HIV escape and attenuation by cytotoxic T lymphocytes.
Much of the current HIV-1 vaccine research focuses on harnessing the cytotoxic T-lymphocyte arm of the immune response. However, HIV-1 appears to have an unerring ability to evade cytotoxic T-lymphocyte responses, through the process of escape mutation, and thus the potential benefit of a cytotoxic T-lymphocyte-based vaccine remains uncertain. This review focuses on several recent studies that question whether escape mutation is always detrimental to the host, and may provide new hope for the success of a cytotoxic T-lymphocyte-based vaccine against HIV. Several recent studies, in both natural HIV-1 infection and the SIV model, have identified examples of cytotoxic T-lymphocyte escape mutants that revert on transmission to individuals lacking the selecting major histocompatibility complex alleles. The obvious implication of these data is that some cytotoxic T-lymphocyte responses can only be evaded through escape mutations that actually reduce the replicative fitness of the virus. In addition, a recent vaccine study in macaques found that the control of immunodeficiency virus to undetectable levels was only achieved in animals that were able to force the virus to make such detrimental escape mutations. These data raise the intriguing possibility that, rather than undermining cytotoxic T-lymphocyte vaccines, escape mutation may be one of the keys to their success. Clearly, not all escape mutations help to control viral replication. We discuss how these new data may assist in the struggle to develop a successful cytotoxic T-lymphocyte-based HIV vaccine, and what they tell us about the responses such a vaccine should aim to elicit.


Consistent detection of bovine papillomavirus in lesions, intact skin and peripheral blood mononuclear cells of horses affected by hoof canker.
Equine hoof canker is a chronic proliferative pododermatitis of as yet unknown aetiology. Like equine sarcoid disease, canker is a therapy-resistant disorder characterised by hyperkeratosis, acanthosis and a marked tendency to recur. There is an association of sarcoid-inducing bovine papillomaviruses of types 1 and 2 (BPV-1, BPV-2) with hoof canker disease. Using PCR-based techniques, we assessed canker tissue, intact skin and/or peripheral blood mononuclear cells (PBMCs) of 25 canker-affected horses for the presence of sarcoid-associated BPV-1 and -2. Conventional PCR revealed BPV-1/-2 DNA in 24/24 canker, 12/13 skin and 10/11 PBMC DNA isolates. Using inverse PCR, full-length BPV episomes were detected in 1/5 canker specimens. Sequencing of viral early and late genes amplified from canker, intact skin and PBMC DNA of 2 cases revealed an overall identity of 98% to BPV-1. Viral DNA loads amounted to ≤16 copies per cell in canker tissue and intact skin, and to ≤0.35 copies per PBMC, as determined by quantitative PCR. Using RT-PCR, the viral major oncogene E5 was shown to be transcribed in 2/4 canker tissue specimens and 5/7 PBMC isolates. Immunocapture PCR from 7 canker and 6 skin extract supernatants revealed capsomere-associated viral DNA in one canker and one skin sample. Hoof tissue, skin and PBMCs collected from 13 individuals with no signs of canker or BPV-related malignancies scored negative throughout the experiments. These findings suggest that the observed presence of BPV-1/-2 in canker-affected horses is not coincidental but indicative of an active contribution to hoof canker disease. The use of antivirals and/or immune modulators may help improving canker therapy.


Evaluation of tetrazolium-based semiautomatic colorimetric assay for measurement of human antitumor cytotoxicity.
A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based colorimetric assay was developed and compared with 51Cr release from different adherent tumor cell targets (human squamous cell carcinoma lines of the head and neck established in our laboratory, melanoma, and colorectal carcinoma) using 5-7-day human lymphokine-activated killer cells and monocyte-depleted peripheral blood lymphocytes as effectors. With adherent tumor cell targets, MTT colorimetry was more sensitive than the 51Cr release assay in measuring the antitumor activity of effectors: median, 4385 (range, 988-8144) versus median, 1061 (range, 582-7294) lytic units (the number of effector cells required to lyse 20% of 5 x 10(3) targets)/10(7) effectors (P less than 0.01). Background effects (without effector cells) were comparable in 4-h assays (9% versus 10%) between MTT colorimetry and 51Cr release. In 24-h assays, MTT colorimetry showed higher antitumor activity (70-100% versus 40-60% lysis at 1:1 effector:target cell ratio) but lower background effects (6% versus 38%) than 51Cr release assay. Thus, MTT colorimetry was more sensitive, did not use radiolabeled targets, required fewer effector cells, and was easier, less expensive, and better adaptable to serial monitoring of effector cell function in cancer patients. This colorimetric assay is especially well suited to adherent tumor cell targets. The use of adherent tumor cell monolayers, as opposed to trypsinized single cell suspensions, provides an opportunity to measure interactions of effector cells with enzymatically unaltered solid tumor targets. Because of the greater sensitivity of the colorimetric assay, the transformation of MTT data into lytic units, as commonly used for 51Cr release assays, required an adjustment to avoid the extrapolation based on the exponential fit equation.


Reliability of toluidine blue vital staining in detection of potentially malignant oral lesions--time to reconsider.
Being simple and inexpensive toluidine blue has been in use for more than two decades for the detection of potentially malignant oral lesions (PMOL's) and malignant lesions. Although there has been concensus that staining often assists in the identification of these lesions, results have been diverse. In most studies false negative were not recorded as biopsies of lesions that did not retain toluidine blue were not performed. Thus the present study attempted to evaluate the efficacy of toluidine blue vital dye for detection of PMOL's. The study included 47 biopsies (TBP:35 and TBN:12), of which 23 cases were confirmed as dysplastic (TBP=17 and TBN=6), 7 as hyperkeratosis (TBP=4 and TBN=3), 8 as epithelial hyperplasia (TBP=6 and TBN=3) and 5 as other benign lesions (TBP=4 and TBN=1). The validity test revealed a senstivity of 73.9% and specificity of 30%. The positive predictive value was 54.8% and negative predictive value of 50%. The study intends to highlight the false negative result (26.1%) which was mainly attributed to mild dysplasia and the false positive (32.6%) which included hyperkeratosis, hyperplasia, lichen planus and traumatic ulcer. The study concludes that toluidine blue staining should not blindly direct the clinician's opinion, and strongly discourages the use of toludine blue as a screening test and the results should be interpreted with caution.


Heme oxygenase inhibition increases blood pressure in pregnant rats.
During normal gestation, the placenta is a relatively hypoxic organ and, as such, is subject to significant oxidative stress. In the preeclamptic patient, inadequate remodeling of the maternal vasculature severely exacerbates placental oxidative stress, which has been shown to be an important component of maternal hypertension. There is emerging evidence that Heme Oxygenase-1 (HO-1) acts as an important regulator of placental and cardiovascular function during normal pregnancy. Here, we have examined the effect of Heme Oxygenase (HO) inhibition in late gestation on maternal blood pressure, angiogenic balance, and placental oxidative stress in pregnant rats. HO activity was inhibited with tin mesoporphyrin (SnMP), which was administered on gestational day 14, and blood pressure was measured on gestational day 19. Placental angiogenic balance and plasma Vascular Endothelial Growth Factor (VEGF) were determined by sandwich enzyme-linked immunosorbent assay. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was measured by lucigenin chemilluminescence. In response to SnMP treatment, maternal mean arterial pressure (MAP) was significantly increased (99±1 vs. 113±2mm Hg; P < 0.05; n = 15 per group). Placental soluble fms-like tyrosine kinase-1 (sFlt-1) (631±47 vs. 648±26 pg/mg; P = 0.76) levels in the placenta were not affected by HO inhibition. Additionally, there was no significant difference in free VEGF in the maternal circulation (287±22 vs. 329±14 pg/ml; P = 0.11). There was, however, a significant decrease in placental VEGF (23±2 vs. 16±1 pg/mg; P < 0.05) and a significant increase in placental NADPH oxidase activity in SnMP-treated rats (2021±238 vs. 3005±301 RLU/min/mg; P < 0.05). Our results demonstrate that HO is an important regulator of blood pressure and an important antioxidant in the developing placenta.


Nevirapine/zidovudine/lamivudine has superior immunological and virological responses not reflected in clinical outcomes in a 48-week randomized comparison with abacavir/zidovudine/lamivudine in HIV-infected Ugandan adults with low CD4 cell counts.
Triple nucleoside reverse transcriptase inhibitor regimens have advantages as first-line antiretroviral therapy (ART), avoiding hepatotoxicity and interactions with anti-tuberculosis therapy, and sparing two drug classes for second-line ART. Concerns exist about virological potency; efficacy has not been assessed in Africa. A safety trial comparing nevirapine with abacavir was conducted in two Ugandan Development of Antiretroviral Therapy in Africa (DART) centres: 600 symptomatic antiretroviral-naïve HIV-infected adults with CD4 counts <200 cells/microL were randomized to zidovudine/lamivudine plus abacavir or nevirapine (placebo-controlled to 24-week primary toxicity endpoint, and then open-label). Documented World Health Organization (WHO) stage 4 events were independently reviewed and plasma HIV-1 RNA assayed retrospectively. Exploratory efficacy analyses are intention-to-treat. The median pre-ART CD4 count was 99 cells/microL, and the median pre-ART viral load was 284 600 HIV-1 RNA copies/mL. A total of 563 participants (94%) completed 48 weeks of follow-up, 25 (4%) died and 12 (2%) were lost to follow-up. The randomized drug was substituted in 21 participants (7%) receiving abacavir vs. 34 (11%) receiving nevirapine (P=0.09). At 48 weeks, 62% of participants receiving abacavir vs. 77% of those receiving nevirapine had viral loads <50 copies/mL (P<0.001), and mean CD4 count increases from baseline were +147 vs. +173 cells/microL, respectively (P=0.006). Nine participants (3%) receiving abacavir vs. 16 (5%) receiving nevirapine died [hazard ratio (HR) 0.55; 95% confidence interval (CI) 0.24-1.25; P=0.15]; 20 receiving abacavir vs. 32 receiving nevirapine developed new or recurrent WHO 4 events or died (HR=0.60; 95% CI 0.34-1.05; P=0.07) and 48 receiving abacavir vs. 68 receiving nevirapine developed new or recurrent WHO 3 or 4 events or died (HR=0.67; 95% CI 0.46-0.96; P=0.03). Seventy-one participants (24%) receiving abacavir experienced 91 grade 4 adverse events compared with 130 events in 109 participants (36%) on nevirapine (P<0.001). The clear virological/immunological superiority of nevirapine over abacavir was not reflected in clinical outcomes over 48 weeks. The inability of CD4 cell count/viral load to predict initial clinical treatment efficacy is unexplained and requires further evaluation.


