Some factors influencing the sensitivity of body temperature to activity in neonates.
In adult humans, core temperature is influenced by activity; the sensitivity of core temperature to such effects shows a phase dependence and is also influenced by the environment and whether the individual is asleep or awake. We have investigated if similar effects are evident in neonates, in whom thermoregulation and the circadian rhythm of core temperature are not fully developed. Eleven full-term, healthy babies were studied singly (light 07:00-19:00) at 2 days of age and again 4 weeks after birth; between these times, they were tended routinely on a communal ward. On study days, 10-minute recordings were made of rectal and skin (abdominal) temperature, heart rate (HR), and behavioral state. Sensitivities of the temperatures to activity ("arousal") were assessed throughout the 24h by measuring the gradient of (temperature/HR). Sensitivities measured at 01:00, 05:00, 09:00, 13:00, 17:00, and 21:00 were used as dependent variables in stepwise regression and linear regression analyses, with "subjects," "light versus dark," "behavioral state," and "difference between time of measurement and the acrophase of the endogenous component of the temperature rhythm" (ignoring sign) as possible predictors. (Acrophases of the temperature rhythms had been estimated from 24h data purified using the behavioral state record.) Light versus dark acted as a significant predictor of the sensitivity of rectal temperature to arousal on day 2 and week 4, the sensitivity increasing in the light, and there was limited evidence for behavioral state acting as a predictor on day 2. Neither factor was a significant predictor when the sensitivity of the babies' skin temperatures to arousal was investigated. There was also some evidence that the difference between the time of measurement and the temperature acrophase acted as a predictor of sensitivity to arousal in both rectal (day 2) and skin (week 4) temperature, with larger differences decreasing the sensitivity. These results indicate that there are masking effects on body temperature due to arousal in neonates, the size of which depends on both internal and external factors. However, this sensitivity of temperature to arousal shows differences from the sensitivity of temperature to physical activity in both adult humans and adult mice. One possible explanation of this result is that temperature regulation and the circadian system are not fully developed in humans at this age.


Transformation of Cryptococcus neoformans by electroporation using a transient CRISPR-Cas9 expression (TRACE) system.
The basidiomycete Cryptococcus neoformans is not only a clinically important pathogen, but also a model organism for studying microbial pathogenesis and eukaryotic biology. One key factor behind its rise as a model organism is its genetic amenability. The widely used methods for transforming the C. neoformans species complex are Agrobacterium-mediated transformation (AMT) for random insertional mutagenesis and biolistic transformation for targeted mutagenesis. Electroporation was introduced to C. neoformans in early 1990s. Although electroporation is economic and yields a large number of transformants, introduced DNA rarely integrates into cryptococcal genome, which limits its use. Biolistic transformation, although costly and inefficient, has been the only method used in targeted mutagenesis in the past two decades. Several modifications, including the use of a donor DNA with split markers, a drug-resistant selection marker, and a recipient strain deficient in non-homologous end joining (NHEJ), have since modestly increased the frequency of genome integration and the rate of homologous replacement of the DNA introduced by electroporation. However, electroporation was not the method of choice for transformation until the recent adoption of CRISPR-Cas9 systems. We have developed a Transient CRISPR-Cas9 coupled with Electroporation System (TRACE), which dramatically facilitates targeted mutagenesis in the Cryptococcus species complex. TRACE combines the high transformation efficiency of electroporation with the high rates of DNA integration due to the transiently expressed CRISPR-Cas9. Here, we briefly discussed the history of electroporation for Cryptococcus transformation and provided detailed procedures for electroporation and the cassettes construction of the TRACE system for various genetic manipulations.


A promoter polymorphism (-77T>C) of DNA repair gene XRCC1 is associated with risk of lung cancer in relation to tobacco smoking.
X-ray repair cross complementing group 1 (XRCC1) is one of the major DNA repair proteins involved in the base-excision repair pathway. Functional Polymorphisms in the XRCC1 gene may lead to decreased DNA repair capacity and thus confer inherited predisposition to cancer risk. In this case-control study of 710 patients with incident lung cancer and 710 cancer-free controls who were frequency matched on age, sex and residential area, we genotyped a novel T>C transition at the promoter region (-77T>C) of XRCC1 and other two common non-synonymous polymorphisms (Arg194Trp and Arg399Gln) to determine their associations with risk of lung cancer. We found that compared with the -77TT wild-type homozygote, the variant genotypes were associated with significantly increased risk of lung cancer [adjusted odds ratio (OR)=1.51; 95% confidence interval (CI)=1.17-1.94 for -77TC; OR=2.98; 95% CI=0.93-9.59 for -77CC; and OR=1.55; 95% CI=1.21-1.98 for -77TC/CC]. By contrast, no significant associations were observed between the other two exonic variants (Arg194Trp and Arg399Gln) and lung cancer risk. Furthermore, we observed a 9.82-fold increased risk (95% CI=5.66-17.02) for heavy smokers carrying the -77C variant (-77TC/CC) and a 4.07-fold increased risk (95% CI=2.85-5.81) for heavy smokers not carrying the variant. However, the interaction between the -77T>C variant and cumulative smoking was not statistically significant (P=0.1560). These findings indicate that the new XRCC1 -77T>C polymorphism may contribute to the aetiology of lung cancer. Further functional studies are warranted to elucidate the underlying molecular mechanisms of the association.


Human histocompatibility antigen associations in subacute cutaneous lupus erythematosus.
We have identified a clinically distinct subset of lupus erythematosus patients marked by the presence of a histologically proven, nonscarring variety of cutaneous LE (subacute cutaneous LE) in which there is a very high frequency of the human leukocyte antigens (HLA) B8 and DR3. Differences in the configuration of their skin lesions allowed a separation of the patients into two clinical subgroups; annular and papulosquamous. HLA-B8 was increased in the annular subgroup (81%, corrected P (Pc) < 0.007) and combined group (65%, Pc < 0.004). HLA-DR3 was present in all 11 of the annular patients (10%, Pc < 0.00008). In addition, HLA-DR3 was present in increased frequencies in the papulosquamous subgroup (60%, Pc < 0.04) and combined group (77%, Pc < 0.00008). Thus, HLA-DR3 positive individuals have a relative risk of 10.8 for developing subacute cutaneous LE of either type and an even greater relative risk (67.1) for the annular variety. The HLA phenotype A1, B8, DR3 was also found more commonly in the annular (73%, P < 0.00008) and combined patient groups (46%, P < 0.004). These HLA associations, which are stronger than ever before reported for any form of LE, did not result from the concurrent presence of subclinical Sjögren's syndrome. Thus, subacute cutaneous LE can now be added to the growing list of HLA-B8, DR3-associated diseases that have autoimmune features.


Nasal administration of amyloid-beta peptide decreases cerebral amyloid burden in a mouse model of Alzheimer's disease.
Progressive cerebral deposition of amyloid-beta (Abeta) peptide, an early and essential feature of Alzheimer's disease (AD), is accompanied by an inflammatory reaction marked by microgliosis, astrocytosis, and the release of proinflammatory cytokines. Mucosal administration of disease-implicated proteins can induce antigen-specific anti-inflammatory immune responses in mucosal lymphoid tissue which then act systemically. We hypothesized that chronic mucosal administration of Abeta peptide might induce an anti-inflammatory process in AD brain tissue that could beneficially affect the neuropathological findings. To test this hypothesis, we treated PDAPP mice, a transgenic line displaying numerous neuropathological features of AD, between the ages of approximately 5 and approximately 12 months with human Abeta synthetic peptide mucosally each week. We found significant decreases in the cerebral Abeta plaque burden and Abeta42 levels in mice treated intranasally with Abeta peptide versus controls treated with myelin basic protein or left untreated. This lower Abeta burden was associated with decreased local microglial and astrocytic activation, decreased neuritic dystrophy, serum anti-Abeta antibodies of the IgG1 and IgG2b classes, and mononuclear cells in the brain expressing the anti-inflammatory cytokines interleukin-4, interleukin-10, and tumor growth factor-beta. Our results demonstrate that chronic nasal administration of Abeta peptide can induce an immune response to Abeta that decreases cerebral Abeta deposition, suggesting a novel mucosal immunological approach for the treatment and prevention of AD.


Prospects for human papillomavirus vaccine development: emerging HPV vaccines.
This review concentrates on recent advances in human papillomavirus vaccine development. Strategies for prophylactic HPV subunit vaccines utilizing recombinantly synthesized, immunogenic virus-like particles are discussed. Therapeutic strategies focusing on the induction of cell-mediated immunity and gene manipulation for the treatment of established HPV-associated disease are also reviewed.


Serum serotonin levels determine the magnitude and type of bystander effects in medium transfer experiments.
Serotonin has been shown to be involved in the production of bystander signals by irradiated cells. In this study we examined the levels of serotonin in 10 different batches of commercially available fetal calf serum and correlated the serotonin levels with the toxicity of medium harvested from irradiated cells (ICCM) using a standard medium transfer colony-forming assay. The serotonin levels in the serum varied widely between batches, and the levels correlated directly with the toxicity of the harvested ICCM. Three serum samples had levels of serotonin below 25 ng/ml, and these did not show medium transfer bystander effects. Exposure of serum samples to normal daylight reduced serotonin levels significantly. We suggest that serum batch variability may underlie much of the observed interlaboratory variation in the ability to produce bystander effects and further suggest that serum batches should be protected from light and prescreened for their ability to produce a bystander effect using a positive control cell line.


A new noise-tolerant and predefined-time ZNN model for time-dependent matrix inversion.
In this work, a new zeroing neural network (ZNN) using a versatile activation function (VAF) is presented and introduced for solving time-dependent matrix inversion. Unlike existing ZNN models, the proposed ZNN model not only converges to zero within a predefined finite time but also tolerates several noises in solving the time-dependent matrix inversion, and thus called new noise-tolerant ZNN (NNTZNN) model. In addition, the convergence and robustness of this model are mathematically analyzed in detail. Two comparative numerical simulations with different dimensions are used to test the efficiency and superiority of the NNTZNN model to the previous ZNN models using other activation functions. In addition, two practical application examples (i.e., a mobile manipulator and a real Kinova JACO2 robot manipulator) are presented to validate the applicability and physical feasibility of the NNTZNN model in a noisy environment. Both simulative and experimental results demonstrate the effectiveness and tolerant-noise ability of the NNTZNN model.


