A stationary  association between El Niño Southern Oscillation (ENSO), the Indian Ocean Dipole (IOD) and epidemics of cholera in Bangladesh has been widely assumed. However, whether or not elements of the local climate that are relevant for cholera transmission have stationary signatures of the IOD on their dynamics over different time scales is still not clear. Here we report results on the time-varying relationships between the various remote and local environmental drivers and cholera incidence in Bangladesh.We performed a cross wavelet coherency analysis to examine patterns of association between monthly cholera cases in the hospitals in Dhaka and Matlab (1983–2008) and indices for both IOD and ENSO. Our results showed that the strength of both the IOD and ENSO associations with cholera hospitalizations changed across time scales during the study period. In Dhaka, 4-year long coherent cycles were observed between cholera and the index of IOD in 1988–1997. In Matlab, the effect of ENSO was more dominant while there was no evidence for an IOD effect on cholera hospitalizations.Our results call for the consideration of non-stationary, possibly non-linear, patterns of association between cholera hospitalizations and climatic factors in cholera epidemic early warning systems. Vibrio cholerae, the bacterium that causes the disease, is known to inhabit riverine, estuarine and coastal ecosystems with specific abiotic  and biotic components  V. cholerae has an increased growth rate in aquatic environments with warmer temperatures, particularly when combined with a high pH and blooms of phytoplankton, aquatic plants or algae Cholera remains a major public health problem in many places, including Bangladesh, India, and a number of countries in Africa and South America V. cholerae multiplication It has been reported that the El Niño-Southern Oscillation (ENSO) plays a role in the interannual variation of endemic cholera in Bangladesh The Indian Ocean Dipole (IOD) is another climate mode that arises from ocean-atmosphere interactions that cause interannual climate variability in the tropical Indian Ocean Wavelet analysis is useful for epidemiological time series analyses mainly because of the non-stationarity of associations between the disease dynamics and exposure covariates Dhaka: The ICDDR,B hospital serves a large urban population within the city of Dhaka and provides free treatment for more than 100,000 cases of diarrhea each year. A surveillance system was established at the ICDDR,B in 1979 to systematically sample children and adults with diarrheal illnesses th patient was enrolled in the surveillance system, and since 1996 every 50th patient has been enrolled. Matlab. Matlab is a rural and riverine delta area situated approximately 57 km southeast of Dhaka. Every hospital visit of patients residing within the area covered by the Health and Demographic Surveillance System (HDSS), which consists of 142 villages with populations over 220,000, was registered in the hospital surveillance system The primary outcome for this study was the monthly number of patients with cholera who attended the International Centre for Diarrhoeal Disease Research, Bangladesh  hospitals in Dhaka and Matlab . Dhaka: V. cholerae. The details of the laboratory procedures were described in a previous study V. cholerae of the serogroups O1 or O139 was identified in the stool specimen, regardless of the presence of other pathogens. From the surveillance data that had been collected over a 26-year period (January 1983 to December 2008) for Dhaka and 28-year period (January 1981 to December 2008) for Matlab, we retrieved the date of the hospital visit and the pathogens identified in each stool specimen. From these data, the monthly cholera cases in Dhaka and Matlab were counted separately and used for analysis.For each diarrhea patient of the ICDDR,B hospitals in Dhaka and Matlab, the stool was microbiologically examined to identify common enteric pathogens including www.jamstec.go.jp/frcgc/research/d1/iod/). The DMI values were calculated using the SST data from the HadISST dataset (http://www.metoffice.gov.uk/hadobs/hadisst/). The base period for calculating anomalies was 1958–2008. These values were standardized to zero mean and unit standard deviation. The strength of the ENSO was measured by monthly average SSTs in the Niño 3 region  (Nino3) in the Pacific Ocean that we derived from the National Oceanic and Atmospheric Administration (NOAA) Climate Prediction Center data (http://www.cpc.ncep.noaa.gov). We examined the association between DMI and monthly average SST in the northern Bay of Bengal to gain some insight into the causal pathways linking the IOD to the number of cholera cases. Mean monthly SSTs in the Bay of Bengal  were derived from the NOAA Optimum Interpolation Sea Surface Temperature dataset The strength of the IOD was measured by the dipole mode index (DMI), defined as the difference in SST between the western  and eastern  tropical Indian Ocean The daily rainfall and maximum and minimum temperature data for Dhaka and Matlab were obtained from the Bangladesh Meteorological Department. The data for the daily river levels of the Brigonga (Millbarrak in Dhaka) and Danagoda (Matlab Bazaar) rivers were obtained from the records maintained by the Bangladesh Water Development Board. The monthly means for the maximum temperature (°C) and maximum river level (m), and the total monthly rainfall (mm) were calculated from the daily records. Missing values (17 [5.4%] in Dhaka and 6 [1.9%] in Matlab) for river levels were imputed using the average values of the same month in the other years over the whole time series.Temporal patterns of association in the time series were studied using the continuous wavelet transform 0 = 6 months , a total smoothing window of 31  and parameter W0 = 6 (dimensionless parameter of the Morlet Wavelet). Further details about cross wavelets and software are described in Maraun and Kurths V. cholerae El Tor O1 biotype strain and one for the O139 Bengal strain.We also estimated the cone of influence, where inferences from the wavelet analysis outside the cone of influence are not valid because of the manipulations that were performed to generate the wavelet spectrum in the absence of data for larger period frequencies We did not stratify the analysis by age, because the seasonal patterns were the same and there was no likely reason to expect different cholera-climate associations between the age groups at interannual time scales.The associations between DMI, Nino3 and monthly average temperature, rainfall and SST in the northern Bay of Bengal were also examined by wavelet coherence analysis. All the analyses were performed with R software (version 2.0).The time series for the number of cholera patients per month, by serogroup and biotype, ambient temperature, rainfall and river level during the study period are shown for Dhaka  and MatlThe time series for Dipole Mode Index (DMI), Nino3 and sea surface temperature (SST) during the same period are shown in Four-year long coherent cycles between O1 cholera and DMI were seen in 1988–1997 in Dhaka  while thV. cholerae were similar to the result for the El Tor O1 biotype strain alone because the frequency of the O1 biotype was dominant during the study period  in the Bay of Bengal The association between Dipole Mode Index (DMI) and cholera hospitalizations that we found in Dhaka was not observed in Matlab, while a consistent association between Nino3 and cholera hospitalizations was observed only in Matlab. The associations that we found between DMI and Nino3 with cholera are broadly consistent with previous finding There are some limitations in this study. First, less severe cases of cholera are less likely to have been included in the surveillance data; however, this should not pose a threat to the validity of the comparison over time, which was the subject of this study. Second, some cases could have been missed because of limitations in the capacity of the hospitals to receive the patients; particularly, during epidemics of cholera. However, in principle, the hospitals accept all patients who visit the hospital. Thus, the capacity of the hospitals should not be an important limitation of this study. Third, the wavelet analysis explores a periodical synchronicity between two time series, and so has no bearing on mechanisms. Further environmental and microbiological studies are necessary to elucidate the causal pathways of the associations.Because of the serious global consequences of cholera and its sensitivity to climate, the World Health Organization has proposed that an early warning system for cholera epidemics using climatic parameters could be developed Figure S1Monthly time series of cholera hospitalizations by (a) serotype O1 & O139 and (b) bio-type El Tor & Classical for Dhaka (January 1983–December 2008).(TIF)Click here for additional data file.Figure S2Monthly time series of cholera hospitalizations by (a) serotype O1 & O139 and (b) bio-type El Tor & Classical for Matlab (November 1981–December 2008).(TIF)Click here for additional data file.Figure S3Cross wavelet coherence of global and local climatic time series with cholera in Dhaka and Matlab. (A) Dipole mode index (DMI) and cholera in Dhaka; (B) DMI and cholera in Matlab; (C) Nino3 and cholera in Dhaka; (D) Nino3 and cholera in Matlab; (E) Sea surface temperature (SST) in the Bay of Bengal (°C) and cholera in Dhaka; (F) SST in the Bay of Bengal (°C) and cholera in Matlab; (G) Rainfall (mm) and cholera in Dhaka; (H) Rainfall (mm) and cholera in Matlab; (I) Temperature (°C) and cholera in Dhaka; (J) Temperature (°C) and cholera in Matlab; (K) River level (m) and cholera in Dhaka; (L) River level (m) and cholera in Matlab. Time series span from January 1983 to December 2008.(TIF)Click here for additional data file.Figure S4Cross wavelet coherence of global and local climatic time series with classical biotype and El Tor cholera in Dhaka. (A) Dipole mode index (DMI) and classical cholera; (B) DMI and El Tor cholera; (C) Nino3 and classical cholera; (D) Nino3 and El Tor cholera; (E) Sea surface temperature (SST) in the Bay of Bengal (°C) and classical cholera; (F) SST in the Bay of Bengal (°C) and El Tor cholera; (G) Rainfall (mm) and classical cholera; (H) Rainfall (mm) and El Tor cholera; (I) Temperature (°C) and classical cholera; (J) Temperature (°C) and El Tor cholera; (K) River level (m) and classical cholera; (L) River level (m) and El Tor cholera. Time series span from January 1983 to December 2008.(TIF)Click here for additional data file.Figure S5Cross wavelet coherence of global and local climatic time series with classical biotype and El Tor cholera in Matlab. (A) Dipole mode index (DMI) and classical cholera; (B) DMI and El Tor cholera; (C) Nino3 and classical cholera; (D) Nino3 and El Tor cholera; (E) Sea surface temperature (SST) in the Bay of Bengal (°C) and classical cholera; (F) SST in the Bay of Bengal (°C) and El Tor cholera; (G) Rainfall (mm) and classical cholera; (H) Rainfall (mm) and El Tor cholera; (I) Temperature (°C) and classical cholera; (J) Temperature (°C) and El Tor cholera; (K) River level (m) and classical cholera; (L) River level (m) and El Tor cholera. Time series span from November 1981 to December 2008, with the exception of river level which begins in January 1983.(TIF)Click here for additional data file.