Prediction of acute myeloid leukaemia risk in healthy individuals.
The incidence of acute myeloid leukaemia (AML) increases with age and mortality exceeds 90% when diagnosed after age 65. Most cases arise without any detectable early symptoms and patients usually present with the acute complications of bone marrow failure1. The onset of such de novo AML cases is typically preceded by the accumulation of somatic mutations in preleukaemic haematopoietic stem and progenitor cells (HSPCs) that undergo clonal expansion2,3. However, recurrent AML mutations also accumulate in HSPCs during ageing of healthy individuals who do not develop AML, a phenomenon referred to as age-related clonal haematopoiesis (ARCH)4-8. Here we use deep sequencing to analyse genes that are recurrently mutated in AML to distinguish between individuals who have a high risk of developing AML and those with benign ARCH. We analysed peripheral blood cells from 95 individuals that were obtained on average 6.3 years before AML diagnosis (pre-AML group), together with 414 unselected age- and gender-matched individuals (control group). Pre-AML cases were distinct from controls and had more mutations per sample, higher variant allele frequencies, indicating greater clonal expansion, and showed enrichment of mutations in specific genes. Genetic parameters were used to derive a model that accurately predicted AML-free survival; this model was validated in an independent cohort of 29 pre-AML cases and 262 controls. Because AML is rare, we also developed an AML predictive model using a large electronic health record database that identified individuals at greater risk. Collectively our findings provide proof-of-concept that it is possible to discriminate ARCH from pre-AML many years before malignant transformation. This could in future enable earlier detection and monitoring, and may help to inform intervention.


L-Sox5 and Sox6 drive expression of the aggrecan gene in cartilage by securing binding of Sox9 to a far-upstream enhancer.
The Sry-related high-mobility-group box transcription factor Sox9 recruits the redundant L-Sox5 and Sox6 proteins to effect chondrogenesis, but the mode of action of the trio remains unclear. We identify here a highly conserved 359-bp sequence 10 kb upstream of the Agc1 gene for aggrecan, a most essential cartilage proteoglycan and key marker of chondrocyte differentiation. This sequence directs expression of a minimal promoter in both embryonic and adult cartilage in transgenic mice, in a manner that matches Agc1 expression. The chondrogenic trio is required and sufficient to mediate the activity of this enhancer. It acts directly, Sox9 binding to a critical cis-acting element and L-Sox5/Sox6 binding to three additional elements, which are cooperatively needed. Upon binding to their specific sites, L-Sox5/Sox6 increases the efficiency of Sox9 binding to its own recognition site and thereby robustly potentiates the ability of Sox9 to activate the enhancer. L-Sox5/Sox6 similarly secures Sox9 binding to Col2a1 (encoding collagen-2) and other cartilage-specific enhancers. This study thus uncovers critical cis-acting elements and transcription factors driving Agc1 expression in cartilage and increases understanding of the mode of action of the chondrogenic Sox trio.


Antioxidant intake and adult-onset wheeze: a case-control study. Aberdeen WHEASE Study Group.
An increase in prevalence of wheezing illness in the UK has coincided with a reduction in the consumption of natural antioxidants, which may modulate the lung's response to oxidant stress, limiting the expression of airway inflammation and respiratory symptoms. The hypothesis that intakes and plasma levels of natural antioxidants would be determinants of adult-onset wheezing illness was tested. A nested case-control study was conducted in 94 cases with adult-onset wheeze and 203 controls aged 39-45 yrs identified in a 30-yr follow-up survey. Antioxidant intake was measured by a food frequency questionnaire, and plasma and red cell measurements of antioxidant status were obtained. Outcome measures were onset of wheeze since age 15 yrs (ever wheeze) and wheeze occurring in the past 12 months (current wheeze). After adjusting for the effects of smoking, socioeconomic status, atopy, family history of atopic disease and total energy intake, intakes of vitamin E (odds ratio (OR) = 4.02 for low compared to high tertile of intake) and plasma levels of ascorbate (OR = 0.98 per unit) and alpha-tocopherol:triglyceride ratio (OR = 0.34 per log(e) unit) were inversely related to adult-onset wheeze. In analyses stratified by social class and smoking, intakes of vitamin C and E and plasma levels of ascorbate and alpha-tocopherol:triglyceride ratio were inversely related to current wheeze in the manual social class and among current smokers. No independent associations of vitamin A, beta-carotene or total plasma antioxidant capacity were found. The results support the hypothesis that deficiencies of vitamins C and E are associated with wheezing symptoms. Smokers in the manual social class are particularly susceptible to these effects.


Prioritizing surveillance activities for certification of yaws eradication based on a review and model of historical case reporting.
The World Health Organization (WHO) has targeted yaws for global eradication. Eradication requires certification that all countries are yaws-free. While only 14 Member States currently report cases to WHO, many more are known to have a history of yaws and some of them may have ongoing transmission. We reviewed the literature and developed a model of case reports to identify countries in which passive surveillance is likely to find and report cases if transmission is still occurring, with the goal of reducing the number of countries in which more costly active surveillance will be required. We reviewed published and unpublished documents to extract data on the number of yaws cases reported to WHO or appearing in other literature in any year between 1945 and 2015. We classified countries as: a) having interrupted transmission; b) being currently endemic; c) being previously endemic (current status unknown); or d) having no history of yaws. We constructed a panel dataset for the years 1945-2015 and ran a regression model to identify factors associated with some countries not reporting cases during periods when there was ongoing (and documented) transmission. For previously endemic countries whose current status is unknown, we then estimated the probability that countries would have reported cases if there had in fact been transmission in the last three years (2013-2015). Yaws has been reported in 103 of the 237 countries and areas considered. 14 Member States and 1 territory (Wallis and Futuna Islands) are currently endemic. 2 countries are believed to have interrupted transmission. 86 countries and areas are previously endemic (current status unknown). Reported cases peaked in the 1950s, with 55 countries reporting at least one case in 1950 and a total of 2.35 million cases reported in 1954. Our regression model suggests that case reporting during periods of ongoing transmission is positively associated with socioeconomic development and, in the short-term, negatively associated with independence. We estimated that for 66 out of the 86 previously endemic countries whose current status is unknown, the probability of reporting cases in the absence of active surveillance is less than 50%. Countries with a history of yaws need to be prioritized so that international resources for global yaws eradication may be deployed efficiently. Heretofore, the focus has been on mass treatment in countries currently reporting cases. It is also important to undertake surveillance in the 86 previously endemic countries for which the current status is unknown. Within this large and diverse group, we have identified a group of 20 countries with more than a 50% probability of reporting cases in the absence of active surveillance. For the other 66 countries, international support for active surveillance will likely be required.


Respiratory health screening using pulmonary function tests and lung sound analysis.
Lung sounds (LS) analysis is a potential source of additional objective, noninvasive and quantitative information on the status of the pulmonary system. We have examined the hypothesis that the addition of lung sounds analysis to spirometry increases the sensitivity of objective population screening, as compared to the use of spirometry alone. Questionnaires, spirometry and lung sounds were obtained in 493 active workers. Lung sounds analysis consisted of the averaged power spectra of breath sounds, measured separately during inspiration and expiration at four standard locations over the trachea and the chest-wall. Of the 493 subjects, 91 had an obstructive lung disease, including 27 with chronic bronchitis identified by a history of prolonged cough and sputum production but with normal spirometry. Twelve additional workers had a restrictive lung disease. Abnormal spirometric results were found in 74 of the patients. Abnormal lung sounds analyses were found in 54 patients, including 14 of the chronic bronchitis cases, so that the overall sensitivity of objective screening tests increased from 71% to 87% by combining the two tests. Thirty three of the subjects considered normal by evaluation of their questionnaire and spirometry had abnormal lung sounds. Of the twenty four who were re-evaluated 12-18 months after the first tests, three had developed a lung or heart disease. We conclude that the combination of spirometry and lung sounds analysis significantly increased the sensitivity of detection of pulmonary diseases by objective tests, and provided an early sign of lung disease that was not detected by spirometry alone.


Phase II study of oral fingolimod (FTY720) in multiple sclerosis: 3-year results.
In a 6-month, placebo-controlled trial, oral fingolimod (FTY720) 1.25 or 5.0 mg, once daily, significantly reduced MRI inflammatory activity and annualized relapse rate compared with placebo in patients with relapsing multiple sclerosis (MS). The objectives were to monitor the 36-month, interim efficacy and safety results of the ongoing extension of this study. In the extension (months 7-36), placebo-treated patients were re-randomized to either dose of fingolimod; fingolimod-treated patients continued at the same dose. During months 15-24, all patients receiving fingolimod 5.0 mg switched to 1.25 mg. Of the 250 patients who entered the extension study, 173 (69%) continued to month 36. Most patients were free from gadolinium-enhanced lesions (88-89%) or new T2 lesions (70-78%) at month 36. Patients receiving continuous fingolimod treatment had sustained low annualized relapse rates of 0.20-0.21, and 68-73% remained relapse-free at month 36. Over 36 months, nasopharyngitis (34%), headache (30%), fatigue (19%) and influenza (18%) were the most commonly reported adverse events. Pulmonary function remained stable and blood pressure was stable after an initial increase (3-5 mmHg) during the first 6 months of fingolimod treatment; serious adverse events included infections and skin cancer. The low MRI and clinical disease activity at 6 months were maintained at 36 months with fingolimod, which was generally well tolerated by most patients. The efficacy and safety of oral fingolimod are being further evaluated in a large phase III MS study programme.


Bovine spongiform encephalopathy: is it time to relax BSE-related measures in the context of international trade?
Bovine spongiform encephalopathy (BSE) has presented serious challenges to both the World Organisation for Animal Health and national governments, in defining and implementing appropriate national control measures, and in agreeing trade rules that permit safe trade in cattle and bovine products. Precautionary trade rules were initially necessary, based upon the science of sheep scrapie, but research into BSE later enabled BSE-specific trade rules to be developed. As a result, current rules on trade are underpinned by a sound body of knowledge on BSE. Declining epidemics in most affected countries confirm the appropriateness of current precautions. Nevertheless, risk is primarily dependent on the prevalence of infection with BSE. In the face of low prevalence scenarios, certain precautionary measures in the Terrestrial Animal Health Code may now be considered excessive. A thorough review is therefore deemed appropriate.