Everyday physical activity and adiposity in Prader-Willi syndrome.
The aim of this study was to assess the impact of Prader-Willi syndrome (PWS) on the level of everyday physical activity and to explore whether the activity level is related to adiposity. Measurements were performed with an accelerometry-based Activity Monitor during two consecutive schooldays in 12 children with PWS (7-16 years of age) and in 12 age- and gender-matched, healthy children. Adiposity was assessed by body mass index standard deviation scores and by percentage body fat (dual energy X-ray absorptiometry). Mean duration of dynamic activities (expressed as percentage of 24 h) was lower in children with PWS than in the comparison group (8.7 [2.5]% and 12.0 [3.1]%, respectively; p = 0.01). Six children with PWS had normal activity levels. Physical activity level was not related to adiposity. The results indicate that, as a group, children with PWS have an inactive lifestyle. However, children with PWS cannot be stereotyped as inactive since half of them had normal activity levels.


Activation of leukotriene synthesis in human neutrophils by exogenous arachidonic acid: inhibition by adenosine A(2a) receptor agonists and crucial role of autocrine activation by leukotriene B(4).
We report here that the apparent inability of isolated human polymorphonuclear leukocytes (PMNs) to efficiently transform arachidonic acid (AA) is the consequence of A(2a) receptor engagement by endogenous adenosine accumulating in incubation media. Indeed, when adenosine is eliminated from PMN suspensions by the addition of adenosine deaminase, or when cells are incubated with adenosine A(2a) receptor antagonists, important quantities (40-80 pmol/10(6) cells) of 5-lipoxygenase products are synthesized by PMN incubated with 1 to 5 microM exogenous AA. The selective A(2a) receptor agonist CGS21680 was a very potent inhibitor of the AA-induced leukotriene (LT) synthesis, showing an IC(50) of approximately 1 nM. The mechanism of AA-induced stimulation of LT synthesis observed in the absence of extracellular adenosine was investigated. In adenosine deaminase-treated PMN, exogenous AA induced Ca(2+) mobilization and the translocation of 5-lipoxygenase to nuclear structures. A time lag of 20 to 60 s (variable between PMN preparations) was observed consistently between the addition of AA and the elevation of intracellular Ca(2+) concentration (and LT synthesis), indicating that AA itself did not trigger the Ca(2+) mobilization in PMN. This AA-induced Ca(2+) mobilization, as well as the corresponding 5-lipoxygenase translocation and stimulation of LT synthesis, was blocked efficiently by the LT synthesis inhibitor MK0591, the LTB(4) receptor antagonists CP105696 and LY223982, and the LTA(4) hydrolase inhibitor SC57461A. These data demonstrate that AA is a highly potent and effective activator of LT synthesis and acts through a mechanism that requires an autocrine stimulatory loop by LTB(4).


MIPSS70+ v2.0 predicts long-term survival in myelofibrosis after allogeneic HCT with the Flu/Mel conditioning regimen.
Although allogeneic hematopoietic cell transplantation (allo-HCT) is the only curative treatment for myelofibrosis (MF), data are limited on how molecular markers predict transplantation outcomes. We retrospectively evaluated transplantation outcomes of 110 consecutive MF patients who underwent allo-HCT with a fludarabine/melphalan (Flu/Mel) conditioning regimen at our center and assessed the impact of molecular markers on outcomes based on a 72-gene next-generation sequencing panel and Mutation-Enhanced International Prognostic Scoring System 70+ v2.0 (MIPSS70+ v2.0). With a median follow-up of 63.7 months, the 5-year overall survival (OS) rate was 65% and the nonrelapse mortality (NRM) rate was 17%. In mutational analysis, JAK2 V617F and ASXL1 mutations were the most common. By univariable analysis, higher Dynamic International Prognostic Scoring System scores, unrelated donor type, and very-high-risk cytogenetics were significantly associated with lower OS. Only CBL mutations were significantly associated with lower OS (hazard ratio [HR], 2.64; P = .032) and increased NRM (HR, 3.68; P = .004) after allo-HCT, but CALR, ASXL1, and IDH mutations did not have an impact on transplantation outcomes. Patient classification per MIPSS70 showed worse OS for high-risk (HR, 0.49; P = .039) compared with intermediate-risk patients. Classification per MIPSS70+ v2.0 demonstrated better OS when intermediate-risk patients were compared with high-risk patients (HR, 0.291) and much lower OS when very-high-risk patients were compared with high-risk patients (HR, 5.05; P ≤ .001). In summary, we present one of the largest single-center experiences of Flu/Mel-based allo-HCT, demonstrating that revised cytogenetic changes and MIPSS70+ v2.0 score predict transplantation outcomes, and thus can better inform physicians and patients in making decisions about allo-HCT.


Magnetic Resonance Imaging Assessment of Lipomatous Soft-tissue Tumors.
To establish the accuracy of magnetic resonance imaging (MRI) in distinguishing between benign and malignant lipomatous tumors; to evaluate the reproducibility of the MRI interpretation assessing the agreement between judgments of two radiologists with the same experience in soft-tissue sarcomas; to identify an association among MRI findings (size, depth, septa, nodules, signal homogeneity) and nature of the lesion. A total of 54 patients (28 men and 26 women), with a mean age of 56 (range=27-84) were included years. All subjects followed-up by the Multidisciplinary Sarcoma Group. The following MRI findings were judged in a blind study by two radiologists: size, localization, septa, nodules and signal homogeneity. A diagnostic indication was then given from among lipoma, atypical lipomatous tumour (ALT) and liposarcoma. Accuracy in distinguishing between benign and malignant lesions, and between lipoma and ALT (Fisher's exact test), inter-operator agreement (Cohen's kappa), association of MRI findings and malignancy of the lesion (Fisher's exact test and odds ratio) were evaluated. The inter-operator agreement was complete (100%). The agreement between diagnostic hypothesis and histological diagnosis was statistically significant (p<0.05). Among the radiological findings taken into account, only septa and signal homogeneity were significantly associated with the malignancy of the lesion (p<0.05). MRI could be helpful in distinguishing lipomatous tumors, allowing biopsy to be avoided in some cases (negative predictive value=100%).


Microdialysis coupled with an embedded systems controller and CMOS image sensor.
Continuous monitoring of specific metabolite and drug levels within a patient's blood can contribute to shorter hospital stays and more successful treatment of both chronic and acute diseases. Intravenous microdialysis is an attractive technology for continuous venous blood sampling that can be used to manage tight glucose control and to sample a large variety of molecules from human blood. In combination with lab-on-a-chip architectures, microdialysis could provide continuous monitoring of important diagnostic and therapeutic substances. Unfortunately, microdialysis is inherently variable and non-transparent, i.e., errors in sampling cannot be detected and corrected in real-time. A portable microdialysis system is presented that gauges membrane diffusive capacity by using a fluorescent tracer, providing a method to track the intrinsic variability. An embedded systems controller and CMOS image sensor is used to measure and wirelessly communicate fluorescent tracer levels. The controller has the capability to generate alarms when probe performance deteriorates, making microdialysis both more accurate and robust for clinical use. The potential to integrate a microparticle-based, turbidimetric vancomycin immunoassay with microdialysis is also demonstrated by using a CMOS image sensor to detect changes in turbidity.


Fatty acid synthase inhibitors modulate energy balance via mammalian target of rapamycin complex 1 signaling in the central nervous system.
Evidence links the hypothalamic fatty acid synthase (FAS) pathway to the regulation of food intake and body weight. This includes pharmacological inhibitors that potently reduce feeding and body weight. The mammalian target of rapamycin (mTOR) is an intracellular fuel sensor whose activity in the hypothalamus is also linked to the regulation of energy balance. The purpose of these experiments was to determine whether hypothalamic mTOR complex 1 (mTORC1) signaling is involved in mediating the effects of FAS inhibitors. We measured the hypothalamic phosphorylation of two downstream targets of mTORC1, S6 kinase 1 (S6K1) and S6 ribosomal protein (S6), after administration of the FAS inhibitors C75 and cerulenin in rats. We evaluated food intake in response to FAS inhibitors in rats pretreated with the mTOR inhibitor rapamycin and in mice lacking functional S6K1 (S6K1(-/-)). Food intake and phosphorylation of S6K1 and S6 were also determined after C75 injection in rats maintained on a ketogenic diet. C75 and cerulenin increased phosphorylation of S6K1 and S6, and their anorexic action was reduced in rapamycin-treated rats and in S6K1(-/-) mice. Consistent with our previous findings, C75 was ineffective at reducing caloric intake in ketotic rats. Under ketosis, C75 was also less efficient at stimulating mTORC1 signaling. These findings collectively indicate an important interaction between the FAS and mTORC1 pathways in the central nervous system for regulating energy balance, possibly via modulation of neuronal glucose utilization.


Low levels of serum calcidiol in an African population compared to a North European population.
To compare vitamin D status in an African population living at 10 degrees N with a Norwegian population living at 60 degrees N. Serum samples from 30 healthy young Ethiopians and 31 full term pregnant women from Addis Ababa were collected in September, and from 24 healthy Norwegians in March and 23 pregnant women from Oslo in February to June. Serum (s) levels of calcidiol and intact parathyroid hormone (iPTH) were measured. The median values for s-calcidiol were significantly lower in Ethiopians compared with Norwegians (young Ethiopians 23.5nmol/l vs young Norwegians 81nmol/l, P<0.001; pregnant Ethiopians 25nmol/l vs pregnant Norwegians 36nmol/l, P<0.05) while those for s-iPTH were significantly higher (young Ethiopians 5.7pmol/l vs young Norwegians 2.4pmol/l, P<0.001; pregnant Ethiopians 4.8pmol/l vs pregnant Norwegians 2.8pmol/l, P<0.02). In spite of abundant availability of ultraviolet radiation, the population from Addis Ababa had a high rate of biochemical vitamin D deficiency compared with the Norwegian group.


[C-reactive protein as an indicator of the severity of war injuries].
During the general reaction to trauma, acute phase proteins are synthetized. The aim of the prospective study was to determine CRP concentrations in sera of war casualties during the first 14 posttraumatic days, and to establish the correlation between these changes and severity of trauma. Subjects were 79 war casualties. 33 blood donors. Injury severity was determined according to ISS and CRP concentrations with immunonephelometric analysis. Blood samples were collected 12 hours after trauma, then on the 1st, 2nd, 5th and 14th posttraumatic day. In war casualties CRP values were significantly increased (56.257.53 mg/dl after 12 hrs, 107.0976.08 on 1st, 144.3570.23 on 2nd, 71.42558.66 on 5th and 37.656.14 on 14th posttraumatic day; p). Significant differences were observed between groups with ISS and ISS12 (p) in first two days and later between group with ISS24 (144.1766.94 mg/dl on 5th and 111.588.5 on 14th posttraumatic day) and others (p). During the acute-phase response to trauma, significant changes in concentration of CRP occur in sera of war casualties. These changes are the most prominent during the first 48 hours, with tendency for normalization after the 5th day. Intensity of these changes depend on the severity of trauma. CRP is valid marker of war wound severity.


