The discourse of dual burden caused through underweight and overweight is well-documented globally but this issue and its connection with women's health in Bangladesh is yet to be explored widely. To enrich the current debate, this study, in the context of Bangladesh, examines the patterns, prevalence, and socioeconomic factors influencing the ever-married women of being underweight and overweight over normal weight. Data used in this study have been extracted from the most recent 2011 Bangladesh Demographic and Health Survey. To achieve results connected with the research objectives, both bivariate and multivariate statistical analyses have been employed. In bivariate analysis, we used seven categories of BMI cutoff points for Asian countries as prescribed by World Health Organization (WHO). Multinomial logistic regression model was constructed to investigate the net effect of socioeconomic factors on underweight, pre-overweight, and overweight over normal weight. The results confirm the co-existence of underweight and overweight among women as we found the prevalence of underweight, normal weight, pre-overweight, overweight, and obesity to be 24.1%, 46.7%, 12.8%, 13.5%, and 2.9% respectively. Compared to the richest, the women from the poorest households were significantly (p<0.001) most likely to be underweight  and least likely to be overweight  over normal weight. The urban women, compared to their rural counterparts, were significantly (p<0.001) less likely to be underweight  and more likely to be overweight  than normal weight. The other socioeconomic grades that were most marked to be underweight and overweight are age, women's education, marital status, age at first childbirth, parity, number of children aged ≤5 years at the household, and food security. The findings confirm the dual burden of both under- and overweight. Systematic and regular monitoring and surveillance of the social trajectory of nutritional status of women and men in Bangladesh is crucial to develop apposite strategy that addresses the persistent and chronic problem of underweight and the emerging problem of overweight. The dual existence of both types of malnutrition among women in Bangladesh must be taken into consideration so that public health interventions may be adopted through appropriate policy. Due to economic situation of developed countries, health concern mainly on overweight receives graver attention. The ‘dual burden’ of health concern caused through both underweight and overweight is significantly important for public health policy in developing nations to address –3. StudiA landmark review of studies on socioeconomic status and obesity supports the view that obesity in the developing world would be essentially a non-communicable disease of the socioeconomic elite . The proBoth lean and obese women carry a risk of adverse pregnancy outcome and overall poor maternal and child health status. Thus, there is growing recognition of a ‘double burden’ of malnutrition among populations in both affluent and less-affluent countries , i.e. thData for this study have been used from the most recent 2011 Bangladesh Demographic and Health Survey (BDHS) . A natioThe 2011 BDHS gathered information on 17,749 ever-married women aged 15-49 years. The women with various missing information and those who were pregnant during the survey were excluded from analysis. After excluding women with missing information (n=407) on the outcome measure and those who were pregnant  during the survey, the final sample for analysis stood at 16,273. 2 (severe underweight), 16·00-16·99 kg/m2 (moderate underweight), 17·00-18·49 kg/m2 (mild underweight), 18·50-22·99 kg/m2 , 23.00-24.99 kg/m2 (pre-overweight), 25.00-29·99 kg/m2 (overweight), and ≥30.00 kg/m2 (obese). Given the identification of a BMI of 23.00 as a public-health cutoff for risk of obesity in Asian populations  shown in j=1, 2, 3, …, k nominal outcomes. In its general form, the probability of an actor i belonging to category j is given by the following formula  for each k=1, 2, 3, …, j. To identify the parameters, it is common to choose one reference category and set the corresponding vector of parameters equal to a vector of zeroes. As we did not find substantial differences in the patterning of the sociodemographic and environmental exposures between the different grades of underweight, we merged the different grades of underweight to one category of BMI <18.50 labelling by ‘underweight’. Similarly, we merged overweight and obesity by labelling as ‘overweight’. The results of the multinomial logistic regression analyses have been shown by odds ratios (ORs) with 95% confidence interval (CI) for easy understanding. The level of significance was set at 10%. Data were analyzed using SPSS (version 21) .where χThe study was based on a nationally-representative anonymous dataset for public use, with no identifiable information on the survey participants. Moreover, the survey followed all protocols prescribed by the WHO and was implemented by National Institute for Population Research and Training (NIPORT) under the Ministry of Health and Family Welfare, Government of Bangladesh. Therefore, no ethics statement is required for this study.Most of the respondents were rural residents and were currently in union, with mean±SD age of 31.2±9.2 years. The mean age at first marriage, mean age at first childbirth, mean number of children ever born per ever-married woman was 15.6±2.9 years, 17.8±3.3 years, and 2.7±1.9 (not shown in The distribution of women across categories of BMI shows that 46.7% were normal, 24.2% were underweight, 12.8% were pre-overweight, and 16.4% were overweight and obese . The assWomen's level of education was significantly and negatively associated with being underweight and was positively associated with pre-overweight and overweight relative to normal weight. The wealth quintiles show familiar fashion in the likelihood of being underweight and overweight. The risk of being underweight increased systematically with the decreases in wealth index. In contrast, the risk of being pre-overweight and overweight decreased significantly with the increase of standard of living index. The higher the food security, the lower the risk of being underweight. However, food security had no longer significant effect of being pre-overweight and overweight.The rural women, compared to their urban counterparts, were more likely to be underweight whereas urban women were more tended to be overweight than their rural counterparts. The women living in the Barisal, Chittagong, Dhaka, Khulna, Rajshahi and Rangpur regions were less likely to be underweight compared to the reference region of Sylhet. Besides, the women from Chittagong, Khulna and Rajshahi regions were more tended to be pre-overweight and overweight than those of the reference region. In addition, compared to the women from Sylhet region, those living in Chittagong and Khulna were