Virtual reality and the role of the prefrontal cortex in adults and children.
In this review, the neural underpinnings of the experience of presence are outlined. Firstly, it is shown that presence is associated with activation of a distributed network, which includes the dorsal and ventral visual stream, the parietal cortex, the premotor cortex, mesial temporal areas, the brainstem and the thalamus. Secondly, the dorsolateral prefrontal cortex (DLPFC) is identified as a key node of the network as it modulates the activity of the network and the associated experience of presence. Thirdly, children lack the strong modulatory influence of the DLPFC on the network due to their unmatured frontal cortex. Fourthly, it is shown that presence-related measures are influenced by manipulating the activation in the DLPFC using transcranial direct current stimulation (tDCS) while participants are exposed to the virtual roller coaster ride. Finally, the findings are discussed in the context of current models explaining the experience of presence, the rubber hand illusion, and out-of-body experiences.


Gender-based differences in the high-risk sexual behaviours of young people aged 15-29 in Melilla (Spain): a cross-sectional study.
Research confirms the existence of gender-based differences regarding the high-risk sexual behaviour (non-use of condoms and casual partners) of young men and women. The objectives were to provide evidence for this association; to analyse the reasons why both sexes have sexual relations with casual partners and to ascertain the motives for condom use or non-use during casual sex. A cross-sectional study was performed on a sample of 900 participants, 524 males and 376 females. All participants were 15-29 (20.93 ± 4.071) years of age and came from four different centres (a university, two secondary schools, and a military base) in Melilla (Spain). The participants were given a socio-demographic survey as well as a psychometric text on high-risk sexual behaviour. The results found gender-based significant differences for sexual relations with penetration (p = 0.001), number of sexual partners (p = 0.001), and sexual relations with casual partners (p = 0.001). In all of these variables, male participants had higher percentages than female participants. Reasons for having casual sexual relations were also different for men and women, differences were found for the items, opportunity (p = 0.001), interest in knowing the other person (p = 0.015), physical excitement (p = 0.056) and drug consumption (p = 0.059). Regarding the reasons for consistent condom use with casual partners, there were differences for the item, my demand of a condom (p = 0.002). For the non-use of condoms with casual partners, differences were found for the items, I do not like to use condoms (p = 0.001) and condoms lessen sensitivity and reduce pleasure (p = 0.009). Men and women were found to have different high-risk sexual behaviours and practices. Of the motives for having sexual relations with casual partners, male participants considered opportunity and interest in knowing the other person to be more important than the female participants. Regarding condom use, the female participants' demand to use a condom was a significant gender-based difference. In contrast to the young women, the male participants mostly justified not using a condom because it lessened sensitivity and reduced pleasure.


Thymidine phosphorylase expression in endometrial carcinomas.
Thymidine phosphorylase (TP) is a potent angiogenic molecule shown to induce endothelial cell migration and proliferation. We investigated the expression of TP in a series of 156 endometrial carcinomas, using immunohistochemical methods. Histopathological parameters of known prognostic significance and the molecular factors of p53, bcl-2 and angiogenesis were also assessed. Thymidine phosphorylase was expressed in cancer cells, stromal fibroblasts and myometrial cells. The pattern of TP staining was nuclear or mixed nuclear/cytoplasmic, and only exceptionally was purely cytoplasmic. An exclusively cytoplasmic staining was documented for the tumour-associated foamy macrophages. Cancer cell reactivity was rather limited; only 3.2% of endometrial carcinomas expressed TP in more than 50% of the neoplastic cell population and only 12% expressed the enzyme in more than 10% of the cancer cells. By contrast, TP reactivity was frequent in the fibroblasts of the tumour supporting stroma and the fibroblasts/myometrial cells at the invading tumour front, where approximately 1/3 of the cases expressed TP in more than 50% of the respective constituent cells. A high TP reactivity in the stromal fibroblasts was significantly associated with the presence of foamy macrophages and an intense lymphocytic response. A high TP reactivity at the invading tumour front was significantly associated with an intense lymphocytic response and the adverse prognostic parameters of high tumour grade, deep myometrial invasion, advanced stage of disease and the non-endometrioid carcinomas. There was no significant association of cancer cell TP reactivity with any of the parameters studied, including nuclear p53 accumulation, cytoplasmic/perinuclear bcl-2 expression, microvessel density (MVD) and prognosis. Similarly, no relationship was established between fibroblastic or fibroblastic/myometrial TP reactivity and MVD. It is concluded that TP is not a major angiogenic factor in endometrial carcinomas. However, a prominent TP activity at the invading tumour front, which is probably induced by cytokines of histiocytic and lymphocytic origin, may promote tumour invasion and progression.


hsp90 is required for heme binding and activation of apo-neuronal nitric-oxide synthase: geldanamycin-mediated oxidant generation is unrelated to any action of hsp90.
It is established that neuronal NO synthase (nNOS) is associated with the chaperone hsp90, although the functional role for this interaction has not been defined. We have discovered that inhibition of hsp90 by radicicol or geldanamycin nearly prevents the heme-mediated activation and assembly of heme-deficient apo-nNOS in insect cells. This effect is concentration-dependent with over 75% inhibition achieved at 20 microm radicicol. The ferrous carbonyl complex of nNOS is not formed when hsp90 is inhibited, indicating that functional heme insertion is prevented. We propose that the hsp90-based chaperone machinery facilitates functional heme entry into apo-nNOS by the opening of the hydrophobic heme-binding cleft in the protein. Previously, it has been reported that the hsp90 inhibitor geldanamycin uncouples endothelial NOS activity and increases endothelial NOS-dependent O(2)() production. Geldanamycin is an ansamycin benzoquinone, and we show here that it causes oxidant production from nNOS in insect cells as well as with the purified protein. At a concentration of 20 microm, geldanamycin causes a 3-fold increase in NADPH oxidation and hydrogen peroxide formation from purified nNOS, whereas the non-quinone hsp90 inhibitor radicicol had no effect. Thus, consistent with the known propensity of other quinones, geldanamycin directly redox cycles with nNOS by a process independent of any action on hsp90, cautioning against the use of geldanamycin as a specific inhibitor of hsp90 in redox-active systems.


Prozone effects in microscopic agglutination tests for leptospirosis in the sera of mice infected with the pathogenic Leptospira interrogans serovar Canicola.
Mice experimentally infected with a pathogenic strain of Leptospira interrogans serovar Canicola produced false negative results (prozone effect) in a microscopic agglutination test (MAT). This prozone effect occurred in several serum samples collected at different post-infection times, but it was more prominent in samples collected from seven-42 days post-infection and for 1:50 and 1:100 sample dilutions. This phenomenon was correlated with increased antibody titres in the early post-infection phase. While prozone effects are often observed in serological agglutination assays for the diagnosis of animal brucellosis and human syphilis, they are not widely reported in leptospirosis MATs.


How droplets nucleate and grow on liquids and liquid impregnated surfaces.
Condensation on liquids has been studied extensively in context of breath figure templating, materials synthesis and enhancing heat transfer using liquid impregnated surfaces. However, the mechanics of nucleation and growth on liquids remains unclear, especially on liquids that spread on the condensate. By examining the energy barriers of nucleation, we provide a framework to choose liquids that can lead to enhanced nucleation. We show that due to limits of vapor sorption within a liquid, nucleation is most favoured at the liquid-air interface and demonstrate that on spreading liquids, droplet submergence within the liquid occurs thereafter. We provide a direct visualization of the thin liquid profile that cloaks the condensed droplet on a liquid impregnated surface and elucidate the vapour transport mechanism in the liquid films. Finally, we show that although the viscosity of the liquid does not affect droplet nucleation, it plays a crucial role in droplet growth.


[Kidney disease in Sant' Andrea Hospital: a biopsy based epidemiologic study].
The aim of this retrospective study is to investigate the prevalence and pathological features of kidney inflammatory nephropathies diagnosed in Sant'Andrea Hospital, from January 2003 to April 2015. In this period, 246 kidney biopsies have been diagnosed in our Hospital. Excluding cases of kidney neoplasms and non-diagnostic samples, 195 cases were reviewed. Primary glomerulonephritis (GN) is the most common diagnosis. Among these, Membranous GN represents the majority of cases (20.4%), followed by IgA Nephropathy (12.7%). The higher prevalence of Membranous GN than IgA Nephropathy represents a difference between our study and national and international kidney biopsies registries. It can be considered a consequence of the average age of patients undergoing renal biopsy in our center (54,1 years). Patients with Diabetic Nephropathy are 1.5%. 10 out of 195 cases (5.1%) show end stage renal disease. This epidemiological study evaluates the prevalence of various kidney diseases in our database, the biopsy policy of SantAndrea Hospital and compares our results with national and international renal biopsies registries.


Dual involvements of cyclooxygenase and nitric oxide synthase expressions in ketamine-induced ulcerative cystitis in rat bladder.
The aims of the present study were to investigate voiding patterns, tissue constituents and the expressions of cyclooxygenase-2 (COX-2) and nitric oxide synthase (NOS) involved in ketamine-induced ulcerative cystitis in rat urinary bladder. Thirty Sprague-Dawley rats were distributed into three groups which received saline or ketamine (25 mg/kg/day) for a period of 14 and 28 days. In each group, cystometry was performed weekly and the concentration of ketamine and its metabolites (norketamine) was assayed. Paraffin-embedded sections were stained with Masson's trichrome stain, and ketamine-induced morphological changes were examined. Western blot analyses were carried out to examine the expressions of COX-2 and different NOS isoforms in bladder tissues. Immunofluorescence study was done to evaluate the expressions of COX-2 and macrophage infiltration (stained with ED-1 macrophage cell surface antigen) within the bladder. Ketamine treatment resulted in bladder hyperactivity and the non-voiding contractions were significantly increased. The urine concentrations of ketamine and norketamine were much higher in ketamine-treated group. Moreover, ulcerated urothelium and mononuclear cell infiltration were noted in ketamine-treated group. These alterations in urodynamic functions and tissue constituents were accompanied by increases in the expression of COX-2. Two NOS isoforms (iNOS and eNOS) were also overexpressed, but no significant change was observed for nNOS. COX-2 positive stained cells were significantly increased. Meanwhile, increased amounts of ED-1 positive stained macrophages were present and most of COX-2 expressed cells were co-stained with ED-1 in the early stage of ketamine treatment. Ketamine treatment affected bladder tissues by enhancing interstitial fibrosis and accelerating macrophages infiltration. Ketamine also initiated the up-regulations of COX-2 and iNOS and eNOS expressions. These up-regulated enzymes might play an important role in contributing to ketamine-induced alterations in micturition patterns and ulcerative cystitis.