The effect of antenatal care follow-up on neonatal health outcomes: a systematic review and meta-analysis.
Neonatal mortality is one of the major public health problems throughout the world and most notably in developing countries. There exist inconclusive findings on the effect of antenatal care visits on neonatal death worldwide. Thus, the aim of this systematic review and meta-analysis was to reveal the pooled effect of antenatal care visits on neonatal death. The present systematic review and meta-analysis was performed using published literature, which was accessed from national and international databases such as, Medline/PubMed, EMBASE, CINAHL, Cochrane Central library, Google Scholar, and HINARI. STATA/SE for windows version 13 software was used to calculate the pooled effect size with 95% confidence intervals (95% CI) of maternal antenatal care visits on neonatal death using the DerSimonian and Laird random effects meta-analysis (random effects model), and results were displayed using forest plot. Statistical heterogeneity was checked using the Cochran Q test (chi-squared statistic) and I 2 test statistic and by visual examination of the forest plot. A total of 18 studies, which fulfilled the inclusion criteria, were included in the present systematic review and meta-analysis. The finding of the present systematic review and meta-analysis revealed that antenatal care visits decrease the risk of neonatal mortality [pooled effect size 0.66 (95% CI, 0.54, 0.80)]. Cochrane Q test (P < 0.001) revealed no significant heterogeneity among included studies, but I 2 statistic revealed sizeable heterogeneity up to 80.5% (I 2 = 80.5%). In the present meta-analysis traditional funnel plot, Egger's weighted regression (P = 0.48) as well as Begg's rank correlation statistic (P = 0.47) revealed no evidence of publication bias. The present systematic review and meta-analysis revealed that antenatal care visits were significantly associated with lower rates of neonatal death. The risk of neonatal death was significantly reduced by 34% among newborns delivered from mothers who had antenatal care visits. Thus, visiting antenatal care clinics during pregnancy is strongly recommended especially in resource-limited settings like countries of sub-Saharan Africa.


Noninvasive brain stimulation for dysphagia after acquired brain injury: a systematic review.
The purpose of this study was to review the best available evidence of noninvasive brain stimulation, including repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) for dysphagia after acquired brain injury. We searched randomized controlled trials that compared noninvasive brain stimulation with control used to improve dysphagia after acquired brain injury. We assessed dysphagia severity rating scales and penetration-aspiration scale as outcomes immediately after intervention. We calculated the pooled estimate of the standardized mean difference (SMD) to combine individual results. We included 8 published studies. Two heterogenous trials of 48 patients showed that rTMS was associated with a significant improvement in the dysphagia severity rating scale score (SMD 2.95). Three homogeneous trials of 88 patients showed a significant effect of rTMS on the penetration-aspiration scale score (SMD 0.77). Two homogeneous trials of 34 patients showed that tDCS was associated with a significant improvement in the dysphagia severity rating scale score (SMD 1.20). The review provided low-quality evidence for the effectiveness of noninvasive brain stimulation in improving dysphagia after acquired brain injury. Further trials of larger sample sizes are needed to determine the most appropriate noninvasive brain stimulation protocol. J. Med. Invest. 63: 153-158, August, 2016.


Factors Associated With Intention of Future Pregnancy Among Women Affected by the Fukushima Nuclear Accident: Analysis of Fukushima Health Management Survey Data From 2012 to 2014.
Little is known about the association between the anxiety toward the effects of radiation on reproduction caused by the Fukushima nuclear accident and the birth rate of people in Fukushima. Therefore, we examined changes and associated factors of future pregnancy intention among mothers in Fukushima Prefecture. Using data from three postal surveys among women who registered their pregnancies in the prefecture (N = 6,751 in 2012, N = 6,871 in 2013, and N = 6,725 in 2014), we analyzed the factors associated with women's intention of future pregnancy using multivariable logistic regression models. The proportion of mothers with pregnancy intention increased from 53.5% in 2012 to 57.9% in 2014, especially among multiparas (P for trend <0.001). Factors inversely associated with pregnancy intention of both groups were older maternal age (adjusted odds ratio [aOR] 0.92 for primipara and 0.87 for multipara), poor subjective health (aOR 0.75 and 0.81, respectively), and presence of depressive symptoms (aOR 0.71 and 0.79, respectively) (P < 0.01 for all items). In addition, not living with husband (aOR 0.24), dissatisfaction with obstetrical care (aOR 0.89) and child abnormalities (aOR 0.72) were inversely associated with pregnancy intention among primiparas, while receiving infertility treatment (aOR 2.05) was positively associated among multiparas (P < 0.01 for all items). A separate analysis of 2012 and 2013 data showed that concern about radiation contamination of breast milk was associated with pregnancy intention among primiparas (aOR 0.61, P < 0.001). Mothers' concern about radiation was associated with lower pregnancy intention, especially among primiparas. Providing quality obstetrical and mental health care and parenting support may be the keys to maintaining the temporal increase in fertility.


Adverse reactions to food constituents: allergy, intolerance, and autoimmunity.
Food allergies and intolerance represent important health concerns to consumers who are predisposed to these illnesses. Unlike many current food safety issues, food sensitivities are complicated by both complex and multiple individual adverse reactions, which can vary from emotional to pathophysiological ailments. In some instances, the underlying mechanisms that result in the development of food allergies or intolerance have marked differences but produce common symptoms. The present-day diagnosis of these disorders can be impeded by intrinsic limitations in generating accurate information from patient history and biochemical, physicochemical, and immunochemical tests. Oral challenge tests represent effective methods for confirming and testing food allergens and food intolerance; however, these procedures are often restricted to clinical trials. It is important to be able to distinguish among food allergy, intolerance, and autoimmune disease in the management of these disorders. The role of food in the development of autoimmune disease may be exemplified by celiac disease, a food-induced enteropathy, requiring exposure to prolamins in wheat, rye, and barley. Various wheat and soy protein sources, including the soy protein isolates used to make infant formulas, have been related to juvenile or insulin-dependent diabetes mellitus (IDDM), a common chronic disease of childhood. Employing food process technologies to eliminate food constituents with potential for intolerance in some individuals is a potentially viable approach for reducing risk to food-related disorders. Finally, the development of food labelling regulations that require the identification of potential food allergens or agents for intolerance in the ingredient declaration on prepackaged food is a positive step toward the prevention of severe adverse reactions in hypersensitive individuals.


Tissue factor expressed by microparticles is associated with mortality but not with thrombosis in cancer patients.
A prothrombotic state is one of the hallmarks of malignancy and a major contributor to morbidity and mortality in cancer patients.Tissue factor (TF) is often overexpressed in malignancy and is a prime candidate in predicting the hypercoagulable state. Moreover, increased number of TF-exposing microparticles (MPs) in cancer patients may contribute to venous thromboembolism (VTE). We have conducted a prospective cohort study to determine whether elevated TF antigen, TF activity and TF associated to MPs (MPs-TF) are predictive of VTE and mortality in cancer patients. The studied population consisted of 252 cancer patients and 36 healthy controls. TF antigen and activity and MPs-TF were determined by ELISA and chromogenic assays. During a median follow-up of 10 months, 40 thrombotic events were recorded in 34 patients (13.5%), and 73 patients (28.9%) died. TF antigen and activity were significantly higher in patients than in controls (p<0.01) mainly in patients with advanced stages, whereas no differences were observed for TF activity of isolated MPs. We did not find a statistically significant association of TF variables with the risk of VTE. Multivariate analysis adjusting for age, sex, type of cancer and other confounding variables showed that TF activity (p<0.01) and MPs-TF activity (p<0.05) were independently associated with mortality. In conclusion, while TF variables were not associated with future VTE in cancer patients, we found a strong association of TF and MPs-TF activity with mortality, thus suggesting they might be good prognostic markers in cancer patients.


Inhibition of Kv4.3 by genistein via a tyrosine phosphorylation-independent mechanism.
The effects of genistein, a protein tyrosine kinase (PTK) inhibitor, on voltage-dependent K(+) (Kv) 4.3 channel were examined using the whole cell patch-clamp techniques. Genistein inhibited Kv4.3 in a reversible, concentration-dependent manner with an IC(50) of 124.78 μM. Other PTK inhibitors (tyrphostin 23, tyrphostin 25, lavendustin A) had no effect on genistein-induced inhibition of Kv4.3. Orthovanadate, an inhibitor of protein phosphatases, did not reverse the inhibition of Kv4.3 by genistein. We also tested the effects of two inactive structural analogs: genistin and daidzein. Whereas Kv4.3 was unaffected by genistin, daidzein inhibited Kv4.3, albeit with a lower potency. Genistein did not affect the activation and inactivation kinetics of Kv4.3. Genistein-induced inhibition of Kv4.3 was voltage dependent with a steep increase over the channel opening voltage range. In the full-activation voltage range positive to +20 mV, no voltage-dependent inhibition was found. Genistein had no significant effect on steady-state activation, but shifted the voltage dependence of the steady-state inactivation of Kv4.3 in the hyperpolarizing direction in a concentration-dependent manner. The K(i) for the interaction between genistein and the inactivated state of Kv4.3, which was estimated from the concentration-dependent shift in the steady-state inactivation curve, was 1.17 μM. Under control conditions, closed-state inactivation was fitted to a single exponential function, and genistein accelerated closed-state inactivation. Genistein induced a weak use-dependent inhibition. These results suggest that genistein directly inhibits Kv4.3 by interacting with the closed-inactivated state of Kv4.3 channels. This effect is not mediated via inhibition of the PTK activity, because other types of PTK inhibitors could not prevent the inhibitory action of genistein.


Perception of Primary School Teachers Towards Voice Problems and Vocal Health-Seeking Behaviors: A qualitative study.
Teachers are the most common occupational group reporting to the voice clinics with dysphonia across the globe. However, less is known about teachers' perceptions about their voice and voice problems. Hence the present study aimed to explore teachers' perception about their teaching voice, voice problems and vocal health-seeking behaviors. A qualitative phenomenological research design using semi-structured interviews were conducted with fifteen female primary school teachers. Participants were asked to express their perception of teaching voice, vocal health care knowledge, and vocal hygiene practices. Using inductive analysis, the researchers converted the raw data into concepts or themes and discussed. The present study results showed that teachers perceive voice as the primary tool for their profession. However, teachers consider voice problems as part of their professional commitments. Despite knowing the harmful effects of voice problems, teachers' pay less attention to their voice problems and perceive them as usual, unavoidable and part of their profession. Due to their demanding work schedule and not getting enough support from the management, they hesitate to consult medical professionals unless it severely affects them. The data obtained from this study can be used to improve the teachers' knowledge regarding vocal health and convince the school management to extend their support for the successful implementation of the vocal health training programs for teachers.