As Breast Cancer and Environment Research Center (BCERC) project leaders, we would like to address what we believe represents inaccuracies and omissions in the recent article by The LIBCSP has been enormously productive, continuing even now, with > 100 scientific publications and $21 million in grant funding using LIBCSP resources. ” 16 times, without noting the impressive contributions of the BCERC Community Outreach and Translation Cores (COTC) projects. Advocacy and COTC in the BCERC since 2003 have resulted in extensive and innovative dissemination of knowledge and new ideas (ew ideas . Mutual As noted by


Sudden cardiac death (SCD) is responsible for 11% of all deaths and 50% of all cardiovascular deaths. Hypertrophic cardiomyopathy (HCM) is the most common cause of SCD among the young patients. SCD is often the first manifestation of the disease. There are 5 major risk factors for SCD in patients with HCM. However, no single risk factor can be used as a screening test in assessment of SCD. Nowadays new non-invasive risk factors, such as T-wave alternation (TWA), are being studied. They may provide important information on the patient’s susceptibility to life threatening arrhythmias.The study involved 33 hypertensive heart disease (HHD) patients and 22 HCM patients. All patients underwent echocardiography test (ECHO) and 24h Holter monitoring with mTWA evaluation using the modified moving average method. The evaluation involved maximum mTWA figures in 24h (mTWA max), mTWA figures at the heart rate of 100/min (mTWA 100) and at 5:00 am (mTWA 5:00).The common group demonstrated a weak direct correlation between the thickness of ventricular septum (IVSth) and mTWA max, as well as a weak reverse correlation between the left ventricular posterior wall thickness (LVPWth) and mTWA 100. These correlation were revealed primarily in HHD patients who demonstrated correlation between mTWA max and IVSth, between mTWA 5:00 and IVSth, LVPWth, as well as between mTWA 100 and LVPWth. The HCM patients did not demonstrate correlation between mTWA and the thickness of left ventricular wall. The patients with non-obstructive HCM (n=15) demonstrated significantly higher mTWA 100 in comparison to the patients with obstructive HCM (n=5). The presence of higher SCD risk factors in the HCM patients (n=6) did not influenced mTWA figures. The mTWA figures in the HHD and HCM patients differed insignificantly, with the only exception of higher mTWA 5:00 in the HHD in comparison to the HCM group.mTWA figures correlate with the degree of left ventricular hypertrophy, however the revealed correlations are weak. The causes of lower MTWA 100 in patients with more intensive hypertrophy (reverse correlation) as well as in patients with obstructive HCM in comparison with non-obstructive HCM call for additional studies.


Bordetella pertussis (DTwP). In contrast, similar studies suggest that many African females and males may have their lives saved each year by the nonspecific immunological benefits of Bacillus Calmette-Guerin (BCG) vaccination. From an immunological point of view, we hypothesise that the adverse effects of DTwP vaccine may occur because of the Th2-polarising effect of the aluminium phosphate adjuvant in the vaccine and because intramuscular administration of the vaccine may cause chronic inflammation at the site of injection. However, the Th1-polarising effect of BCG is likely to be beneficial. Sexual dimorphism affecting immune functions and vitamin A supplementation may influence both the deleterious and beneficial nonspecific effects of immunisation.A number of mainly observational studies suggest that many African females below the age of one year die each year from the nonspecific effects of vaccination with diphtheria-tetanus toxoids and killed (whole-cell)  The paper discusses the immunology behind the reported nonspecific effects of DTwP and BCG vaccination: increased childhood mortality rates after DTwP and decreased mortality rates after BCG vaccination given during the course of the WHO recommended vaccine programs implemented in Guinea-Bissau and other low-income countries , 2. BothBordetella pertussis vaccine. A typical DTwP vaccine dose (0.5 mL) from the Serum Institute of India contains diphtheria toxoid (25 Lf), tetanus toxoid (5 Lf), and pertussis toxoid (4 IU), aluminium phosphate (1.5 mg), and a preservative, thiomersal (0.01%). Aluminium phosphate acts primarily as an antigen-adsorbing and Th2-polarizing adjuvant γ and TNFIn spite of numerous studies, there is still only weak evidence for any effect of micronutrient supplementation on vaccine efficacy (reviewed in ). Howeve The deleterious effect of DTwP vaccination in infant females and the beneficial outcomes of BCG vaccination might reflect nonspecific immune-polarizing and nonintended regulatory effects of vaccines and sex hormones in early childhood. The data from West Africa on the nonspecific effects of DTwP and BCG vaccination in young children discussed here suggest (1) replacement of intramuscular DTwP vaccination with cutaneous vaccination or development of a DTwP vaccine live bacterial carrier such as BCG  which maIn low-income countries with high rates of childhood mortality, the nonspecific effects of routine vaccinations are likely to be of increased importance due to the high disease burden. The nonspecific vaccine effects should therefore be a high-priority research area and be considered in the future planning of immunization programs in these countries to enhance the beneficial impact of immunizations on child survival.The author has no relevant affiliations or financial involvement with any organization of entity with financial interest in or financial conflict with the subject matter discussed in this paper. No writing assistance was utilized in the production of this paper.


Transient receptor potential channels are important mediators of thermal and mechanical stimuli and play an important role in neuropathic pain. The contribution of hereditary variants in the genes of transient receptor potential channels to neuropathic pain is unknown. We investigated the frequency of transient receptor potential ankyrin 1, transient receptor potential melastin 8 and transient receptor potential vanilloid 1 single nucleotide polymorphisms and their impact on somatosensory abnormalities in neuropathic pain patients. Within the German Research Network on Neuropathic Pain (Deutscher Forscbungsverbund Neuropathischer Schmerz) 371 neuropathic pain patients were phenotypically characterized using standardized quantitative sensory testing. Pyrosequencing was employed to determine a total of eleven single nucleotide polymorphisms in transient receptor potential channel genes of the neuropathic pain patients and a cohort of 253 German healthy volunteers. Associations of quantitative sensory testing parameters and single nucleotide polymorphisms between and within groups and subgroups, based on sensory phenotypes, were analyzed. Single nucleotide polymorphisms frequencies did not differ between both the cohorts. However, in neuropathic pain patients transient receptor potential ankyrin 1 710G>A  was associated with the presence of paradoxical heat sensation (p = 0.03), and transient receptor potential vanilloid 1 1911A>G  with cold hypoalgesia (p = 0.0035). Two main subgroups characterized by preserved (1) and impaired (2) sensory function were identified. In subgroup 1 transient receptor potential vanilloid 1 1911A>G led to significantly less heat hyperalgesia, pinprick hyperalgesia and mechanical hypaesthesia  and transient receptor potential vanilloid 1 1103C>G  to cold hypaesthesia (p = 0.002), but there was absence of associations in subgroup 2. In this study we found no evidence that genetic variants of transient receptor potential channels are involved in the expression of neuropathic pain, but transient receptor potential channel polymorphisms contributed significantly to the somatosensory abnormalities of neuropathic pain patients. Neuropathic pain arises after lesions or diseases of the somatosensory nervous system and involves multiple somatosensory phenomena. In addition to spontaneous pain, these include negative and positive symptoms and signs such as hypaesthesia, hypoalgesia, thermal and mechanical hyperalgesia and allodynia The understanding of the molecular basis of neuropathic pain has been expanded by the cloning and characterization of the transient receptor potential (TRP) family of voltage-gated ion channels. Among these the TRP vanilloid 1 (TRPV1), TRP melastin 8 (TRPM8) and TRP ankyrin 1 (TRPA1) subfamilies have been found to play important roles in transduction and sensitisation in primary afferent somatosensory neurons Beside these physiological functions, TRPs are thought to play an important role in the generation of neuropathic pain, as demonstrated by in vitro and in vivo animal studies and neurophysiological and psychophysical studies in human experimental pain models and pain patients However, the contribution of a genetic variability in TRP channels on pain perception and pain generation in neuropathic pain patients is unknown. In this study we describe the results of an association analysis of polymorphisms in TRPV1, TRPM8 and TRPA1 genes with somatosensory signs of neuropathic pain patients. The aims were (1) to determine the frequencies of TRP variants in a large group of neuropathic pain patients of different aetiology and a cohort of healthy volunteers and (2) to determine the relationship of TRP gene variants and somatosensory function in neuropathic pain patients. Within this study we could demonstrate significant associations between the somatosensory function and TRP gene variants in neuropathic pain patients but no differences in the TRP frequencies compared with healthy volunteers.All subjects gave their written informed consent and the study was approved by the ethics committee of the Medical Faculty of the Christian-Albrechts-University Kiel and subsequently the Ethic Committees of the other participating DFNS centers and performed according to the Declaration of Helsinki.www.neuro.med.tu-muenchen.de/dfns/) were included in the study . The patients suffered from complex regional pain syndrome, postherpetic neuralgia, peripheral nerve injury, trigeminal neuropathy, polyneuropathy, central pain and neuropathies that did not fulfill diagnostic criteria of the above mentioned syndromes as diagnosed by expert physicians. Patient characteristics are listed in A total of 371 patients with chronic neuropathic pain syndromes, all Caucasians, who participated in the German Research Network on Neuropathic Pain  established by the DFNS to determinate somatosensory function and signs of pain Whole genomic DNA was extracted from venous blood samples obtained from patients and volunteers using the Qiagen Gentra Puregene Blood Kit . The final concentration of gDNA was determined spectrophotometrically. Two SNPs in TRPV1, six in TRPM8 and three in TRPA1 genes were identified by literature review in the context of neuropathic pain and selected for detailed association analysis . Except Before statistical analysis the QST values that did not show normal distribution  were transformed logarithmically In order to identify major subpopulations of patients who are characterized by a typical combination of signs, a cluster analysis was performed in those patients only from whom complete QST results were available  and for bi-allelic polymorphisms each subtest and genotype were calculated using analysis of variance (ANOVA). For dominant and recessive models the t-test was used, if applicable. To correct for multiple testing each test was performed using 100,000 permutations.The frequency distribution of all polymorphisms investigated was in Hardy-Weinberg equilibrium. Moreover, after correcting for multiple comparisons, the frequency of all SNPs did not differ significantly between neuropathic pain patients, healthy controls and data from the HapMap project .First, within the group of patients who suffered from paradoxical heat sensation (PHS) heterozygous and homozygous carriers of the TRPA1 710G>A variant were significantly under-represented as compared to neuropathic pain patients without PHS  . Second,Hierarchical cluster analysis detected two main clusters . ClusterZ-scores of all QST tests were stratified to all TRP variants investigated in the entire sample and in both clusters. After correcting for multiple comparisons, two TRPV1 polymorphisms, 1911A>G and 1103C>G, were identified that had a significant relationship with the somatosensory function in cluster 1 .The TRPV1 1911A>G polymorphism was significantly associated with altered heat pain thresholds (HPT). Neuropathic pain patients being heterozygote or with wild type TRPV1 1911G genotype (AA or AG) tended to show heat hyperalgesia whereas TRPV1 1911G homozygotes exhibited significantly higher, i.e. normal heat pain thresholds . Homozygous variant carriers (GG) exhibited cold hypaesthesia compared with heterozygous or wild type carriers Click here for additional data file.