A 15.6 frames per second 1-megapixel multiple exposure laser speckle contrast imaging setup.
A multiple exposure laser speckle contrast imaging (MELSCI) setup for visualizing blood perfusion was developed using a field programmable gate array (FPGA), connected to a 1000 frames per second (fps) 1-megapixel camera sensor. Multiple exposure time images at 1, 2, 4, 8, 16, 32 and 64 milliseconds were calculated by cumulative summation of 64 consecutive snapshot images. The local contrast was calculated for all exposure times using regions of 4 × 4 pixels. Averaging of multiple contrast images from the 64-millisecond acquisition was done to improve the signal-to-noise ratio. The results show that with an effective implementation of the algorithm on an FPGA, contrast images at all exposure times can be calculated in only 28 milliseconds. The algorithm was applied to data recorded during a 5 minutes finger occlusion. Expected contrast changes were found during occlusion and the following hyperemia in the occluded finger, while unprovoked fingers showed constant contrast during the experiment. The developed setup is capable of massive data processing on an FPGA that enables processing of MELSCI data in 15.6 fps (1000/64 milliseconds). It also leads to improved frame rates, enhanced image quality and enables the calculation of improved microcirculatory perfusion estimates compared to single exposure time systems.


Quantum criticality in an Ising chain: experimental evidence for emergent E8 symmetry.
Quantum phase transitions take place between distinct phases of matter at zero temperature. Near the transition point, exotic quantum symmetries can emerge that govern the excitation spectrum of the system. A symmetry described by the E8 Lie group with a spectrum of eight particles was long predicted to appear near the critical point of an Ising chain. We realize this system experimentally by using strong transverse magnetic fields to tune the quasi-one-dimensional Ising ferromagnet CoNb2O6 (cobalt niobate) through its critical point. Spin excitations are observed to change character from pairs of kinks in the ordered phase to spin-flips in the paramagnetic phase. Just below the critical field, the spin dynamics shows a fine structure with two sharp modes at low energies, in a ratio that approaches the golden mean predicted for the first two meson particles of the E8 spectrum. Our results demonstrate the power of symmetry to describe complex quantum behaviors.


Microcin-E492-insensitive mutants of Escherichia coli K12.
Mutations in three Escherichia coli K12 genes, tonB, exbB and the newly discovered semA, reduce sensitivity to the low Mr polypeptide antibiotic microcin E492. The products of the tonB and exbB genes were previously shown to be involved in the uptake of siderophore-complexed iron and in the action of a number of colicins. Strains mutated at or close to semA (collectively referred to as sem mutations) remained fully sensitive to these colicins, and grew as well as wild-type strains under conditions of iron starvation. Expression of a number of sem-lacZ operon fusions was not affected by iron limitation, and sem mutations did not affect the production of iron-regulated outer membrane proteins which are known or thought to be involved in iron uptake. Hfr conjugation and P1 phage transduction experiments indicated that semA is located close to pabB at 40 min on the E. coli K12 chromosome. This places semA close to the mng locus, wherein mutations result in decreased manganese sensitivity. However, strains carrying the semA mutation exhibited increased manganese sensitivity.


Joint UK societies' 2014 consensus statement on renal denervation for resistant hypertension.
Resistant hypertension continues to pose a major challenge to clinicians worldwide and has serious implications for patients who are at increased risk of cardiovascular morbidity and mortality with this diagnosis. Pharmacological therapy for resistant hypertension follows guidelines-based regimens although there is surprisingly scant evidence for beneficial outcomes using additional drug treatment after three antihypertensives have failed to achieve target blood pressure. Recently there has been considerable interest in the use of endoluminal renal denervation as an interventional technique to achieve renal nerve ablation and lower blood pressure. Although initial clinical trials of renal denervation in patients with resistant hypertension demonstrated encouraging office blood pressure reduction, a large randomised control trial (Symplicity HTN-3) with a sham-control limb, failed to meet its primary efficacy end point. The trial however was subject to a number of flaws which must be taken into consideration in interpreting the final results. Moreover a substantial body of evidence from non-randomised smaller trials does suggest that renal denervation may have an important role in the management of hypertension and other disease states characterised by overactivation of the sympathetic nervous system. The Joint UK Societies does not recommend the use of renal denervation for treatment of resistant hypertension in routine clinical practice but remains committed to supporting research activity in this field. A number of research strategies are identified and much that can be improved upon to ensure better design and conduct of future randomised studies.


Prenatal undernutrition and postnatal growth are associated with adolescent thymic function.
The fetal and early infant origins of a number of adult cardiovascular and metabolic diseases have received considerable attention, but the long-term consequences of early environments for human immune function have not been reported. We investigated the effects of pre- and postnatal environments on thymic hormone production in adolescents participating in an ongoing longitudinal study in the Philippines. Prospective data collected at birth, during y 1 of life, in childhood and in adolescence were used to predict plasma thymopoietin concentration in 14- to 15-y-old adolescents (n = 103). Thymopoietin concentration was compared for small-for-gestational-age and appropriate-for-gestational-age individuals while controlling for a range of postnatal exposures. Prenatal undernutrition was significantly associated with reduced thymopoietin production in interaction with the duration of exclusive breast-feeding (P = 0.006). Growth in length during y 1 of life was positively associated with adolescent thymopoietin production (P = 0.002). These associations remained significant after adjusting for a range of potentially confounding variables. These findings provide support for the importance of fetal and early infant programming of thymic function, and suggest that early environments may have long-term implications for immunocompetence and adult disease risk.


Electricity production and microbial biofilm characterization in cellulose-fed microbial fuel cells.
Converting biodegradable materials into electricity, microbial fuel cells (MFCs) present a promising technology for renewable energy production in specific applications. Unlike typical soluble substrates that have been used as electron donors in MFC studies, cellulose is unique because it requires a microbial consortium that can metabolize both an insoluble electron donor (cellulose) and electron acceptor (electrode). In this study, electricity generation and the microbial ecology of cellulose-fed MFCs were analyzed using a defined co-culture of Clostridium cellulolyticum and Geobacter sulfurreducens. Fluorescent in situ hybridization and quantitative PCR showed that when particulate MN301 cellulose was used as sole substrate, most Clostridium cells were found adhered to cellulose particles in suspension, while most Geobacter cells were attached to the electrode. By comparison, both bacteria resided in suspension and biofilm samples when soluble carboxymethyl cellulose was used. This distinct function-related distribution of the bacteria suggests an opportunity to optimize reactor operation by settling cellulose and decanting supernatant to extend cellulose hydrolysis and improve cellulose-electricity conversion.


Fiber-connected UWB sensor network for high-resolution localization using optical time-division multiplexing.
A fiber-connected ultra-wideband (UWB) sensor network for high-resolution localization which consists of a central station and several sensor nodes is proposed and demonstrated. To make the central station easily identify the received UWB pulses from different sensor nodes, optical time-division multiplexing (OTDM), realized by inserting a certain length of optical fiber between every two sensor nodes, is implemented. Due to the OTDM technology, the UWB pulses received by different sensors are mapped into different time slots, so neither parameter estimation nor clock synchronization is required in the UWB sensor node. All complex signal processing is completed in the central station, which greatly improve the localization accuracy and simplify the system. A proof-of-concept experiment for two-dimensional localization is demonstrated. Spatial resolution as high as 3.9 cm is achieved.


Hypoxia-inducible factor-1 signalling promotes goblet cell hyperplasia in airway epithelium.
Goblet cell hyperplasia is a common feature of chronic obstructive pulmonary disease (COPD) airways, but the mechanisms that underlie this epithelial remodelling in COPD are not understood. Based on our previous finding of hypoxia-inducible factor-1α (HIF-1α) nuclear localization in large airways from patients with COPD, we investigated whether hypoxia-inducible signalling could influence the development of goblet cell hyperplasia. We evaluated large airway samples obtained from 18 lifelong non-smokers and 13 former smokers without COPD, and 45 former smokers with COPD. In these specimens, HIF-1α nuclear staining occurred almost exclusively in COPD patients in areas of airway remodelling. In COPD patients, 93.2 ± 3.9% (range 65-100%) of goblet cells were HIF-1α positive in areas of goblet cell hyperplasia, whereas nuclear HIF-1α was not detected in individuals without COPD or in normal-appearing pseudostratified epithelium from COPD patients. To determine the direct effects of hypoxia-inducible signalling on epithelial cell differentiation in vitro, human bronchial epithelial cells (HBECs) were grown in air-liquid interface cultures under hypoxia (1% O(2)) or following treatment with a selective HIF-1α stabilizer, (2R)-[(4-biphenylylsulphonyl)amino]-N-hydroxy-3-phenyl-propionamide (BiPS). HBECs grown in hypoxia or with BiPS treatment were characterized by HIF-1α activation, carbonic anhydrase IX expression, mucus-producing cell hyperplasia and increased expression of MUC5AC. Analysis of signal transduction pathways in cells with HIF-1α activation showed increased ERK1/2 phosphorylation without activation of epidermal growth factor receptor, Ras, PI3K-Akt or STAT6. These data indicate an important effect of hypoxia-inducible signalling on airway epithelial cell differentiation and identify a new potential target to limit mucus production in COPD.


