal, auditory degradation, that is, hearing impairment, affects the neural mechanisms of WM when audibility has been ensured via amplification. Using an adapted auditory Sternberg paradigm, we varied the orthogonal factors memory load and background noise level, while the electroencephalogram was recorded. In each trial, participants were presented with 2, 4, or 6 spoken digits embedded in one of three different levels of background noise. After a stimulus-free delay interval, participants indicated whether a probe digit had appeared in the sequence of digits. Participants were healthy older adults (62-86 years), with normal to moderately impaired hearing. Importantly, the background noise levels were individually adjusted and participants were wearing hearing aids to equalize audibility across participants. Irrespective of hearing loss (HL), behavioral performance improved with lower memory load and also with lower levels of background noise. Interestingly, the alpha power in the stimulus-free delay interval was dependent on the interplay between task demands (memory load and noise level) and HL; while alpha power increased with HL during low and intermediate levels of memory load and background noise, it dropped for participants with the relatively most severe HL under the highest memory load and background noise level. These findings suggest that adaptive neural mechanisms for coping with adverse listening conditions break down for higher degrees of HL, even when adequate hearing aid amplification is in place.


School-based identification of asthma in a low-income population.
The increase in the prevalence, morbidity, and mortality of asthma among children over the last decade has been well documented, especially among low-income minority children. Hypotheses for the increases in morbidity and mortality include limited access to primary care services and the failure to recognize the presence and severity of asthma. The University of Miami Pediatric Mobile Clinic (Mobile Clinic) Asthma Intervention Program is designed to identify underserved asthmatic children at school and offer them culturally sensitive care. Nine elementary schools with low income, predominantly Hispanic and African-American populations regularly served by the Mobile Clinic, were chosen for study participation. All 5,800 students who were enrolled in kindergarten through third grade were given letters at the time of registration by their homeroom teachers about the asthma program. Caretakers who returned the questionnaire and reported that the student had asthma symptoms were invited to have the student undergo a medical evaluation in the Mobile Clinic. Over a 2-year period, caretakers of 423 students (7.3% of all students) expressed an interest in further evaluating their child's respiratory health. Of these, we enrolled and evaluated 154 in the Mobile Clinic's Asthma Intervention Program. The Mobile Clinic physicians identified 145 of the enrollees as having asthma. These results indicate that in elementary schools serving predominantly low-income minority populations, a large fraction of the asthmatic population (estimated prevalence, 6-10%) can be identified by a school-based letter. Further, in a subset of asthmatic students (children of interested caretakers), there is good agreement between caretaker responses and physician diagnosis of asthma. Since school attendance is mandatory, school-based methods may be an effective method for identifying low-income children with asthma.


Immunization as a factor affecting the course of septicemic anthrax.
Immunization of guinea pigs with alum-precipitated antigen increased resistance approximately 1600 times. The growth rate of Bacillus anthracis, in the septicemic phase, was the same in both normal and immunized animals; however, the number of bacilli per milliliter of blood at death was decreased by 75 percent in immunized animals. Neither the enhancement of virulence by treatment with egg yolk nor the size of the challenge dose affected the growth rate or the number of bacilli per milliliter of blood at death. Mean time to death for the treatments tested varied from approximately 20 to 100 hours. Nonimmunized animals challenged with a strain of low virulence had a terminal number of bacilli per milliliter of blood and a growth rate in the blood the same as that for the highly virulent strain


Impact of Elexacaftor/Tezacaftor/Ivacaftor on Healthcare Resource Utilization and Associated Costs Among People With Cystic Fibrosis in the US: A Retrospective Claims Analysis.
Cystic fibrosis (CF) is a life-limiting genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA) is a CFTR modulator (CFTRm) that targets the underlying cause of CF. Based on safety and efficacy demonstrated in clinical trials, ELX/TEZ/IVA is approved in the US for the treatment of CF in people aged ≥ 2 years who have ≥ 1 F508del-CFTR mutation or a CFTR mutation that is responsive to ELX/TEZ/IVA based on in vitro data. While ELX/TEZ/IVA demonstrated unprecedented improvements in lung function and dramatic reductions in pulmonary exacerbations (PEx) and associated hospitalizations in clinical trials, a limited number of studies have examined the impact of ELX/TEZ/IVA on healthcare resource utilization (HCRU) and associated costs in a real-world setting. The aim of this retrospective study was to evaluate changes in PEx, HCRU, and associated non-CFTRm healthcare costs following initiation of ELX/TEZ/IVA among people with CF aged ≥ 12 years in the US. We evaluated the rates of PEx, HCRU, and associated costs before and after initiation of ELX/TEZ/IVA in people with CF aged ≥ 12 years using data from the Merative MarketScan® Commercial Claims and Encounters Database and the Merative Multi-State Medicaid Database from April 21, 2019 to December 31, 2020. Because the study period included time following the onset of the COVID-19 pandemic, we limited our primary analysis to the period prior to the pandemic (October 21, 2019 to March 12, 2020). Outcomes following the onset of the pandemic (March 13 to December 31, 2020) were examined in an exploratory analysis. In both commercially insured and Medicaid-insured people with CF, ELX/TEZ/IVA was associated with reductions in PEx, hospitalizations, and associated costs prior to the COVID-19 pandemic, and these reductions were maintained following the onset of the pandemic. These findings suggest that ELX/TEZ/IVA reduces the burden and costs associated with PEx and hospitalizations in people with CF.