The intestine and the intestinal immune system have evolved through a symbiotic homeostasis under which a highly diverse microbial flora is maintained in the gastrointestinal tract while pathogenic bacteria are recognized and eliminated. Disruption of the balance between the immune system and the gut microbiota results in the development of multiple pathologies in humans. Inflammatory bowel diseases (IBD) have been associated with alterations in the composition of intestinal flora but whether these changes are causal or result of inflammation is still under dispute. Various chemical and genetic models of IBD have been developed and utilized to elucidate the complex relationship between intestinal epithelium, immune system and the gut microbiota. In this review we describe some of the most commonly used mouse models of colitis and Crohn's disease (CD) and summarize the current knowledge of how changes in microbiota composition may affect intestinal disease pathogenesis. The pursuit of gut-microbiota interactions will no doubt continue to provide invaluable insight into the complex biology of IBD. The lower gastrointestinal tract of healthy adult humans contains more than 100 trillion bacteria  and the impact of microbiota on these pathologies Figure .+ and CD8+ T lymphocytes, regulatory T cells, neutrophils and macrophages, resembling the pathogenic events in human colitis. Mucosal macrophages may prime the local inflammatory response through both phagocytosis of DSS and activation by bacteria products. The contribution of macrophage polarization phenotype to the development of CAC has been described using this model including the demonstration that Akt2 deficient mice are partly protected from DSS-induced colitis because of a macrophage phenotype shift from M1 to M2 in the colonic mucosa . DSS administered to the drinking water in repeated cycles triggers a state of chronic intestinal inflammation by binding to medium-chain-length fatty acids present in the mouse colon, inducing disruption of colonic epithelial barrier  by cytochrome P450 . DMH is metabolized in liver and its derivatives induce the production of diazonium by gut epithelial cells. The aforementioned metabolite exerts mutagenic effects through oxidative stress and methylation events  initiates acute T cell-mediated, IL-12 driven intestinal inflammation  and develops numerous adenomas. Exposure of Apcmin/+ mice to DSS alone mimics CAC and results in accelerated tumorigenesis  gene in humans are critically involved in familial adenomatous polyposis (FAP) and represent an early genetic aberration in sporadic colorectal cancer  can be adoptively transferred into immunodeficient SCID or RAG1/2−/− mice, where they traffic to the intestine and induce gut inflammation. Recipient mice repopulated with CD4+CD45RBLo T cells or total CD4+ T lymphocytes do not develop colitis, despite their ability to colonize the host gut. This phenomenon is attributed to the presence of CD25+FoxP3+ regulatory T cells within the CD4+CD45RBLo population  conditions  results in spontaneous colitis which is transmissible to wild-type animals (which express T-bet) upon cross-fostering or co-housing , which harbors 8 genes and 5 micro RNAs and confers protection against H. hepaticus-induced chronic colitis and inflammation-driven colon cancer  activity. Resident Treg produce IL-10 which inhibits Th1 cells and monocyte effector functions associated with inflammation. Suppression of Treg activity thereby unleashes inflammation, leading to a switch in the differentiation program of Ly6Chi monocytes from anti-inflammatory M2 macrophages to inflammatory dendritic cells and M1 macrophages in the colon   microbes or inhibitors of specific microbes and/or their products which “normalize” the intestinal flora and can improve human health. As the current repertoire of probiotics is limited, further studies to explore the potential of fecal microbiota transplantation (FMT) therapy, the infusion of fecal bacteria from a healthy individual into a recipient patient, for the treatment of intestinal disorders are warranted. FMT has demonstrated tremendous efficacy in treating refractory The microbiome plays an important role in immunity and energy metabolism and will thus be important to determine if the microbial gut ecology may also impact on non-gastrointestinal diseases, including obesity, cancer and neurological disorders.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Early diagnosis of macrophage activations syndrome (MAS) in systemic juvenile idiopathic arthritis (sJIA) may be challenging because it may mimic the clinical features of the underlying disease or be confused with an infectious complication. However, the diagnostic value of the guidelines for hemophagocytic lymphohistiocytosis (HLH) (1) or sJIA-associated MAS (2) has seldom been examined.To investigate the sensitivity and specificity of diagnostic guidelines for HLH and sJIA-associated MAS in patients with sJIA who developed MAS.The study sample included 155 children with sJIA who had MAS (diagnosed and treated as such by the attending physician) and 2 control groups with potentially “confusable” conditions, including active sJIA without MAS (n=303) and a systemic febrile infection requiring hospitalization (n=191). Diagnostic guidelines for HLH and sJIA-associated MAS were applied to all MAS and control patients. Because no patient had NK-cell activity and soluble CD25 determination available and bone marrow aspirate was performed in only a few patients, these 3 criteria were excluded from HLH guidelines. HLH criteria were, therefore, met when at least 4 of the 5 remaining variables were present. sJIA-associated MAS criteria were met when at least 2 laboratory criteria or at least 1 laboratory criterion and 1 clinical criterion were present. Sensitivity and specificity of guidelines in discriminating patients with MAS from control patients were assessed.The table shows the comparison of sensitivity and specificity of diagnostic guidelines.The diagnostic guidelines for sJIA-associated MAS revealed strong sensitivity and specificity, whereas HLH guidelines were highly specific, but lacked sensitivity. Sensitivity of HLH was mostly hampered by the excessive stringent threshold for cytopenia and hypofibrinogenemia, and the infrequent occurrence of splenomegaly in patients with MAS.