Blue pigment colors from wall painting churches in danger (Portugal 15th to 18th century): identification, diagnosis, and color evaluation.
Samples of blue wall paint layers from selected 15th to 18th century religious mural paintings from southern Portugal (Alentejo) have been analyzed using a multi-analytical methodology involving the combination of in situ visible spectro-colorimetry with microanalytical techniques such as optical and scanning electron microscopy and Raman spectroscopy. In situ analyses and micro-sampling were carried out in nine different churches, many in an advanced state of deterioration. The objectives of this study were: (a) to identify and compare the pigments that were used in the blue paint layers across the Alentejo region and through time by analysis of the elemental and mineralogical composition and pictorial techniques, and (b) to correlate the data between the actual color of the paint layer and the state of conservation of the pigments. For the paintings dated from the 16th century forward, the results show a generalized use of smalt blue. To a lesser extent, natural azurite was used despite the geological richness of the region in copper and pyrite ores. In only one painting was an optical blue made of carbon black and lime found. The pigments, pure or mixed with red and yellow ochres, were coarsely ground and used in different concentrations to create three-dimensional effects. These parameters as well as the presence of iron oxides in underlayer paints influence the colorimetric coordinates in the more transparent smalt blue paint layers. The state of conservation of the pigments plays an important role in the alteration of the paint color. A clear example of this is the fading of the smalt blue in several paintings due to lixiviation processes.


Associations between online food outlet access and online food delivery service use amongst adults in the UK: a cross-sectional analysis of linked data.
Online food delivery services facilitate 'online' access to food outlets that typically sell lenergy-dense nutrient-poor food. Greater online food outlet access might be related to the use of this purchasing format and living with excess bodyweight, however, this is not known. We aimed to investigate the association between aspects of online food outlet access and online food delivery service use, and differences according to customer sociodemographic characteristics, as well as the association between the number of food outlets accessible online and bodyweight. In 2019, we used an automated data collection method to collect data on all food outlets in the UK registered with the leading online food delivery service Just Eat (n = 33,204). We linked this with contemporaneous data on food purchasing, bodyweight, and sociodemographic information collected through the International Food Policy Study (analytic sample n = 3067). We used adjusted binomial logistic, linear, and multinomial logistic regression models to examine associations. Adults in the UK had online access to a median of 85 food outlets (IQR: 34-181) and 85 unique types of cuisine (IQR: 64-108), and 15.1% reported online food delivery service use in the previous week. Those with the greatest number of accessible food outlets (quarter four, 182-879) had 71% greater odds of online food delivery service use (OR: 1.71; 95% CI: 1.09, 2.68) compared to those with the least (quarter one, 0-34). This pattern was evident amongst adults with a university degree (OR: 2.11; 95% CI: 1.15, 3.85), adults aged between 18 and 29 years (OR: 3.27, 95% CI: 1.59, 6.72), those living with children (OR: 1.94; 95% CI: 1.01; 3.75), and females at each level of increased exposure. We found no association between the number of unique types of cuisine accessible online and online food delivery service use, or between the number of food outlets accessible online and bodyweight. The number of food outlets accessible online is positively associated with online food delivery service use. Adults with the highest education, younger adults, those living with children, and females, were particularly susceptible to the greatest online food outlet access. Further research is required to investigate the possible health implications of online food delivery service use.


A Qualitative Examination of Health Barriers and Facilitators Among African American Mothers in a Subsidized Housing Community.
Although African American families are at particular risk for obesity and its associated health comorbidities, few interventions have directly targeted low-income members of this group living in subsidized public housing. Using a consensual qualitative research approach, we conducted 11 interviews with African American mothers living in two public housing communities to enhance understanding of their perceived barriers and facilitators to health. Five primary domains emerged, including barriers (access, financial, personal, and neighborhood concerns), resources (personal and community), current behaviors (diet, physical activity, and program participation), definition of health (mental well-being, physical well-being, and health behaviors), and needs/interests in programming (health behavior-specific programs, non-health-related programs, child-focused programming, and qualities of programs and their leaders). Results demonstrate the complex interaction among social, environmental, and personal factors on health behaviors for this priority population, and highlight the need for community members' involvement in the development of community-based obesity prevention programming.


Bilateral absence of adrenal glands: a case series that expands the spectrum of associations and highlights the difficulties in prenatal diagnosis.
Adrenal agenesis (AA) defined by the complete absence of development of adrenals is a rare anomaly, which is documented mainly in experimental animals and less frequently in human subjects in the literature. This study was aimed at describing the varied phenotype of this condition in two stillborn and one termination of pregnancy fetuses and two neonates, the associated abnormalities and the difficulties encountered to achieve the prenatal diagnosis. Five cases with AA diagnosed at post-mortem examination were selected and their characteristics were analyzed. The detection of this unusual condition has usually been made as an incidental discovery at post-mortem examination. None of the cases described in this series had been diagnosed at prenatal ultrasonography. Respiratory distress was the commonest clinical presentation in the liveborn. Maternal diabetes was associated with one case. Anomalies of kidneys, lungs, spleen, and blood vessels were associated with two of the cases. No gonadal abnormalities were detected in any of the cases. These cases illustrate the varied clinical presentation of this rare condition and confirm the difficulty in achieving a prenatal diagnosis.


Underwater Noise from a Wave Energy Converter Is Unlikely to Affect Marine Mammals.
Underwater noise was recorded from the Wavestar wave energy converter; a full-scale hydraulic point absorber, placed on a jack-up rig on the Danish North Sea coast. Noise was recorded 25 m from the converter with an autonomous recording unit (10 Hz to 20 kHz bandwidth). Median sound pressure levels (Leq) in third-octave bands during operation of the converter were 106-109 dB re. 1 μPa in the range 125-250 Hz, 1-2 dB above ambient noise levels (statistically significant). Outside the range 125-250 Hz the noise from the converter was undetectable above the ambient noise. During start and stop of the converter a more powerful tone at 150 Hz (sound pressure level (Leq) 121-125 dB re 1 μPa) was easily detectable. This tone likely originated from the hydraulic pump which was used to lower the absorbers into the water and lift them out of the water at shutdown. Noise levels from the operating wave converter were so low that they would barely be audible to marine mammals and the likelihood of negative impact from the noise appears minimal. A likely explanation for the low noise emissions is the construction of the converter where all moving parts, except for the absorbers themselves, are placed above water on a jack-up rig. The results may thus not be directly transferable to other wave converter designs but do demonstrate that it is possible to harness wave energy without noise pollution to the marine environment.


Tissue response, macrophage phenotype, and intrinsic calcification induced by cardiovascular biomaterials: Can clinical regenerative potential be predicted in a rat subcutaneous implant model?
The host immune response to an implanted biomaterial, particularly the phenotype of infiltrating macrophages, is a key determinant of biocompatibility and downstream remodeling outcome. The present study used a subcutaneous rat model to compare the tissue response, including macrophage phenotype, remodeling potential, and calcification propensity of a biologic scaffold composed of glutaraldehyde-fixed bovine pericardium (GF-BP), the standard of care for heart valve replacement, with those of an electrospun polycarbonate-based supramolecular polymer scaffold (ePC-UPy), urinary bladder extracellular matrix (UBM-ECM), and a polypropylene mesh (PP). The ePC-UPy and UBM-ECM materials induced infiltration of mononuclear cells throughout the thickness of the scaffold within 2 days and neovascularization at 14 days. GF-BP and PP elicited a balance of pro-inflammatory (M1-like) and anti-inflammatory (M2-like) macrophages, while UBM-ECM and ePC-UPy supported a dominant M2-like macrophage phenotype at all timepoints. Relative to GF-BP, ePC-UPy was markedly less susceptible to calcification for the 180 day duration of the study. UBM-ECM induced an archetypical constructive remodeling response dominated by M2-like macrophages and the PP caused a typical foreign body reaction dominated by M1-like macrophages. The results of this study highlight the divergent macrophage and host remodeling response to biomaterials with distinct physical and chemical properties and suggest that the rat subcutaneous implantation model can be used to predict in vivo biocompatibility and regenerative potential for clinical application of cardiovascular biomaterials.


Conceptual modeling for identification of worst case conditions in environmental risk assessment of nanomaterials using nZVI and C60 as case studies.
Conducting environmental risk assessment of engineered nanomaterials has been an extremely challenging endeavor thus far. Moreover, recent findings from the nano-risk scientific community indicate that it is unlikely that many of these challenges will be easily resolved in the near future, especially given the vast variety and complexity of nanomaterials and their applications. As an approach to help optimize environmental risk assessments of nanomaterials, we apply the Worst-Case Definition (WCD) model to identify best estimates for worst-case conditions of environmental risks of two case studies which use engineered nanoparticles, namely nZVI in soil and groundwater remediation and C(60) in an engine oil lubricant. Results generated from this analysis may ultimately help prioritize research areas for environmental risk assessments of nZVI and C(60) in these applications as well as demonstrate the use of worst-case conditions to optimize future research efforts for other nanomaterials. Through the application of the WCD model, we find that the most probable worst-case conditions for both case studies include i) active uptake mechanisms, ii) accumulation in organisms, iii) ecotoxicological response mechanisms such as reactive oxygen species (ROS) production and cell membrane damage or disruption, iv) surface properties of nZVI and C(60), and v) acute exposure tolerance of organisms. Additional estimates of worst-case conditions for C(60) also include the physical location of C(60) in the environment from surface run-off, cellular exposure routes for heterotrophic organisms, and the presence of light to amplify adverse effects. Based on results of this analysis, we recommend the prioritization of research for the selected applications within the following areas: organism active uptake ability of nZVI and C(60) and ecotoxicological response end-points and response mechanisms including ROS production and cell membrane damage, full nanomaterial characterization taking into account detailed information on nanomaterial surface properties, and investigations of dose-response relationships for a variety of organisms.


Human mitochondrial DNA deletions associated with mutations in the gene encoding Twinkle, a phage T7 gene 4-like protein localized in mitochondria.
The gene products involved in mammalian mitochondrial DNA (mtDNA) maintenance and organization remain largely unknown. We report here a novel mitochondrial protein, Twinkle, with structural similarity to phage T7 gene 4 primase/helicase and other hexameric ring helicases. Twinkle colocalizes with mtDNA in mitochondrial nucleoids. Screening of the gene encoding Twinkle in individuals with autosomal dominant progressive external ophthalmoplegia (adPEO), associated with multiple mtDNA deletions, identified 11 different coding-region mutations co-segregating with the disorder in 12 adPEO pedigrees of various ethnic origins. The mutations cluster in a region of the protein proposed to be involved in subunit interactions. The function of Twinkle is inferred to be critical for lifetime maintenance of human mtDNA integrity.