significantly at increased risk of overweight.The present study, to our knowledge, examines for the first time the sociodemographic and environmental distribution of nutritional status of ever-married women with mean age of 31.2±9.2 years, using the Asia-specific cutoff points of BMI. The findings document the co-existence of dual burden of underweight and overweight among women in Bangladesh, using data of the most recent nationally-representative population sample survey covering all regions of Bangladesh. Findings show that, in 2011, the prevalence of underweight, normal weight, and overweight was 24.1%, 46.7%, and 29.2% respectively, suggesting that the incidence of overweight exceeded underweight for the first time. The incidence of underweight has decreased substantially and continually during the period from 2004 to 2011.There is an apparent socioeconomic, demographic and environmental distribution of causal patterns of nutritional status, with younger women, women from low socioeconomic position, and rural residents experiencing a greater risk of underweight, and the elder women, those from high socioeconomic status, and urban residents experiencing the greatest risk of being pre-overweight, overweight, and obese. Our findings are consistent with earlier studies conducted on women and men from the neighbouring country India ,29–32 anThe increased likelihood of being underweight and decreased risk of being overweight among the younger women, compared to their elder counterparts, and decreased risk of underweight among women who were currently in union may be partly attributed to physical activity, cultural norms, and food practising. Biological phenomena are also concerned with undernutrition of young women compared to their elders. The ever-married women who were not currently in union during the survey were possibly older than were those who were currently in union. The martially-disrupted women in Bangladesh may have less physical activity than those who live with husbands because of limited outdoor activities due to specific climatic and social circumstances. Our findings are supported by earlier studies ,33. StudFindings reveal a strong significant association between BMI and women's level of education and wealth quintile. The higher the level of education and the wealth quintile, the lower was the risk of being underweight. In contrast, the grades of overweight increased significantly with increase in women's level of education and wealth quintile. The increased risk of grades of overweight for women in more flattering social position is partially attributed to the capability of boasting up more-than-adequate food supplies and lesser physical activities than women belonging to poor socioeconomic status. The higher incidence of overweight and obesity in higher socioeconomic groups may be partly explained by the possible indifference to body-shapes. These differences may contribute to explaining the strong observed association between high socioeconomic positions and overweight .Moreover, these days people with increased income are willing to pay for a larger number of high-calorie beverages to respond to the elite marketing, which substantially contribute to gain weight. Thus, in spite of socioeconomic variability in population, the increased consumption of high-calorie beverages may explain some of the positive relationship between wealth quintile and BMI . ConsistThe unavailability of food is strongly associated with underweight body-shape of women and not significantly with overweight. Whereas obesity is associated with privileged circumstances in low-income countries , it is mThe association between socioeconomic status and BMI in low-income settings, like Bangladesh suggests that rural people is likely to be subjected to the changing patterns of food availability, food composition, and consumption behaviour. Findings reveal that rural women were less likely to be overweight than those in urban areas. This finding is similar to that of an Indian study . MoreoveThe study has several strengths and limitations that urge explanations. The first limitation is that, we used BMI as the only measure of overweight. Although BMI is a reliable indicator of body fatness and a low BMI is likely to be a valid proxy for chronic energy deficiency, BMI does not distinguish between body fat and lean body mass. Waist-circumference  and waisBangladesh is facing the dual burden of underweight and overweight. The findings suggest that the nutritional status of women is related to individual, socioeconomic and environmental level. The nutritional status of individuals and societies, at a given point in time, is likely to reflect the cumulative synergy between physiologic endowments and the social environment . It is lConflict of interest: Authors declare no conflicts of interest.


Objective. To compare quality, quantity, and trends of food supply from 1984 to 2009 and degree of food westernization in Taiwan with Asian countries and world continents by using food balance data. Methods. We compiled data from food balance sheets of Taiwan and Food and Agriculture Organization, including five continents and three most populated countries each in Eastern, Southern, and Southeastern Asia over the period 1984–2009. Quantity of food supply per capita was referenced to Taiwan food guides. The population-weighted means of food supply from Europe, North America, South America, and Australia and New Zealand continents in terms of energy and nutrient distributions, animal/plant sources, and sugar/alcohol contribution were used as indicators of westernization. Trends of food supply per capita of six food groups were plotted, and linear regression was applied to evaluate food changes. Findings. Taiwan's food supply provided sufficient quantity in food energy, with the lowest cereals/roots supply and rice to wheat ratio, but the highest meat and oil supplies per capita among the 10 studied Asian countries. Taiwan food supply showed the most westernization among these countries. Conclusion. Food supply of Taiwan, although currently sufficient, indicated some security problems and high tendency of diet westernization. There is a need in every country for reliable sources of dietary data that can provide information for monitoring and identifying dietary and nutritional status of citizens. The food balance sheet (FBS) assembled by the Food and Agriculture Organization (FAO) of the United Nations  providesAccording to the Global Burden of Disease Study 2010 , the hig“Western diet,” although without clear definition, is quite commonly used in describing the dietary pattern that corresponds to a diet low in dietary fiber but high in total fat, saturated fat, animal fat, and additive sugar , and theTaiwan has gone through rapid economic growth and is now one of the newly industrialized Asian economies . The disCompared to the dietary patterns observed in the 1993–1996 NAHSIT, those observed