[Cerebral mapping and attention-deficit hyperactivity syndrome].
The clinical features of the attention-deficit hyperactivity disorder are characterized by attention deficit, an impulsive style of behavior and cognition and excessive motor activity. The incidence is estimated to be between 3 and 5% of the school population, and it is considered that there are one or two children with attention deficit in all children s classes during the early years of schooling. Electroencephalogram measurements using spectral analysis gives fresh information about the cerebral electrogenesis of children with attention-deficit hyperactivity disorder. In this study we diagnosed 21 children as having attention-deficit hyperactivity disorder and compared them with a control group of 13 healthy children of the same average age, at rest and whilst carrying out various tasks. Greater values of delta relative amplitude were found in the attention-deficit hyperactivity disorder group in their basal EEG at the F7, F8, FP1, FP2, F3, F4 and C3 sites. During visual testing differences were observed in the alpha band, also in the frontal region (F7, FP1, FP2 and F4) with the children of the control group showing the higher values. The level of significance used was 0.05. The cortical distribution of the dominant frequencies was different also. In addition, we present a method for cerebral mapping by means of spline functions developed by Duchon, which allow cerebral bioelectrical activity to be mapped in three dimensions.


Free Radicals Within the Antarctic Vortex: The Role of CFCs in Antarctic Ozone Loss.
How strong is the case linking global release of chlorofluorocarbons to episodic disappearance of ozone from the Antarctic stratosphere each austral spring? Three lines of evidence defining a link are (i) observed containment in the vortex of ClO concentrations two orders of magnitude greater than normal levels; (ii) in situ observations obtained during ten high-altitude aircraft flights into the vortex as the ozone hole was forming that show a decrease in ozone concentrations as ClO concentrations increased; and (iii) a comparison between observed ozone loss rates and those predicted with the use of absolute concentrations of ClO and BrO, the rate-limiting radicals in an array of proposed catalytic cycles. Recent advances in our understanding of the kinetics, photochemistry, and structural details of key intermediates in these catalytic cycles as well as an improved absolute calibration for ClO and BrO concentrations at the temperatures and pressures encountered in the lower antarctic stratosphere have been essential for defining the link.


Hepatitis C virus infection reduces hepatocellular polarity in a vascular endothelial growth factor-dependent manner.
Hepatitis C virus (HCV) infection leads to progressive liver disease, frequently culminating in fibrosis and hepatocellular carcinoma. The mechanisms underlying liver injury in chronic hepatitis C are poorly understood. This study evaluated the role of vascular endothelial growth factor (VEGF) in hepatocyte polarity and HCV infection. We used polarized hepatoma cell lines and the recently described infectious HCV Japanese fulminant hepatitis (JFH)-1 cell culture system to study the role of VEGF in regulating hepatoma permeability and HCV infection. VEGF negatively regulates hepatocellular tight junction integrity and cell polarity by a novel VEGF receptor 2-dependent pathway. VEGF reduced hepatoma tight junction integrity, induced a re-organization of occludin, and promoted HCV entry. Conversely, inhibition of hepatoma expressed VEGF with the receptor kinase inhibitor sorafenib or with neutralizing anti-VEGF antibodies promoted polarization and inhibited HCV entry, showing an autocrine pathway. HCV infection of primary hepatocytes or hepatoma cell lines promoted VEGF expression and reduced their polarity. Importantly, treatment of HCV-infected cells with VEGF inhibitors restored their ability to polarize, showing a VEGF-dependent pathway. Hepatic polarity is critical to normal liver physiology. HCV infection promotes VEGF expression that depolarizes hepatoma cells, promoting viral transmission and lymphocyte migration into the parenchyma that may promote hepatocyte injury.


Clinical value of NGAL, L-FABP and albuminuria in predicting GFR decline in type 2 diabetes mellitus patients.
Neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid binding protein (L-FABP) are emerging as excellent biomarkers in the urine and plasma for the early prediction of acute and chronic kidney injury. The aims of this prospective study were to determine the role of albuminuria, and that of serum and urine levels of NGAL and L-FABP as predictors of a decline in the glomerular filtration rate (GFR) in patients with type 2 diabetes. A longitudinal cohort study with one hundred forty type 2 diabetic patients was conducted. Serum and urine levels of NGAL and L-FABP, and the urine albumin excretion rate were determined. The correlation between the kidney injury biomarkers and rate of GFR decline was analyzed. The eGFR of study subjects decreased significantly as the study progressed (86.4±31.1 vs. 74.4±27.3 ml/min/1.73 m(2), P<0.001), and the urine albumin excretion rate increased significantly (264.9±1060.3 vs. 557.7±2092.5 mg/day, P = 0.009). The baseline urine albumin excretion rate and serum L-FABP level were significantly correlated with baseline eGFR (P<0.05). The results of regression analysis for the correlations between the rate of eGFR change and the baseline levels of NGAL and L-FABP, and the urine albumin excretion rate showed that only the urine albumin excretion rate was significantly correlated with the rate of eGFR change (standardized coefficients: -0.378; t: -4.298; P<0.001). Tubular markers, such as NGAL and L-FABP, may not be predictive factors associated with GFR decline in type 2 diabetic patients.


Uninephrectomy in Rats on a Fixed Food Intake Potentiates Both Anorexia and Circulating Cytokine Subsets in Response to LPS.
Recent human studies have suggested that mild reduction in kidney function can alter immune response and increase susceptibility to infection. The role of mild reduction in kidney function in altering susceptibility to bacterial lipopolysaccharide (LPS) responses was investigated in uninephrectomized rats compared to Sham-operated controls rats 4 weeks after surgery. Throughout the 4 weeks, all rats were maintained under mild food restriction at 90% of ad libitum intake to ensure the same caloric intake in both groups. In comparison to Sham, uninephrectomy (UniNX) potentiated LPS-induced anorexia by 2.1-fold. The circulating anorexigenic cytokines granulocyte-macrophage colony stimulating factor, interferon-γ, tumor necrosis factor-α, and complement-derived acylation-stimulating protein were elevated after LPS in UniNX animals compared to Sham animals. Interleukin(IL)1β and IL6 pro-inflammatory cytokines were transiently increased. Anti-inflammatory cytokines IL4 and IL10 did not differ or had a tendency to be lower in UniNX group compared to Sham animals. LPS-induced anorexia was associated with increased anorexigenic neuropeptides mRNA for pro-opiomelanocortin, corticotrophin-releasing factor, and cocaine-amphetamine-regulated transcript in the hypothalamus of both Sham and UniNX groups, but at higher levels in the UniNX group. Melanocortin-4-receptor mRNA was markedly increased in the UniNX group, which may have contributed to the enhanced anorexic response to LPS of the UniNX group. In summary, UniNX potentiates pro-inflammatory cytokine production, anorexia, and selected hypothalamic anorexigenic neuropeptides in response to LPS.


Fish gut-associated bacterial communities in a tropical lagoon (Aghien lagoon, Ivory Coast).
Aghien lagoon (Ivory Coast) is a eutrophic freshwater lagoon that harbors high biomasses of phytoplankton. Despite Increasing interest in fish gut microbiomes diversity and functions, little data is currently available regarding wild species from tropical west African lakes. Here, gut-associated bacterial communities are investigated in four fish species that are consumed by locale populations, namely the Cichlidae Hemichromis fasciatus, Tilapia guineensis and Sarotherodon melanotheron, and the Claroteidae Chrysichthys nigrodigitatus. Species-related differences are identified, that can be attributed to host phylogeny and diet. Important variations throughout the year are observed in T. guineensis and C. nigrodigitatus. This result emphasized the importance of time-series sampling and comparison with environmental variables even in tropical regions, that are not often conducted in wild populations. Effects of environmental factors (anthropogenic or not) on the microbiota and potential outcomes for fish health and populations sustainability need to be further explored. Interestingly, fish appear as major reservoirs of bacterial diversity, suggesting that they could contribute to the overall stability and resilience of bacterial communities present in the Aghien lagoon.


On entropy rate for the complex domain.
We derive the entropy rate formula for a complex Gaussian random process by using a widely linear model. The resulting expression is general and applicable to both circular and noncircular Gaussian processes, since any second-order stationary process can be modeled as the output of a widely linear system driven by a circular white noise. Furthermore, we demonstrate application of the derived formula to an order selection problem. We extend a scheme for independent and identically distributed (i.i.d.) sampling to the complex domain to improve the estimation performance of information-theoretic criteria when samples are correlated. We show the effectiveness of the approach for order selection for simulated and actual functional magnetic resonance imaging (fMRI) data that are inherently complex valued.


Interactions between Chemotherapy, Endocrine Therapy and Radiation.
In an investigation by the Swedish Cancer Society, an expert group described the present status, critical issues and future aspects and potentials for each of nine major areas of radiation therapy research. This report deals with interactions between chemotherapy, endocrine therapy, other anti-tumour drugs and radiation.


In vitro behaviour of human osteoblasts on dentin and bone.
This investigation studied how the behaviour of isolated osteoblasts on standard tissue culture polystyrene compared with cells cultured on cut surfaces of dentin, a natural calcified material. Cellular attachment, viability and growth were monitored in parallel cultures of human osteosarcoma cell lines (MG63, HOS TE85, SaOS-2) and primary human osteoblast-like cells (HOBs). Culture plastic was either left untreated or roughened with abrasive paper of various grit sizes (4000-1200 grit) in order to obtain a level of roughness comparable to that of the dentin slices. Cell counting and intracellular BCECF staining showed that after an initial incubation of 2 h, the primary cells attached and spread out more quickly on the different substrates than the three cell lines. The primary cells also showed a stronger mitochondrial staining and viability on dentin. During subsequent culture morphological differences appeared with the cells on dentin displaying more cellular extensions. All three cell lines proliferated more slowly on dentin than on plastic. In contrast, the primary HOBs were not significantly affected in their growth by the different substrates. Total and specific alkaline phosphatase (AP) activity of the cell lines was not significantly affected by the different substrates after short-term adhesion, but it was increased for the primary cells on the dentin. However, after 2-3 days of culture, AP was decreased on the dentin slices for both the cell lines and primary HOBs. Plasma treatment of the roughened plastic did not alter cellular viability or AP activity, suggesting that grinding of the surface did not affect the property of the culture plastic to support cell attachment and growth. In conclusion, the results show that not only do osteoblastic cells behave differently on a natural calcified substrate surface than on standard culture plastic, but also that differences were evident between the various cell types, in particular the primary HOB versus the continuous cell lines.