Prevalence of caseous lymphadenitis and usage of caseous lymphadenitis vaccines in sheep flocks.
To estimate the prevalence of caseous lymphadenitis (CLA), determine the current usage of vaccines against CLA and to measure the effectiveness of these vaccines on sheep farms. A survey was undertaken on 223 sheep flocks in New South Wales, Victoria and Western Australia. The prevalence of CLA was measured by conventional inspection techniques at abattoirs in lines of sheep that could be traced back to a farm. Managers of the flocks were sent a questionnaire about their vaccine practices, management practices and knowledge of CLA. The average prevalence of CLA in adult sheep in these flocks was 26% and varied from 20% in Western Australia to 29% in New South Wales. About 43% of sheep producers used CLA vaccines; only 12% used them as recommended. Awareness of CLA was highest in Western Australia. More producers would use CLA vaccine if they knew the prevalence of CLA in their flock and producers obtained most information about CLA from vaccine resellers. Only 10 to 15% of producers are currently achieving effective CLA control through the use of recommended CLA vaccination programs. In Western Australian flocks more than 25% of effectively vaccinated ewes will be sent to abattoirs in the 2 to 3 years after this study. However, large decreases in the prevalence of CLA can be achieved by about 70% of producers by either making adjustments to their vaccination programs or buying a vaccine with a CLA component. Two or three key facts on effective CLA vaccination could be made available at the point of sale of vaccines and from abattoirs that reported the prevalence of CLA to farmers.


How life history influences the responses of the clam Scrobicularia plana to the combined impacts of carbamazepine and pH decrease.
In the present study, the bivalve Scrobicularia plana, collected from two contrasting areas (pristine location and mercury contaminated area), was selected to assess the biochemical alterations imposed by pH decrease, carbamazepine (an antiepileptic) and the combined effect of both stressors. The effects on oxidative stress related biomarkers after 96 h exposure revealed that pH decrease and carbamazepine induced alterations on clams, with greater impacts on individuals from the contaminated area which presented higher mortality, higher lipid peroxidation and higher glutathione S-transferase activity. These results emphasize the risk of extrapolating results from one area to another, since the same species inhabiting different areas may be affected differently when exposed to the same stressors. Furthermore, the results obtained showed that, when combined, the impact of pH decrease and carbamazepine was lower than each stressor acting alone, which could be related to the defence mechanism of valves closure when bivalves are under higher stressful conditions.


Distribution of IL-5 receptor-positive B cells. Expression of IL-5 receptor on Ly-1(CD5)+ B cells.
mAb to murine IL-5R were prepared by means of fusion between mouse myeloma cells and spleen cells from a rat immunized with membrane-enriched fractions of IL-5-dependent early B cell line (T88-M). Two mAb (H7 and T21) were selected for their competitive inhibition of receptor binding by 35S-labeled IL-5 and of IL-5 biologic activities. The number of binding sites recognized by the mAb on different cell lines correlated with IL-5 responsiveness. Most surface IgM+ peritoneal B cells were H7+ and more than 70% were also Ly-1(CD5)dull+, and responded to IL-5 for polyclonal IgM production in a high frequency. A significant proportion of splenic B cells reacted with these mAb, although lower number (one-log less) than peritoneal B cells and a small proportion of H7dull+ splenic B cells seems to be Ly-1(CD5)dull+, 1 of 200 splenic B cells responded to IL-5 for IgM production. These results suggest that IL-5R+ B cells may consist of a subpopulation of B cells. Intriguingly, lymphoid populations of bone marrow cells were stained with H7 and T21, whereas myeloid populations were brightly stained with only T21. Finally, both H7 and T21 mAb specifically precipitated a protein of a Mr 60,000 from 125I-labeled cell lysates of IL-5R+ T88-M cells. The IL-5R with similar size (Mr 55,000 to 60,000) was precipitated from the cell lysates of peritoneal B cells. T21 mAb but not H7 mAb precipitated a protein of a Mr 110,000 from the cell lysates of bone marrow cells.