in the 2005–2008 NAHSIT showed some changes that should be beneficial to health , while there were also some detrimental changes  . Those dFBS assembled by the Council of Agriculture of Taiwan was compiled with FAO food balance data of the whole world, six continents , three most populated countries in Southeastern Asia , three most populated countries in Eastern Asia , and three most populated countries in Southern Asia . Taiwan had public accessible food balance data published from 1984 to 2010, while the FAO had the most updated international data published till 2009. As we calculated the 3-year moving averages, we hereby show data over the period of 1985 to 2008.We examined Taiwan FBS data item by item to assign the codes of similar food from the FAO food balance data. For those food subgroup items without similar classification, we compared the total quantities rather than any single subgroup item. For example, Taiwan FBS listed green leaves, roots, bulbs and tubers, flowers and fruits, and mushroom as subgroups of the vegetables group, while FAO assigned the vegetable subgroup items as tomatoes, onions, beans, peas, aquatic plants, and other vegetables; therefore, we compared the whole vegetables group only, not the vegetable subgroup items.The information on total energy  and protein and fat (g) available per capita per day was compiled from FAO data and Taiwan FBS. We calculated the carbohydrate availability (g) by subtracting the sum of energy from fat and protein from the total energy and then dividing the number by 4. All single food items were further classified into one of the six food groups  as categorized by the Taiwan food guides. Except for the vegetables group, we calculated servings of food of their belonging food groups by one major nutrient quantity. In other words, items in the cereals/roots group that contain 15 g of carbohydrate were counted as one serving each, and it was 7 g of protein for soy/fish/meat/egg, 15 g of carbohydrate for fruits, 5 g of fat for oil/nuts, and 8 g of protein for dairy. For the vegetables group, an edible 100 g portion was counted as one serving .The reference food needs per capita retrieved from the Taiwan food guides for adults were 12 servings of cereals/roots, 1 serving of dairy, 4 servings of soy/fish/meat/egg, 3 servings of vegetables, 2 servings of fruits, and 6 servings of oil/nuts. We used food availability in servings of the studied countries or continents as the indicator of food quantity.We used the nutritional indices calculated as per capita food energy supplied from animal food , total and animal protein (g), total and animal fat (g), contribution of animal-derived energy (%), contribution of animal protein (%), contribution of animal fat (%), contributions of protein and fat to total energy (%), and energy from sugar and alcohol  as the indicators of food quality. This analysis was done on the 2007 data because we believe the stability before the 2008 economic crisis would give more reliable results. In addition, we determined the changes in food supply by using the slope  obtained from the linear regression model.For cereals/roots supply, all four Eastern Asian countries  dropped steadily with the annual mean changes of −0.12, −0.24, −0.29, and −0.10 servings/capita/day, respectively. Taiwan and China showed similar rates of decrease, while Korea had faster and Japan had slower rates. Starting from 2003, Eastern Asian countries have had the mean cereals/roots supply falling below that of South Eastern Asian countries. Among all compared Asian countries, Taiwan had the lowest supply of cereals/roots in any single year throughout the study period.Soy/fish/meat/egg supplied by most of the world continents kept an increasing trend, but Japan and Taiwan have been showing a decreasing trend since 1996. Albeit with a steady annual decrease of 0.08 servings/capita/day of meat, Taiwan had the highest meat supply per capita among Asian countries since 1990, and the number was even higher than that of any continent since 1986 and was twice that recommended by the Taiwan food guides.Asian and western continents had obvious differences in dairy supply. Except for Japan, all of the Asian countries had mean dairy supply less than 1 serving/capita/day, much lower than that in Europe, North America, and Australia and New Zealand continents. In Taiwan, starting from 0.79 servings/capita/day in 1994, milk supply had decreased annually at a rate of 0.02 servings/capita/day. Till 2009, it decreased to as low as 0.58 servings/capita/day, which was 58% of the minimal recommended level.Eastern Asia had a much faster increasing trend in vegetables supply than Southeastern Asia since 1990, with major contribution from the sharp increase of vegetable supply in China and steady high supply in Korea. All the Southeastern Asian countries had vegetables supply less than 2 servings/capita/day. Taiwan had approximately 3 servings/capita/day, which barely reached the reference vegetables needs for healthy adults.Taiwan had the second highest fruit supply next to the Philippines, while it already fell below the recommended 2 servings/capita/day and kept an annual drop of 0.02 servings/capita/day. Korea and China, although low on fruit supply throughout the study period, showed significant annual increases of 0.03 and 0.05 servings/capita/day, respectively, which contributed significantly to the annual increase of 0.03 servings/capita/day in the whole world. It is worthy to note that all the world continents had fruit supply less than 2 servings/capita/day, the referenced fruit supply for healthy adults.The oil/nuts supply had steadily increased in most of the Asian countries during the years before 1995 and thereafter showed a steady state in Taiwan, Japan, and Philippines. Taiwan had the highest edible oil supply among Asian countries, and the mean supply of 12.4 servings/capita/day observed in 2009 was twice the recommended oil/nuts serving of Taiwan food guides.Sugar energy supply in most of the Asian countries  exceeded 200 Kcal/capita/day. When alcohol was added, the energy supply usually exceeded 300 Kcal. Japan  and Taiwan  had continuous annual dropping, while China  and Korea  had their annual increases in sugar + alcohol energy  in the recent 10 years. The sugar + alcohol energy plotted in Using FBS data in 2007, we found that the population-weighted data of Europe, North America, and Australia and New Zealand continents indicated a total energy of 3,300 Kcal/capita/day, with 25.5% of energy from animal food, 33.2% from fat, 11.8% from protein, 13.5% from sugar, and 4.3% from alcohol. The total food protein was 97 g/capita/day, with 54.5 g from animal food, and the total food fat was 122 g/capita/day, with 60.6 g from animal food. Sugar and alcohol contributed 446 and 142 Kcal/capita/day, respectively. We used these data as reference indices of food westernization. Among the indices, Taiwan had the highest food