Pain is a common problem for patients admitted to the ICU, causing patient discomfort, agitation and accidental self-extubation. For this reason the recognition of pain and its severity is extremely important. Several pain scores and protocols are in use. We aimed to elucidate current practice of pain measurements and treatment in Dutch ICUs.In March 2011, a questionnaire was sent to all Dutch adult ICUs irrespective of the number of ICU beds with active follow-up by telephone calls to optimize the participation rate.A total of 84 ICUs (84/107) returned the survey, representing a response rate of 87%. Most ICUs are community teaching hospitals and nonteaching hospitals (85%) in comparison to academic hospitals (15%). Most ICUs (94%) use a standardized pain score in the group of patients who are capable of verbal communication: the Visual Analogue Scale (57%), Numerical Rating Scale (48%) and Faces Pain Scale (5%) being the most frequently used scores. In the group of patients who are unable to communicate, ICUs less frequently use pain scores (19%), with the Critical-Care Pain Observation Tool (6%) and Behaviour Pain Scale (5%) being used most frequently. Measurement of pain was considered most important for patients with burn wounds (67%), trauma patients (64%), postoperative patients (57%) and those who receive end-of-life care (64%). Barriers to use pain measurements included the patient's inability to communicate (82%), interference with pain assessment due to sedation (79%), hemodynamic instability (64%), insufficient dosages of analgesics (60%) and the unavailability of a standard pain scoring system (51%). In addition, guidelines for management of sedation and analgesics from the Netherlands Association for Intensive Care (NVIC) had been read by only 20% of the respondents. Factors that were mentioned to be useful in contributing to an improvement in pain assessment and effective pain control included adequate analgesic dosage (87%), utilization of protocols and directives (86%), enthusiastic and motivated personnel (81%) and the utilization of standardized pain measurement tools.Most Dutch ICUs measure pain frequently (94%) in patients who are able to communicate. However, in the group of patients who cannot communicate only 19% of the Dutch ICUs use a standardized pain score. This finding applied to both academic and nonacademic ICUs, which suggests that efforts should be put into implementing pain measures in Dutch ICUs.


Critical transitions between alternative stable states have been shown to occur across an array of complex systems. While our ability to identify abrupt regime shifts in natural ecosystems has improved, detection of potential early-warning signals previous to such shifts is still very limited. Using real monitoring data of a key ecosystem component, we here apply multiple early-warning indicators in order to assess their ability to forewarn a major ecosystem regime shift in the Central Baltic Sea. We show that some indicators and methods can result in clear early-warning signals, while other methods may have limited utility in ecosystem-based management as they show no or weak potential for early-warning. We therefore propose a multiple method approach for early detection of ecosystem regime shifts in monitoring data that may be useful in informing timely management actions in the face of ecosystem change. Transitions between alternative states, i.e., regime shifts, have been shown to occur across an array of complex systems Several early-warning indicators have been proposed to describe the temporal dynamics of complex systems close to a critical transition Pseudocalanus acuspes and Acartia spp., two key indicator species significantly contributing to the reorganization of the Baltic Sea ecosystem  and sprat (Sprattus sprattus) Pseudocalanus acuspes and Acartia spp. serve as suitable indicators for the ecosystem regime shift in the Central Baltic Sea. Furthermore, long-term temporally and spatially resolved monitoring programs Pseudocalanus acuspes and Acartia spp. biomass covering the period 1960–2008. Since the primary aim of the study is to investigate and evaluate potential early-warning signals, we chose to focus on and present results on method performance during the time-period immediately preceding and following the regime shift . We used data representing spring as this is the main reproductive season of the copepods We used Pseudocalanus acuspes and Acartia spp. within a 10, 15 and 20-year sliding window, following the approach by Dakos et al. We measured temporal variance  and autocorrelation at lag-1 (AR(1)) in detrended time series of average spring biomass of f′′) as a proxy for statistically significant acceleration in the rate of change (slope) of ecological time series a priori specified degree of smoothing In a second set of methods we applied statistical methods for assessing recent trends in the zooplankton time series. Although not specifically designed for early detection of regime shifts, the idea of using trend analysis as an early-warning signal lies in the possibility of detecting a slight increase in the rate of change (either in an upward or downward trend) in advance of a critical transition in an ecological time series. The approach is based on fitting non-linear Generalized Additive Models (GAM) The third approach was not primarily designed for early detection either, but rather for the identification and detection of regime shift. It is based on the evaluation of statistical time series models including structural breaks and combines several statistical indicators into a so-called “shiftogram” per se but to assess how under the constraints of real ecological time series similar to ours, different approaches may or may not work, and which assumptions and drawbacks may pose the greatest challenges in terms of early-warning detection and applicability to management.The full set of early-warning methods were assessed in terms of (i) the potential for detecting early-warnings signals in the selected indicator time series, (ii) how far in advance early-warning signals could potentially be detected, (iii) associated methodological assumptions and drawbacks influencing early-detection and (iv) applicability to real ecosystem management in terms of data requirements. While the trend analysis and shiftogram approach may quantitatively evaluate the first two criteria , no predefined reference levels exist to objectively assess the performance of our ecological indicators of critical slowing down, nor the possibility to theoretically crash test the methods against a simulated (modeled) spatio-temporal data set Pseudocalanus acuspes  strongly increased two years before the regime shift in 1988  one year ahead of the regime shift  correlation showed p-values exceeding their upper confidence intervals one year in advance of the regime shift for both species . This mate-1980s , may simContrary to temporal indicators of slowing down, spatial approaches for detecting changes in correlation and variance patterns Pseudocalanus acuspes and Acartia spp. prior to the regime shift (f′ (slope) and a positive f′′ (acceleration). As in the previous example, no significant trend or change point was detected when excluding also 1987.The trend analysis me shift . When reme shift , while wme shift , as illuThe trend analysis seems to be highly dependent on the length of time series and the numbers of regression splines used during fitting of the generalized additive models GAMs; . These aPseudocalanus acuspes both the Akaike’s information criterion (AICC) and the p-joint significance test showed a gradual transition towards a local minimum  3–4 years before the regime shift  transitions or more gradual changes strongly influence the shape  of the local minimum and hence the degree to which these transitions can be detected sufficiently in advance; a difference illustrated by the abrupt decrease in tia spp.  and the  acuspes . In addiThe versatility of methods for early detection of regime shifts in ecological time series provide an important toolbox for scientists and ecosystem managers. As learned from our example, no ‘one-size-fits all’ solution to deriving and interpreting spatio-temporal patterns announcing critical transitions exists Hence, we suggest that a multiple method approach may provide a sound scientific basis for detecting and evaluating early-warning signals and thus provide timely advice for immediate management actions in the face of future ecosystem changes Figure S1Map of the Baltic Sea and its location within Northern Europe. The central part of the Baltic Sea encompasses three deep (<70 m) basins important for marine biota, the Bornholm Basin (BB), the Gdansk Deep (GD) and the Gotland Basin (GB); largely corresponding to the International Council for the Exploration of the Sea (ICES) official sub-divisions 25, 25 and 28, respectively (thin lines). Furthermore, these basins are part of a long-term spatially and temporally disaggregated zooplankton monitoring program in the Baltic Sea.(JPG)Click here for additional data file.Figure S2Temporal variance of Pseudocalanus acuspes (circles) and Acartia spp. (triangles) estimated by standard deviations (SD) and the first-order autocorrelation coefficient (AR(1)) of detrended time-series for a sliding window of 10 , 15  and 20  years. Vertical dashed bars mark the timing of the Central Baltic Sea regime shift in the late 1980s.(TIF)Click here for additional data file.Figure S3The degree of spatial correlation for Pseudocalanus acuspes (A) and Acartia spp. (B) estimated as the mean significance  of spatial correlation coefficients derived from a Moran’s I test across 6 (black), 8 (grey) and 10 (black) randomly assigned stations (after 1000 resamples). Vertical dashed bars mark the timing of the Central Baltic Sea regime shift in the late 1980s.(TIF)Click here for additional data file.Figure S4Smoothed indicator time-series of Pseudocalanus acuspes and Acartia spp. with GAM df = 10  and df = 20  from 1960–1987. Bootstrapped confidence intervals are shown by grey lines. Acceleration in the rate of change (slope) in each year are shown by statistically significant second derivatives (f′′’), where black and white dots represent major downward- and upward trends, respectively.(PPTX)Click here for additional data file.Table S1Test results for recent trends and changes in trends over 3-year periods before the regime shift in 1988 using intersection–union tests. P-values from a χ2 goodness-of-fit test indicate whether the GAM fits satisfactory to the entire time-series. Significant negative (−) or positive (+) time trends in the rate of change (f′), as well acceleration (+) or deceleration (−) of the current trend (f′′) are shown. Hence, an increase in a rate of decline is indicated by negative f′ and a positive f′′.(DOC)Click here for additional data file.


The survival rates for oncology patients admitted to the ICU have improved significantly. The prognostic influence of the pre-admission oncological and treatment history is being questioned, the most significant impact being related to acute physiological status. In this survey, we sought to evaluate the awareness of overall mortality rates in critically ill cancer patients among intensivists in the UK.We surveyed intensive care lead clinicians in December 2011 in order to establish: a profile of the hospital and ICU they work in; their estimate of overall ICU mortality for critically ill cancer patients; the value of six outcome indicators in predicting mortality in two subgroups of oncological candidates for ICU admission; and the local management of acutely deteriorating cancer patients potentially requiring ICU care.The ICU mortality rates estimated by survey respondents differed from those reported in the literature: for solid tumor 21% (SEM 3) versus 10 to 23%, for metastatic solid tumor 38% (SEM 4) versus 23%, hematological malignancy 45% (SEM 3) versus 33 to 43% with allograft transplant 54.8% (SEM 5) versus 39 to 50% and autograft transplant 56% (SEM 5) versus 44%. Regarding the management of cancer patients, there were conflicts reported between teams . Few units had established triage policies for the acutely ill cancer patient (5%) and it was also not common that plans were made prior to the patient's deterioration . Figure The awareness of improved outcome in critically ill cancer patients differs among physicians, and in general estimated mortalities were far higher than those reported in the literature.