Modification of yeast ribosomal proteins. Phosphorylation.
Two-dimensional polyacrylamide-gel electrophoretic analysis of yeast ribosomal proteins labelled in vivo with 32PO43- revealed that the proteins S2 and S10 of the 40S ribosomal subunit, and the proteins L9, L30, L44 and L45 of the 60S ribosomal subunit, are phosphorylated in vivo. Most of the phosphate groups appeared to be linked to serine residues. Teh number of phosphate groups per molecule of phosphorylated protein species ranged from 0.01 to 0.79. Since most of the phosphorylated ribosomal proteins appear to associate with the pre-ribosomal particles at a very late stage of ribosome assembly, phosphorylation is more likely to play a role in the functioning of the ribosome than in its assembly.


Loss of eyes in zebrafish caused by mutation of chokh/rx3.
The vertebrate eye forms by specification of the retina anlage and subsequent morphogenesis of the optic vesicles, from which the neural retina differentiates. chokh (chk) mutant zebrafish lack eyes from the earliest stages in development. Marker gene analysis indicates that retinal fate is specified normally, but optic vesicle evagination and neuronal differentiation are blocked. We show that the chk gene encodes the homeodomain-containing transcription factor, Rx3. Loss of Rx3 function in another teleost,medaka, has also been shown to result in an eyeless phenotype. The medaka rx3 locus can fully rescue the zebrafish mutant phenotype. We provide evidence that the regulation of rx3 is evolutionarily conserved, whereas the downstream cascade contains significant differences in gene regulation. Thus, these mutations in orthologous genes allow us to study the evolution of vertebrate eye development at the molecular level.


Phylogeography of the Marine Otter (Lontra felina): historical and contemporary factors determining its distribution.
The evolutionary history of a species can be revealed by phylogeographical analysis; nevertheless, not only historical but also contemporary processes can imprint on the distribution of genetic diversity. We report on the phylogeny of Lontra ssp. in South America, and the role of spatial heterogeneity in shaping the distribution and population structure of the endangered marine otter, Lontra felina. Analyzing a total of 2261 bp of mitochondrial DNA (mtDNA) revealed the recent divergence of L. felina from L. provocax. A strong population structure (Φ(st) = 0.83, P < 0.0001) and a significant pattern of isolation by distance were described for L. felina (n = 168) across a wide geographical distribution (13°53'S to 43°36'S). Lontra felina mtDNA phylogeny is composed of 2 main clades: a clade from Peru and another composed of Chilean haplotypes. Northern populations show different divergent lineages and higher genetic diversity when compared with more recently colonized southern populations. Furthermore, long sandy beaches seem to act as barriers to dispersal, creating 2 evolutionary significant units in agreement with subspecies previous description, and at least 5 different management units (MUs). At a fine spatial scale, the size of rocky seashore patches, the distance between patches and anthropogenic factors also play important roles in species gene flow.


[T lymphocyte subsets combined with inflammatory indicators for auxiliary diagnosis and prognosis evaluation of sepsis].
Objective: To explore the application value of T lymphocyte subsets combined with procalcitonin (PCT), C-reactive protein (CRP), neutrophil to lymphocyte ratio (NLR) and white blood cell count (WBC) in the auxiliary diagnosis and prognosis evaluation of sepsis. Methods: In a retrospective study, seventy-two patients with sepsis diagnosed and treated in Tianjin First Central Hospital from June 2018 to April 2021 were selected as the research objects, and included in the sepsis group were 46 males and 26 females, aged 68 (57.3, 80.3) years. In addition, 111 patients with local infection admitted to hospital during the same period were included in the local infection group, including 62 males and 49 females, aged 68 (51, 77) years. Sepsis patients were divided into survival group (43 cases) and death group (29 cases) according to the 28-day outcome. CD3+, CD4+, CD8+, CD4+/CD8+ ratio were detected by flow cytometry within 24 h after admission, PCT was detected by ELISA, CRP was detected by immunoturbidimetry, blood routine examination, blood lactic acid (Lac) and oxygen partial pressure (PO2) were detected by instrumental method. Multivariate Logistic regression analysis was used to evaluate the correlation between each indicator and sepsis, and receiver operating characteristic curve (ROC) was drawn to evaluate the diagnostic value of each indicator for sepsis. Multivariate Logistic regression analysis and Kaplan Meier survival analysis were used to evaluate the prognostic value of each index for patients with sepsis. Results: Peripheral blood CD3+, CD4+, CD8+, CD4+/CD8+ ratio and PLT in sepsis group were significantly lower than those in local infection group(Z=-8.184,P<0.001;Z=-7.210,P<0.001;Z=-5.936,P<0.001;Z=-2.700,P=0.007;Z=-6.381,P<0.001); PCT, CRP, NLR and Lac levels were significantly higher than those in local infection group(Z=-8.262,P<0.001;Z=-3.094,P=0.002;Z=-9.004,P<0.001;Z=-4.770,P<0.001). Multivariate Logistic regression model showed that PCT, NLR, CD3+, CD8+, CD4+/CD8+ were independent risk factors for sepsis. According to ROC curve analysis, AUC of sepsis patients diagnosed by each indicator were 0.862, 0.894, 0.858, 0.760 and 0.618, respectively. The cut-off values were 3.075 ng/ml, 10.715, 44.935×109/L, 27.463×109/L and 0.750, respectively. The NLR sensitivity was 80.6%, and the CD3+ specificity was 94.6%. The AUC of combined detection of PCT and NLR was 0.947, sensitivity was 87.5% and specificity was 91.9%. The combined detection AUC of PCT, NLR, CD3+, CD4+/CD8+ was 0.958, the sensitivity and specificity were 90.3% and 91.0% respectively(P<0.001). PCT and Lac in death group were significantly higher than those in survival group(Z=-2.302,P=0.021;Z=-3.095,P=0.002);Peripheral blood CD4+/CD8+ levels were significantly lower than those in survival group(Z=-3.691,P<0.001),Multivariate Logistic regression model showed that CD4+/CD8+ ratio was an independent risk factor for 28 d mortality in patients with sepsis (P<0.001). The ROC curve showed that the AUC was 0.758, and the Youden index reached the maximum when the cut-off value was 1.27, the sensitivity and specificity were 79.3% and 60.5%, respectively. Compared with patients with CD4+/CD8+ ≥1.27, 28-day mortality was significantly increased in patients with CD4+/CD8+<1.27 (P=0.032). Conclusion: The combined detection of PCT, NLR, CD3+ and CD4+/CD8+ can improve the auxiliary diagnostic efficiency of sepsis, and the ratio of CD4+/CD8+ in peripheral blood may have certain predictive value for the prognosis of sepsis.


Hydrogen-bonding catalysis and inhibition by simple solvents in the stereoselective kinetic epoxide-opening spirocyclization of glycal epoxides to form spiroketals.
Mechanistic investigations of a MeOH-induced kinetic epoxide-opening spirocyclization of glycal epoxides have revealed dramatic, specific roles for simple solvents in hydrogen-bonding catalysis of this reaction to form spiroketal products stereoselectively with inversion of configuration at the anomeric carbon. A series of electronically tuned C1-aryl glycal epoxides was used to study the mechanism of this reaction based on differential reaction rates and inherent preferences for S(N)2 versus S(N)1 reaction manifolds. Hammett analysis of reaction kinetics with these substrates is consistent with an S(N)2 or S(N)2-like mechanism (ρ = -1.3 vs ρ = -5.1 for corresponding S(N)1 reactions of these substrates). Notably, the spirocyclization reaction is second-order dependent on MeOH, and the glycal ring oxygen is required for second-order MeOH catalysis. However, acetone cosolvent is a first-order inhibitor of the reaction. A transition state consistent with the experimental data is proposed in which one equivalent of MeOH activates the epoxide electrophile via a hydrogen bond while a second equivalent of MeOH chelates the side-chain nucleophile and glycal ring oxygen. A paradoxical previous observation that decreased MeOH concentration leads to increased competing intermolecular methyl glycoside formation is resolved by the finding that this side reaction is only first-order dependent on MeOH. This study highlights the unusual abilities of simple solvents to act as hydrogen-bonding catalysts and inhibitors in epoxide-opening reactions, providing both stereoselectivity and discrimination between competing reaction manifolds. This spirocyclization reaction provides efficient, stereocontrolled access to spiroketals that are key structural motifs in natural products.


Squamous differentiation in carcinoma of the endometrium: a critical appraisal of adenoacanthoma and adenosquamous carcinoma.
Although squamous differentiation within endometrial carcinomas has long been recognized by pathologists, its biologic significance has been the subject of continued debate. While some authors have found a worsened prognosis for women who have tumors with squamous elements, others have reported the prognosis to be better than conventional endometrial adenocarcinomas. Persisting confusion and disagreement about the use of the terms adenoacanthoma and adenosquamous carcinoma have complicated the issue. In this article we review the literature on the pathogenesis of squamous differentiation in the endometrium and discuss the histologic features, prevalence, and biologic behavior of adenocarcinoma with squamous differentiation. We conclude that keratin is a constituent of normal and neoplastic endometrial epithelial cells, and that overt squamous differentiation occurs by a mechanism that is currently unknown. Squamous differentiation is present in about 25% of endometrial adenocarcinomas, a frequency that appears to have been constant for the past 50 years. The squamous component of endometrial carcinomas may histologically appear benign, malignant, or indeterminant, and in the majority of instances closely parallels the differentiation of the glandular component. Endometrial adenocarcinomas with malignant-appearing squamous elements usually have poorly differentiated glandular components and have a prognosis identical to that of poorly differentiated adenocarcinoma without squamous differentiation. Endometrial adenocarcinomas with benign-appearing squamous elements are usually associated with well-differentiated glandular components and have a prognosis identical to that of typical well-differentiated adenocarcinoma. Some endometrial adenocarcinomas contain foci of squamous differentiation that appear neither clearly benign nor malignant. These often are associated with moderately differentiated glandular components; the prognosis for these women is not yet clearly defined. At the present time we are unable to attribute any prognostic significance to the presence of squamous differentiation in endometrial carcinomas. Because of the confusion and semantic arguments that revolve about the use of the terms adenoacanthoma and adenosquamous carcinomas, we recommend that those terms be abandoned and be replaced by the single term adenocarcinoma with squamous differentiation. As for any other endometrial tumor, the pathologist should provide information on histological grade, depth of myometrial invasion, and presence of vascular involvement or spread to the cervix in order to guide the gynecologist in determining appropriate therapy.