energy from animal food, contribution of animal food to total energy, total food protein, total food fat, contribution of fat to total energy, and contribution of protein to total energy. In addition, next to Japan, Taiwan had the second highest food protein from animal sources, animal protein in total protein, and protein energy in total energy. In fact, Taiwan also had the second highest animal fat, next to China. Judging from the data above, we believe Taiwan had the most westernized food patterns among the Asian countries studied. Nonetheless, in Taiwan, the contribution of sugar + alcohol  to total energy was less than 53% of that in the western reference, indicating a less severe loading of those two components in Taiwan's dietary environment in comparison with western countries.FBS data represent the abundance or availability of food supply to people at the population level, without taking into consideration the issue of distribution among subgroups of the population. It is generally believed that FBS data overestimate food consumption compared with other dietary assessment methods. The difference may arise from household or business waste and spoilage, as well as food used for purposes other than human consumption. Thus, if FBS data showed a food supply that is equal to or lower than the recommended quantity, it should be regarded as “not adequate.” From our analysis, vegetables supply of Taiwan would be regarded as barely adequate, fruits as suboptimally adequate, and dairy as not adequate, while soy/fish/meat/egg and oil/nuts were adequate. Cereals/roots supply dropped steadily and had been not much higher than the minimal recommended level.The data of food supply in Taiwan during the studied period showed that food energy had increased and then decreased, and similar trends were observed in soy/fish/meat/egg and dairy supplies; cereals/roots supply showed a constant decreasing trend; oil/nuts had a steep increase and then stayed constant; and vegetables and empty calorie  supplies had slightly decreasing trends.In comparison with other Asian countries, Taiwan had the highest food energy from animal food, protein supply, fat supply, contribution of fat to total energy, protein food servings , total oil servings, and plant oil servings but had the lowest contribution of carbohydrate to total energy, cereal servings, and rice-to-wheat ratio.When the population-weighted food supply data of Europe, Australia and New Zealand, and North America Continents were selected as references to evaluate the degree of “westernization” of food pattern in Asian countries, Taiwan showed more westernization than the other Asian countries. On the other hand, Taiwan had some unique characteristics that would be beneficial to health, including higher supplies of soybean and seafood; relatively ample fruits and vegetables ; a higher plant oil to animal fat ratio; and a sugar + alcohol supply that was not too high. Because the food environment interacts with dietary habits of the population, those beneficial factors remaining in the Taiwan food supply system are worthy of further studies. Specifically, the impacts on health from the mixture of westernized factors and beneficial factors in Taiwan should be investigated.From the evaluation of FBS over the period of 1984 to 2009, we found that food supply in Taiwan provided low dairy, suboptimal adequate fruits, barely adequate vegetables, constantly decreasing cereals/roots, and sufficient meats and oil/nuts. Compared to those in other Asian countries, Taiwan food supply provided higher animal food, meat servings, and animal protein and fat, but lower cereals/roots servings. It had the lowest plant-to-animal energy ratio, rice servings, and rice-to-wheat ratio, but the highest soy/fish/meat/egg supply. While food westernization in Taiwan is evident, there are also some characteristics that are beneficial to health such as higher supplies of soybean and seafood, relatively ample fruits and vegetables, a higher plant oil to animal fat ratio, and a sugar + alcohol supply that was not too high. These characteristics may alter the health impacts from pure western diet.The understanding and continuous monitoring of food environment of Taiwan are helpful for food and nutrition research, policy, and education. Further investigation on food supply to health issues is also needed.


Posidonia oceanica, widely used as a biological quality element in the context of the European Union Water Framework Directive (WFD). The good quality of a water body and the apparent health of a species, whether or not an ecosystem engineer such as P. oceanica, is not always indicative of the good structure and functioning of the whole ecosystem. A key point of the recent Marine Strategy Framework Directive (MSFD) is the ecosystem-based approach. Here, on the basis of a simplified conceptual model of the P. oceanica ecosystem, we have proposed an ecosystem-based index of the quality of its functioning, compliant with the MSFD requirements. This index (EBQI) is based upon a set of representative functional compartments, the weighting of these compartments and the assessment of the quality of each compartment by comparison of a supposed baseline. The index well discriminated 17 sites in the north-western Mediterranean  covering a wide range of human pressure levels. The strong points of the EBQI are that it is easy to implement, non-destructive, relatively robust, according to the selection of the compartments and to their weighting, and associated with confidence indices that indicate possible weakness and biases and therefore the need for further field data acquisition.Biotic indices, which reflect the quality of the environment, are widely used in the marine realm. Sometimes, key species or ecosystem engineers are selected for this purpose. This is the case of the Mediterranean seagrass  Human activities can deeply alter the environment, species composition and functioning of ecosystems. These alterations can be tracked by the use of biotic indices, i.e. species or groups of species whose function, population, or status reflect the environmental quality. Thus, biotic indices are monitored for changes in presence and abundance. The occurrence of an organism in a specific environment indicates that overall, its biological requirements are satisfied, whereas its disappearance suggests a change in the environment. Species are also monitored for changes in biochemistry, physiology or behaviour induced by environmental conditions. Biotic indices are used in terrestrial, freshwater and marine habitats, because they enable the quality of an environment to be characterized in an integrated way (i) assess the quality of a water body, (ii) assess processes such as currents, sedimentation and climate under natural and anthropogenic forcing, and (iii) monitor the status of species or communities of interest, either emblematic species, indicators of some ecosystemic