Numerous viruses are responsible for respiratory infections; however, both their distribution and genetic diversity, in a limited area and a population subgroup, have been studied only rarely during a sustained period of time.A 2-year surveillance program of children presenting with acute respiratory infections (ARIs) was carried out to characterize the viral etiology and to assess whether using gene amplification and sequencing could be a reliable approach to monitor virus introduction and spread in a population subgroup.Using multiplex RT-PCR, 15 different respiratory viruses were detected within the 486 nasopharyngeal positive samples collected among 817 children aged <9 years old who presented with ARI during October 2006 to September 2008. A single virus was detected in 373 patients (45.7%), and two to four viruses in 113 patients (13.8%). The most frequent causative viruses were respiratory syncytial virus (RSV) (24.7%), human bocavirus (24.5%), and human rhinovirus (HRV) (15%). RSV was more prevalent in winter and among young infants. Cases of seasonal influenza A and B viruses were reported mainly in January and August. An increase in adenovirus infection was observed during the spring of the second year of the study. Sequence analyses showed multiple introductions of different virus subtypes and identified a high prevalence of the newly defined HRV-C species. A higher viral incidence was observed during the winter of 2008, which was unusually cold.This study supports the usefulness of multiplex RT-PCR for virus detection and co-infection, and for implementation of a molecular monitoring system for endemic and epidemic viral respiratory infections.


This review provides the available evidence on sexual dysfunctions in India. Most of the studies have concentrated on male sexual dysfunction and hardly a few have voiced the sexual problems in females. Erectile dysfunction (ED), premature ejaculation (PME) and combinations of ED and PME appear to be main dysfunctions reported in males. Dhat syndrome remains an important diagnosis reported in studies from North India. There is a paucity of literature on management issues with an emergent need to conduct systematic studies in this neglected area so that the concerns of these patients can be properly dealt with. Human sexuality is inherently related to some of the social and public health problems in India. These problems may involve contraceptive use, child abuse, sex education, legal issues of homosexuality and AIDS. These health problems have a significant impact on existing health infrastructure and budget. These problems also need to look within the context of poverty, stressful living situations, diverse cultural belief systems, quackery, ignorance and inadequate health services. However, there is little recognition of how these health problems are related to human sexuality and their dysfunctions. There is a need to understand how sexual attitudes, beliefs, and values act and influence these problems. Our cultural perspective can also shape the experience and understanding of these disorders. There isIn this review, our aim is to present sexual dysfunction from the Indian perspective. Available data, based clinical studies from India, are reviewed and important findings highlighted. Our presentation is limited to sexual dysfunction only and paraphilias will be not discussed.et al. (1959). conVerma and his colleagues (1998) analyzedA file review of 178 male patients with sexual dysfunction by Avasthi and his colleagues (2003) revealedGupta and his colleagues (2004) attempteKendurkar and his colleagues (2008) assessedAs compared to male sexual dysfunction, a few Indian studies are available in the area of female sexual dysfunction. This area remains largely unexplored. Agarwal (1977) reportedIn the review by Kulhara and Avasthi (1995), there waKar and Koola (2007) conducteIn the study among 100 consecutive women attending the Department of Pediatrics for the care of non-critical children in a tertiary care teaching hospital, Avasthi and his colleagues (2008), found 17In another cross-sectional survey of 149 married women in a medical outpatient clinic of a tertiary care hospital, Singh and his colleagues (2009), reportedet al. (2005),[et al. (2007).[Studies pertaining to Dhat syndrome, a culture bound syndrome, have mostly defined clinical features. This “semen loss”-related psychological distress has been extensively reviewed by Prakash 2007), and Avas07, and A. (2005), and Dhik. (2007).Wig (1960), coined tBehere and Natraj (1984) and BhatMost studies found erectile dysfunction (22-62%) and premature ejaculation (22-44%) as commonly associated psychosexual dysfunctions, while depressive neurosis (40-42%), anxiety neurosis (21-38%), somatoform/hypochondriasis (32-40%) as the most reported psychiatric disorders in patients diagnosed with Dhat syndrome.Chadda and Ahuja (1990) could not find any abnormality on urine examination except oxaluria (10%) and phosphaturia (6%). On follow-up of these patients, Behere and Natraj (1984) found that majority of the patients recovered (66%), while the rest either improved (22%) or were unchanged (12%).Behere and Natraj (1984) and BhatRegarding management of Dhat syndrome, Wig (1960) suggestePrakash and Meena (2007), providedChadda and Ahuja (1990) advocateThis review highlights the available evidence in the field of psychosexual medicine in India. It is important to mention that all studies were from a hospital setting and none from community. Only a few studies explored female sexual dysfunction. Very few studies spoke about management issues. Dhat syndrome could be an important diagnostic entity to be researched. There is a strong need to perform studies in these areas.


Studies of spontaneous EEG and visual or somatosensory evoked high frequency oscillations indicate that the abnormal fluctuations of cortical responsivity over time in relation to the migraine attack could be due to abnormal thalamic control. Here we searched for possible structural changes in the thalamus of migraineurs by mean of acquiring diffusion tensor magnetic resonance imaging (MRI). This MRI technique provides quantitative data on water molecular motion, as a marker of tissue structure. Materials & Method – Seventeen untreated migraine without aura (MO) patients underwent MRI scan (3-Tesla Siemens Gyroscan) during (n=7) and between attacks (n=10) and were compared to a group of 14 healthy volunteers (HV). We examined fractional anisotropy (FA) and mean diffusivity (MD) in the thalamus.Between attacks MO patients had a significantly higher FA and lower MD values in the bilateral thalami when compared to HV (p<0.05). During attacks, all MRI quantitative measurements in migraineurs were similar to those found in HV. In MO patients, FA of the right thalamus was positively correlated with the number of days since the last migraine attack .The higher thalamic FA values noted between attacks in MO patients may be related to a decrease in regional branching and crossing of fibers, which normalizes during an attack. Whether these changes could be considered as the anatomical counterpart of the cyclic functional fluctuations previously observed with the neurophysiology in migraine remains to be determined.


