People of South Asian descent have an increased risk of cardiovascular disease (CVD) and diabetes, but little is known about the microcirculation in South Asian people despite evidence that this plays an important role in the aetiology of CVD. We compared the retinal microcirculation in a population-based sample of 287 middle-aged adults (144 European 143 South Asian) matched for age and sex. Retinal photographs were taken and analysed using a validated semi-automated program and microvascular measures were compared. Blood pressure, anthropometry and fasting bloods were also measured. South Asians had significantly fewer arteriolar and venular vessels and bifurcations. Arterioles and venules were longer and venules were also more tortuous in South Asians. These differences were not explained by adjustment for traditional risk factors including blood pressure, body mass index, diabetes or measures of insulin resistance. People of South Asian descent have rarefaction of the retinal microcirculation compared to age-sex matched individuals of European descent. Reduced microvascular density could contribute to the elevated risk of CVD and impaired glucose tolerance in South Asian people. People of South Asian descent, whether resident in the Indian subcontinent The microcirculation is increasingly recognized as an important and early determinant of CVD Abnormalities of the retinal microvasculature are commonly observed in the general population, and are associated with target organ damage including left ventricular remodelling; In view of the high rates of diabetes and CVD in South Asians we investigated whether abnormalities of the retinal microcirculation were more prevalent in this ethnic group, and explored whether any differences observed could be explained by conventional CVD risk factors, including abnormalities of glycaemic control.The study was conducted according to the principles expressed in the Declaration of Helsinki; approved by the Imperial College Healthcare NHS Trust local ethics committee and all participants gave written informed consent.2 test). Of Europeans and South Asians invited to participate 47% and 48% respectively agreed to participate, 17% and 22% respectively refused to participate and 35% and 30% respectively failed to respond.Participants were recruited from the London Life Science Prospective Population Study (LOLIPOP), a multi-ethnic population-based study of chronic disease and its determinants that includes over 25000 individuals identified from 58 primary care lists in West London. All participants attended Imperial College Healthcare NHS Trust after an overnight fast having completed a questionnaire detailing demographic data, health behaviours and medical history. Ethnicity was confirmed with the participant, based on self -assessment and place of birth of all 4 grandparents. Height, weight, waist and hip circumference were measured according to a standard protocol. rd order, with diameters ≥10 pixels (approximately 56 µm) were measured. Arteriolar and venular diameters were measured at a series of intensity cross-sections at 2-pixel intervals along its length. At each cross-section, the vessel diameter was measured using a sliding linear regression filter technique and an average calculated for each vessel. Retinal photographs were taken using a Zeiss FF 450 plus fundus camera (Carl Zeiss Ltd.) with a 30° field of view following mydriasis with tropicamide (1%) eye drops and additional phenylephrine (2.5%) eye drops, if necessary. Images were captured on a Basler digital camera and field 1 M of the right eye was subsequently analysed offline as described previously, Population characteristics are presented as means ± SD, or median  for skewed data. Retinal measures are given as means . Statistical comparisons of unadjusted data were made using a Student’s t-test after log or square root transformation as appropriate. Other comparisons were made following covariate adjustment by multivariate regression analysis with transformation of data as appropriate. P<0.05 was considered statistically significant. All statistical analyses were performed using Stata/IC 11.0 .Two thirds of participants were male, and the mean age was approximately 62 years . Height nd and 3rd order arterioles (Table2).Compared to Europeans, South Asians had significantly fewer arteriolar vessels and bifurcations, and arterioles were longer, and consequently LDR was higher, although arteriolar diameters did not differ significantly . The difSouth Asians also had fewer venular bifurcations; venules were longer and more tortuous, and venular LDR was higher . Since cIn all participants there was a positive relationship between systolic blood pressure and arteriolar narrowing, assessed as arteriolar LDR ; this relationship did not differ by ethnicity (p interaction systolic blood pressure×ethnicity = 0.3). The number of arteriolar bifurcations was also negatively correlated with blood pressure  and pulse wave velocity . Diabetes was associated with wider arterioles  pixels, n = 239 vs. 17.5  pixels in people with diabetes, n = 48; p = 0.03); this difference also did not differ by ethnicity (p interaction diabetes×ethnicity = 0.4). Diabetes was also associated with significantly fewer arteriolar , n = 48 vs. 9.4 , n = 239; p = 0.01) and venular  ); n = 48 vs. 11.8 , n = 239; p = 0.03) bifurcations.1c, HOMA, or fasting glucose) as covariates for arteriolar measures, and age, sex, blood pressure, BMI, diabetes  and smoking for venular parameters. These covariates were chosen a priori on the basis of previous studies We proceeded to explore possible explanations for these ethnic differences in the retinal microcirculation through multivariate analysis, using age, sex, blood pressure, heart rate, diabetes  arterioles and is unexplained by blood pressure, diabetes, insulin resistance, or other cardiovascular risk factors known to influence the retinal microvascular architecture.et al.et al.As far as we are aware no other studies have compared the retinal microcirculation between Europeans and South Asian adults. Owen et al.,et al.et al.et al. et al.Others have studied different ethnic groups and found inter-ethnic differences in the retinal microcirculation of adults. In the Multi-ethnic Study of Atherosclerosis (MESA) retinal arteriolar calibre was larger in African Americans and Hispanics than in Whites and Chinese, while venular calibre was largest in African Americans, intermediate in Hispanics and Chinese and smallest in whites. Our study has a number of limitations, it is cross sectional and therefore questions of causality cannot be resolved. However it is worth noting that abnormalities in the retinal microcirculation are already apparent in youth and related to low birth weight and post natal growth. In summary, we have shown rarefaction and elongation of the vascular segments of the retinal microvasculature in people of South Asian descent compared with those of European descent in the UK. Abnormalities of the microvasculature could contribute to the elevated risk of CVD in South Asians.


Escherichia coli isolates collected from urine samples from urology services in the Euregio Meuse-Rhine, the border region of the Netherlands (n = 176), Belgium (n = 126) and Germay (n = 119). Significant differences in resistance between the three regions were observed. Amoxicillin-clavulanic acid resistance ranged from 24% in the Netherlands to 39% in Belgium (p = 0.018), from 20% to 40% (p<0.004) for the fluoroquinolones and from 20% to 40% (p = 0.018) for the folate antagonists. Resistance to nitrofurantoin was less than 5%. The prevalence of ESBL producing isolates varied from 2% among the Dutch isolates to 8% among the German ones (p = 0.012) and were mainly CTX-M 15. The prevalence of MDR isolates among the Dutch, German and Belgian isolates was 11%, 17% and 27%, respectively (p< = 0.001 for the Belgian compared with the Dutch isolates). The majority of the MDR and ESBL producing isolates belonged to ST131. This study indicates that most antibiotics used as first choice oral empiric treatment for UTIs  are not appropriate for this purpose and that MDR strains such as CTX-M producing ST131 have spread in the entire Euregion. Our data stress the importance of ward specific surveillance to optimize empiric treatment. Also, prudent use of antibiotics and further research to alternative agents are warranted.We determined the prevalence and spread of antibiotic resistance and the characteristics of ESBL producing and/or multi drug resistant (MDR)  Escherichia coli, the most prevalent causative agent of urinary tract infections (UTIs) E. coli clones increased not only in the hospitals but also in the general population E. coliE. coli ST131 E. coli strains will guide physicians in their choice of adequate empiric treatment. Since such ward specific resistance data for patients visiting the urology services are hardly available we conducted a surveillance to determine the prevalence of (multi drug) resistant E. coli isolates including ESBL producing strains collected from patients visiting a urology service in the Euregion Meuse-Rhine.The increase of antimicrobial resistance is a major concern worldwide. Surveillance studies showed an increase in resistance for both Gram positive and Gram negative bacteria All bacterial isolates in this study were collected and analysed anonymously. Therefore, consent from the patient was not required and ethical approval was waivered. This is in agreement with the code for proper use of human tissue as formulated by the Dutch Federation of Medical Scientific Societies and the policy of the Medical Ethics Committee of het Maastricht University Medical Centre (MUMC).During a 6 month collection period between 2009 and 2011 unique, unrelated, non duplicate, consecutive isolates from urine samples from patients attending the urology services were collected.E. coli isolates were included in this collection: 176, 126 and 119 from the Dutch, Belgian and German urology services, respectively. The following hospitals participated in the study: MUMC – Maastricht, Atrium Medical Centre – Heerlen and Viecuri Medical Centre – Venlo in the Netherlands, General Hospital Vesalius – Tongeren, Jessa Hospital – Hasselt and Centre Hospitalier Universitaire – Liège in Belgium, and Hermann-Josef Hospital – Erkelenz, St. Antonius Hospital – Eschweiler and St. Marien Hospital – Düren in Germany. Only one isolate per patient was included. Clinical data were not available. The isolates were collected and identified at the local laboratories according to standard microbiological methods, stored and sent to the MUMC for susceptibility testing.A total of 421 E. coli ATCC 35218 and ATCC 25922 were used as control strains. The MIC data were analysed using breakpoints defined by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) Quantitative susceptibility testing was performed using a microbroth dilution with Mueller-Hinton II cation-adjusted broth  and micro titre plates with freeze-dried antibiotics . The minimal inhibitory concentration (MIC) was defined as the lowest concentration showing no growth after 18 hours of incubation at 35°C. blaTEM or blaCTX-M were amplified with PCR and specific primers Putative ESBL producing isolates (i.e. MIC for cefotaxime or ceftazidime > = 2 mg/L) were confirmed as an ESBL producer with a combination disk diffusion test E. coli MLST web site All MDR isolates and ESBL producing isolates were further analyzed with PFGE The appropriateness of amoxicillin-clavulanic acid, piperacillin-tazobactam, cefuroxime, ceftazidime, cefixime, ceftibuten, ciprofloxacin, nitrofurantoin, trimethoprim-sulfamethoxazole and gentamicin for empiric treatment was determined, These agents are considered representatives to other antibiotics in their class.As criterion for appropriateness of empiric treatment for complicated UTI, a 10% resistance cutoff value was used A Pearson’s chi square test or Fischer’s exact test was performed to determine statistically significant differences of resistance between the three different countries (subregions) of the Euregion Meuse-Rhine . A modified false discovery rate (FDR) method developed by Benjamini and Yekutieli was used as correction for multiple testing Significant differences in prevalence of antibiotic resistance between the three regions in the Euregion were found for