processes or indicators of pollution. Sometimes, key species and ecosystem engineers Posidonia oceanicaBiotic indices are widely used in the marine realm to (i) Biological diversity is maintained, (ii) Introduced species are at levels that do not adversely alter the ecosystems, (iii) Populations of all exploited fish and shellfish are within safely biological limits, (iv) All elements of the food webs occur at levels capable of ensuring the long-term abundance of the species, (v) Human-induced eutrophication is at a minimum, (vi) Sea-floor integrity is at a level that ensures that the structure and functions of the ecosystems are safeguarded, (vii) Permanent alteration of hydrographical conditions does not adversely affect marine ecosystems, (viii) Concentration of contaminants are at levels not giving rise to pollution effects, (ix) Contaminants in fish and other seafood do not exceed levels established by Community legislation, (x) Properties and quantities of marine litter do not cause harm to the coastal and marine environment, and (xi) Introduction of energy is at levels that do not adversely affect the marine environment. The EU MSFD established a framework within which Member States agreed to take the appropriate measures to achieve or maintain good environmental status in the marine realm by the year 2020 at the latest.In the European Union (EU), the so called Habitats Directive of 1992 (92/43/ECC) listed habitats and species that are used to designate areas (‘Natura 2000 sites’), where they are strictly protected. While the Habitats Directive also considered the marine realm, more recently, the EU Marine Strategy Framework Directive (MSFD: 2008/56/EC) established a framework for conservation in the field of marine environmental policy. The MSFD is considered to be the environmental pillar of the Integrated Maritime Policy adopted in 2010 by the European Commission (2010/477/EU). This directive established eleven criteria, based on the descriptors set out in the MSFD, to determine ‘good environmental status’: A key point of the MSFD is the ecosystem-based approach. Most previous strategies only dealing with Biological Quality Elements (BQE: species and/or communities) used in the European Union Water Framework Directive (WFD) were not indicative of ecosystem status, but merely indicative of the environmental status of water bodies According to the MSFD, ‘good environmental status’ means that the marine environment is at a level that allows uses and activities by current and future generations, i.e. the structure, functions and processes of the constituent marine ecosystems, together with the associated physiographic, geographic, geological and climatic factors, allow those ecosystems to function fully and to maintain their resilience to human induced environmental change. Overall, marine species and marine habitats are protected, human-induced decline of biodiversity is prevented and diverse biological components function in balance.(i) how the status of an ecosystem can be measured and how much its current status differs from ‘reference conditions’, and (ii) which parameters we need to monitor to ascertain these targets. The Posidonia oceanica seagrass meadow was chosen for this attempt because: (i) it is widely present in almost the whole of the European Mediterranean; (ii) it is the only marine ecosystem considered as ‘priority habitat’ by the EU Habitats Directive; (iii) its functioning is relatively well known (iv) like many seagrass ecosystems in the world ocean, P. oceanica meadows have been impacted or lost under the influence of direct and indirect effects of human activities and are therefore regarded as threatened While the ecosystem-based approach constitutes the background and the guidelines of the MSFD, the pristine state of an ecosystem is not clearly defined. Obviously, such a notion is very complex and prone to anthropocentrism; reference conditions, as observed in areas distant from known human impact, may constitute a more realistic notion. Here we try to define: Posidonia oceanica (L.) Delile is a seagrass species  endemic to the Mediterranean Sea P. oceanica dies, the matte persists (hereafter ‘dead matte’), since the decay of the rhizomes proceeds extremely slowly P. oceanica is the engineer of an ecosystem of major importance in the Mediterranean Sea P. oceanica ecosystem has been proposed P. oceanica ecosystem in the north-western Mediterranean Sea used in the present study .P. oceanica rhizomes.Multicellular Photosynthetic Organisms (MPOs) epibiotic on P. oceanica leaves (box 2).MPO leaf epibiota (box 3).Dissolved Organic Carbon (DOC).Pelagic microbial loop.Filter- and suspension-feeder leaf epibiota (box 4).P. oceanica rhizomes: the bivalve Pinna nobilis (box 5) and other species, e.g. bryozoans and ascidians (box 6).Filter- and suspension-feeder benthic epibiota on P. oceanica leaves and some broken rhizomes) (box 7).The litter detritus .Detritus feeders 1, e.g. Amphipoda, Isopoda (crustaceans) and Holothuria spp. (sea cucumber) (box 8).Detritus feeders 2 and 3, e.g. Amphipoda, Isopoda (crustaceans) and Bacteria, Archaea, Fungi and heterotrophic stramenopiles (BAFHS) such as Labyrinthulomycota and Oomycota.Decomposers, namely The benthic microbial loop.The matte endofauna, e.g. annelids and mollusks.Sarpa salpa (teleost), Paracentrotus lividus (sea urchin), Idotea spp. and Pisa spp. (crustaceans) (box 9).Herbivores 1, e.g. Jujubinus spp. and Rissoa spp. (gastropods).Herbivores 2, e.g. Amphipoda (crustaceans), Diplodus spp., Sparus aurata, Labrus spp. and Symphodus spp.), cephalopods and seastars  (box 10).Predatory teleosts .Piscivorous teleosts, e.g. Spicara spp. and Chromis chromis (box 12).Planktivorous teleosts of the water column, e.g. Phalacrocorax aristotelis ssp. desmarestii and Pandion haliaetus (box 13).Sea birds, e.g. Plankton (photosynthetic plankton and zooplankton) and non-living Particulate Organic Matter (POM).