Evolutionary conserved brainstem circuits encode category, concentration and mixtures of taste.
Evolutionary conserved brainstem circuits are the first relay for gustatory information in the vertebrate brain. While the brainstem circuits act as our life support system and they mediate vital taste related behaviors, the principles of gustatory computations in these circuits are poorly understood. By a combination of two-photon calcium imaging and quantitative animal behavior in juvenile zebrafish, we showed that taste categories are represented by dissimilar brainstem responses and generate different behaviors. We also showed that the concentration of sour and bitter tastes are encoded by different principles and with different levels of sensitivity. Moreover, we observed that the taste mixtures lead to synergistic and suppressive interactions. Our results suggest that these interactions in early brainstem circuits can result in non-linear computations, such as dynamic gain modulation and discrete representation of taste mixtures, which can be utilized for detecting food items at broad range of concentrations of tastes and rejecting inedible substances.


Virtual reality for stroke rehabilitation.
Virtual reality and interactive video gaming have emerged as recent treatment approaches in stroke rehabilitation. In particular, commercial gaming consoles have been rapidly adopted in clinical settings. This is an update of a Cochrane Review published in 2011. To determine the efficacy of virtual reality compared with an alternative intervention or no intervention on upper limb function and activity. To determine the efficacy of virtual reality compared with an alternative intervention or no intervention on: gait and balance activity, global motor function, cognitive function, activity limitation, participation restriction and quality of life, voxels or regions of interest identified via imaging, and adverse events. Additionally, we aimed to comment on the feasibility of virtual reality for use with stroke patients by reporting on patient eligibility criteria and recruitment. We searched the Cochrane Stroke Group Trials Register (October 2013), the Cochrane Central Register of Controlled Trials (The Cochrane Library 2013, Issue 11), MEDLINE (1950 to November 2013), EMBASE (1980 to November 2013) and seven additional databases. We also searched trials registries and reference lists. Randomised and quasi-randomised trials of virtual reality ("an advanced form of human-computer interface that allows the user to 'interact' with and become 'immersed' in a computer-generated environment in a naturalistic fashion") in adults after stroke. The primary outcome of interest was upper limb function and activity. Secondary outcomes included gait and balance function and activity, and global motor function. Two review authors independently selected trials based on pre-defined inclusion criteria, extracted data and assessed risk of bias. A third review author moderated disagreements when required. The authors contacted investigators to obtain missing information. We included 37 trials that involved 1019 participants. Study sample sizes were generally small and interventions varied. The risk of bias present in many studies was unclear due to poor reporting. Thus, while there are a large number of randomised controlled trials, the evidence remains 'low' or 'very low' quality when rated using the GRADE system. Control groups received no intervention or therapy based on a standard care approach. Intervention approaches in the included studies were predominantly designed to improve motor function rather than cognitive function or activity performance. The majority of participants were relatively young and more than one year post stroke. results were statistically significant for upper limb function (standardised mean difference (SMD) 0.28, 95% confidence intervals (CI) 0.08 to 0.49 based on 12 studies with 397 participants). there were no statistically significant effects for grip strength, gait speed or global motor function. Results were statistically significant for the activities of daily living (ADL) outcome (SMD 0.43, 95% CI 0.18 to 0.69 based on eight studies with 253 participants); however, we were unable to pool results for cognitive function, participation restriction, quality of life or imaging studies. There were few adverse events reported across studies and those reported were relatively mild. Studies that reported on eligibility rates showed that only 26% of participants screened were recruited. We found evidence that the use of virtual reality and interactive video gaming may be beneficial in improving upper limb function and ADL function when used as an adjunct to usual care (to increase overall therapy time) or when compared with the same dose of conventional therapy. There was insufficient evidence to reach conclusions about the effect of virtual reality and interactive video gaming on grip strength, gait speed or global motor function. It is unclear at present which characteristics of virtual reality are most important and it is unknown whether effects are sustained in the longer term.


Antibodies Against Hypocretin Receptor 2 Are Rare in Narcolepsy.
Recently, antibodies to the hypocretin receptor 2 (HCRTR2-Abs) were reported in a high proportion of narcolepsy patients who developed the disease following Pandemrix® vaccination. We tested a group of narcolepsy patients for the HCRTR2-Abs using a newly established cell-based assay. Sera from 50 narcolepsy type 1 (NT1) and 11 narcolepsy type 2 (NT2) patients, 22 patients with other sleep disorders, 15 healthy controls, and 93 disease controls were studied. Cerebrospinal fluid (CSFs) from three narcoleptic patients were subsequently included. Human embryonic kidney cells were transiently transfected with human HCRTR2, incubated with patients' sera for 1 hr at 1:20 dilution and then fixed. Binding of antibodies was detected by fluorescently labeled secondary antibodies to human immunoglobulin G (IgG) and the different IgG subclasses. A nonlinear visual scoring system was used from 0 to 4; samples scoring ≥1 were considered positive. Only 3 (5%) of 61 patients showed a score ≥1, one with IgG1- and two with IgG3-antibodies, but titers were low (1:40-1:100). CSFs from these patients were negative. The three positive patients included one NT1 case with associated psychotic features, one NT2 patient, and an NT1 patient with normal hypocretin CSF levels. Low levels of IgG1 or IgG3 antibodies against HCRTR2 were found in 3 of 61 patients with narcolepsy, although only 1 presented with full-blown NT1. HCRTR2-Abs are not common in narcolepsy unrelated to vaccination.


Spatial correlates of firing patterns of single cells in the subiculum of the freely moving rat.
Hippocampal lesions cause spatial learning deficits, and single hippocampal cells show location-specific firing patterns, known as place fields. This suggests the hippocampus plays a critical role in navigation by providing an ongoing indication of the animal's momentary spatial location. One question that has received little attention is how this locational signal is used by downstream brain regions to orchestrate actual navigational behavior. As a first step, we have examined the spatial firing correlates of cells in the dorsal subiculum as rats navigate in an open-field, pellet-searching task. The subiculum is one of the few major output zones for the hippocampus, and it, in turn, projects to numerous other brain areas, each thought to be involved in various learning and memory functions. Most subicular cells showed a robust locational signal. The patterns observed were different from those in the hippocampus, however, in that cells tended to fire throughout much of the environment, but showed graded, location-related rate modulation, such that there were some localized regions of high firing and other regions with relatively low firing. There were slight quantitative differences between the proximal (adjacent to the hippocampus) and distal (farther from the hippocampus) subicular regions, with distal cells showing slightly higher average firing rates, spatial signaling, and firing field size. This was of interest since these two regions have different efferent connections. Examination of spike trains allowed classification of cells into bursting, nonbursting, and theta (putative interneuron) categories, and this is similar to subicular cell types identified in vitro. Interestingly, the bursting and nonbursting types did not differ detectably in spatial firing properties, suggesting that differences in intrinsic membrane properties do not necessitate differences in coding of environmental inputs. The results suggest that the subiculum transmits a robust, highly distributed spatial signal to each of its projection areas, and that this signal is transmitted in both a bursting and nonbursting mode.


Mechanisms involved in Escherichia coli and Serratia marcescens removal during activated sludge wastewater treatment.
Wastewater treatment reduces environmental contamination by removing gross solids and mitigating the effects of pollution. Treatment also reduces the number of indicator organisms and pathogens. In this work, the fates of two coliform bacteria, Escherichia coli and Serratia marcescens, were analyzed in an activated sludge process to determine the main mechanisms involved in the reduction of pathogenic microorganisms during wastewater treatment. These bacteria, modified to express green fluorescent protein, were inoculated in an activated sludge unit and in batch systems containing wastewater. The results suggested that, among the different biological factors implied in bacterial removal, bacterivorous protozoa play a key role. Moreover, a representative number of bacteria persisted in the system as free-living or embedded cells, but their distribution into liquid or solid fractions varied depending on the bacterium tested, questioning the real value of bacterial indicators for the control of wastewater treatment process. Additionally, viable but nonculturable cells constituted an important part of the bacterial population adhered to solid fractions, what can be derived from the competition relationships with native bacteria, present in high densities in this environment. These facts, taken together, emphasize the need for reliable quantitative and qualitative analysis tools for the evaluation of pathogenic microbial composition in sludge, which could represent an undefined risk to public health and ecosystem functions when considering its recycling.


NIDDM, impaired glucose tolerance, and pulmonary function in older adults. The Rancho Bernardo Study.
To determine whether NIDDM or plasma glucose level in subjects without diabetes is associated with reduced pulmonary function in 525 men and 714 women, 51-95 years of age. The analysis was based on data from a community-based study, the Rancho Bernardo Study. Between 1984 and 1987, 82% of community-dwelling residents had an oral glucose tolerance test. Between 1988 and 1991, 80% had lung function assessed by spirometry (forced expiratory volume in 1 s [FEV1] and forced vital capacity [FVC]). In analyses adjusted for age, height, and cigarette smoking, pulmonary function was not associated with known or newly diagnosed NIDDM in men or women. However, FEV1 and FVC were each independently reduced in men with diabetes of 10 or more years' duration. Fasting plasma glucose (FPG) levels were correlated with FEV1 and FVC in men without diabetes. No associations were found in women. The overall absence of an association of NIDDM with pulmonary function in these older adults may reflect survival bias and the small number of subjects with severe diabetes or diabetes of prolonged duration. The apparent relation of FPG levels to FEV1 and FVC suggests that any effect of glycemia precedes diabetes and contradicts any putative causal role for duration and severity of glycemia, however. More epidemiological studies are needed to provide further information about the relationship between NIDDM and lung function.