Aberrant composition of gut microbiota of allergic infants: a target of bifidobacterial therapy at weaning?
Recent data have outlined a relationship between the composition of the intestinal microflora and allergic inflammation, and demonstrated the competence of probiotics in downregulation of such inflammation. Our aims were to characterise the relationship between gut microbes and the extent of allergic sensitisation and to assess whether the efficacy of bifidobacterial supplementation in the treatment of allergy could relate to modulation of the intestinal microbiota. This randomised study included 21 infants with early onset atopic eczema of whom eight were intolerant (highly sensitised group (HSG)) and 13 tolerant (sensitised group (SG)) to extensively hydrolysed whey formula (EHF). In the SG, six were weaned to EHF without (placebo group (PG)) and seven to EHF with Bifidobacterium lactis Bb-12 supplementation (bifidobacteria treated group (BbG)). The faecal microflora of infants in the HSG was analysed only before weaning whereas in the SG the faecal microflora was analysed both before and after weaning. Infants in the HSG had greater numbers of lactobacilli/enterococci than those in the SG. Serum total IgE concentration correlated directly with Escherichia coli counts in all infants and with bacteroides counts in the HSG, indicating that the presence of these bacteria is associated with the extent of atopic sensitisation. The effect of supplementation was characterised as a decrease in the numbers of Escherichia coli and protection against an increase in bacteroides numbers during weaning. These data indicate that bifidobacterial supplementation appears to modify the gut microbiota in a manner that may alleviate allergic inflammation. Further studies are needed to confirm this conclusion.


Description of strain 3CB-1, a genomovar of Thauera aromatica, capable of degrading 3-chlorobenzoate coupled to nitrate reduction.
A Gram-negative bacterium, strain 3CB-1, isolated from a 3-chlorobenzoate enrichment culture inoculated with a sediment sample is capable of degrading various aromatic compounds and halogenated derivatives with nitrate as electron acceptor. Compounds capable of serving as carbon and energy sources include 3-chlorobenzoate, 3-bromobenzoate, 2-fluorobenzoate, 4-fluorobenzoate, benzoate, 3-hydroxybenzoate, 4-hydroxybenzoate, 3-aminobenzoate, protocatechuate, m-cresol and p-cresol. Oxygen, nitrate and nitrite were used as electron acceptors for growth. Cells are Gram-negative short rods with peritrichous flagellation. The predominant fatty acids are cis-9-hexadecenoic acid (16:1 omega 7c), hexadecanoic acid (16:0), octadecanoic acid (18:0), octadecenoic acid (18:1), 3-hydroxydecanoic acid (10:0 3OH) and dodecanoic acid (12:0). The sequence of the 16S rRNA gene, as well as the fatty acid composition, indicate that the strain is a member of the genus Thauera in the beta-subclass of the Proteobacteria and very close to Thauera aromatica. DNA-DNA hybridization and nutrient screening indicate that strain 3CB-1 is a genomovar of Thauera aromatica with the proposed name Thauera aromatica genomovar chlorobenzoica.


Mechanism of the melibiose porter in membrane vesicles of Escherichia coli.
The melibiose transport system of Escherichia coli catalyzes sodium--methyl 1-thio-beta-D-galactopyranoside (TMG) symport, and the cation is required not only for respiration-driven active transport but also for binding of substrate to the carrier in the absence of energy and for carrier-mediated TMG efflux. As opposed to the proton--beta-galactoside symport system [Kaczorowski, G. J., & Kaback, H. R. (1979) Biochemistry 18, 3691], efflux and exchange of TMG occur at the same rate, implying that the rates of the two processes are limited by a common step, most likely the translocation of substrate across the membrane. Furthermore, the rate of exchange, as well as efflux, is influenced by imposition of a membrane potential (delta psi; interior negative), suggesting that the ternary complex between sodium, TMG, and the porter may bear a net positive charge. Consistently, energization of the vesicles leads to a large increase in the Vmax for TMG influx, with little or no change in the apparent Km of the process. It is proposed that the sodium gradient (Na+out < Na+in) and the delta psi (interior negative) may affect different steps in the overall mechanism of active TMG accumulation in the following manner: the sodium gradient causes an increased affinity for TMG on the outer surface of the membrane relative to the inside and the delta psi facilitates a reaction involved with the translocation of the positively charged ternary complex to the inner surface of the membrane.