(i) a steady trend of the parameter, from very good to a low quality state; (ii) an upward/downward slope parameter, when the very good state corresponds to intermediate values.Whenever possible, non-destructive methods were chosen to measure the parameters of the status of the functional compartments, as suggested by −1; higher sedimentation rates result in the death of the buried leaf bundle in situ measures (growth of the rhizome corresponding to the last 8 dead leaf bases) are recommended. The obtained value was multiplied by 1.5 in order to take into account the fact that the rhizome continues to grow slowly during the following two years .box 1, . This coP. oceanica ecosystem. It was estimated by the number of shoots per square meter, measured within a small square frame  and animals, share the same habitat (the leaf surface) and are usually co-consumed by the same species. In addition, some consumers eat simultaneously epibiota and the supporting leaf , ascidians , gastropods and bivalves (such as the fan mussel Pinna nobilis) Pinna nobilis (box 5) density was estimated along 20 transects 10-m long and 1-m wide. Filter- and suspension-feeders other than P. nobilis (box 6) are indicators either of: (i) high level of organic matter in the water  low level of organic matter . For colonial species, the number of colonies was taken into account, as suggested by P. nobilis; box 6) on rhizomes index, for a given site, was estimated as the arithmetical mean between HOM and LOM indices..box 8, . DetrituParacentrotus lividus and the teleost Sarpa salpa, at 5 meters depth or, if the meadow is not present at this depth, at the upper limit of the meadow. They consume P. oceanica leaves, together with their epibiota, if present P. oceanica leaves, such as the spider crabs Pisa spp. and the isopod Idotea hecticaP. lividus was assessed within 1-m2 quadrats, with 30 replicates randomly localized. The census only considered individuals >3 cm (test diameter without spines), because small individuals can be hidden within the matte, generating bias in the census. A second proxy of the macro-herbivore pressure was the grazing index, i.e. the percentage of intermediate and adult leaves . Bite scars were carefully distinguished from broken leaves, the latter being related to hydrodynamism. Most bite scars are due to the S. salpa browsing, and they are easy to distinguish from those due to other macro-herbivores Paracentrotus index and the grazing index.box 9, . Macro-hes sensu exhibitiP. oceanica ecosystem  during the warm season (summer-autumn), preferentially in uniform beds (at least a long way from rocky substrates). All teleosts were counted within ten linear and 5-m wide transects, each census lasting 5 minutes. Around 50 m were covered so that each transect represents a surface area of nearly 250 m2. Total length (at the nearest 2 cm) of individuals and the number of individual per species were noted. The Specific Relative Diversity Index (SRDI) is the mean number of species met with per transect. These data enable calculation of (i) predator biomass , (ii) top predator biomass , (iii) planktivorous teleost biomass (box 12) and (iv) the Specific Relative Diversity Index (SRDI). The planktivorous teleost biomass is divided into 2 categories: the exclusive zooplankton feeders  and the omnivorous feeders, which consume both zooplankton and POM . Some top predators  are active only during night time while Serranus spp. are active by day and thus more present in our visual counts. Some predators (e.g. Symphodus rostratus), though diurnal, are often hidden within the P. oceanica leaf canopy. As a result, their counts were underestimated to a greater or lesser extent. The parameter ranges within the status scale , but benthic teleosts  account for 35% of the diet Diplodus spp. (13%) and Sarpa salpa (11%) via the distance of the nearest Phalacrocorax spp. and Pandion haliaetus, respectively, nesting sites from the study site. As far as shags are concerned, the mean maximum foraging range is 16 km box 13, . Most seP. oceanica ecosystem from the pelagic habitat: plankton, POM and planktivorous teleosts. Outputs are also to be considered: (i) ca. 15–30% of the net primary production (NPP) corresponds to roots, rhizomes and dead sheaths buried and sequestrated within the matte (ii) ca. 6 to 50% of the NPP represents dead leaves that are exported as detritus towards beaches and adjacent habitats (iii) a number of organisms  leave the meadow, either temporarily, to feed in adjacent habitats, or permanently, to reach their adult quarters Some of the above-mentioned compartments correspond to inputs into the The rationale governing our ecosystem-based approach is trying to quantify and assess each compartment (box) of the conceptual model by means of a set of parameters, to balance their relative weighting and by using a simple algorithm to calculate a rank for the ecosystem status within a given area, matching the five Ecological Statuses of the Water Framework Directive (WFD), from bad to high. A great variety of parameters are available for the assessment of each compartment (box). Many of them are redundant. Others have been poorly used so that data are not available for most areas. For this reason a limited set of relevant parameters was selected for a restricted set of compartments.Each parameter was assessed by means of a semi-quantitative scale (4 through 0), from very good (4) to very low (0). Calibration of the scale was based upon the available literature e.g. , includii is the weighting of the box i, Si the status of the box i, Smax the highest possible grade ( = 4) for a box and i is the number of the box (1 through 13).Boxes were balanced, according to their relative weighting (W) in the ecosystem functioning, from 5 (highest weighting) to 1 (lowest weighting). The general principle in the ranking of the weighting of a box was that the boxes localized at the very base (bottom up control by primary producers), the herbivores (box 9) and the boxes localized at the very top (top down control via cascade effect) of the model were regarded as of major importance (with the exception of box 13), while intermediate ones were less weighted. Wasp-waist control has not been evidenced in that ecosystem Five ecological status classes, from high to bad, according to the practice of the WFD, were delineated: bad (EBQI<3.5), poor (3.5≥EBQI<4.5), moderate (4.5≥EBQI<6), good (6.0≥EBQI<7.5) and high (EBQI≥7.5).Since the box weightings were supported by partly subjective arguments (see above), we aimed to analyse the effect of the weighting choice on the EBQI and the ranking of the sites. In order to achieve our aim, we perturbed each weighting value and determined the new ranking obtained with the perturbed weightings. The perturbation on each weighting was obtained as follows. We first defined the maximum amplitude of the perturbation for all weightings and we then defined, for each weighting, a random perturbation, according to a uniform law between 0 and the maximum amplitude of perturbations. We then added or subtracted this perturbation term to or from the corresponding weighting. If the new weighting was less than 1, it was then set as equal to 1. If the new weighting was greater than 5, it was then set as equal to 5. We ended up with weightings between 1 and 5, close to the original ones if the maximum perturbation amplitude was low, and randomly selected otherwise. This has been calculated as follows: the perturbation method described previously was repeated 1,000 times and, for each site, we determined whether the rank of the site was the same as the initial one or if it had changed. An index of similarity was produced for each site, which was equal to 100% when the rank of the site was always unchanged after perturbation and 0% if it always changed.