several antimicrobial agents tested including the fluoroquinolones, and folate antagonists . OverallPutative ESBL producing isolates were found in 10, 14 and 9 isolates among the Dutch, Belgian and German isolates, respectively. ESBL production was confirmed for 20 isolates  resulting in a prevalence of ESBL producing isolates of 1.7%, 5.5% and 7.6% among the Dutch, Belgian and German isolates, respectively (p = 0.043 for the Dutch and German isolates). The confirmed ESBLs were mainly CTX-M 15 .E. coli ST 131 strain was the most prevalent one (63%) of which 53% was an ESBL producer divided over 3 different pulsotypes  in the Dutch subregion to 27% (n = 34) in the Belgian subregion (p< = 0.001) .The PFGEOnly pulsotype A was demonstrated in all three subregions. The other pulsotypes were only observed in one subregion.For empiric treatment of a complicated UTI, the prevalence of resistance should not exceed 10%. Taken into account this cutoff value several antibiotic agents are no longer appropriate for  empiric treatment . These iE. coli isolates collected from nine urology services in the Euregio Meuse-Rhine. We found significant differences in resistance between the Dutch, Belgian and German isolates. Although, overall the prevalence of resistance was highest among the Belgian isolates and lowest among the Dutch ones, the prevalence of ESBL producing isolates was highest among the German isolates.This study provides current data of the antimicrobial resistance of E. coli ST131 More importantly, this study also indicates that most antibiotics used as first choice oral empiric treatment for UTIs  E. coli isolates were collected from urine samples from patients of both gender and all ages visiting the urology services in the participating hospitals of this cross border multi centre surveillance study to prevent selection bias. Since all isolates were tested in one centre, the results were not influenced by differences in methodology. Unfortunately, additional patient and clinical data were not available. Since we cannot specify the underlying illness of the patients, this should be taken into consideration when initiating empiric treatment.The E. coli from the EARSS annual report 2009, which consist mainly of isolates from invasive disease Comparing the resistance rates of Moreover, differences in antibiotic use Compared with prevalence of resistance among general practice (GP) patients E. coli ST131 clone is emerging During the last years (multi drug) resistant strains have become a major healthissue. National and international surveys show an increase in nosocomial and community acquired infections involving ESBL and/or carbapenemase producing isolates E.coli ST131 and CTX-M type ESBLs, suggesting a spread of this clone in the entire Euregion.Concluding, ward specific surveillance of antimicrobial resistance is important to monitor resistance over time on a regional, national and international level since we demonstrated significant differences in antibiotic susceptibility between the three subregions in the Euregion Meuse-Rhine. We also found a high prevalence of Due to the high prevalence of resistance many antibiotics including amoxicillin-clavulanic acid, the fluoroquinolones and the folate antagonists are no longer suitable for empiric treatment of complicated UTI, therefore, further research to other possible agents is needed.


Burden of disease estimates are widely used for priority setting in public health and disability-adjusted life years are a powerful “currency” nowadays. However, disability weights, which capture the disability incurred by a typical patient of a certain condition, are fundamental to such burden calculation and their determination remains a widely debated issue.A cross-sectional epidemiological survey was conducted in the recently established Taabo health demographic surveillance system (HDSS) in south-central Côte d'Ivoire, to provide new, population-based evidence on the disability caused by schistosomiasis and soil-transmitted helminthiasis. Parasitological results from stool, urine, and blood examinations were juxtaposed to quality of life (QoL) questionnaire results from 187 adults. A multivariable linear regression model with stepwise backward elimination was used to identify significant associations, considering also sociodemographic characteristics obtained from the Taabo HDSS database.Plasmodium spp., Trichuris trichiura, Schistosoma haematobium and Schistosoma mansoni were 39.0%, 18.2%, 2.7%, 2.1% and 2.1%, respectively. S. mansoni and T. trichiura infections of any intensity reduced the participants' self-rated QoL by 16 points : 4–29 points) and 13 points (95% CI: 1–24 points), respectively, on a scale from 0 (worst QoL) to 100 points (best QoL). The only other statistically significant effect was a 1-point (95% CI: 0.1–2 points) increase on the QoL scale per one unit increase in a calculated wealth index.Prevalences for hookworm, We found consistent and significant results on the negative effects of schistosomiasis and soil-transmitted helminthiasis on adults' self-rated QoL, also when taking sociodemographic characteristics into account. Our results warrant further investigation on the disability incurred by helmintic infections and the usefulness of generic QoL questionnaires in this endeavor. In public health, "burden" estimates should capture the human suffering caused by certain health states, and the estimates are often used for priority setting. However, such "burden" estimates need to assess not only the number of affected people by certain conditions, but also the disability incurred by the average patient, and the determination of the degree of disability remains a widely debated issue. In order to provide new, population-based evidence on the disability caused by infections with parasitic worms, we administered a quality of life (QoL) questionnaire to 187 adults in rural Côte d'Ivoire and concurrently examined them for parasitic worm infections. We also considered sociodemographic characteristics in our analysis. In comparison with their non-infected counterparts, infected people reported a 13–16 points lower QoL on a scale from 0 (worst QoL) to 100 points (best QoL). At the same time, a one unit increase in a calculated wealth index revealed a 1-point increase in the participants' QoL. The results are consistent and warrant further investigation on the disability induced by parasitic worm infections and the usefulness of QoL questionnaires in this endeavor. Efforts are underway for a comprehensive revision of the global burden due to major diseases, injuries, and risk factors An undeniable merit of the global burden of disease concept is the renewed interest in descriptive epidemiology and population health measurement. Not surprisingly though, the concept also stimulated considerable controversies. Amongst other issues, criticism about the disability weights, which should measure the disability caused by a certain condition on a continuous scale from 0  to 1 (death), was raised as the original disability weights were solely based on expert opinion In order to provide new, setting-specific, population-based evidence on the disability caused by schistosomiasis and soil-transmitted helminth infections, we conducted a cross-sectional survey among adults in the Taabo health demographic surveillance system (HDSS), in south-central Côte d'Ivoire. Generic quality of life (QoL) questionnaires and standardized, quality-controlled parasitological methods were applied and the results juxtaposed, taking into consideration readily available sociodemographic data from the Taabo HDSS database as potential confounders. To our knowledge, only few studies employed generic QoL questionnaires to assess disability attributable to helminth infections The study protocol was approved by the institutional research commissions of the Swiss Tropical and Public Health Institute  and the Centre Suisse de Recherches Scientifiques en Côte d'Ivoire . Ethical clearance was provided by the ethics committee of Basel  and the Comité National d'Ethique et de la Recherche (CNER) in Côte d'Ivoire (reference no. 1086 MSHP/CNER).The Taabo HDSS was set-up in mid-2008, located in the Taabo area in the south-central part of Côte d'Ivoire. While establishing the Taabo HDSS, district and village authorities and the general public were informed about its purpose, operational procedures, potential risks, and benefits. The present study was carried out in June 2010, readily embedded in the second cross-sectional epidemiological survey pursued once every year. Written informed consent was obtained from all participants. It was emphasized that participation was voluntary, and hence people could withdraw anytime without further obligation. All results were coded and kept confidential. At the end of the study, all people living in the Taabo HDSS were invited for deworming with albendazole  and ivermectin (∼200 µg/kg using a dose pole) irrespective of participants' infection status Schistosoma haematobium has been observed after praziquantel administration The Taabo HDSS covers most of the rural Sous-Préfecture Taabo. Its main office is located in Taabo Cité, 160 km northwest of Abidjan. The region's tropical climate follows a seasonal pattern with a long dry season between November and April and two rainy seasons, a long one between April and July and a shorter one in September and October Vital statistics  and the health of some 38,500 individuals registered in the Taabo HDSS are monitored longitudinally. People are mainly engaged in subsistence farming of manioc, yams, and banana, while cacao and coffee are farmed as cash crops For the present study, sociodemographic data on individual and household level were obtained from the Taabo HDSS database. Individual data included sex, age, relationship with the head of household, education, and main occupation. Household-level data contained information on the households' location, the number of household members, housing construction material, availability of certain facilities, and the possession of equipment.All collaborators, including local health personnel, were trained and informed about the purpose, procedures, potential risks, and benefits of the cross-sectional survey and the deworming. Subsequently, key informants, field enumerators, and supervisors of the Taabo HDSS and the local health personnel informed all heads of households to visit, together with their families, a predefined, nearby survey location on a specified date and time to receive anthelmintic treatment. In addition, approximately 7% of all households in the Taabo HDSS were selected by stratified random sampling. These households were visited the day before the treatment and two plastic containers were distributed to all household members for collection of a lemon-sized fresh morning stool and a urine sample the next day.On the day of treatment, people not selected for in-depth clinical and parasitological examinations received albendazole and ivermectin and they could continue with their daily chores . People Ascaris lumbricoides, Trichuris trichiura, and hookworm) and Schistosoma mansoni. The sum of the helminth-specific egg counts of the two Kato-Katz thick smears were multiplied by a factor 12 to obtain infection intensities, as expressed in eggs per gram of stool (EPG). Urine samples were vigorously shaken, 10 ml drawn up into a syringe and pressed through a meshed nylon filter with a pore size of 20 µm . Next, filters were placed on a microscope slide and, after adding a drop of Lugol, examined for S. haematobium eggs under a light microscope. All parasitological examinations were performed by experienced laboratory technicians. For quality control, ∼5% of all microscope slides were re-examined by a senior technician. In case of disagreement, slides were read a third time and the result discussed among the technicians until agreement was reached. Thin and thick blood films were stained with Giemsa and examined for Plasmodium parasitemia.Blood, stool, and urine samples were transferred to the laboratory of the hospital in Taabo Cité and worked up the same day using standardized, quality-controlled techniques as described elsewhere We used previously employed questionnaires in Côte d'Ivoire The initial version of the questionnaire after translation into French was discussed with the field enumerators