Sheep chromosome 3 (Oar3) has the largest number of QTLs reported to be significantly associated with resistance to gastro-intestinal nematodes. This study aimed to identify single nucleotide polymorphisms (SNPs) within candidate genes located in sheep chromosome 3 as well as genes involved in major immune pathways. A total of 41 SNPs were identified across 38 candidate genes in a panel of unrelated sheep and genotyped in 713 animals belonging to 22 breeds across Asia, Europe and South America. The variations and evolution of immune pathway genes were assessed in sheep populations across these macro-environmental regions that significantly differ in the diversity and load of pathogens. The mean minor allele frequency (MAF) did not vary between Asian and European sheep reflecting the absence of ascertainment bias. Phylogenetic analysis revealed two major clusters with most of South Asian, South East Asian and South West Asian breeds clustering together while European and South American sheep breeds clustered together distinctly. Analysis of molecular variance revealed strong phylogeographic structure at loci located in immune pathway genes, unlike microsatellite and genome wide SNP markers. To understand the influence of natural selection processes, SNP loci located in chromosome 3 were utilized to reconstruct haplotypes, the diversity of which showed significant deviations from selective neutrality. Reduced Median network of reconstructed haplotypes showed balancing selection in force at these loci. Preliminary association of SNP genotypes with phenotypes recorded 42 days post challenge revealed significant differences (P<0.05) in fecal egg count, body weight change and packed cell volume at two, four and six SNP loci respectively. In conclusion, the present study reports strong phylogeographic structure and balancing selection operating at SNP loci located within immune pathway genes. Further, SNP loci identified in the study were found to have potential for future large scale association studies in naturally exposed sheep populations. Haemonchus contortus, Teledorsagia circumcincta, Trichostrongyles, Nematodirus sp. impose severe constraints on sheep and goat production especially those reared by marginal farmers under low external input system. These parasites incur heavy losses to farmers in terms of body weight loss, direct cost of anthelminthic drugs, loss due to mortality, etc. For example, annual treatment cost for Haemonchus contortus alone had been estimated to be 26 million USD in Kenya, 46 million USD in South Africa and 103 million USD in India Haemonchus contortus, Trichostrongylus colubriformis, Teledorsagia circumcincta and various Nematodirus species. Considerable variation has been reported among sheep breeds on their ability to resist gastro-intestinal nematodes (GIN). For example, indigenous sheep breeds like Red Maasai 2 = 0.149, Avikalin sheep 2 = 0.41, Armidale sheep Assessment of livestock health conditions in developing countries for identification of priority diseases to be targeted for control, revealed helminth infections as one of the most important problems in sheep and goat Exploration of genetic variation either within specific regions of genome or more specifically in candidate genes involved in innate and adaptive immune pathways may help to identify a set of DNA markers significantly associated with parasite resistance characteristics. The former approach in terms of quantitative trait locus (QTL) analysis is a powerful method to understand genotype-phenotype relationship. Several QTL studies on parasite resistance characteristics have been reported in sheep. A quick evaluation of Animal QTL database Haemonchus contortus parasite).One of the important objectives of QTL studies is to identify underlying causative gene polymorphisms associated with the trait. Different QTLs reported in chromosome 3 for parasite resistance characteristics were found to be distributed all over the chromosome with varying overlapping regions. Hence, different candidate genes within chromosome 3 along with genes involved in immune related KEGG pathways (KEGG-Kyoto Encyclopedia of Genes and Genomes) could be important targets for establishing underlying causative variations. It is expected that the potential causative polymorphisms within candidate genes are members of the same overarching KEGG pathway that lead to the phenotypic expression on parasite resistance characteristics in each population. Further, the extent of genetic diversity and population sub-structure at such polymorphic loci are critical for such a genotype-phenotype association study. Population stratification has been demonstrated to result in false positive associations in various species including humans Haemonchus contortus and blood samples were collected from jugular vein under the supervision of qualified veterinarians for extraction of DNA and assessment of blood parameters. 42 days post challenge, animals were dewormed to clear parasites from the gut after the experiment. The experimental challenge in both locations did not involve animals from any endangered or protected species/breeds. Blood sample collection for DNA extraction and genotyping from remaining breeds were performed by local veterinarians in respective countries following good animal practice.All procedures for artificial challenge experiment at different locations in Argentina and Indonesia were approved respectively by the Institutional Committee for Care and Use of Experimental Animals of the National Institute of Agricultural Technology (CICUAE-INTA), Argentina (protocol number 35/2010) and Institutional Research Animal Facility, Bogor Agricultural University, Indonesia following the guidelines described in their institutional manuals. Experimental animals were challenged with infective L3 larvae of in vitro conditions. All animals were maintained together during the entire trial in dry lot or under conditions of minimum additional parasite challenge. Body weight (BW), fecal egg count (FEC) and packed cell volume (PCV) were recorded at 0, 28, 35 and 42 days after artificial infection. 136 animals from Corriedale (66), Pampinta (34), Indonesian Fat Tail (17) and Indonesian Thin Tail breeds of sheep (19) were utilized for artificial challenge at experimental stations located in Argentina and Indonesia respectively.A total of 713 unrelated sheep from 22 different breeds/populations were utilized for diversity analysis in the present study. The sheep breeds/populations were distributed in three macro-environmental geographical locations including Asia (tropics), Europe (temperate) and South America (tropics) with the assumption that level of parasite load and infections vary significantly across these regions http://www.mbio. ncsu.edu/bioedit/bioedit.html). 44 novel SNPs were identified within the candidate genes under study for which competitive allele specific PCR (KASPar) assay based on FRET chemistry were developed for genotyping . Briefly, two forward primers one specific to each allele were designed with the respective proprietary tail sequence complementing the FAM or HEX fluorescence reporting system. A common reverse primer was designed for each genotyping assay. Thermal cycling parameters and recycling conditions were followed as per manufacturer's recommendations and are available on request. Endpoint allele discrimination module incorporated within the BioRad CFX96  was utilized for calling the genotypes based on fluorescent intensity recorded for each of the two alleles. The emission data of all the samples in the plate were plotted in X and Y axis respectively for each allele and the genotypes were called based on distinct clustering. Quality of allele calling was confirmed by comparing the genotypes derived from KASPar assay with the available sequence data on individuals from the panel of unrelated animals. 38 out of 44 assays passed quality control and were subsequently utilized for genotyping large number of animals. Additionally, ten toll like receptor (TLR) genes were selected for in silico mining of SNP variations from sequences available at NCBI-GenBank database. A total of 14 non-synonymous SNPs within coding DNA regions of TLR genes were identified for development of genotyping assays  using SPSS version 13.0. The first three principal components were used to draw the scattergram so as to understand underlying genetic structure and relationship of different breeds in three dimensional geometric space. The extent of population sub-structure was further explored using STRUCTURE with the assumption of different clusters, K = 1–15, 20, 25 and 30. Five replicate runs were performed for each K under admixture model without a priori population information. The number of burn in periods and MCMC repeats used for all the runs were 50000 and 100000 respectively. To identify the optimal ‘K’, the second order rate of change of L(K) with respect to ‘K’ was calculated by following the procedure reported elsewhere Basic diversity indices like allele frequency, genotype frequency, expected heterozygosity and test for Hardy Weinberg equilibrium were calculated using PEAS  www.stat.washington.edu/stephens) ST values using ARLEQUIN version 3.1 Thirteen SNP loci within candidate genes involved in different immune related KEGG pathways and located in chromosome 3 were used to reconstruct haplotypes from unphased genotypic data. Reconstruction of haplotypes and estimation of haplotype frequencies were performed using PHASE for Windows, version 2.1 , body weight change (BWC) and packed cell volume change (PCVC) measured 42 days post infective L3 larvae challenge. The data on FEC were subjected to log transformation before applying the program of least squares, LSMLMW A total of 243 sequences were generated by targeted re-sequencing of selected candidate genes  to 0.315 (Hamdani) while the mean expected heterozyosity varied from 0.237 (Pampinta) to 0.315 (Indonesian Thin Tail). Among different geographical regions, mean observed heterozygosity was highest in South West Asian sheep populations (0.309) followed by European populations [0.296] while South East Asian populations had the least mean observed heterozygosity (0.270). This is consistent with the fact that the diversity remains higher around the centre of domestication while decreasing with increasing geographic distance ST  to 0.469 (Pampinta/Mouflon). 21.3% of total genetic variation was found to be due to between breed differences while 77.3% was due to within breed differences. The values are much higher than that reported for European sheep (13.1%) ST values observed could be understood from the fact that the samples were derived from wide geographic locations in the present study . Further, phylogenetic analysis of pair-wise allele sharing distance revealed two major clusters with most of South Asian, South East Asian and South West Asian breeds clustering together while the European and South American sheep breeds clustered together separately  were plotted on a three dimensional scattergram to evaluate the genetic structure of sheep breeds  on 136 animals was used for least squares analysis under a complete fixed effect model. The effect of location of experimental stations  was not found to have significant influence on fecal egg count and packed cell volume while significant effect was observed with respect to body weight change (P<0.01). Although the locations are wide apart geographically, uniform protocol was followed across different experimental stations in terms of age of lambs selected for experiment, deworming and data recording schedule, however some differences did exist in terms of quality of pasture available for grazing, etc. Higher observed body weight change in animals challenged at Aguil Experimental Station (AES), Argentina was due to better growth performance of Pampinta lambs. Higher body weight achieved by Pampinta lambs were due to their genetic differences in growth rate (average pre-weaning weight gain of 295 g/day) and weight gain (average weaning weight of 33.4 kg) as compared to other breeds like Corriedale (218 g/day and 24.6 kg) To evaluate the potential utility of the SNP loci for future association study on a large number of samples, a pilot analysis was performed with the phenotypes generated in four breeds after artificial challenge with infective L3 larvae of In conclusion, the present study reports strong phylogeographic structure in sheep across Asia, Europe and South America and balancing selection operating at SNP loci located within immune pathway genes. Although the present association analysis is preliminary in nature, the SNP loci on chromosome 3 and those within immune pathway genes indicated their potential for future large scale association studies in naturally exposed populations.Figure S1http://www.animalgenome.org/cgi-bin/QTLdb/index).(a–c) QTLs related to gastro-intestinal nematode resistance in sheep (d) Chromosome-wise distribution of QTLs related to parasite resistance traits in sheep and number of SNP loci investigated in the present study (e) Quantitative trait loci (QTL) map of chromosome 3 related to parasite resistance traits in sheep Click here for additional data file.Figure S2Distribution of allele sharing distance (IBS) between pairs of individuals. Distance was plotted separately where pairs were drawn from within the same breed (blue bars) and from across the breeds (red bars).(TIF)Click here for additional data file.Figure S3Bayesian clustering of 713 sheep based on genotype data at (a) 18 non-neutral SNP loci (b) 23 neutral SNP loci under assumption of 2 to 6 clusters without a priori population information. The breed names are given below the box plot and the geographical origin indicated above the box plot with the individuals of different breeds separated by vertical black lines.(TIF)Click here for additional data file.Table S1Details of SNPs identified in silico within TLR genes of sheep.(DOCX)Click here for additional data file.Table S2ST (lower triangle) and allele sharing distance (upper triangle) among different sheep breeds.Pairwise F .(DOCX)Click here for additional data file.Table S3Global F-Statistics among different sheep populations at 41 SNP loci.(DOCX)Click here for additional data file.Table S4Results of Ewens Watterson Neutrality test at different SNP loci in various sheep breeds .(DOCX)Click here for additional data file.Table S5Analysis of molecular variance among different sheep breeds based on (right) genotypes at 18 non-neutral SNP loci and (left) genotypes at 23 neutral SNP loci.(DOCX)Click here for additional data file.