(i) that data for one or several compartments may be missing or of poor quality in some sites, (ii) the reliability of available data may be different between compartments and sites, and (iii) it is worth drawing the attention of managers and scientists to those compartments that are poorly known and which merit further field studies. The grade of each considered compartment was given by its CI (0 through 4) and by its weighting (1 through 5), and therefore they are graded from 0 through 20. The grades of all considered compartments were added up, which gave the final mark of the CI at a given site. For practical purposes, the CI was converted to a scale from 0 to 4:i is the weighting of the box i, CIi the Confidence Index of the box i, CImax the highest possible Confidence Index ( = 4) for a compartment (box) and i is the number of the box (1 through 13).For each compartment (box) status at each site, a Confidence Index (CI) was proposed . The reaP. oceanica meadow, from its upper limit down to the lower limit, covering in the order of a dozen to several hundred hectares. The sites are localized in the north-western Mediterranean Sea, including continental France , Corsica, Balearic Islands and Spanish Catalonia. This area is considered as homogenous and is consistent with the marine subregions, as defined in the MSFD. These sites also cover of a wide range of human pressures, from lesser impact, within well implemented Marine Protected Areas (MPAs), such as the Port-Cros National Park, to highly impacted areas due to different disturbance and/or stress sources : Sitges.Poor (3.5≥EBQI<4.5): Niolon, Gulf of Giens and Porquerolles Island northern coast.Moderate (4.5≥EBQI<6): Villefranche Bay, Saint-Cyr Bay, Plateau des Chèvres (Marseilles), Prado Bay (Marseilles), Valincu Gulf, Tossa de Mar, Scandola (Elbu Bay) and Peyrefite Bay.Good (6.0≥EBQI<7.5): Espardell and Porquerolles Island southern coast.High (EBQI≥7.5): Bagaud Pass (Port-Cros Island), Medes Islands and Port-Cros Island (southern coast).P. oceanica  have been proposed and are currently routinely used, generally for the purpose of monitoring the ecological status of a water body, e.g. POMI s = 0.469, p value = 0.11) or when only PREI data are concerned . This result is not unexpected: EQRs mainly assess the health of the seagrass, linked to e.g. the water quality, while EBQI measures the status of the ecosystem, linked not only to water quality but also to e.g. the overfishing. The contrasting ranks of Porquerolles Island (northern coast) through EBQI (poor) and EQR  based upon Are some of the parameters used  redundant with other boxes or with the EBQI? Should such a redundancy exist, this could mean either that the box alone is sufficient to estimate the whole ecosystem's status, or that some boxes are useless. Obviously, the higher weighting is given to a box, the stronger is the probability of such a correlation. Some boxes (e.g. box 5 ‘Pinna’ and box 10 ‘predators’) are significantly correlated with others and/or with the EBQI . HoweverIs the EBQI robust, i.e. only slightly influenced by arbitrary choices in its conception and biases? Possible arbitrary choices are e.g. choice of the considered compartments (boxes) and the weighting (1 through 5) of the boxes. As far as the choice of the compartments taken into account is considered, it should be first emphasized that the choice is anything but arbitrary; boxes encompass the whole spectrum of an ecosystem functioning, from primary producers, herbivores, filter-feeders, suspension-feeders and detritus-feeders to top predators . It is wIs the EBQI excessively time-consuming or easy and rapid to implement? The acquisition of the parameters requires SCUBA diving field work. Considering that it is possible to work for around 1 hour at 15 m depth, we estimate that 6 dives involving 2 scientific divers are necessary to acquire all of the data for the assessment of one site . For safWithin the northern Mediterranean, the study sites are spread within three eco-regions, namely Corsica, Provence and French Riviera and Languedoc-Catalonia. Is the EBQI biased by regional environmental conditions, e.g. a higher mean temperature in Corsica and more water turbidity in Languedoc-Catalonia? The present study cannot answer this question as the sites were not randomly selected but chosen according to data availability. However, the EBQI range within each one of the three eco-regions does not exhibit obvious differences , as far P. oceanica, is not always indicative of the good structure and functioning of the whole ecosystem, a network of compartments, fluxes, functions, inputs and exportations. The most original contribution of the new MSFD European Directive is to provide an ecosystem-based approach to assess the ecological status of marine regions. The present approach constitutes a contribution towards this goal, and focuses on the P. oceanica seagrass meadow.In the context of European Directives (Habitats Directive and WFD) and national regulations, a number of species and indices, based upon one or several species, have been designated to assess the quality of water bodies, the health status of emblematic species and habitats and to establish Marine Protected Areas (e.g. PMN P. oceanica meadow. This index (EBQI) is based upon a set of representative functional compartments, the weighting of these compartments and the assessment of each compartment quality by comparison of a supposed baseline based upon the available literature. The strong points of the EBQI are: (i) It is easy to implement, not too time-consuming and therefore relatively cheap; (ii) It is non-destructive, which is particularly important, dealing with protected species (e.g. P. oceanica and Pinna nobilis); sampling only concerns old leaves (not the shoot) and the litter, i.e. shed dead leaves; (iii) It is relatively robust, according to the selection of the compartments and to their weighting; (iv) It is associated with confidence indices CI  which indicate possible weakness and biases and therefore the need for further field data acquisition; (v) It can draw the attention of managers to sites whose CI is high and where routine monitoring can therefore be implemented, and conversely those with a low CI indicating a lack of knowledge.Here, we have developed, applied and tested an ecosystem-based index of the structure and functioning of the most emblematic and best-known Mediterranean coastal ecosystem, the The Integrated Coastal Zone Management (ICZM) contributes towards maximizing the benefits provided by the coastal zone and minimizing conflicts and the harmful effects of activities upon each other e.g. . ICZM, t(i) The baselines used to assess the compartments, which may prove to be biased either by the poor knowledge or availability of totally non-human impacted areas, or by climate change which renders the supposed baseline obsolete; (ii) The probable need for testing and/or adapting the EBQI in areas distant from the study area -the north-western Mediterranean- and more generally to other seagrass