[Acute poisoning with carbon dioxide: report of 2 fatal cases].
Carbon dioxide (CO2), a very high density gas, tends to stratify at the lowest levels of the atmosphere. It can be produced by neutral geothermal emissions, fermentative processes or by human and industrial activity. When carbon dioxide concentrations rise to a very high level in a confined and poorly ventilated space, the anoxic hazard is a very important cause of severe accidents that can involve workers and rescuers. At CO2 levels higher than 20% there is a very high risk of a fatal accident, also considering the odourless feature of this gas. Two fatal accidents in workers are described which occurred during inspection of a concrete well, built as a part of sewerage network in a rural area. In the weeks after the accident, composition and concentration of gases inside the well were analysed. We also considered the influence of an organic fertilizer called "pollina" which was found on the ground around the concrete well, in order to ascertain whether fermentation could alter the gas composition inside the well. Samples of air and water were collected in the well and samples of the organic fertilizer (pollina) on the ground surrounding the concrete well were also taken. Different quantities of organic fertilizer (pollina) with or without water were incubated in airtight glass bottles and placed in a dark room at 20 degrees C temperature; analysis of air inside the glass bottles was performed after 7 and 18 days of incubation. All the samples of air and water were analysed by gas-chromatographic-mass-spectrometry. Analysis of the air samples collected inside the well after 2, 16 and 18 weeks after the accident showed a low concentration of O2 (range 4.2-9%), a high concentration of CO2 (range 5.9-12.3%), a normal level of N2 (range 78-85%) and a concentration of N2O between 0.03 and 0.19%. In water collected 2 weeks after the accident at the bottom of the concrete well, CO2 and N2O concentrations of respectively 222 mg/L and 2 mg/L were measured. In the bottles with "pollina" we found, at different times of incubation, high concentrations of CO2 (highest value 25.2%), low levels of O2 (lowest value 0.5%) and negligible concentrations of N2O (< 0.015%). All these findings suggest that the atmosphere inside the concrete well was altered by the fermentative processes of pollina. The death of the two workers, caused by a poorly oxygenated atmosphere with a high concentration of carbon dioxide, can be classified under the confined space hypoxic syndrome (CSHS).


Morning preference is associated with subjective happiness among Japanese female workers: A moderation analysis by sleep characteristics from the SLEPT study.
Research on mental health in working populations has predominantly focussed on negative outcomes, while studies on the positive aspects of life remain very limited, especially in Japan. Although morningness has recently been considered a factor for being happy, the role of sleep characteristics as it relates to the association between morningness and happiness has not been substantiated. The aim of this study is to investigate the associations between diurnal preference and level of subjective happiness in healthy, full-time, daytime workers in Japan. We also tested the moderation effect of sleep parameters on the relationship between diurnal preference and subjective happiness. This is an exploratory analysis from the cross-sectional data of the SLeep Epidemiology Project at the University of Tsukuba (SLEPT Study). Subjective happiness was evaluated using a single-item question. Diurnal preference was assessed using the Morningness-Eveningness Questionnaire (MEQ), in which higher scores indicate greater tendency to have morning preference. The participants underwent assessment of sleep parameters for 7 consecutive days using a waist-worn actigraphy device and kept a sleep diary. Sleep parameters investigated were subjective sleep quality, sleep disturbance, daytime sleepiness, weekend oversleep, total sleep time, sleep onset time, wake time, and sleep efficiency. A total of 205 males (average 42.6 ± 10.4 years) and 272 females (41.1 ± 9.8 years) were eligible for analysis. Hierarchical liner regression analysis was used to show the relationships of subjective happiness with MEQ score, and the sleep parameters. Further, moderation analysis was conducted by adding the interaction between MEQ score and the sleep parameters. After adjusting for age, psychological distress, self-rated health, and occupational stress, we found that subjective happiness was significantly associated with higher MEQ score and sleep efficiency but only in female. The moderating role of sleep parameters was not found. We discussed the implications of the obtained results and a possible strategy to maintain and improve subjective happiness of female workers who have evening preference.


Factors affecting RIG-I-Like receptors activation - New research direction for viral hemorrhagic fevers.
Viral hemorrhagic fever (VHF) is a term referring to a group of life-threatening infections caused by several virus families (Arenaviridae, Bunyaviridae, Filoviridae and Flaviviridae). Depending on the virus, the infection can be mild and can be also characterized by an acute course with fever accompanied by hypervolemia and coagulopathy, resulting in bleeding and shock. It has been suggested that the course of the disease is strongly influenced by the activation of signaling pathways leading to RIG-I-like receptor-dependent interferon production. RIG-I-like receptors (RLRs) are one of two major receptor families that detect viral nucleic acid. RLR receptor activation is influenced by a number of factors that may have a key role in the differences that occur during the antiviral immune response in VHF. In the present study, we collected data on RLR receptors in viral hemorrhagic fevers and described factors that may influence the activation of the antiviral response. RLR receptors seem to be a good target for VHF research, which may contribute to better therapeutic and diagnostic strategies. However, due to the difficulty of conducting such studies in humans, we suggest using Lagovirus europaeus as an animal model for VHF.