and supervisors of the Taabo HDSS. These field enumerators and supervisors are locals, who live in the different communities of the Taabo HDSS, are able to read, write, and speak French as well as the local languages Baoulé, Dioula, or Senufo. The questionnaire was further adapted based on their comments, then pre-tested in a nearby village and again refined in order to obtain the finally applied version . During Data were double-entered and cross-checked in EpiInfo version 3.5.1  and analyzed in STATA version 10.1 . For convenience, the myriad of main occupations obtained from the Taabo HDSS database were categorized into primary economic sector , secondary economic sector , and tertiary economic sector , with housewives included in the primary sector as they are usually involved in (subsistence) farming. Socioeconomic household data were used to calculate an asset-based wealth index and deduce the inhabitants' socioeconomic status, according to an approach put forth in a World Bank publication p<0.05) variations in the mean QoL scores assessed by the different interviewers . Furthermore, to assess the internal consistency and validity of the resulting QoL scores, we used Cronbach's alpha and a univariable linear regression model with the calculated overall QoL scores as outcome and the QoL ratings directly expressed by the participants in the final question of the questionnaire as explanatory variable  and parasitological findings  as explanatory variables, considering also potential clustering of the results in interviewers and residential areas. A stepwise backward elimination procedure of non-significant explanatory factors was adopted to identify those variables most significantly influencing the participants' scores on QoL. In each iteration, the explanatory variable with the highest p-value was eliminated as long as the Akaike information criterion (AIC) was decreasing and the likelihood ratio test indicated no statistically significant association between the eliminated explanatory variable and the QoL scores. Categories of the same explanatory variable were combined, based on expert knowledge and logical deduction, before eventually eliminating the respective variable.Wilcoxon rank sum and Kruskal-Wallis test, χOnly participants with signed written informed consent and complete data records  were included in the final analysis. Participants with completed written informed consent and questionnaire, but incomplete parasitological results were included in an attrition analysis.p = 0.926). About half  of the participants were heads of household.Overall, 255 adults were invited to participate in the study . Seven dp = 0.663), women reported a significantly lower educational level than men (p = 0.046). Differences in occupational sector categorization showed no statistically significant difference between sex (p = 0.295) and age groups (p = 0.218) with most participants working in the primary sector (mainly subsistence farming) and only very few in the secondary sector. Not surprisingly, higher educated people were more often working in the tertiary rather than in the primary or secondary sectors (p<0.001).The educational level and the main sector of occupation are summarized in p = 0.377) or age group (p = 0.060), but positively associated with the educational level and working in the tertiary sector (both p<0.001).Results from the socioeconomic analysis are shown in Plasmodium infection are summarized in Plasmodium spp., T. trichiura, S. haematobium and S. mansoni prevalences of 39.0%, 18.2%, 2.7%, 2.1% and 2.1%, respectively. Most helminth infections were of light intensity and no heavy infections were diagnosed at all. With the exception of a higher prevalence of Plasmodium spp. infection in people aged 60 years and above (p = 0.028), no significant differences occurred in the prevalence and intensity of helminth and Plasmodium infection with regard to sex, age group, educational level, and occupational sector. Regarding participants' socioeconomic status, hookworm prevalence (p<0.001) and infection intensity (p = 0.002) as well as T. trichiura prevalence (p = 0.025) were significantly lower in wealthier participants.Sex- and age-specific prevalence and intensity of helminth infection and p = 0.104). The mean summary score on QoL was 63.9 (range: 21.2–93.9) and Cronbach's alpha (0.805) indicated a good internal consistency of the QoL scores. The univariable linear regression model with the calculated QoL summary scores as outcome, and the QoL ratings directly expressed by the participants as explanatory variable, indicated a statistically significant positive correlation (p<0.001). The calculated scores on the four different domains and the summary score on QoL are illustrated in Thirteen different field enumerators and supervisors of the Taabo HDSS were involved in interviewing the study participants, with no statistically significant inter-observer variation in reported mean QoL scores (p = 0.043), those belonging to the wealthiest quintile (p = 0.006), and people with no hookworm infection (p = 0.002).The mean domain and overall QoL scores in relation to sociodemographic and parasitological variables are shown in p = 0.034) and S. mansoni and T. trichiura infections of any intensity  remained as the only three statistically significant explanatory variables for the overall QoL scores in the multivariable linear regression model with backward elimination (p = 0.067), working in the secondary or tertiary sectors (p = 0.094), and hookworm infection of any intensity (p = 0.061). Whether age and wealth were used as continuous variables  or as categorical variables  influenced these findings only insofar as the individuals' occupation was also eliminated in the latter models.The wealth index : 4–29 points) and 13 points (95% CI: 1–24 points) lower on a scale from 0 to 100 than the reported QoL of non-infected individuals. The only other statistically significant effect was a 1-point (95% CI: 0.1–2 points) increase in QoL per one unit increase in the wealth index. Other important explanatory variables that remained in our multivariable linear regression model after applying a stepwise backward elimination procedure were sex, indicating a 4 points (95% CI: −7–0.2 points) decrease in QoL of females; occupation, indicating a 4 points (95% CI: −0.7–8 points) increase in QoL of those working mainly in the secondary or tertiary sectors; and hookworm infections, indicating a 4 points (95% CI: −8–0.2 points) decrease in QoL of those infected.We present an analysis from a QoL questionnaire survey conducted alongside the 2010 cross-sectional epidemiological survey and deworming campaign in the Taabo HDSS in south-central Côte d'Ivoire. Results of a multivariable linear logistic regression model revealed that adults' QoL is reduced considerably among those infected with different species of helminths, regardless of the intensity of infection. Indeed, we found that the perceived QoL among adults infected with Our results have to be interpreted with caution, but raise many interesting issues. As a first critical point, it has to be considered that the sampling of the current study depended on a stratified random sampling. Starting in mid-2009, our team pursued a yearly cross-sectional epidemiological survey among approximately 7% of the people who were under demographic and health surveillance in the Taabo HDSS. The present study was linked to the June 2010 cross-sectional survey, using a sub-sample . Given our sampling approach and in view of operational and financial considerations, no formal sample size calculation was made for the present study.Second, our final sample size of 187 individuals was relatively small and the compliance rate of 73.3% suboptimal. However, somewhat higher drop-out rates had to be expected as the participants were adults, many of whom were illiterate. Compared to school-aged children, adults seemed to be somewhat reluctant or ashamed to provide any stool or urine samples. Importantly though, the attrition analysis revealed no statistically significant differences in the available indicators between the included and excluded adults, and hence no selection bias seems to have been introduced by the drop-outs.Third, the absence of a statistically significant inter-observer variation suggests that our questionnaire results are reliable. Cronbach's alpha as well as the highly significant positive correlation between the calculated summary scores on QoL based on all questions and the QoL ratings directly expressed by the participants in the final question of the questionnaire indicate internal consistency and validity of the QoL scores.Fourth, the parasitological diagnosis was based on single stool and urine samples with duplicate Kato-Katz thick smear examinations and single urine filtration, respectively. There is a large body of work demonstrating that multiple sampling or a combination of diagnostic methods result in more accurate diagnosis S. mansoni reported a statistically non-significant, but consistent decrease in the mean environmental (p = 0.251), psychological, physical, and social wellbeing . This remarkable consistency increases the plausibility of the presented findings. However, with regard to Plasmodium spp. infections the picture was less clear. Participants with positive malaria RDTs reported higher mean scores on environmental and physical wellbeing and had higher overall QoL score. These counterintuitive findings might at least partially be explained by false positive results of the RDTs because of delayed clearance of the circulating antigen Fifth, the mean domain and overall QoL scores displayed in Sixth, the effects of the sociodemographic determinants on the mean domain and overall QoL scores were somewhat less clear in the univariable comparison as summarized in S. haematobium, S. mansoni, and T. trichiura in our study sample and even though the probably most obvious confounders  were included in our analyses, we cannot rule out an effect of other potential confounders . Furthermore, we did not consider infection intensities or explore interaction terms for combined infections in our multivariable linear regression model as more complex modeling was not possible due to the comparatively small numbers.Seventh, there were only a few people infected with S. mansoni infections can be interpreted as a disability of 16%. This disability estimate is slightly below previously presented results from QoL surveys on advanced, chronic S. japonicum in the People's Republic of China, which revealed a mean disability of 19% Schistosoma infection and 10% for advanced renal or hepatic infection T. trichiura infections and the 4 points reduction for hookworm infections are also considered as disabilities of 13% and 4%, they are in the range of the disability weights listed by WHO  Eighth, the here presented decrease of 16 points on the 0 to 100 QoL scale due to In conclusion, we found consistent and significant results on the effect of schistosomiasis, soil-transmitted helminthiasis, and sociodemographic determinants on adults' QoL in rural Côte d'Ivoire. It is conceivable that helminth-infected adults in the present study suffered from advanced chronic infections and therefore reported notable losses in QoL. Our results warrant further investigation on the disability induced by helmintic infections and further probing of the usefulness and applicability of generic QoL questionnaires in this regard. Future studies should adhere to a more rigorous sampling strategy and sample size calculation, optimally in a randomized trial design, which allows for an improved control of potential confounders and the assessment of interactions due to combined infections. Furthermore, they should consider additional qualitative research to further explore the local residents' concept about QoL, additional verification of the QoL questionnaire's reliability and validity Click here for additional data file.Text S1Questionnaire for evaluating the health state of individuals (in French).(DOC)Click here for additional data file.Text S2Questionnaire for evaluating the health state of individuals .(DOC)Click here for additional data file.Table S1Explanatory variables and indicators of the multivariable linear regression model at each step of the backward elimination procedure.(DOC)Click here for additional data file.