Script training and generalization for people with aphasia.
To examine the effects and generalization of a modified script training intervention, delivered partly via videoconferencing, on dialogue scripts that were produced by 2 individuals with aphasia. Each participant was trained on 2 personally relevant scripts. Intervention sessions occurred 3 times per week, with a combination of in-person meetings and videoconferencing, and lasted for 3 weeks per script. This study followed a multiple baseline design across scripts. Measures of accuracy, grammatical productivity, speaking rate, and articulatory fluency were obtained during baseline, intervention, and maintenance phases. Generalization probes were administered by challenging participants to engage in a conversation about their script topic with conversation partners who did not follow the script. Both participants showed improvement on all dependent variables for both scripts during and after the intervention phase. Generalization samples showed improved grammatical morpheme use and increased rate of speech over prebaseline samples. There is evidence that script training intervention can improve accuracy, grammatical productivity, speaking rate, and articulatory fluency in script production tasks as well as in more functional conversational tasks. Videoconferencing is a viable method of conducting script training intervention when it is supported by face-to-face intervention sessions, slight modifications to the procedure, and an emphasis on self-cuing skills.


Bottom-up sensory processing can induce negative BOLD responses and reduce functional connectivity in nodes of the default mode-like network in rats.
The default mode network is a large-scale brain network that is active during rest and internally focused states and deactivates as well as desynchronizes during externally oriented (top-down) attention demanding cognitive tasks. However, it is not sufficiently understood if salient stimuli, able to trigger bottom-up attentional processes, could also result in similar reduction of activity and functional connectivity in the DMN. In this study, we investigated whether bottom-up sensory processing could influence the default mode-like network (DMLN) in rats. DMLN activity was examined using block-design visual functional magnetic resonance imaging (fMRI) while its synchronization was investigated by comparing functional connectivity during a resting versus a continuously stimulated brain state by unpredicted light flashes. We demonstrated that the BOLD response in DMLN regions was decreased during visual stimulus blocks and increased during blanks. Furthermore, decreased inter-network functional connectivity between the DMLN and visual networks as well as decreased intra-network functional connectivity within the DMLN was observed during the continuous visual stimulation. These results suggest that triggering of bottom-up attention mechanisms in sedated rats can lead to a cascade similar to top-down orienting of attention in humans and is able to deactivate and desynchronize the DMLN.


A fully integrated microfluidic platform for highly sensitive analysis of immunochemical parameters.
We present a novel fully integrated centrifugal microfluidic platform for highly sensitive immunoassays in point-of-care settings. The platform consists of a disposable cartridge containing structures for assay processing, a porous membrane and all dried reagents required for the analysis. Additionally, a blister containing a washing buffer is connected to a new aliquoting structure enabling the serial aliquoting of washing buffer for repetitive bound-free separation steps. The proof-of-concept for two immunoassays is shown in the cartridge with each requiring only 30 μL of whole blood or plasma as the sample material. The detection of the cardiac marker Troponin T with a functional sensitivity of 7.55 ng L-1 (cv = 10%) within 11 minutes is shown based on samples from ten donors which were measured with six breadboard instruments to prove the platform capability for highly sensitive measurements at diagnostic relevant concentrations. Furthermore an assay for the cardiac marker NT-proBNP (five donors, six instruments) with a time-to-result of 12 minutes demonstrates that high-titer analytes (43 to 16.566 ng L-1) can be measured as well. A method comparison of our platform with a state-of-the-art laboratory analyzer proves an excellent correlation of the measured analyte concentrations. All results are obtained from injection moulded cartridges and all components of the platform are compatible for mass production.


The content of soyasaponin and soyasapogenol in soy foods and their estimated intake in the Japanese.
Soyasaponins have been reported to promote various health functions. However, the total soyasaponin and soyasapogenol content in soy products and the daily intake remain to be fully elucidated. We developed a high-performance liquid chromatography coupled with tandem mass spectrometric (HPLC-MS/MS) method to evaluate the content of group A and B soyasaponins and soyasapogenols. The total soyasaponin content was measured after pretreatment converted soyasaponins to soyasapogenols. The total soyasaponin content in soy foods was 200-1800 nmol g(-1), although that of soy sauce was 2-7 nmol g(-1). The soyasapogenol to total soyasaponin ratio was 30-50% in long-term matured miso. The majority of the soyasapogenol detected was soyasapogenol B rather than soyasapogenol A, resulting in speculation that further steps are required to liberate aglycones from glycoside-conjugated soyasaponins in soyasapogenol A. We estimated the daily intake of total soyasaponins and soyasapogenols by the Japanese, which was 50.3 and 0.59 μmol, respectively. The soyasapogenol content and the soyasapogenol to total soyasaponin ratio was considerably low in most soy products, except for long-term maturated miso. The major source of the daily intake of soyasaponins and soyasapogenols were tofu and miso, respectively.


Development of a single-gene, signature-tag-based approach in combination with alanine mutagenesis to identify listeriolysin O residues critical for the in vivo survival of Listeria monocytogenes.
Listeriolysin O (LLO) is a pore-forming toxin of the cholesterol-dependent cytolysin (CDC) family and a primary virulence factor of the intracellular pathogen Listeria monocytogenes. LLO mediates rupture of phagosomal membranes, thereby releasing bacteria into the growth-permissive host cell cytosol. Several unique features of LLO allow its activity to be precisely regulated in order to facilitate phagosomal escape, intracellular growth, and cell-to-cell spread. To improve our understanding of the multifaceted contribution of LLO to the pathogenesis of L. monocytogenes, we developed a screen that combined saturation mutagenesis and signature tags, termed in vivo analysis by saturation mutagenesis and signature tags (IVASS). We generated a library of LLO mutant strains, each harboring a single amino acid substitution and a signature tag, by using the previously described pPL2 integration vector. The signature tags acted as molecular barcodes, enabling high-throughput, parallel analysis of 40 mutants in a single animal and identification of attenuated mutants by negative selection. Using the IVASS technique we were able to screen over 90% of the 505 amino acids present in LLO and identified 60 attenuated mutants. Of these, 39 LLO residues were previously uncharacterized and potentially revealed novel functions of the toxin during infection. The mutants that were subsequently analyzed in vivo each conferred a 2- to 4-orders of magnitude loss in virulence compared to wild type, thereby validating the screening methods. Phenotypic analysis of the LLO mutant library using common in vitro techniques suggested that the functional contributions of some residues could only have been revealed through in vivo analysis.


Obesity enhances eosinophilic inflammation in a murine model of allergic asthma.
Obesity is associated with deterioration in asthma outcomes. Although airways eosinophil accumulation is characteristic of lung allergic diseases, little is known about the influence of obesity on the allergic eosinophil trafficking from bone marrow to lung tissues, and recruitment to airways lumen. Here, we have assessed the effects of diet-induced obesity on allergic eosinophilic inflammation in mice, examining eosinophil trafficking from bone marrow to airways, and production of T(H)1/T(H)2 cytokines. C57BL/6 mice fed for 10 weeks with standard chow or high-fat diet were sensitized and challenged with ovalbumin. At 24-96 h post-ovalbumin challenge, bronchoalveolar lavage (BAL) fluid, lung tissue and bone marrow were examined. The high-fat-fed mice exhibited increased body weight and epididymal fat, glucose intolerance and alterations in lipid profile compared with the lean mice. Obesity markedly elevated serum leptin and lowered adiponectin levels. Ovalbumin challenge in obese mice promoted a markedly higher eosinophil accumulation in bone marrow and connective tissue surrounding the bronchial and bronchiolar segments. Eosinophil number in BAL fluid of obese mice was lower at 24 and 48 h. Levels of interleukin (IL)-5, eotaxin, tumour necrosis factor-alpha and IL-10 in BAL fluid of obese mice were significantly higher than in lean mice. Diet-induced obesity enhanced eosinophil trafficking from bone marrow to lung tissues, and delayed their transit through the airway epithelium into the airway lumen. Consequently, eosinophils remain longer in lung peribronchiolar segments due to overproduction of T(H)1/T(H)2 cytokines and chemokines.


Revision after shoulder replacement for acute fracture of the proximal humerus.
Background and purpose - For more than half a century, stemmed hemiarthroplasty (SHA) has been used in the treatment of comminuted and displaced fractures of the proximal humerus. Reverse shoulder arthroplasty (RSA) has been increasingly popular in cases where it is difficult to obtain satisfactory fixation of the tuberosities. We report revision rates and reasons for revision after shoulder arthroplasty for acute fractures of the proximal humerus. Patients and methods - This study was based on a common dataset from the Nordic Arthroplasty Register Association (NARA), which includes data reported to the national shoulder arthroplasty registries in Denmark, Sweden, and Norway. We included 6,756 shoulder arthroplasties performed for acute fractures between 2004 and 2013. Results - There were 6,112 SHAs (90%) and 565 RSAs (8.4%). The cumulative arthroplasty survival rate after 5 years was 0.96 for both SHA and RSA. The relative risk of revision of RSA was 1.4 (95% CI: 0.9-2.2) with SHA as reference. For both types of arthroplasty, the most common reason for revision was infection (SHA 0.8%, RSA 2.1%). The relative risk of revision due to infection was 3.1 (95% CI: 1.6-5.9) for RSA with SHA as reference. The relative risk of revision for patients who were less than 75 years of age was 2.8 (95% CI: 2.0-3.8) compared to older patients. Interpretation - Revision after shoulder arthroplasty for acute fractures was rare. Survival rates were similar between SHA and RSA, but RSA had a statistically significant and clinically relevant higher risk of revision because of infection.


Outcomes following detection of low level plasma HIV RNA in HIV-infected patients previously virologically suppressed on antiretroviral therapy: a retrospective observational study.
Progressively sensitive assays for plasma HIV RNA have led to increased detection of plasma HIV RNA between 20 and 200 copies/ml, known as low level viremia (LLV) when recurrent or persistent, in HIV-infected patients on antiretroviral therapy (ART). The aim of this study was to determine outcomes following initial detection of LLV in an Australian cohort. A retrospective study using the HIV Service Database (Alfred Hospital) included all patients on ART who recorded plasma HIV RNA 20-200 copies/mL following prior virological suppression (viral load (VL) HIV RNA <20 copies/mL) over 2 years (2010 to 2012), with follow-up to June 2013. Factors associated with subsequent virological outcome were assessed via univariate and multivariate analysis. Of 919 patients managed by The Alfred HIV service, 207 (22.5%) met inclusion criteria. Mean age was 48.8 years, 91.3% were male. During follow-up, 54% patients recorded no further HIV RNA 20-200 copies/mL (viral blip); 39% had recurrent or persistent VL 20-200 copies/mL (LLV); and 7% progressed to virological failure with VL >200 copies/mL. Factors associated with LLV included co-morbid type 2 diabetes, shorter prior virological suppression and lower nadir CD4 cell count. Clinician management of VL 20-200 copies/mL was generally conservative, with infrequent requests for genotypic analysis (3.3% cases) or change in ART (<1% cases). LLV following virological suppression is common, and occurred as an isolated viral blip in half the patients. Those patients with persistent or recurrent LLV had higher rates of type 2 diabetes, shorter prior virological suppression and lower nadir CD4 cell count.