Tetrahydrobiopterin and dysfunction of endothelial nitric oxide synthase in coronary arteries.
The L-arginine/nitric oxide pathway plays a key role in the regulation of arterial tone. Biosynthesis of nitric oxide requires activation of nitric oxide synthase in the presence of tetrahydrobiopterin as a cofactor. Biochemical studies demonstrated that activation of purified nitric oxide synthase at suboptimal concentrations of tetrahydrobiopterin leads to production of hydrogen peroxide. The present experiments were designed to determine whether in coronary arteries inhibition of tetrahydrobiopterin synthesis may favor nitric oxide synthase-catalyzed production of hydrogen peroxide. Primary branches of canine left anterior descending artery were incubated for 6 hours in minimum essential medium in the presence or in the absence of the tetrahydrobiopterin synthesis inhibitor 2,4-diamino-6-hydroxypyrimidine (DAHP; 10(-2) mol/L). Arterial rings were suspended for isometric tension recording. Production of cGMP was measured by radioimmunoassay. Experiments were performed in the presence of indomethacin (10(-5) mol/L). During contractions to the thromboxane A2/prostaglandin H2 receptor agonist U46619 (10(-7) mol/L), calcium ionophore A23187 (10(-9) to 10(-6) mol/L) caused endothelium-dependent relaxations. A nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (3 x 10(-4) mol/L), significantly inhibited these relaxations. In DAHP-treated arteries, relaxations to A23187 and its stimulating effect on cGMP production were significantly reduced in the presence of catalase (1200 U/mL). By contrast, catalase did not exert any effect in rings incubated in the absence of DAHP. Furthermore, the inhibitory effect of catalase on A23187-induced relaxations was abolished when coronary arteries were incubated in the presence of DAHP plus a liposoluble analogue of tetrahydrobiopterin, 6-methyltetrahydropterin (10(-4) mol/L). The present study suggests that hydrogen peroxide may be a mediator of endothelium-dependent relaxations in coronary arteries depleted of tetrahydrobiopterin. This initially compensatory response, triggered by a dysfunctional nitric oxide synthase, may represent an important mechanism underlying oxidative vascular injury.


Development of a prediction model to identify undiagnosed chronic obstructive pulmonary disease patients in primary care settings in China.
At present, a large number of chronic obstructive pulmonary disease (COPD) patients are undiagnosed in China. Thus, this study aimed to develop a simple prediction model as a screening tool to identify patients at risk for COPD. The study was based on the data of 22,943 subjects aged 30 to 79 years and enrolled in the second resurvey of China Kadoorie Biobank during 2012 and 2013 in China. We stepwisely selected the predictors using logistic regression model. Then we tested the model validity through P-P graph, area under the receiver operating characteristic curve (AUROC), ten-fold cross validation and an external validation in a sample of 3492 individuals from the Enjoying Breathing Program in China. The final prediction model involved 14 independent variables, including age, sex, location (urban/rural), region, educational background, smoking status, smoking amount (pack-years), years of exposure to air pollution by cooking fuel, family history of COPD, history of tuberculosis, body mass index, shortness of breath, sputum and wheeze. The model showed an area under curve (AUC) of 0.72 (95% confidence interval [CI]: 0.72-0.73) for detecting undiagnosed COPD patients, with the cutoff of predicted probability of COPD=0.22, presenting a sensitivity of 70.13% and a specificity of 62.25%. The AUROC value for screening undiagnosed patients with clinically significant COPD was 0.68 (95% CI: 0.66-0.69). Moreover, the ten-fold cross validation reported an AUC of 0.72 (95% CI: 0.71-0.73), and the external validation presented an AUC of 0.69 (95% CI: 0.68-0.71). This prediction model can serve as a first-stage screening tool for undiagnosed COPD patients in primary care settings.


Yellow Fever Virus: Knowledge Gaps Impeding the Fight Against an Old Foe.
Yellow fever (YF) was one of the most dangerous infectious diseases of the 18th and 19th centuries, resulting in mass casualties in Africa and the Americas. The etiologic agent is yellow fever virus (YFV), and its live-attenuated form, YFV-17D, remains one of the most potent vaccines ever developed. During the first half of the 20th century, vaccination combined with mosquito control eradicated YFV transmission in urban areas. However, the recent 2016-2018 outbreaks in areas with historically low or no YFV activity have raised serious concerns for an estimated 400-500 million unvaccinated people who now live in at-risk areas. Once a forgotten disease, we highlight here that YF still represents a very real threat to human health and economies. As many gaps remain in our understanding of how YFV interacts with the human host and causes disease, there is an urgent need to address these knowledge gaps and propel YFV research forward.


Signal design and courtship presentation coincide for highly biased delivery of an iridescent butterfly mating signal.
Sensory drive theory contends that signaling systems should evolve to optimize transmission between senders and intended receivers, while minimizing visibility to eavesdroppers where possible. In visual communication systems, the high directionality afforded by iridescent coloration presents underappreciated avenues for mediating this trade-off. This hypothesis predicts functional links between signal design and presentation such that visual conspicuousness is maximized only under ecologically relevant settings and/or to select audiences. We addressed this prediction using Hypolimnas bolina, a butterfly in which males possess ultraviolet markings on their dorsal wing surfaces with a narrow angular reflectance function. Males bearing brighter dorsal markings are increasingly attractive to females, but also likely more conspicuous to predators. Our data indicate that, during courtship (and given the ritualized wingbeat dynamics at these times), males position themselves relative to females in such a way as to simultaneously maximize three components of known or putative signal conspicuousness: brightness, area, and iridescent flash. This suggests that male signal design and display have coevolved for the delivery of an optimally conspicuous signal to courted females. More broadly, these findings imply a potential signaling role for iridescence itself, and pose a novel example for how signal design may coevolve with the behavioral context of display.


ASKP1240, a fully human anti-CD40 monoclonal antibody, prolongs pancreatic islet allograft survival in nonhuman primates.
A strategy for inhibiting CD40 has been considered as an alternative approach for immunosuppression because of undesirable effects of anti-CD154 monoclonal antibodies (mAbs). Previously, we demonstrated that ASKP1240, which is a fully human anti-CD40 mAb, significantly prolonged kidney and liver allograft survival in cynomolgus monkeys without causing thromboembolic complications. Herein, we evaluated the effect of ASKP1240 on pancreatic islet transplantation (PITx) in cynomolgus monkeys. Diabetes was induced by total pancreatectomy, and islet allografts were transplanted into the liver. Following PITx (8201-12 438 IEQ/kg), blood glucose levels normalized promptly in all animals. Control islet allografts were rejected within 9 days (n = 3), whereas ASKP1240 (10 mg/kg) given on postoperative days 0, 4, 7, 11 and 14 (induction treatment, n = 5) significantly prolonged graft survival time (GST) to >15, >23, 210, 250 and >608 days, respectively. When ASKP1240 (5 mg/kg) was administered weekly thereafter up to post-PITx 6 months (maintenance treatment, n = 4), GST was markedly prolonged to >96, >115, 523 and >607 days. During the ASKP1240 treatment period, both anti-donor cellular responses and development of anti-donor antibodies were abolished, and no serious adverse events were noted. ASKP1240 appears to be a promising candidate for immunosuppression in clinical PITx.


High-resolution Greenland ice core data show abrupt climate change happens in few years.
The last two abrupt warmings at the onset of our present warm interglacial period, interrupted by the Younger Dryas cooling event, were investigated at high temporal resolution from the North Greenland Ice Core Project ice core. The deuterium excess, a proxy of Greenland precipitation moisture source, switched mode within 1 to 3 years over these transitions and initiated a more gradual change (over 50 years) of the Greenland air temperature, as recorded by stable water isotopes. The onsets of both abrupt Greenland warmings were slightly preceded by decreasing Greenland dust deposition, reflecting the wetting of Asian deserts. A northern shift of the Intertropical Convergence Zone could be the trigger of these abrupt shifts of Northern Hemisphere atmospheric circulation, resulting in changes of 2 to 4 kelvin in Greenland moisture source temperature from one year to the next.


Lrp5 functions in bone to regulate bone mass.
The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis.


Spatially congruent sites of importance for global shark and ray biodiversity.
Many important areas identified for conservation priorities focus on areas of high species richness, however, it is unclear whether these areas change depending on what aspect of richness is considered (e.g. evolutionary distinctiveness, endemicity, or threatened species). Furthermore, little is known of the extent of spatial congruency between biodiversity measures in the marine realm. Here, we used the distribution maps of all known marine sharks, rays, and chimaeras (class Chondrichthyes) to examine the extent of spatial congruency across the hotspots of three measures of species richness: total number of species, evolutionarily distinct species, and endemic species. We assessed the spatial congruency between hotspots considering all species, as well as on the subset of the threatened species only. We consider three definitions of hotspot (2.5%, 5%, and 10% of cells with the highest numbers of species) and three levels of spatial resolution (1°, 4°, and 8° grid cells). Overall, we found low congruency among all three measures of species richness, with the threatened species comprising a smaller subset of the overall species patterns irrespective of hotspot definition. Areas of congruency at 1° and 5% richest cells contain over half (64%) of all sharks and rays and occurred off the coasts of: (1) Northern Mexico Gulf of California, (2) USA Gulf of Mexico, (3) Ecuador, (4) Uruguay and southern Brazil, (5) South Africa, southern Mozambique, and southern Namibia, (6) Japan, Taiwan, and parts of southern China, and (7) eastern and western Australia. Coarsening resolution increases congruency two-fold for all species but remains relatively low for threatened measures, and geographic locations of congruent areas also change. Finally, for pairwise comparisons of biodiversity measures, evolutionarily distinct species richness had the highest overlap with total species richness regardless of resolution or definition of hotspot. We suggest that focusing conservation attention solely on areas of high total species richness will not necessarily contribute efforts towards species that are most at risk, nor will it protect other important dimensions of species richness.


Transcription factor NFE2L2/NRF2 modulates chaperone-mediated autophagy through the regulation of LAMP2A.
Chaperone-mediated autophagy (CMA) is a selective degradative process for cytosolic proteins that contributes to the maintenance of proteostasis. The signaling mechanisms that control CMA are not fully understood but might involve response to stress conditions including oxidative stress. Considering the role of CMA in redoxtasis and proteostasis, we sought to determine if the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid derived 2, like 2) has an impact on CMA modulation. In this work, we identified and validated 2 NFE2L2 binding sequences in the LAMP2 gene and demonstrated in several human and mouse cell types that NFE2L2 deficiency and overexpression was linked to reduced and increased LAMP2A levels, respectively. Accordingly, lysosomal LAMP2A levels were drastically reduced in nfe2l2-knockout hepatocytes, which also displayed a marked decrease in CMA activity. Oxidant challenge with paraquat or hydrogen peroxide, or pharmacological activation of NFE2L2 with sulforaphane or dimethyl fumarate also increased LAMP2A levels and CMA activity. Overall, our study identifies for the first time basal and inducible regulation of LAMP2A, and consequently CMA activity, by NFE2L2. ACTB: actin, beta, ARE: antioxidant response element; ATG5: autophagy related 5; BACH1: BTB domain and CNC homolog 1; ChIP: chromatin immunoprecipitation; CMA: chaperone-mediated autophagy; DHE: dihydroethidium; DMF: dimethyl fumarate; ENCODE: Encyclopedia of DNA elements at the University of California, Santa Cruz; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GBA: glucosylceramidase beta; GFP: green fluorescent protein; HMOX1: heme oxygenase 1; H2O2: hydrogen peroxide; HSPA8/HSC70: heat shock protein family A (Hsp70) member 8; KEAP1: kelch like ECH associated protein 1; LAMP2A: lysosomal associated membrane protein 2A; LAMP2B: lysosomal associated membrane protein 2B; LAMP2C: lysosomal associated membrane protein 2C; LAMP1: lysosomal associated membrane protein 1; MAFF: MAF bZIP transcription factor F; MAFK: MAF bZIP transcription factor K; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PQ: paraquat; PI: protease inhibitors; qRT-PCR: quantitative real-time polymerase chain reaction; RNASE: ribonuclease A family member; SFN: sulforaphane; SQSTM1/p62: sequestosome 1; TBP: TATA-box binding protein.