Neuraminidase (NA) is a prominent surface antigen of Influenza viruses, which helps in release of viruses from the host cells after replication. Anti influenza drugs such as Oseltamivir target a highly conserved active site of NA, which comprises of 8 functional residues  to restrict viral release from host cells, thus inhibiting its ability to cleave sialic acid residues on the cell membrane. Reports on the emergence of Oseltamivir resistant strains of H1N1 Influenza virus necessitated a search for alternative drug candidates. Pleconaril is a novel antiviral drug being developed by Schering-Plough to treat Picornaviridae infections, and is in its late clinical trials stage. Since, Pleconaril was designed to bind the highly conserved hydrophobic binding site on VP1 protein of Picorna viruses, the ability of Pleconaril and its novel substituted derivatives to bind highly conserved hydrophobic active site of H1N1 Neuraminidase, targeting which oseltamivir has been designed was investigated.310 novel substituted variants of Pleconaril were designed using Chemsketch software and docked into the highly conserved active site of NA using arguslab software. 198 out of 310 Pleconaril variants analyzed for docking with NA active site were proven effective, based on their free binding energy.Pleconaril variants with F, Cl, Br, CH3, OH and aromatic ring substitutions were shown to be effective alternatives to Oseltamivir as anti influenza drugs. In-silico computational techniques and a target site in the protein can be selected [Most of the early antiviral drugs were discovered after screening large number of possible drug compounds using trial and error method. Lately, this approach has been largely replaced by rational drug design, in which, a target viral protein is identified for the drug . A detaiselected . In inflselected . Ever siselected ,5. The rselected .Pleconaril Figure  is a novMolecular modeling tools help in generating new candidate drug molecules within a short span of time. After generation of new possible drug candidates, the drug and target protein interaction dynamics can be predicted by carrying out a docking analysis. The knowledge so derived is used to predict the strength of association or binding affinity between the two molecules based on scoring functions.A database of potential drug molecules and target protein structure serve as inputs for docking analysis. The success of the docking analysis is determined by scoring function and search algorithm , which hIn our present study, a database of 310 novel substituted Pleconaril variants was built by altering the side chains and substituting different aromatic rings into the original Pleconaril molecule. Molecular docking analysis was performed to visualize the interaction of each of these variants with target molecule. An attempt was made to identify Pleconaril variants with best NA binding ability.3BEQ) was obtained from Protein Data Bank (PDB) [The crystal structure of NA of 1918 Spanish flu (A/Brevig Mission/1/18 H1N1) virus (PDB ID: nk (PDB)  with a rnk (PDB) . Before 65028) [1684) [Oseltamivir (Compound ID 65028)  and Plec) [1684) . Using A) [1684)  softwareUsing Arguslab 4.0.1  softwareThe highly conserved active site of NA, which comprises of 8 functional residues  targeting which oseltamivir has been designed was considered as active site for docking analysis.® Discovery Studio 3.0 Visualizer software [Docking between receptor and ligands was performed using "Dock a Ligand" option of arguslab 4.0.1 software. A spacing of 0.4 Å between the grid points was used. "ArgusDock" was selected as docking engine. "Regular precision" was selected in docking precision menu, "Dock" was chosen as calculation type, "Flexible" for the ligand and "AScore" was used as the scoring function. A maximum of 150 poses were allowed to be analyzed, binding site box was set to 25 × 25 × 25 angstroms to encompass the entire active site. Each docking run was repeated three times to get best results. Resulted docked molecules were saved in .pdb format and all the docking images were generated using AccelrysAll the 310 variants of Pleconaril molecule were analyzed for binding with the active site of NA. 198 out of these were found to have optimum binding efficiency, based on the binding energy calculations in comparison with Oseltamivir.Further investigations showed that Oseltamivir formed 6 hydrogen bonds with TYR 406, GLU 277, ARG 224 Figure , whereasPleconaril derivative (Numbered 3 in Table In-silico investigation in modifying the Pleconaril molecule for anti-influenza activity, further In-vivo validation and conformation of the present findings is required.Computer aided drug designing and molecular docking analysis are highly effective in creating and analyzing new candidate drug molecules. 198 out of 310 Pleconaril variants analyzed for docking with NA active site were proven effective. Pleconaril variants with F, Br, CH3, Cl, OH and aromatic ring substitutes showed higher levels of NA binding ability. Several interactions such as hydrogen bonds, hydrophobic, hydrophilic interactions, electrostatics and Vanderwaal forces are thought to have played an important role in stabilizing the drug and target complexes based on the theoretical modeling. Thus, based on the above results we propose Pleconaril variants numbered 3, 4 and 11 (Figure The authors declare that they have no competing interests.SHB conceptualized the idea, carried out the work including acquisition of the data, analysis of the final results and drafting the manuscript. RNP was involved in supervising the work designed, analysis and interpretation of the data as well as drafting the manuscript. Both authors read and approved the final manuscript.