Differential effects of paclitaxel and docetaxel on gap junctions affects their cytotoxicities in transfected HeLa cells.
Gap junctions (GJs) enhance the cytotoxicity of specific cancer chemotherapeutic drugs and therefore, the inhibition of functional GJs may represent a mechanism by which the toxicity of chemotherapeutics in cancer cells can be reduced. In the present study, the effects and mechanisms of paclitaxel and docetaxel on GJ intercellular communication (GJIC) and the modulation of drug cytotoxicity were investigated in HeLa cells that were stably transfected with the connexin (Cx) 32 expression plasmid. Paclitaxel, but not docetaxel, was observed to inhibit dye‑coupling through junctional channels. Gating closure rather than the alteration of Cx32 expression or its membrane localization was responsible for the inhibitory action of paclitaxel on GJ function following short‑term exposure. The results revealed that the cytotoxicity of paclitaxel or docetaxel increased in the presence of functional GJs compared with that observed when GJIC was suppressed. In addition, paclitaxel‑induced downregulation of GJIC decreased the cytotoxicity of paclitaxel in the presence of functional GJs compared with that of docetaxel, which did not affect Cx32 channels. These observations demonstrated that the differential effects of paclitaxel and docetaxel on GJIC may affect the cytotoxicity of chemotherapeutic drugs. The present study provides a promising new approach to select antineoplastics and improve drug efficacy in carcinoma cells that form GJs.


Effects of different carbon sources on trichothecene production and Tri gene expression by Fusarium graminearum in liquid culture.
Fusarium head blight caused by Fusarium graminearum is a disease of cereal crops that not only reduces crop yield and quality but also results in contamination with trichothecenes such as nivalenol and deoxynivalenol (DON). To analyze the trichothecene induction mechanism, effects of 12 carbon sources on the production of DON and 3-acetyldexynivalenol (3ADON) were examined in liquid cultures incubated with nine strains of 3ADON-producing F. graminearum. Significantly high levels of trichothecene (DON and 3ADON) production by sucrose, 1-kestose and nystose were commonly observed among all of the strains tested. On the other hand, the levels of trichothecene biosynthesis induced by the other carbon sources were strain-specific. Tri4 and Tri5 expressions were up-regulated in the sucrose-containing medium but not in glucose. Trichothecene accumulation in the sucrose-containing medium was not repressed by the addition of glucose, indicating that trichothecene production was not regulated by carbon catabolite repression. These findings suggest that F. graminearum recognizes sucrose molecules, activates Tri gene expression and induces trichothecene biosynthesis.


[Spectral discrimination method information divergence combined with gradient angle].
The present paper proposes a spectral discrimination method combining spectral information divergence with spectral gradient angle (SID x tan(SGA(pi/2)) which overcomes the shortages of the existing methods which can not take the whole spectral shape and local characteristics into account simultaneously. Using the simulation spectra as input data, according to the interferogram acquirement principle and spectrum recovery algorithm of the temporally and spatially modulated Fourier transform imaging spectrometer (TSMFTIS), we simulated the distortion spectra recovery process of the TMSFTIS in different maximum mix ratio and distinguished the difference between the recovered spectra and the true spectrum by different spectral discrimination methods. The experiment results show that the SID x tan(SGA(pi/2)) can not only identify the similarity of the whole spectral shapes, but also distinguish local differences of the spectral characteristics. A comparative study was conducted among the different discrimination methods. The results have validated that the SID x tan(SGA(pi/2)) has a significant improvement in the discriminatory ability.


The association of particular types of Proteus with chronic suppurative otitis media.
During a period of 12 months, 57 strains of Proteus were isolated from the ears of 38 unrelated patients with chronic suppurative otitis media. Each strain was identified, typed for bacteriocin production and sensitivity, and tested for Dienes compatibility. The majority of the strains (79%) were P. mirabilis; all but one of the remainder were P.vulgaris. Although 19 different bacteriocin production/sensitivity types were found, two rare types, P. mirabilis P7/S5,12 and P. vulgaris P0/S9, were associated with 47% of these infections. This was confirmed by Dienes typing. Patients with bilateral ear disease carried a different strain in each ear. There was no evidence that persistence of infection had arisen because of the development of antibiotic resistance. Although there was some evidence that persistence may have been the result of reinfection, the isolation of these rare types of Proteus from so many patients with chronic suppurative otitis media may indicate that they play an important role in the pathogenesis of the disease. Most of the Proteus isolates were of "non-faecal" types and it is believed that infection had arisen by a route other than the faecal-aural one.


The influence of ultrasound frequency and power, on the algal species Microcystis aeruginosa, Aphanizomenon flos-aquae, Scenedesmus subspicatus and Melosira sp.
We report on the effectiveness of sonication on controlling the growth of four problematic algal species which are morphologically different and from three algal divisions. Two cyanobacterial species Microcystis aeruginosa (unicellular) and Aphanizomenon flos-aquae (filamentous), one green alga Scenedesmus subspicatus (colonial) and lastly a diatom species Melosira sp. (filamentous) were subjected to ultrasound of selected low to high frequencies ranging from 20 to 1144 kHz. Microcystis aeruginosa and Scenedesmus subspicatus highest cell removal rates were 16 +/- 2% and 20 +/- 3% when treated with the same ultrasound frequency of 862 kHz but differing energy levels of 133 and 67 kWh m(-3), respectively. Aphanizomenon flos-aquae best removal rate was 99 +/- 1% after 862 kHz and 133 kWh m(-3) of energy, with Melosira sp. achieving its highest cell removal at 83% subsequent to ultrasound of 20 kHz and 19 kWh m(-3). Microcystis aeruginosa and Scenedesmus subspicatus are considered non-susceptible species to ultrasound treatment from a water treatment perspective due to their low cell removal rates; however, photosynthetic activity reduction of 65% for Microcystis aeruginosa does indicate the possible utilization of ultrasound to control bloom growth, rather than bloom elimination. Conversely, Aphanizomenon flos-aquae and Melosira sp. are deemed species highly susceptible to ultrasound. Morphological differences in shape (filamentous/non-filamentous) and cell wall structure (silica/peptidoglycan), and presence of gas vacuoles are probable reasons for these differing levels of susceptibility to ultrasound.


Measurement of 68Zn and 70Zn in human blood in reference to the study of zinc metabolism.
A method based on radiochemical neutron activation analysis has been developed for measurement of the stable isotopes 68Zn and 70Zn in human plasma and red cells. The method has been applied to measure the extent of 70Zn enrichment in plasma samples from four healthy adult volunteers who had each consumed a single 3.2 mg dose of 70ZnCl2 in fasting state. It is shown that measurements of 70Zn/68Zn ratio in plasma and red cell sample sizes of 2 ml can be made with precisions of 10% or less depending on the degree of enrichment achieved. The limiting factor in the precision of these measurements appears to be related to the counting statistics of the 386 kev photopeak (71mZn) which improves with increased enrichment. Clinical feasibility trials on these subjects with regard to plasma and red cell 70Zn enrichment have been carried out over a 24-h postadministration period. The results are consistent with kinetic studies reported in literature with radiozinc, and show that this method can be used to study kinetics of plasma appearance of 70Zn after oral administration of the isotope under physiological intake conditions. The present results further indicate that under these experimental conditions, enrichment of red cells can be measured after 24 h or longer postadministration periods, but short-term measurements yield marginal results. This new method provides an alternative approach to the use of radiolabeled zinc for the study of human zinc metabolism and appearance of isotopic zinc in plasma after oral administration under physiological intake conditions. It can be applied to all human population groups as a noninvasive and safe method which does not require safety considerations arising from human use of radiotracers. However, its application is both more expensive and limited in terms of sample throughput as compared with the radiotracer technique.


Sexual, Reproductive Health Needs, and Rights of Young People in Slum Areas of Kampala, Uganda: A Cross Sectional Study.
Young people in Uganda face various sexual and reproductive health risks, especially those living in urban slums. The aim of this study was to examine factors associated with comprehensive categories of sexual and reproductive health, including sexual behaviours; sexual education and access to contraceptive services; family planning; prevention of STDs; sexual consent as a right; gender based violence; as well as HIV testing, counselling, disclosure and support. The study was cross-sectional in design and was carried out in July 2014 in Makindye and Nakawa Divisions of Kampala City, Uganda. Using systematic random sampling, data were collected on 663 participants aged between 13 and 24 years in Kampala's urban slums. Sixty two percent of participants reported having ever had sex and the mean age of sexual debut was 16 years (95%CI: 15.6, 16.4 years, range: 5-23 years). The odds of reporting ever having had sexual intercourse were higher among respondents living alone (OR: 2.75; 95%CI: 1.35, 5.61; p<0.01) than those living in a nuclear family. However, condom use was only 54%. The number of sexual partners in the last 12 months preceding the survey averaged 1.8 partners (95%CI: 1.7, 1.9; range 1-4) with 18.1% reporting an age gap of 10 years or older. More than three quarters (80.6%) of sexually active participants reported that their first sexual encounter was consensual, suggesting that most young people are choosing when they make their sexual debut. Low prevalence of willing first sexual intercourse was associated with younger age (OR = 0.48, 95%CI: 0.25, 0.90, p<0.05), having a disability (OR = 0.40, 95%CI: 0.16, 0.98, p<0.05), living with non-relatives (OR = 0.44, 95%CI: 0.16, 0.97, p<0.05), and being still at school (OR = 0.29, 95%CI: 0.12, 0.67, p<0.01). These results remained significant after adjusting for covariates, except for disability and the age of participants. The proportion of unwilling first sexual intercourse was significantly higher among women for persuasion (13.2% vs. 2.4%, p<0.001), being tricked (7.1% vs 2.9%, p<0.05) and being forced or raped (9.9% vs 4.4%, p<0.05) than men. A high level of sexual abuse emerged from the data with 34.3% affirming that it was alright for a boy to force a girl to have sex if he had feelings for her; 73.3% affirming that it was common for strangers and relatives to force young females to have sexual intercourse with them without consent; 26.3% indicating that it was sometimes justifiable for a boy to hit his girlfriend, as long as they loved each other. This study has explored current sexual practice among young people in a specific part of urban Kampala. Young people's sexual and reproductive health remains a challenge in Uganda. To address these barriers, a comprehensive and harmonised sexual and reproductive health system that is youth friendly and takes into account local socio-cultural contexts is urgently needed.