The stimulation of Na+, K+-ATPase activity in the medulla of the rat kidney by [arginine] vasopressin and its analogues.
Total and Na+, K+-ATPase activities have been measured in sections (10 micron thick) from blocks of rat kidney cultured for 5 h at 37 degrees C, pH 7.5, in Glasgow Eagle's Minimum Essential Medium. Synthetic [arginine]vasopressin [( Arg]VP) stimulated ATPase activity in the thick tubular cells of the renal outer medulla over the concentration range 0.01-10 fmol/l, but failed to affect ATPase activity in the proximal and distal convoluted tubules of the cortex. The increase in medullary total ATPase activity induced by [Arg]VP and its analogues was wholly due to stimulation of Na+, K+-ATPase activity. Stimulation of medullary ATPase activity was blocked by [Arg]VP antiserum. The [Arg]VP analogues desmopressin, [lysine]vasopressin, [arginine]vasotocin and oxytocin stimulated medullary Na+, K+-ATPase activity, the three last-named analogues being considerably less potent than [Arg]VP.


Blue-shifted contra-directional coupling between a periodic and conventional dielectric waveguides.
The interaction between a periodic and conventional dielectric waveguides is investigated theoretically for a two-dimensional model system. A modified coupled-mode theory is formulated for the considered system and found to agree well with rigorous numerical calculations. It is shown that in a certain wavelength range the contra-directional coupling between the two waveguides can be achieved with high efficiency. But the spectrum of the coupling efficiency is blue-shifted and thus the strongest coupling does not occur in the case when two individual waveguides have the same propagation constant. For such a contra-directional coupling system, the coupling efficiency grows with the coupling length and it tends to 100% (excluding insertion loss) when the coupling length is larger than a certain value, and the coupling window can be largely broaden by reducing the distance between the coupled waveguides.


Detection of human adaptation during the past 2000 years.
Detection of recent natural selection is a challenging problem in population genetics. Here we introduce the singleton density score (SDS), a method to infer very recent changes in allele frequencies from contemporary genome sequences. Applied to data from the UK10K Project, SDS reflects allele frequency changes in the ancestors of modern Britons during the past ~2000 to 3000 years. We see strong signals of selection at lactase and the major histocompatibility complex, and in favor of blond hair and blue eyes. For polygenic adaptation, we find that recent selection for increased height has driven allele frequency shifts across most of the genome. Moreover, we identify shifts associated with other complex traits, suggesting that polygenic adaptation has played a pervasive role in shaping genotypic and phenotypic variation in modern humans.


Impact of changing drug treatment and malaria endemicity on the heritability of malaria phenotypes in a longitudinal family-based cohort study.
Despite considerable success of genome wide association (GWA) studies in identifying causal variants for many human diseases, their success in unraveling the genetic basis to complex diseases has been more mitigated. Pathogen population structure may impact upon the infectious phenotype, especially with the intense short-term selective pressure that drug treatment exerts on pathogens. Rigorous analysis that accounts for repeated measures and disentangles the influence of genetic and environmental factors must be performed. Attempts should be made to consider whether pathogen diversity will impact upon host genetic responses to infection.We analyzed the heritability of two Plasmodium falciparum phenotypes, the number of clinical malaria episodes (PFA) and the proportion of these episodes positive for gametocytes (Pfgam), in a family-based cohort followed for 19 years, during which time there were four successive drug treatment regimes, with documented appearance of drug resistance. Repeated measures and variance components analyses were performed with fixed environmental, additive genetic, intra-individual and maternal effects for each drug period. Whilst there was a significant additive genetic effect underlying PFA during the first drug period of study, this was lost in subsequent periods. There was no additive genetic effect for Pfgam. The intra-individual effect increased significantly in the chloroquine period.The loss of an additive genetic effect following novel drug treatment may result in significant loss of power to detect genes in a GWA study. Prior genetic analysis must be a pre-requisite for more detailed GWA studies. The temporal changes in the individual genetic and the intra-individual estimates are consistent with those expected if there were specific host-parasite interactions. The complex basis to the human response to malaria parasite infection likely includes dominance/epistatic genetic effects encompassed within the intra-individual variance component. Evaluating their role in influencing the outcome of infection through host genotype by parasite genotype interactions warrants research effort.


Using sound-taste correspondences to enhance the subjective value of tasting experiences.
The soundscapes of those places where we eat and drink can influence our perception of taste. Here, we investigated whether contextual sound would enhance the subjective value of a tasting experience. The customers in a chocolate shop were invited to take part in an experiment in which they had to evaluate a chocolate's taste while listening to an auditory stimulus. Four different conditions were presented in a between-participants design. Envisioning a more ecological approach, a pre-recorded piece of popular music and the shop's own soundscape were used as the sonic stimuli. The results revealed that not only did the customers report having a significantly better tasting experience when the sounds were presented as part of the food's identity, but they were also willing to pay significantly more for the experience. The method outlined here paves a new approach to dealing with the design of multisensory tasting experiences, and gastronomic situations.


ERAMOSA controls lateral branching in snapdragon.
Plant forms display a wide variety of architectures, depending on the number of lateral branches, internode elongation and phyllotaxy. These are in turn determined by the number, the position and the fate of the Axillary Meristems (AMs). Mutants that affect AM determination during the vegetative phase have been isolated in several model plants. Among these genes, the GRAS transcription factor LATERAL SUPPRESSOR (Ls) plays a pivotal role in AM determination during the vegetative phase. Hereby we characterize the phylogenetic orthologue of Ls in Antirrhinum, ERAMOSA (ERA). Our data supported ERA control of AM formation during both the vegetative and the reproductive phase in snapdragon. A phylogenetic analysis combined with an analysis of the synteny of Ls in several species strongly supported the hypothesis that ERA is a phylogenetic orthologue of Ls, although it plays a broader role. During the reproductive phase ERA promotes the establishment of the stem niche at the bract axis but, after the reproductive transition, it is antagonized by the MADS box transcription factor SQUAMOSA (SQUA). Surprisingly double mutant era squa plants display a squa phenotype developing axillary meristems, which can eventually turn into inflorescences or flowers.


GLP-1 receptor activation modulates appetite- and reward-related brain areas in humans.
Gut-derived hormones, such as GLP-1, have been proposed to relay information to the brain to regulate appetite. GLP-1 receptor agonists, currently used for the treatment of type 2 diabetes (T2DM), improve glycemic control and stimulate satiety, leading to decreases in food intake and body weight. We hypothesized that food intake reduction after GLP-1 receptor activation is mediated through appetite- and reward-related brain areas. Obese T2DM patients and normoglycemic obese and lean individuals (n = 48) were studied in a randomized, crossover, placebo-controlled trial. Using functional MRI, we determined the acute effects of intravenous administration of the GLP-1 receptor agonist exenatide, with or without prior GLP-1 receptor blockade using exendin 9-39, on brain responses to food pictures during a somatostatin pancreatic-pituitary clamp. Obese T2DM patients and normoglycemic obese versus lean subjects showed increased brain responses to food pictures in appetite- and reward-related brain regions (insula and amygdala). Exenatide versus placebo decreased food intake and food-related brain responses in T2DM patients and obese subjects (in insula, amygdala, putamen, and orbitofrontal cortex). These effects were largely blocked by prior GLP-1 receptor blockade using exendin 9-39. Our findings provide novel insights into the mechanisms by which GLP-1 regulates food intake and how GLP-1 receptor agonists cause weight loss.


Increased prevalence of carotid artery atherosclerosis in rheumatoid arthritis is artery-specific.
Cardiovascular (CV) morbidity and mortality are increased in rheumatoid arthritis (RA). Prior investigations of the association of RA with measures of carotid atherosclerosis have yielded conflicting results. We compared carotid intima-media thickness (IMT) of both the common carotid (CCA) and proximal internal carotid (bulb-ICA) arteries, and plaque prevalence, between RA and non-RA participants. Subjects with RA were participants in a cohort study of subclinical CV disease in RA. Non-RA controls were selected from the Multi-Ethnic Study of Atherosclerosis. Both groups underwent B-mode ultrasonography of the right and left CCA and bulb-ICA. Linear regression was used to model the association of RA status with CCA and bulb-ICA-IMT, and logistic regression for the association of RA status with plaque. We compared 195 RA patients to 198 non-RA controls. CV risk factors were similarly distributed, except for a higher prevalence of hypertension in the RA group. Mean adjusted bulb-ICA-IMT was higher in RA patients than controls (1.16 vs 1.02 mm, respectively; p < 0.001), while mean adjusted CCA-IMT did not differ significantly. After adjusting for CV risk factors, the odds of plaque were significantly increased in RA participants compared to controls (OR 2.41, 95% CI 1.26-4.61). The association of gender, age, smoking, and hypertension with bulb-ICA-IMT and plaque did not significantly differ by RA status. Interleukin 6 was strongly associated with bulb-ICA-IMT and plaque in controls but not in RA patients. In the RA group, shared epitope was associated with an increased prevalence of plaque. Compared to controls, RA was associated with a higher prevalence and higher severity of atherosclerosis in the bulb-ICA but not the CCA. Our data suggest that future studies in RA that utilize carotid artery measurements should include assessment of the bulb-ICA.