Five one-day weighed food records (WFR) were administered over one month followed by the DQESV2. Estimates for nutrients  and fruit and vegetable servings were compared between methods using correlation coefficients, 95% limits of agreement, and quintile classifications. One week later, a second DQESV2 was completed by n = 77 of the participants to assess reproducibility using intra-class correlations (ICC) and weighted kappa. Comparing methods, all nutrients and fruit and vegetable servings showed significant positive correlations (P<0.05) except protein intake in males; over 60% of participants were within one quintile classification except total fat and dietary fibre intakes in males ; and differences in nutrient and food intakes between methods were all within +/−20% of the mean WFR values except alcohol intake in females. Between first and second administrations of the DQESV2 all ICC coefficients were positive (P<0.01) and weighted kappa coefficients ranged from 0.54 for fruit servings (including fruit juice) in males to 0.91 for protein intake in females. Over a one month period, the DQESV2 demonstrated good reproducibility for the studied nutrients and for fruit and vegetable servings and provided a valid measure of the studied nutrients, except alcohol in females, and of fruit servings (including fruit juice) in both genders, at the group level in this young adult population.This research assessed the relative validity and reproducibility of the Dietary Questionnaire for Epidemiological Studies (DQESV2) over one month in young adults, given the lack of concise and convenient instruments for assessing recent dietary intake in this population. Participants were recruited from a large Australian university (N = 102; 35% male; age 18–34 years; body mass index 16–37 kg/m The food frequency questionnaire (FFQ) is a method of dietary assessment that measures an individual’s usual consumption of food items or food groups within a specified time period In the 1980’s the Cancer Council Victoria developed a 121 item FFQ to measure dietary intake in men and women aged 40 to 69 years, participating in the Melbourne Collaborative Cohort study Compared with older adults, young adults consume the lowest amount of fruit and vegetables Participants were recruited via student newsletters and advertisements posted around the main university campus during August, 2011. Prospective participants were screened for the following exclusion criteria: weight reducing or medical diet; pregnant; aged <18 or >34 years. Body weight was measured to nearest 0.1 kg using digital scales in light clothing, without shoes, having removed accessories e.g. watches, wallets. Standing height was measured to nearest 0.1 cm using a stadiometer, standing with a straight spine, gaze forward. All participants were reimbursed for their participation. This study was conducted according to the guidelines laid down in the Declaration of Helsinki and all procedures involving human subjects/patients were approved by the University of Sydney Human Research Ethics Committee . Written informed consent was obtained from all participants.The WFR was selected as the reference method to validate the DQESV2 ‘Over the last 1 month, on average, how often did you eat the following foods?’ Participants were not provided with a specific definition of the number of days in a month. Respondents were also invited to complete a second DQESV2 one week later, to study the reproducibility of the questionnaire. A dietitian collected and checked all questionnaires for errors and missing responses at the time of administration.The DQESV2 was self-administered two weeks after completing the last one-day WFR to minimise the effect of recalling intake on responses. The semi-quantitative DQESV2 is categorised into three parts: 1) ten questions about the quantity and type of commonly consumed items which are used to provide additional detail for some of 74 food frequency items; 2) four questions based on a series of portion size photos for different food types used to scale intake data; 3) a list of 74 food/beverage items categorised under “cereal foods, sweets and snacks” (21 items), “dairy products, meat and fish” (15 items), “fruit” (13 items) and “vegetables ” (25 items) with ten frequency responses ranging from “never” to “3 or more times per day”; followed by three additional questions to quantify intake of alcoholic beverages. Further detail on the DQESV2 instrument is provided by Hodge et al. total energy, protein, total fat, saturated fat, carbohydrate, sugars, dietary fibre, and alcohol). To ensure comparability, participants’ WFR were also analysed using the NUTTAB 1995 database in the nutrient analysis software Foodworks Professional Edition version 6 , from which, total daily intakes for the selected nutrients were obtained through an average of the five days of records (WFR Mean).Completed DQESV2 forms were sent to Cancer Council Victoria for analyses using specialist software based on the Australian NUTTAB 1995 food composition database, which yielded total daily intakes for the selected nutrients . The number of daily fruit servings and vegetable servings were then calculated separately for both the DQESV2 and WFR Mean, by dividing whole fruit by 150 g, whole vegetables by 75 g, and fruit or vegetables juices by 130.9 grams (i.e. 125 mL * 1.046 specific gravity) rep:BMR). Statistical methods commonly used in validation studies of FFQs include correlation as well as measures of agreement such as weighted kappa on quintiles of intake and the Bland and Altman methods t *SD of the difference to assess the acceptability of agreement which is based on clinical judgement of the LOA rather than set thresholds or cut-points Difference = a+β(Average) was calculated for each nutrient and fruit and vegetable servings, with the beta-coefficient (β) value for the slope used to indicate the degree of over- or under-estimation (agreement or difference) over the level of intake (average). Quintiles of intake from each method (WFR Mean and DQESV2) were calculated for each nutrient and fruit and vegetable servings, with the agreement between these quintile classifications compared using the weighted kappa statistic. To test the reproducibility of the DQESV2, nutrient and fruit and vegetable servings from the first and second DQESV2 administrations were compared using intra-class correlations (ICC) and the weighted kappa statistic for quintile classifications, including 95% confidence intervals, to provide measures of both consistency and agreement between repeat administrations All statistical analyses were performed using SPSS , except weighted kappa statistics which were computed using SAS version 9.2 . The basal metabolic rate (BMR) of study participants was calculated using Schofield equations based on body weight, age and gender 2 (range 16–37 kg/m2). Of the seven excluded participants, three did not follow the study protocol, two lost contact and two withdrew. Cut-offs for the top and bottom 2.5% of the distribution for the EIrep:BMR ratio were >2.10 and <0.68 for the five-day average of the WFR (WFR Mean), compared with >2.15 and <0.53 for the DQESV2. A total of seven participants were outside these cut-offs, ranging from 0.56 to 2.42 in the WFR Mean and 0.41 to 2.25 for the DQESV2. As the exclusion of these participants from the analysis did not influence the findings, all participants were included in the analyses presented. Due to observed differences in validity and reproducibility findings between genders, the following results are presented separately for males and females.From a total of 109 participants recruited, 102 (93.6%) completed the study and were included in the analyses. Of these, sixty six participants were female (64.7%) and thirty six were male (35.3%), the mean (SD) age was 23.5 (4.1) years (range 18–34 years) and BMI 23.0 (3.3) kg/mP<0.05), except protein intake in males.Difference = a+β(Average). Larger beta coefficients indicated poorer agreement with increasing level of intake (P<0.001), as shown in Through visual inspection of the Bland and Altman plots (i.e. the difference between methods (WFR Mean – DQESV2) plotted against the average ([WFR Mean+DQESV2]/2), a tendency toward poorer agreement in nutrient intakes between methods was observed with higher levels of intake, i.e. the difference between the DQESV2 and WFR Mean estimates became larger as nutrient intake increased. f intake . Most cof intake  and protAgreement within DQESV2 quintile classifications in the nutrients and fruit and vegetable servings is presented in P<0.01), ranging from 0.54 for fruit servings (including fruit juice) in males to 0.91 for protein intake in females. Weighted kappa coefficients ranged from 0.43 (moderate agreement) for dietary fibre in males to 0.74  for protein intake in females.A total of 77 participants completed a second DQESV2. These participants were not statistically different in age, gender or BMI to those who did not complete a second DQESV2. P<0.05), except protein intake in males. Correlation coefficients reported in this study were substantially higher than those found in an earlier study assessing the validity of an Australian FFQ against food records in 9–16 year old children over a six month period This study aimed to examine the relative validity and reproducibility of a semi-quantitative food frequency questionnaire (DQESV2) against five-days weighed food records, over a one month period in young adults. Comparing the two dietary assessment methods, all nutrients, fruit servings and vegetable servings were found to be significantly correlated  and eggplant , which if included may have improved estimates from the DQESV2. The DQESV2 slightly over-estimated fruit intake at the group level in males by around half a serving per day, although this over-estimation was attenuated when fruit juice was included. It is unlikely that this over-estimation was due to the scaling question “How many pieces of fresh fruit do you usually eat per day?”, as the median response to this question, 1–2 pieces per day, was consistent with the mean 1.9 total daily equivalent serves of fruit (excluding fruit juice) reported in the frequency section of the DQESV2. Worth noting however was that, similar to vegetables, there were also a number of fruits that were reported as consumed in the WFRs that were not included in the DQESV2, such as blueberries, cranberries, raspberries, cherries, grapes, guava, kiwi fruit, lychees, passion fruit, plum, prune, quince, and dried fruits. Of these, the most frequently consumed were kiwi fruit , grapes  and blueberries , which if included, may have increased the degree to which the DQESV2 over-estimated fruit intake.Food groups have not previously been reported in validation studies of the DQESV2. Increasingly it is being recognised that taking a reductionist nutrient-centric approach to study diet-disease relationships is less useful than a food-based approach. The findings of the present study were similar to another Australian study that used a different FFQ  in 96 subjects who completed 6 two-day weighed food records over a one-year period, where fruit and vegetable intake was shown to be correctly classified to within one quartile for 79% and 69% of subjects, respectively P<0.01), ranging from 0.54 for fruit servings (including fruit juice) in males to 0.91 for protein intake in females. These coefficients are substantially higher than those found from repeat administration of an FFQ one year apart on a sample of Australian women 22 to 79 years of age Reproducibility of the DQESV2 has not been previously reported. This study found all reproducibility correlation coefficients to be significant , cream, oils, and some bakery items . The DQESV2 asks about butter and margarine spreads but not oils, and this may be why total fat intake among females was underestimated in the DQESV2 while estimates for saturated fats were more comparable between methods. As the DQESV2 only asks about slices of bread, participants may not have included other types of breads consumed (e.g. flat breads or bagels). Further, as the DQESV2 asks about specific brands of breakfast cereals, participants might not have included other cereal brands, despite them having similar nutritional content. These factors might explain why energy, carbohydrate and dietary fibre intakes were under-estimated by the DQESV2. It is possible the participants had a self interest in dieting for weight management  which could have led to some of the young adults under-eating during the study, although this should have been captured in both dietary assessment methods. Participants were also tertiary education students from a largely urban area of New South Wales, Australia, and hence this limits the applicability of validity and reproducibility findings to young adults outside of this population. Finally, this study only assessed the specific nutrients energy, protein, total fat, saturated fat, carbohydrate, sugars, dietary fibre and alcohol as well as fruit and vegetable servings, which are the focus of our proposed intervention studies.One of the strengths of the current study is that five one-day WFR were used for comparison which is the gold standard for measuring dietary intake In summary, the DQESV2 is appropriate for use in this population group for estimating intake of the studied nutrients, except alcohol intake among females, and of fruit servings when fruit juice is included, at the group level over a one month period. While the agreement between methods was poorer for alcohol generally, it is not clear which was the better method. The DQESV2 cannot be recommended for estimating vegetable servings or for estimating fruit servings at the individual or group level, unless servings of fruit juice are included, due to the poor agreement found between methods. However, the DQESV2 demonstrated good reproducibility for all nutrients studied and for fruit and vegetable servings among both males and females from this population. The ability to use the easily-administered DQESV2 to assess diet in this population will be beneficial, particularly given the recent availability of an electronic version of this questionnaire.