To the Editor: The severe acute respiratory syndrome (SARS) outbreak of 2002–03 and the subsequent implication of bats as reservoir hosts of the causative agent, a coronavirus (CoV), prompted numerous studies of bats and the viruses they harbor. A novel clade 2c betacoronavirus, termed Middle East respiratory syndrome (MERS)–CoV, was recently identified as the causative agent of a severe respiratory disease that is mainly affecting humans on the Arabian Peninsula . Details about the bat sample are available in the RNA-dependent RNA polymerase (RdRp) gene . PCR results were positive for 5 (8%) of the 62 specimens. PCR amplicons for 4 positive specimens yielded alphacoronavirus sequences related to recently described bat alphacoronaviruses from South Africa . During 2011–2012, fecal pellets were collected from 62 bats representing 13 different species in the KwaZulu-Natal and Western Cape Provinces of South Africa and stored in RNARdRp fragment generated by the screening PCR to 816 nt, as described . A partial spike gene–based phylogeny using this sequence yielded the same topology as that using the partial RdRp sequence  are urgently needed to elucidate the emergence of MERS-CoV. Such studies should focus on the Arabian Peninsula and Africa.Neoromicia zuluensis bats, and spike gene phylogeny of the 2c betacoronavirus clade.Description of bat sampling, screened bat species, distribution of


Verapamil is an off-label first-line drug in prevention of cluster headache (CH) but little is known about the optimal rate of dose escalation. The safety profile of slow dose increment in 80mg intervals has been previously documented. It is possible to achieve the effective dosage earlier with fast escalation, although the safety and tolerability are of concerns.To compare the electrocardiographic abnormalities and other adverse events between fast  and slow titration (by 80mg) of verapamil in CH treatment.Electrocardiograms (ECGs) at baseline and every two weeks in parallel with the increment of the drug were performed in patients attending our clinic. Medical records and ECGs performed in CH patients between 2007 and 2011 were retrospectively reviewed.Of 169 patients, 80 followed the fast and 89 the slow regimen. The slow regimen group differed from the fast regimen group in terms of: higher proportion of chronic CH , duration of verapamil use  and maximum dosage of verapamil achieved . Eighty-three (49%) patients showed ECG changes: bradycardia in 60 (36%), first-degree atrioventricular block (AVB) in 22 (13%), second-degree AVB in 1(1%), third-degree AVB in 2 (1%), junctional rhythm in 7 (4%) and bundle branch block in 3 (2%) patients. The rate of arrhythmias did not differ between two groups. Verapamil was stopped only in second- and third-degree AVB. The rate of drug discontinuation due to arrhythmias or non-ECG adverse events did not differ between groups.The occurrence of verapamil-related serious cardiac arrhythmias was rare in patients adopting fast titration regimen. Our data support the safety and tolerability of rapid escalation of verapamil in patients with CH. Regular ECG assessment is essential throughout the therapy.


Enzyme-replacement therapy (ERT) with alglucosidase alfa has improved the lifespan of patients with classic infantile Pompe disease, although ERT is not effective in a subset of patients who mount an immune response to the exogenous enzyme. We studied the development of antibodies in response to ERT and its effect on clinical outcomes in 11 patients with classic infantile Pompe disease treated with ERT since 1999 for a median of 4.2 years (range 3 months to 12 years).We determined the endogenous acid alpha-glucosidase expression  and regularly assessed antibody formation in all 11 patients, all of whom developed antibodies. The patients who lacked any endogenous acid alpha-glucosidase production  did not develop a substantially different antibody titer than those who produced an inactive form of the enzyme (n=8). However, the patient’s age at the start of ERT proved to be important: none of the four patients who started ERT before 2 months of age developed titers of more than 1:6,250. Gross motor function and cardiac dimension improved less in patients with high titers and in CRIM-negative patients. The three CRIM-negative patients have died, whereas the eight CRIM-positive patients survived.Antibody formation is common. High antibody titers and a CRIM-negative status are associated with a poorer clinical outcome. Earlier initiation of ERT may prevent the formation of a severe immune response.


The perceptual inference of the three-dimensional (3D) external world from two-dimensional (2D) retinal input of the left and right eye is a fundamental problem  or changing disparity over time (CDOT) are both binocular inputs that may have improved discrimination performance in this task. However, motion detection and therefore IOVD is relatively insensitive to velocity changes (Gottsdanker, Early motion and disparity processing in the human visual system show tuning characteristics that supplement each other. Motion processing tends to have high temporal but relatively coarse spatial resolution whereas disparity processing has high spatial and relatively limited temporal resolution Tyler, . It has In an influential paper Weiss et al.  demonstrA geometric-statistical model based on motion and disparity constraints provides a flexible computational framework for binocular 3D motion perception that can capture arbitrary 3D trajectories of moving features and objects. For example, motion and disparity information needs to be combined to disambiguate local motion direction of a single line or edge moving in depth (see Figure The observations by Pierce et al.  demonstr


Severe hypoglycemia (blood glucose level (BGL) <40 mg/dl) is independently associated with an increased risk of mortality in critically ill patients. The impact of milder hypoglycemia (BGL <70 mg/dl) on outcome is less clear.Prospectively collected data from two observational cohorts in the USA and in the Netherlands and from the prospective GLUCONTROL trial were analyzed. Hospital mortality was the primary endpoint.n = 1,098) and strict glycemic protocol ) and 914 patients who participated in the GLUCONTROL trial  and intensive insulin therapy arm ). The percentage of patients with hypoglycemia varied widely among the different cohorts. Patients with hypoglycemia experienced higher mortality than did those without hypoglycemia within each subgroup , even after stratification by severity of illness or diabetic status. Multivariable logistic regression analysis revealed that hypoglycemia had a greater impact on the mortality of surgical patients than of medical patients. The impact of hypoglycemia on mortality occurred independently of mean glucose level during ICU stay or glycemic variability.We analyzed data from 3,262 patients admitted to Stamford Hospital (ST), 2,063 patients admitted to three institutions in the Netherlands (NL; loose glycemic protocol (L, Even a single episode of mild hypoglycemia was associated with a significantly increased risk of mortality in heterogeneous cohorts of critically ill patients, independently of severity of illness, diabetic status, diagnostic category and glycemic variability.


The aim of genetic tests performed by Genetic Counseling at Centre of Oncology was to screen for carriers of germline mutations in BRCA1 and 2 genes who are at high risk of developing breast or/and ovarian cancer.Among the patients qualified to genetic tests there are the patients who do not meet the Amsterdam criteria of HBOC syndrome.In Genetic Counseling at Centre of Oncology we find germline mutations in BRCA1 and 2 genes in about 5% of patients. Beginning in January 2003 until now we found mutations in BRCA genes in 247 families. Not all families belong to the group of high risk of developing breast or ovarian cancer. Among them there were 23 families which did not meet pedigree and clinical criteria.Analysis of tumors which were developed by the members of families carrying germline mutations in BRCA1/2 genes revealed mainly breast (415) and ovarian (170) cancers. There were also tumors which were not typical for this mutation. The most frequent not typical tumors in the families with germline BRCA1/2 mutations were: lung cancer (38), intestinal tract tumors (26) – including stomach cancer, liver cancer (20), pancreas cancer (15) and also hematopoietic tumors – among them leukemias (17), prostate cancer (14) and other.These findings justify performing genetic tests for germline mutations in BRCA1/2 genes not only for the patients who meet pedigree and clinical criteria typical for HBOC syndrome.