Stability analysis and synthesis for an affine fuzzy system via LMI and ILMI: discrete case.
This paper develops a stability analysis and controller synthesis methodology for a discrete affine fuzzy system based on the convex optimization techniques. In analysis, the stability condition under which the affine fuzzy system is quadratically stable is derived. Then, the condition Is recast in the formulation of Linear Matrix Inequalities (LMI) and numerically addressed. The emphasis of this paper, however, is on the synthesis of fuzzy controller based on the derived stability condition. In synthesis, the stabilizability condition turns out to be in the formulation of nonconvex matrix inequalities and is solved numerically in an iterative manner. Discrete iterative LMI (ILMI) approach is proposed to obtain the feasible solution for the synthesis of the affine fuzzy system. Finally, the applicability of the suggested methodology is demonstrated via some examples and computer simulations.


Maintenance and Loss of Duplicated Genes by Dosage Subfunctionalization.
Whole-genome duplications (WGDs) have contributed to gene-repertoire enrichment in many eukaryotic lineages. However, most duplicated genes are eventually lost and it is still unclear why some duplicated genes are evolutionary successful whereas others quickly turn to pseudogenes. Here, we show that dosage constraints are major factors opposing post-WGD gene loss in several Paramecium species that share a common ancestral WGD. We propose a model where a majority of WGD-derived duplicates preserve their ancestral function and are retained to produce enough of the proteins performing this same ancestral function. Under this model, the expression level of individual duplicated genes can evolve neutrally as long as they maintain a roughly constant summed expression, and this allows random genetic drift toward uneven contributions of the two copies to total expression. Our analysis suggests that once a high level of imbalance is reached, which can require substantial lengths of time, the copy with the lowest expression level contributes a small enough fraction of the total expression that selection no longer opposes its loss. Extension of our analysis to yeast species sharing a common ancestral WGD yields similar results, suggesting that duplicated-gene retention for dosage constraints followed by divergence in expression level and eventual deterministic gene loss might be a universal feature of post-WGD evolution.


Early biomarkers of cadmium exposure and nephrotoxicity.
As the risks of cadmium (Cd)-induced kidney disease have become increasingly apparent, much attention has been focused on the development and use of sensitive biomarkers of Cd nephrotoxicity. The purpose of this review is to briefly summarize the current state of Cd biomarker research. The review includes overviews of the toxicokinetics of Cd, the mechanisms of Cd-induced proximal tubule injury, and mechanistic summaries of some of the biomarkers (N-acetyl-β-D-glucosamidase; β(2)-microglubulin, metallothionein, etc.) that have been most widely used in monitoring of human populations for Cd exposure and nephrotoxicity. In addition, several novel biomarkers (kidney injury molecule-1, α-glutathione-S-transferase and insulin) that offer the potential for improved biomonitoring of Cd-exposed populations are discussed.


Roles of TOG and jelly-roll domains of centrosomal protein CEP104 in its functions in cilium elongation and Hedgehog signaling.
Primary cilia are generated through the extension of the microtubule-based axoneme. Centrosomal protein 104 (CEP104) localizes to the tip of the elongating axoneme, and CEP104 mutations are linked to a ciliopathy, Joubert syndrome. Thus, CEP104 has been implicated in ciliogenesis. However, the mechanism by which CEP104 regulates ciliogenesis remains elusive. We report here that CEP104 is critical for cilium elongation but not for initiating ciliogenesis. We also demonstrated that the tumor-overexpressed gene (TOG) domain of CEP104 exhibits microtubule-polymerizing activity and that this activity is essential for the cilium-elongating activity of CEP104. Knockdown/rescue experiments showed that the N-terminal jelly-roll (JR) fold partially contributes to cilium-elongating activity of CEP104, but neither the zinc-finger region nor the SXIP motif is required for this activity. CEP104 binds to a centriole-capping protein, CP110, through the zinc-finger region and to a microtubule plus-end-binding protein, EB1, through the SXIP motif, indicating that the binding of CP110 and EB1 is dispensable for the cilium-elongating activity of CEP104. Moreover, CEP104 depletion does not affect CP110 removal from the mother centriole, which suggests that CEP104 functions after the removal of CP110. Last, we also showed that CEP104 is required for the ciliary entry of Smoothened and export of GPR161 upon Hedgehog signal activation and that the TOG domain plays a critical role in this activity. Our results define the roles of the individual domains of CEP104 in its functions in cilium elongation and Hedgehog signaling and should enhance our understanding of the mechanism underlying CEP104 mutation-associated ciliopathies.


Community-academic partnerships in implementation research.
Community-academic partnerships(CAPs) are a critical component of implementing and sustaining evidence-based practices (EBPs) in community settings; however, the approaches used and mechanisms of change within CAPs have not been rigorously studied. The first step to advancing the science of CAP is to operationally define and contextualize the approaches used in CAP as part of the implementation process. Our research group has gleaned valuable lessons about the best ways to develop, support, and nurture community partnerships within the context of implementation. In this article, we share these lessons learned and relate them to implementation strategies that are most relevant to community-partnered implementation endeavors. The implementation strategies most relevant to CAPs are as follows: (a) building a coalition, (b) conducting local consensus discussions, (c) identifying barriers and facilitators to implementation, (d) facilitating interactive problem solving, (e) using an advisory board or workgroup, (f) tailoring strategies, (g) promoting adaptability, and (h) auditing and providing feedback. We offer suggestions for future research to systematically evaluate these strategies, with an eye toward advancing the science of CAP and implementation science and the goal of guiding future research and improving the implementation of EBPs in community settings.


Catecholamines increase monocyte TNF receptors and inhibit TNF through beta 2-adrenoreceptor activation.
Postinjury deficits in monocyte tumor necrosis factor receptors (moTNFR) activity may alter beneficial functions during an inflammatory response. Several counter-regulatory hormones elicited during inflammation may modulate tumor necrosis factor (TNF) activity, but little is known about their influence on moTNFR. Also, catecholamines inhibit TNF production, but the adrenoreceptor mechanism of this effect has not been fully clarified. To determine the effect of catecholamines and corticosteroids on moTNFR, whole blood was coincubated for up to 8 (moTNFR) or 24 h (cytokines) in the presence of lipopolysaccharide (100 ng/ml) and 1) epinephrine (Epi, 10(-6) M), dexamethasone (Dex, 10(-6) M) or both (EpiDex, 10(-6) M) to assess the expression of total moTNFR, moTNFR-I, and moTNFR-II. 2) Epi and norepinephrine (EpiNE, 10(-6) M) and the alpha 1 + 2-, beta 1 + 2-, beta 1-, or beta 2-adrenergic antagonists were used to assess the role of such adrenoreceptors on total moTNFR and TNF production, and N6,2'-O-dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) alone or in combination with the phosphodiesterase inhibitor Ro-20-1724/000, to study the cAMP-dependent pathway on total moTNFR. We found that Epi upregulated total moTNFR and moTNFR-II. Dex did not significantly influence total moTNFR or moTNFR-II. Also, EpiNE increased total moTNFR and inhibited TNF by a beta 2-dependent mechanism. DBcAMP (10(-5) M) modestly enhanced total moTNFR. This suggests a common mechanism for acutely enhancing moTNFR and attenuation of soluble TNF appearance during conditions of severe stress.


Epigenome-wide association of DNA methylation markers in peripheral blood from Indian Asians and Europeans with incident type 2 diabetes: a nested case-control study.
Indian Asians, who make up a quarter of the world's population, are at high risk of developing type 2 diabetes. We investigated whether DNA methylation is associated with future type 2 diabetes incidence in Indian Asians and whether differences in methylation patterns between Indian Asians and Europeans are associated with, and could be used to predict, differences in the magnitude of risk of developing type 2 diabetes. We did a nested case-control study of DNA methylation in Indian Asians and Europeans with incident type 2 diabetes who were identified from the 8-year follow-up of 25 372 participants in the London Life Sciences Prospective Population (LOLIPOP) study. Patients were recruited between May 1, 2002, and Sept 12, 2008. We did epigenome-wide association analysis using samples from Indian Asians with incident type 2 diabetes and age-matched and sex-matched Indian Asian controls, followed by replication testing of top-ranking signals in Europeans. For both discovery and replication, DNA methylation was measured in the baseline blood sample, which was collected before the onset of type 2 diabetes. Epigenome-wide significance was set at p<1 × 10(-7). We compared methylation levels between Indian Asian and European controls without type 2 diabetes at baseline to estimate the potential contribution of DNA methylation to increased risk of future type 2 diabetes incidence among Indian Asians. 1608 (11·9%) of 13 535 Indian Asians and 306 (4·3%) of 7066 Europeans developed type 2 diabetes over a mean of 8·5 years (SD 1·8) of follow-up. The age-adjusted and sex-adjusted incidence of type 2 diabetes was 3·1 times (95% CI 2·8-3·6; p<0·0001) higher among Indian Asians than among Europeans, and remained 2·5 times (2·1-2·9; p<0·0001) higher after adjustment for adiposity, physical activity, family history of type 2 diabetes, and baseline glycaemic measures. The mean absolute difference in methylation level between type 2 diabetes cases and controls ranged from 0·5% (SD 0·1) to 1·1% (0·2). Methylation markers at five loci were associated with future type 2 diabetes incidence; the relative risk per 1% increase in methylation was 1·09 (95% CI 1·07-1·11; p=1·3 × 10(-17)) for ABCG1, 0·94 (0·92-0·95; p=4·2 × 10(-11)) for PHOSPHO1, 0·94 (0·92-0·96; p=1·4 × 10(-9)) for SOCS3, 1·07 (1·04-1·09; p=2·1 × 10(-10)) for SREBF1, and 0·92 (0·90-0·94; p=1·2 × 10(-17)) for TXNIP. A methylation score combining results for the five loci was associated with future type 2 diabetes incidence (relative risk quartile 4 vs quartile 1 3·51, 95% CI 2·79-4·42; p=1·3 × 10(-26)), and was independent of established risk factors. Methylation score was higher among Indian Asians than Europeans (p=1 × 10(-34)). DNA methylation might provide new insights into the pathways underlying type 2 diabetes and offer new opportunities for risk stratification and prevention of type 2 diabetes among Indian Asians. The European Union, the UK National Institute for Health Research, the Wellcome Trust, the UK Medical Research Council, Action on Hearing Loss, the UK Biotechnology and Biological Sciences Research Council, the Oak Foundation, the Economic and Social Research Council, Helmholtz Zentrum Munchen, the German Research Center for Environmental Health, the German Federal Ministry of Education and Research, the German Center for Diabetes Research, the Munich Center for Health Sciences, the Ministry of Science and Research of the State of North Rhine-Westphalia, and the German Federal Ministry of Health.