Understanding diversity patterns and the mechanisms underlying those patterns along elevational gradients is critically important for conservation efforts in montane ecosystems, especially those that are biodiversity hotspots. Despite recent advances, consensus on the underlying causes, or even the relative influence of a suite of factors on elevational diversity patterns has remained elusive.We examined patterns of species richness, density and range size distribution of birds, and the suite of biotic and abiotic factors  that governs diversity along a 4500-m elevational gradient in the Eastern Himalayan region, a biodiversity hotspot within the world's tallest mountains. We used point count methods for sampling birds and quadrats for estimating vegetation at 22 sites along the elevational gradient. We found that species richness increased to approximately 2000 m, then declined. We found no evidence that geometric constraints influenced this pattern, whereas actual evapotranspiration (a surrogate for primary productivity) and various habitat variables  accounted for most of the variation in bird species richness. We also observed that ranges of most bird species were narrow along the elevation gradient. We find little evidence to support Rapoport's rule for the birds of Sikkim region of the Himalaya.This study in the Eastern Himalaya indicates that species richness of birds is highest at intermediate elevations along one of the most extensive elevational gradients ever examined. Additionally, primary productivity and factors associated with habitat accounted for most of the variation in avian species richness. The diversity peak at intermediate elevations and the narrow elevational ranges of most species suggest important conservation implications: not only should mid-elevation areas be conserved, but the entire gradient requires equal conservation attention. Biodiversity varies geographically, and understanding why is one of the fundamental questions in biogeography, macroecology, and conservation ecology. Perhaps the best- studied pattern in species richness is the latitudinal gradient in diversity - the decline (for most taxa) in richness with increasing distance from the equator Patterns in diversity along elevational gradients might vary among taxa, regions, and spatial scales The most frequently documented correlates and drivers of elevational patterns of diversity include contemporary climate  are important drivers for such a pattern In this study, we examine the elevational gradient in bird diversity in the Sikkim region of the Eastern Himalaya, home to the tallest mountains in the world. In particular, our aims are to document, describe, and explain the elevational gradient in bird diversity in the Eastern Himalaya. First, we describe the pattern along this extensive gradient . Then, we evaluate a suite of biotic and abiotic factors that might be correlated with bird diversity, focusing on geometric constraints, temperature, precipitation, potential evapotranspiration (PET), actual evapotranspiration (AET), plant species richness, tree density, shrub density and basal area of tree. These parameters broadly represent MDE, energy, productivity and habitat diversity. Finally, we assessed the range size distribution pattern of birds along the elevation gradient by examining the elevational range size of each bird species and the applicability of Rapoport's rule.The study area is in the Eastern Himalayan Mountains . Elevation in this region ranges from c.300 m to above 8000 m. The study sites were located in the Teesta Valley which consists of rough hilly terrain and varies in elevation from 300–5500 m. Both climate  and vegetation type  vary with elevation, within a distance of ∼150 km. The lower and middle valleys (<2000 m) are hot and humid with annual precipitation exceeding 2500 mm, while elevations above 2500 m are relatively drier and colder with substantially less rainfall (<1000 mm). At the high elevation sites, precipitation is in the form of snowfall, and most of the alpine region remains under snow for almost 7–8 months a year.Six major vegetation zones occur in the study area. These are Tropical semi-deciduous forests (<900 m), Tropical moist and broad-leaved forests (900–1800 m), Temperate broad-leaved forests (1800–2800 m), Temperate coniferous and broad-leaved forests (2800–3800 m), Sub-alpine (3800–4500 m) and Alpine vegetation (>4500 m) To quantify variation in the richness and abundance of birds along this elevational gradient, at 22 sites, we used the open width point count method along transects 2/4Л, where Л  = 3.14.We also sampled the trees and shrubs at each of the 22 sites. Along each transect used for sampling birds at each site, we placed 10 20 m × 10 m quadrats for enumeration of trees. Plants with GBH (girth at breast height) >20 cm were considered as trees. For estimating shrub density, two 5 m × 5 m sub-quadrats were placed diagonally within each of the 20 × 10 m quadrats. Thus, for each site, we recorded the richness and density of trees, the richness and density of shrubs, and the GBH of trees. We also estimated basal area of trees for each site using the formula: Basal Area  =  (GBH)We obtained rainfall and temperature data from seven locations at different elevations in the study region from Indian Meteorological Department. Based on these data, rainfall and temperature were estimated for all locations using regression equations, as is often done in these types of analyses 2 = 0.975, p<0.01rainfall =  -0.7909(elevation) + 4046.1, R2 = 0.983, p<0.01.temperature  = -0.0062(elevation) + 29.85, R 2]1/2 with L = 300 + 25T + 0.05T3, where P = mean annual precipitation and T = mean annual temperature We calculated potential evapotranspiration (PET) using the formula [PET =  mean annual bio-temperature (i.e. temperature > 0°C) x 58.93] see . PET is 2C, where D = bird density (numbers/ha), n = total number of birds observed in all counts within the specific radius, r = specific radius (m) , C = total number of counts conducted and Л = 3.14. We also estimated individual-based rarefied richness, which accounts for variation in the number of individuals sampled. We rarefied to the lowest number of individuals detected in any one survey (n = 260). Because some sites were more frequently sampled than others, we also used sample-based rarefaction (rarefied to lowest number of counts conducted (n = 72 point counts) for any site. Since the individual and sample-based rarefaction results were qualitatively similar, we report only the results from the individual-based rarefaction  at each site from one season (summer) and rarefied to lowest number of individuals detected at any one site (n = 15). We found that the pattern of this two-step rarefaction procedure did not differ qualitatively from either total bird species richness see  or the m2) in each model to relate each of the response variables  to elevation. We compared AIC values to determine whether the linear or quadratic model best accounted for variation in each of the response variables. Because spatial autocorrelation can inflate errors in the statistical analyses of ecological data I coefficients with the software SAM version 4.0  regressions. Because the relationship between any estimate of richness and elevation need not be linear, we also used a quadratic term . However, the pattern was not clearly hump-shaped. Instead, below about 2000 m, there was no systematic variation in rarefied richness with elevation, but above 2000 m, rarefied richness declined with elevation  and density  peaked at mid-elevations . Additionally, bird species richness did not correlate with the MDE predicted richness , rarefied richness and density as a function of six environmental factors are shown in The rThe spatial correlogram for species richness  indicateElevational range profiles of the birds of Eastern Himalaya showed that most species occupied very narrow elevational ranges along the gradient . Ninety Along one of the most extensive elevational gradients in the world, we found that the species richness of birds in Eastern Himalaya displayed a distinct mid-elevation peak in species richness and density. Such a pattern is frequently documented in birds see , small mOne idea is that perhaps the geometry of bounded domains or available area accounts for the mid-elevation peak in richness found here Various processes such as climate, productivity, habitat heterogeneity and mass effects have been proposed to explain elevational distributions of species Many studies have examined how productivity might influence diversity Evolutionary and historical events such as geographic isolation, dispersal, speciation and endemism could shape elevational diversity in montane regions Variation in species richness and density are rarely, if ever, wholly explained by a single factor The decline in both species richness and density of birds above 2500 m is striking, and suggests an abrupt change in some factor or suite of factors that limits birds. The stature of the forest decreases dramatically at about this point, and the climatic conditions become increasingly severe beyond 2500 m in the region; both of these changes could cause declines in abundance and size distribution of invertebrates and scarcity of other food items for birds Rapoport's rule  occurred at all 22 sites. Most occurred at only a few sites, suggesting that range sizes are extremely limited in this system, probably by a combination of dispersal ability, particular habitat associations, competition, or environmental tolerance In sum, along one of the longest elevational gradients in the world, we found that bird species richness and density showed hump-shaped relationship with elevation, peaking at approximately 2000 m, in the Eastern Himalayan region. The variation in richness and density was correlated strongly with both productivity and habitat rather than geometric constraints. The small elevational ranges of species along the gradient suggest that conservation efforts should consider the entire gradient rather than just portions of it.Figure S1Species accumulation curves of birds. Curves based on number of point counts in different elevational transects in Sikkim, Eastern Himalaya. Numbers in the figures indicate elevation (m) of the sampling site.(TIF)Click here for additional data file.Figure S2Elevational variation of rarefied bird species richness. Species richness observed when rarefied to 72 point counts along elevational transects in Sikkim, Eastern Himalaya.(TIF)Click here for additional data file.Figure S3Elevational variation of rarefied bird species richness. Species richness observed when rarefied to 15 individuals from six point counts from each site along elevational transects in Sikkim, Eastern Himalaya.(TIF)Click here for additional data file.