Normal development of the mammalian embryo requires epigenetic reprogramming of the genome. The level of cytosine methylation of CpG-rich (5meC) regions of the genome is a major epigenetic regulator and active global demethylation of 5meC throughout the genome is reported to occur within the first cell-cycle following fertilization. An enzyme or mechanism capable of catalysing such rapid global demethylation has not been identified. The mouse is a widely used model for studying developmental epigenetics. We have reassessed the evidence for this phenomenon of genome-wide demethylation following fertilisation in the mouse. We found when using conventional methods of immunolocalization that 5meC showed a progressive acid-resistant antigenic masking during zygotic maturation which gave the appearance of demethylation. Changing the unmasking strategy by also performing tryptic digestion revealed a persistence of a methylated state. Analysis of methyl binding domain 1 protein (MBD1) binding confirmed that the genome remained methylated following fertilisation. The maintenance of this methylated state over the first several cell-cycles required the actions of DNA methyltransferase activity. The study shows that any 5meC remodelling that occurs during early development is not explained by a global active loss of 5meC staining during the cleavage stage of development and global loss of methylation following fertilization is not a major component of epigenetic reprogramming in the mouse zygote. The mechanism that is currently considered to best describe 5meC reprogramming should result in an almost complete loss of methylation (<1%) by the time the embryo reaches the blastocyst stage. Analysis of around 1000 CpG islands (CGIs) within ovulated eggs shows that 15% are methylated de novo methylation occurs later, upon epiblast formation (D6.5)The dominant paradigm describing the processes of epigenetic reprogramming in the embryo holds that global active demethylation of 5meC occurs within the first cell-cycle. This demethylation acts preferentially on DNA inherited from the male while passive demethylation of the maternally derived genome occurs over subsequent mitoses Reports of an active process of global 5meC demethylation of the zygotic genome within hours of fertilisation in some species . This re-analysis did not find evidence for extensive active loss of methylation in zygotes or progressive loss due to an absence of maintenance methylation across the first several rounds of cell division. Rather, it was found that the reported loss of methylation immediately after fertilisation was accounted for by changes in the conformation or structure of chromatin that resulted in antigenic masking of 5meC.Mouse zygotes, 2-cell, 4-cell and 8-cell embryos were collected directly from the female reproductive tract from B6CBF1 strain female mice . The embryos were fixed and immunostained with anti-5meC. Zygotes were collected at various times after mating and staged according to the maturation of their pronuclei , is an independent measure of the global levels of CpG methylation. Western blot analysis showed two dominant molecular weight forms of MBD1 present in oocytes and the early stages of development . ImmunolBrief acid-treatment of cells has been the most widely used method of 5meC antigen retrieval reported. Altering the duration and concentration of HCl treatment did not change the pattern of 5meC staining in PN5 zygotes (not shown) showing that the difference between anti-5meC and anti-MBD1 staining was not due to increased acid-sensitive masking of 5meC. To determine whether other forms of masking may have occurred, fixed and acid-treated zygotes were subjected to extensive tryptic digestion. This caused a loss of MBD1 staining from nuclei 1–7 and aTo further assess this antigenic masking, condensing chromosomes of the zygote and 2-cell embryo were triple stained for DNA (blue), 5meC (green) and MBD1 (red). Acid-treated chromosomes from the zygote had some segments of undecorated DNA and large segments decorated by MBD1, but were largely devoid of staining by anti-5meC 1. By conIt is reported that further passive demethylation of the embryonic genome occurs during the cell-cycles following fertilisation due to limited maintenance methylation of the newly synthesized DNA Asymmetric anti-5meC staining of the male and female pronucleus after acid-pretreatment has been reported These analyses were all undertaken on embryos from hybrid (B6CBF1) mice. To assess whether the strain of mouse influenced the patterns of anti-5meC staining we also examined staining in an inbred (C57BL/6j) and an outbred (Swiss Quackenbush) strain embryos. Zygotes were collected directly from the reproductive tract fixed and stained for anti-5meC and PI. Hybrid embryos were collected and processed at the same time to act as controls. This analysis showed in acid-treated zygotes there was a similar progressive loss of anti-5meC staining of both PN4 and PN5 pronuclei in all three strains . Hybrid  mice were used in most experiments. In the experiments reported in 2 for the periods indicated in individual experiments. Embryos were treated with the DNA methyltransferases inhibitor N-Phthalyl-L-tryptophan  Embryos were collected from the reproductive tract in Hepes-buffered modified human tubal fluid medium (Hepes-mHTF) Immunofluorescence of zygotes was performed as previously described Unmasking of the 5meC antigen was performed by brief exposure to HCl as previously described Embryos from each treatment were processed at the same time and in parallel for each experimental replicate. All treatments were exposed to the same preparations and dilutions of all reagents including primary and secondary antibodies. Similarly all preparations from an experiment were examined microscopically within the same session, and used identical microscope and camera settings. All image analysis was performed in an identical manner for all embryos within an experiment. All preparations were performed by the same experienced operator throughout the study. Semi-quantitative analysis of staining was independently confirmed by two experienced observers.Western blot analysis on embryo and oocyte extracts was performed as previously described


To assess and compare attitudes of general practitioners in different European countries towards screening and early interventions in alcohol use disorders.A total of 2435 general practitioners (GPs) from 9 European countries  were surveyed. The questionnaire included questions on demographic, education and training on alcohol, received by general practitioners, as well as their attitudes towards management of alcohol problems. In addition, the Shortened Alcohol and Alcohol Problems Perception Questionnaire (SAAPPQ) was used.Seventy seven per cent of GPs declared that they placed ‘somewhat high’ or ‘very high’ their priority on disease prevention of the general practitioners; 54% reported having received 4 or more hours of education and training on managing alcohol problems, and 43% reported managing seven or more patients for alcohol problems in the previous year. GPs who reported higher levels of alcohol-related CME training were more likely to report regularly asking their patients about alcohol use (chi-square (3)=14.9, p=0.002). Moreover, there was a significant association between experience of alcohol-related CME and the number of patients managed for hazardous drinking (chi-square (5)=83.6, p<0.0005). Busyness (64%) and lack of training (52%) were considered most important barriers and readily available support services (84%) most effective facilitator of early intervention.The recommendations coming from the results of the ODHIN study for improving the delivery of early alcohol intervention and the management of alcohol problems in general practice are: provide better training and infrastructure. The main barriers and facilitators of early interventions did not change throughout last 16 years.


The human protein encoded by the FOXO1 gene functions as a transcription factor of insulin signaling key genes. Human uncoupling proteins 3 (UCP3) are mitochondrial proteins that are involved in the control of energy metabolism and the pathophysiology of obesity. In this study we investigated the role of genetic variation in the FOXO1 and UCP3 gene in susceptibility to non-alcoholic fatty liver disease (NAFLD) and relevant metabolic traits.We genotyped nine single nucleotide polymorphisms (SNPs) for association analyses in children . Body mass index (BMI), waist and hip circumference, blood pressure, fasting blood glucose (FBG), insulin (FIN), lipid profiles were measured and performed B-ultrasound examination in all the subjects.In the NAFLD group, FOXO1A and UCP3 allele were significantly more frequent in both association studies. There was a significant difference in the overall distribution of the genotype frequencies , (FOXO1 rs2721068), and there was a significant difference  in the distribution of the haplotype , might be good NAFLD markers.In conclusion, our study suggests a effect of UCP3 haplotype on NAFLD development and relevant intermediate phenotypes which predispose for NAFLD.


Patients with atrial fibrillation (Afib) routinely undergo a transesophageal echocardiogram (TEE) for evaluation of the left atrial appendage (LAA) to rule out thrombus prior to undergoing cardioversion or pulmonary vein isolation (PVI). Cardiac MRI (CMR) is now increasingly used for evaluation of these patients for defining pulmonary vein anatomy prior to PVI.We hypothesized that 2D and 3D non-contrast and contrast CMR is as effective as TEE in evaluating for LAA thrombus while providing simultaneous comprehensive non-invasive evaluation of the pulmonary vein anatomy within a single exam.Afib Pts  underwent TEE and non-contrast and contrast CMR prior to undergoing an initial PVI procedure. CMR was performed on 1.5T GE scanner and two blinded CMR experts analyzed the images. The CMR images were analyzed under two categories: 1) the 2D non-contrast cine images showing LAA in 2 chamber and orthogonal views 2) 3D atrial contrast source-images acquired during pulmonary vein angiogram. CMR variables evaluated were the presence or absence of LAA thrombus, quality of images and the results were compared with the results of TEE in a blinded fashion.In 62% of pts (n=65) the CMR and TEE studies were performed on the same day, in 37 % of patients (n=39) the mean interval between the two studies was 7 days and in the remaining 6 patients the mean interval was 3+1months. All pts (n=110) were analyzed for the evaluation of LAA thrombus (avg time: 35 min). All (100%) of patients were in Afib and in all (100%) the images were of diagnostic quality (good correlation between the two CMR observers with only one grade difference when in disagreement). Thrombus was absent in 108 of 110 pts on TEE and CMR and present on 2 TEE and CMR images (100% concordance). In 9 cases the 2D CMR images were indeterminate where 3D contrast images were most helpful in the final rule out of LAA thrombus.CMR offers a comparable and equally specific alternative to TEE for the complete non-invasive evaluation of LAA thrombus in patients with Afib without the obligate need for sedation, radiation or nephrotoxicity. In one single examination a CMR exam can provide LAA anatomy, rule out thrombus and display the pulmonary veins in patients undergoing a PVI procedure.


Despite the availability of an effective vaccine, Hepatitis B virus (HBV) infection still remains a foremost health problem. Undoubtedly, finding the key routes of hepatitis transmission from the point of prevention in every country, specifically in endemic regions, is of high priority. Such efforts are especially important given that many infected patients with hepatitis are asymptomatic 2]. Iran. Iran2].HBV is transmitted through both vertical and horizontal routes. Although vertical routes have both been very common in Turkey and Iran, nowadays the horizontal routes are much more important . In otheOther studies recommend the extension of HBV vaccinations in infant and high risk group, screening during pregnancy, and implementing additional strategies Such as adding the injection of hepatitis B immunoglobulin (HBIG) to the routine vaccination in neonates of mother HBs Ag positive, extension of vaccination in adults 13]14][[14][13][[[14][13]


Cross-priming serves to activate cytotoxic T lymphocytes for immune defense against viruses and tumors and plays an important role in vaccinations. Only certain dendritic cell (DC) subsets can cross-present. Several cell surface markers have been described that more or less specifically and sensitively characterize these subsets. The cell-biological mechanism(s) why a DC subset can cross-present are less clear. Theoretically, the task of cross-presentation can be divided into several mechanistic steps: (1) Antigen uptake by various endocytosis mechanisms, (2) Intracellular antigen routing into distinct organelles including the crossing of organelle membranes, (3) Antigen processing into peptides, (4) Peptide loading onto MHC molecules, and (5) Transport of these complexes to the cell surfaces for presentation to T cells. Each of these steps is dependent on numerous parameters, not only the DC subtype, but also the nature of the antigen or the presence of further signals that impact DC function or signify the presence of danger or infection.This research topic contains 10 articles by leading experts in the field of antigen presentation that cover our current knowledge on the molecular mechanisms underlying cross-presentation (Chopin et al.,


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