The aim of the study was to assess the prevalence of fruits and vegetable consumption and associated factors among Southeast Asian in-school adolescents. Data were collected by self-report questionnaire from nationally representative samples  of school children aged 13 to 15 years in five Southeast Asian countries. Overall, 76.3% of the 13 to 15 year-olds had inadequate fruits and vegetables consumptions (less than five servings per day); 28% reported consuming fruits less than once per day and 13.8% indicated consuming vegetables less than once per day. In multivariable analysis, lack of protective factors and being physically inactive were associated with inadequate fruits and vegetable consumption, and sedentary behaviour and being overweight was protective of inadequate fruits and vegetable consumption. The results stress the need for intervention programmes aimed at increased consumption of fruits and vegetables, targeting proximal factors such as the family environment and distal factors by aiming at integrating other risk factors such as physical activity into health promotion among adolescents.  High fruit and vegetable intake can promote health and prevent chronic diseases such as heart diseases and certain types of cancer ,2. The hIn spite of the importance of an adequate intake of fruits and vegetables during adolescence, large population groups, including children and adolescents in most Western countries ,6, AsianTo develop effective interventions to increase fruit/vegetable intake, an understanding of etiological processes and the identification of potentially modifiable correlates is needed. Few studies have examined fruit/vegetable intake and its correlates among adolescents in Southeast Asian countries. Factors that have been found to be correlated with fruit and vegetable intake in previous studies of adolescents include age, gender, socioeconomic position, preferences, parental intake, parental modeling, family rules and parental encouragement and home availability/accessibility ,14. Theshttp://www.who.int/chp/gshs/methodology/en/index.html. For all countries national samples were included. The aim of the GSHS is to collect data primarily for students from age 13 to 15 years. A two-stage cluster sample design was used to collect data to represent all students in grades 6, 7, 8, 9, and 10 in the country. At the first stage of sampling, schools were selected with probability proportional to their reported enrollment size. In the second stage, classes in the selected schools were randomly selected and all students in selected classes were eligible to participate irrespective of their actual ages. School-aged youth completed a self-administered questionnaire within the selected school. Data collection was conducted by trained survey administrators during one regular class period. Student privacy was protected through anonymous and voluntary participation, and informed consent was obtained as appropriate from the students, parents and/or school officials.This study involved secondary analysis of existing data from the Global School-Based Health Survey (GSHS) from five Southeast Asian countries . Details and data of the GSHS can be accessed at The GSHS 10 core questionnaire modules address the leading causes of morbidity and mortality among children and adults worldwide: tobacco, alcohol and other drug use; dietary behaviors; hygiene; mental health; physical activity; sexual behaviors that contribute to HIV infection, other sexually-transmitted infections, and unintended pregnancy; unintentional injuries and violence; hygiene; protective factors and respondent demographics .Fruits and Vegetables Consumption and Hunger: Fruits: “During the past 30 days, how many times per day did you usually eat fruit, such as ‘country specific examples’?”. Response options were 1 = I did not eat fruit during the past 30 days, 2 = less than one time per day, 3 = one time per day to 7 = five or more times per day. Vegetables: “During the past 30 days, how many times per day did you usually eat vegetables, such as ‘country specific examples’?”. Response options were 1 = I did not eat vegetables during the past 30 days, 2 = less than one time per day, 3 = one time per day to 7 = five or more times per day. The United States Department of Health and Human Services [Hunger: A measure of hunger was derived from a question reporting the frequency that a young person went hungry because there was not enough food at home in the past 30 days  . Services  recommenBody Mass Index (BMI) Measurement and Overweight Classification: Height and body weight were based on self-reports. BMI was calculated as weight/height2 (kg/m2). The international age-and gender-specific child BMI cut-points were used to define overweight and obesity [2 were classified as overweight.  obesity . These cSubstance Use Variables: Smoking cigarettes: During the past 30 days, on how many days did you smoke cigarettes?  . Alcohol use: during the past 30 days, on how many days did you have at least one drink containing alcohol. Response options were from 1 = zero days to 7 = all 30 days; Coded 1 = 1 or 2 to all 30 days, and 0 = zero days. Drugs: During your life, how many times have you used drugs, such as glue, benzene, marijuana, cocaine, or mandrax? Response options were from 1 = zero times to 4 = ten or more times; Coded 1 = one or two to ten or more times, and 0 = zero times.Physical Activity: Leisure time physical activity was assessed by asking participants: “Physical activity is any activity that increases your heart rate and makes you get out of breath some of the time. Physical activity can be done in sports, playing with friends, or walking to school. Some examples of physical activity are running, fast walking, biking, dancing, football. Do not include your physical education or gym class.” “During the past 7 days, on how many days were you physically active for a total of at least 60 minutes per day?” and “During a typical or usual week, on how many days are you physically active for a total of at least 60 minutes per day?” Leisure time sedentary behaviour was assessed by asking participants about the time they spend mostly sitting when not in school or doing homework: “How much time do you spend during a typical or usual day sitting and watching television, playing computer games, talking with friends, or doing other sitting activities.” Sedentary behaviour was defined as three or more hours sitting in a day.Psychological Distress: Psychological distress was assessed with 5 items. Loneliness: “During the past 12 months, how often have you felt lonely?”  . Suicide ideation: “During the past 12 months, did you ever seriously consider attempting suicide?” . No close friends:”How many close friends do you have?” . Anxiety or worried: During the past 12 months, how often have you been so worried about something that you could not sleep at night?  . Sadness: During the past 12 months, did you ever feel so sad or hopeless almost every day for two weeks or more in a row that you stopped doing your usual activities? (response option 1 = yes and 2 = no) . A psychological index was created by adding up all five items, and recoding the sum into low = no psychological distress, medium = 1 item of psychological distress and high = 2 or more psychological distresses endorsed.Protective Factors: Protective factors were assessed with five items on peer support at school, parental or guardian supervision, connectedness, and bonding. Peer support at school was assessed with the question “During the past 30 days, how often were most of the students in your school kind and helpful?” Parental or guardian supervision “During the past 30 days, how often did your parents or guardians check to see if your homework was done”? Parental or guardian connectedness “During the past 30 days, how often did your parents or guardians understand your problems or worries?” and Parental or guardian bonding “During the past 30 days, how often did your parents or guardians really know what you were doing with your free time?” Response options to these questions were from 1 = never to 5 = always, coded 1 = never or rarely and 0 = sometimes to always. All four protective factor items were added up, and recoded into low, medium and high lack of protective factors.In order to compare study samples across countries each country sample was restricted to the age group 13 to 15 years, younger and older participants were excluded from the analyses. For each country, the overall response rates ranged from 81 to 95% see .i.e., cluster sampling owing to the sampling of school classes. Associations between distal factors , proximal factors (protective factors) and other risk behaviour  among school children were evaluated calculating odds ratios (OR). Logistic regression was used for evaluation of the impact of explanatory variable for inadequate fruits and vegetables (<5 servings per day) (binary dependent variable). In the analysis, weighted percentages are reported. The reported sample size refers to the sample that was asked the target question. The two-sided 95% confidence intervals are reported. The p-value less or equal to 5% is used to indicate statistical significance. Both the reported 95% confidence intervals and the p-value are adjusted for the multistage stratified cluster sample design of the study.Data analysis was performed using STATA software version 10.0 . This software has the advantage of directly including robust standard errors that account for the sampling design, The total sample included 16,084 school children aged 13 to 15 years from five Southeast Asian countries. There were slightly more male (51.2%) than female (48.8%) school children. In all most children 76.3% consumed less than the recommended five servings of fruits and/or vegetable, this was the same among boys and in girls. Twenty-eight percent of the participants reported consuming less than once fruits per day and 13.8% indicated consuming less than once vegetables per day. The mean number of fruits consumed per day was 1.3, far below the recommended two fruits a day target. The mean number of vegetable consumption was 1.9, also far below the recommended three vegetables a day target. The mean daily servings of fruits and/or vegetables per day was 3.2, far below the recommended five servings a day target. Indian and Myanmar school children had the most insufficient. In terms of mean daily servings of fruits and/or vegetables, Thai school children had the highest consumption (3.7), followed by Indonesian (3.2) and Sri Lankan (3.1) school children, and the lowest consumption was among Myanmar (2.9) and Indian (3.0) school children see .In univariate analysis lack of protective factors and being physically inactive were associated with inadequate fruits and vegetable consumption and substance use , sedentary behaviour and being overweight or obese were protective of inadequate fruits and vegetable consumption. In multivariable analysis lack of protective factors  and being physically inactive  were associated with inadequate fruits and vegetable consumption and sedentary behaviour  and being overweight  were protective of inadequate fruits and vegetable consumption see .The current study explores fruits and vegetable consumption and its correlates among in-school adolescents from nationally representative samples in five Southeast Asian countries. Overall, low fruits and vegetable consumption were reported, as 76.3% of the sampled population consumed less than the recommended five servings of fruits and/or vegetable, 28% reported consuming less than once fruits per day and 13.8% indicated consuming less than once vegetables per day. Studies from other low and middle income countries and high income countries seem to confirm low fruits and vegetable consumption levels among adolescents, e.g., in seven African countries 77.5% of adolescent learners  had eateThis study found cross-national variations in the prevalence of fruits and vegetable consumption. In terms of mean daily servings of fruits and/or vegetables Thai school children had the highest consumption (3.7), followed by Indonesian (3.2), Sri Lankan (3.1), Indian (3.0) and Myanmarian (2.9) school children. Possible reasons for such country differences could be fruits and vegetable availability, income and urbanization rate. Availability data on fruits and vegetables in the study countries seem to confirm a relationship between consumption and availability for Thailand and Sri Lanka but not for India, Indonesia and Myanmar see . The WHOet al. [In our study Thai school children had the highest consumption of fruits and vegetables, which may be due to larger availability of fruits and vegetables and greater income  ,14,19, aet al.  reviewed [et al.  found th [et al. ,18,19,22The cross-national data found inadequate fruits and vegetable consumption in five Southeast Asian countries, pointing to the conclusion that programmes are needed to improve fruit and vegetable consumption of the adolescent population. Interventions to improve health-related behaviours should be tailored to the most important determinants or mediators of these behaviours including targeting proximal factors such as the family environment for the promotion of healthy eating behaviours among adolescents. The risk factors identified were consistent with the problem behaviour theory in which inadequate fruits and/or vegetable consumption are shared with other problem behaviours such as physical inactivity, which indicates that health promotion programmes should be broadened to include these factors collectively in health interventions for adolescents.This study had several limitations. Firstly, the GSHS only enrolls adolescents who are in school. School-going adolescents may not be representative of all adolescents in a country as the occurrence of fruits and vegetable consumption may differ between the two groups. As the questionnaire was self-completed, it is possible that some study participants may have miss reported either intentionally or inadvertently on any of the questions asked. Intentionally miss reporting was probably minimized by the fact that study participants completed the questionnaires anonymously. Further, the assessment of correlates of fruits and vegetable consumption was limited and other factors such as preferences, parental intake, parental modeling, family rules and parental encouragement, home availability/ accessibility, school level factors such as nutrition education, availability and policy of healthy and unhealthy foods, community and or neighbourhood level factors, and national level factors such as price levels, policy, guidelines, supply, and exposure to mass media and commercials should be included in other studies among adolescents ,14,19. F


To examine the role of both protein kinase C (PKC)-β and vascular endothelial growth factor receptor (VEGFR)-2 in malignant pleural mesothelioma (MPM) using respective inhibitors, enzastaurin and KRN633.Caenorhabditis elegans, was treated with enzastaurin to determine its suitability to screen for highly potent kinase inhibitors.MPM cell lines, control cells, and a variety of archived MPM tumor samples were used to determine the protein expression levels of PKC-β, VEGFR-2, VEGF, and p-AKT. Effects of enzastaurin and KRN633 on phosphorylation status of key signaling molecules and viability of the mesothelioma cells were determined. The common soil nematode, C. elegans with enzastaurin resulted in clear phenotypic changes and the worms were hypermotile with abnormal pattern and shape of eggs, suggesting altered fecundity.PKC-β1, PKC-β2 and VEGFR-2/KDR were overexpressed in MPM cell lines and MPM tumor tissues. Enzastaurin treatment resulted in significant loss in viability of VEGF induced cell proliferation; however, the effect of KRN633 was much less. Enzastaurin also dramatically decreased the phosphorylation of PKC-β, its downstream target p-AKT, and surprisingly, the upstream VEGFR-2. The combination of the two drugs at best was additive and similar results were obtained with respect to cell viability. Treatment of C. elegans can be used to screen for the next generation inhibitors as treatment with enzastaurin resulted in clear phenotypic changes that could be assayed.PKC-β1 and VEGFR-2 are both excellent therapeutic targets in MPM. Enzastaurin was better at killing MPM cells than KRN633 and the combination lacked synergy. In addition, we show here that  Caenorhabditis elegans, can be potentially used to screen (in vivo) for the next generation inhibitors as treatment with enzastaurin resulted in clear phenotypic changes that could be assayed.Malignant pleural mesothelioma (MPM) is a devastating disease and the prognosis is relatively poor despite recent advances in chemotherapy. There is a clear and urgent requirement for the identification of novel mesothelioma targets for the development of highly efficacious therapeutics. In this paper, we have shown that both protein kinase C (PKC-β1 and -β2 isoforms) and vascular endothelial growth factor receptor (VEGFR)-2 are overexpressed in different types of mesotheliomas, thereby making them excellent combinatorial therapeutic targets. Enzastaurin, in comparison to KRN633, was highly effective in arresting key phosphorylation events and cell survival; however, the combination of the two was at best additive. As PKC appears to be the most attractive primary chemotherapeutic target, we subjected it to further study. Here, we have shown that the common soil nematode, µM and is orally absorbed. Early pre-clinical studies have shown that enzastaurin is active against lung cancer. The cell lysates were cleared by centrifugation at 14,000 g for 15 minutes. The protein concentrations were determined by using the Bradford assay. Protein lysates (60–80 µg) were separated by 7.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions and transferred to PVDF membranes . The membranes were blocked in 5% non-fat dried milk freshly made in Tris-Buffered Saline Tween-20 (TBST). Proteins were detected by immunoblotting using an enhanced chemiluminescense kit  according to manufacturer’s protocol.Cells were exposed to varying concentrations of enzastaurin for the indicated time periods as described above. The cells were washed twice with ice-cold phosphate buffered saline (PBS), and whole cell lysates were prepared in RIPA lysis buffer [50 mM Tris (pH 8.0), 150 mM NaCl, 10% glycerol, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS and 0.42% NaF containing protease inhibitor cocktail  was dose dependent. The Met5A control cells showed a robust cell survival at a wide range of enzastaurin concentrations (data not shown), as compared with H2052, MSTO, H513, H2373, H2452, H2691, and H2461 malignant mesothelioma cell lines. Of the cell lines examined, H2461 showed the highest sensitivity to enzastaurin, whereas H2052 was relatively less sensitive.We examined the role of enzastaurin, an inhibitor of PKC-β in normaWe investigated the effect of VEGFR-2 inhibitor, KRN633, and enzastaurin for their antiproliferative activity in normal and malignant cell lines. To determine the role of VEGF induced tyrosine phosphorylation in H2461 mesothelioma cells that expressed PKC-β1 and PKC-β2, we pre-treated serum-starved mesothelioma H2461 and Met5A cells with or without the indicated concentrations of KRN633 (4 hours) and enzastaurin for 24 hours . Cells wTo assess the signaling events associated with malignant mesothelioma cells, we determined the phosphorylation status of VEGFR-2, PKC-β2, and AKT in the presence of KRN633 and/or enzastaurin in normal mesothelial Met5A cell and H2461 cells . Our resC. elegans. We therefore determined the effect of enzastaurin treatment on development and the morphology of C. elegans. At lower concentrations of enzastaurin (1–5 µM), there was no behavioral or morphological alterations observed in C. elegans larvae or adult hermaphrodites (data not shown). However, at higher concentrations, nematodes demonstrated uncoordinated hyperactivity movement and defects in body morphology [Figure An orthologue of PKC-β is expressed in y Figure –d. In adPKC-β2 gene in MPM (9 cell lines and 33 tumor tissues), there was no amplification seen of PKC-β2; however, in two different tumor tissues, there were synonymous single nucleotide variants (SNVs), 119/T7TC: Pro and 1770C7CT: Gly. Both of these SNVs were in the ATP binding domain (data not shown).MPM is a difficult disease to treat. Even with the best of therapies, the prognosis is quite poor.–2 In ordC. elegans nematodes, there are four isoforms of PKC, of which the PKC1 isoform is specifically expressed in neurons.[C. elegans model could be used for the study of PKC mediated signaling and for the purpose of establishing a model that is amenable for high throughput screening, we showed specific enzastaurin induced changes in overall morphology, motility and egg laying pattern. It would be important in generating transgenic worms that overexpress constitutively active form of PKC-β1 and use them for screening for potent inhibitors. Also, as we have shown previously for inducing a potential lung cancer model, C. elegans can be exposed to nicotine[C. elegans to asbestos fibers. Chrysotile has been used more than any other type and accounts for about 95% of the asbestos found in buildings in the USA. We have preliminarily examined the toxic effect of this fiber in control (N2) C. elegans. C. elegans was cultured in M9 medium for 2 weeks with or without chrysotile fibers (asbestos), after which survival and rates of developmental defects were estimated. Chrysotile reduced survival and induced locomotion defects. It would be important now to determine the effects on the PKC (and other molecule) axis with this model and as well determine the effects of drugs such as enzastaurin.The classification for MPM is based upon morphology and consists of EPI, biphasic, and SAR histology. Our data neurons. In order nicotine or smokeC. elegans model can be used to investigate PKC signaling, especially in the context of drug screening.In conclusion, we have unambiguously shown that VEGFR/PKC/AKT signaling axis is highly active in MPM and that the signals mediated by this pathway are essential for the survival of MPM. We also demonstrate here that PKC inhibitor, enzaustarin, is more effective in killing MPM cells compared to KRN633 and the combination of the two drugs is additive. Most importantly, Dr. Ravi Salgia, is an expert in the area of thoracic oncology, translational research and basic science research. His main research interest is to arrive at novel targeted therapeutics to enhance the quality of life and survival for patients with cancer. He and his lab have recently identified several novel receptor tyrosine kinases that are abnormal in lung cancer. He is an expert in signal transduction, as related to growth factor receptors and oncogenes/ tumor suppressor genes, and is developing novel inhibitors based on these pathways. He is also an expert in tumor and body fluid biomarkers, and is applying this knowledge to develop new serum/tumor tissue tests for thoracic oncology.