ecosystems; (iii) The conceptual model which constitutes the basis for the present ecosystem-based approach is obviously oversimplified. For example, small-sized predatory ‘invertebrates’ were not taken into consideration. Some of them belong to the matte endofauna box, which encompasses detritus feeders, suspension-feeders and predators. Other are creeping or clinging organisms on the P. oceanica leaves. However, considering them would have made the model more complex and weighed down the assessment of ecosystem status. Moreover, other compartments are likely to provide redundant information, so that no improvement in accuracy is to be expected.The weak points of the EBQI are: (i) problems of applicability due to practical and conceptual difficulties, affecting most of the indices related to ecosystem function, (ii) the failure of many indices using e.g. the taxonomic composition of the community to connect its condition with the stressors, and (iii), as far as indices based upon the sub-individual level are concerned (e.g. multi-biomarkers), their poor strength to assess the ecological integrity of the habitat. They concluded that the most promising approach would be to aggregate indices with complementary strengths. In a sense, the EBQI could be considered as such an aggregative index. Although the goal of the EBQI is not to compete with the already available biotic indices utilizing the P. oceanica seagrass meadow for assessing the quality of a water body, but to assess the status of the proper seagrass ecosystem, the comparison of the EBQI with some other biotic indices will be of relevance. An attempt has been made here, using some published data; a more accurate comparison would require data acquisition at the same sites as those used for the EBQI, and is therefore beyond the scope of the present work. In addition, the correlation between the EBQI and anthropogenic gradients, including overfishing, will be useful, in order to assess which human impacts are of major relevance for the status of the ecosystem. This will constitute the next step.A number of attempts to review and compare the biotic indices using benthic (e.g. macrophytes) and pelagic marine, lagoon and estuarine organisms, have been performed P. oceanica ecosystem at a given site, due e.g. to the health of the seagrass, the clearness of the water (and even the beauty of the landscape and the seascape) confirms the usefulness of the EBQI index, based upon the whole ecosystem rather than upon only the seagrass. Other anthropogenic impacts, e.g. overfishing, are putatively more important than the above mentioned parameters for the structure and functioning of the ecosystem. Finally, the EBQI that is proposed here for the P. oceanica ecosystem could constitute a model for similar indices designed for other ecosystems, such as coralligenous outcrops, underwater caves, soft bottoms and sublittoral reefs.Overall, the Ecosystem Based Quality Index (EBQI) is easy to implement, relatively robust and does not seem to be redundant with existing indices based upon the seagrass itself. In addition, the non-congruence of the EBQI, i.e. the quality of the ecosystem functioning, with the empirical idea one may have of the


Diabetes mellitus (DM) is a common endocrinopathy in cats. Feline DM is similar to human type 2 diabetes (T2D), with obesity as a major risk factor in both cats and humans. Genetics play an important role in the development of obesity and T2D in humans.The aim was to describe the incidence of feline diabetes in Sweden, and the association with different epidemiologic risk factors.We used data from a cohort of 504,688 cats in a Swedish insurance company from 2009 to 2013, contributing with 1,229,699 cat-years at risk (CYAR). Overall incidence rates (IR) and IRs stratified on age, breed and gender were estimated.The IR of diabetes was twelve cases per 10,000 CYAR. Male cats had twice as high IR as females. A significant association with breed was seen, with the Burmese, Russian Blue, Norwegian Forest Cat, and Abyssinian breeds at increased risk. Also domestic cats were at higher risk compared to purebred cats. IRs of DM was highest in cats 12 to 15 years old.Our results identified several cat breeds associated with an increased risk of DM. Also male gender and middle age were risk factors for DM. A link has been shown between obesity and DM in cats. A higher incidence of obesity has been reported in domestic cats as compared to purebreds, in male cats, and in middle-aged cats; these groups are similar to those in which we identified a higher occurrence of DM.


Phelsuma.Color traits in animals play crucial roles in thermoregulation, photoprotection, camouflage, and visual communication, and are amenable to objective quantification and modeling. However, the extensive variation in non-melanic pigments and structural colors in squamate reptiles has been largely disregarded. Here, we used an integrated approach to investigate the morphological basis and physical mechanisms generating variation in color traits in tropical day geckos of the genus Phelsuma species is generated by complex interactions between, on the one hand, chromatophores containing yellow/red pteridine pigments and, on the other hand, iridophores producing structural color by constructive interference of light with guanine nanocrystals. More specifically, we show that 1) the hue of the vivid dorsolateral skin is modulated both by variation in geometry of structural, highly ordered narrowband reflectors, and by the presence of yellow pigments, and 2) that the reflectivity of the white belly and of dorsolateral pigmentary red marks, is increased by underlying structural disorganized broadband reflectors. Most importantly, these interactions require precise colocalization of yellow and red chromatophores with different types of iridophores, characterized by ordered and disordered nanocrystals, respectively. We validated these results through numerical simulations combining pigmentary components with a multilayer interferential optical model. Finally, we show that melanophores form dark lateral patterns but do not significantly contribute to variation in blue/green or red coloration, and that changes in the pH or redox state of pigments provide yet another source of color variation in squamates.Combining histology, optics, mass spectrometry, and UV and Raman spectroscopy, we found that the extensive variation in color patterns within and among Phelsuma lizards as models in evolutionary developmental biology.Precisely colocalized interacting pigmentary and structural elements generate extensive variation in lizard color patterns. Our results indicate the need to identify the developmental mechanisms responsible for the control of the size, shape, and orientation of nanocrystals, and the superposition of specific chromatophore types. This study opens up new perspectives on  Indeed, color traits play crucial roles in thermoregulation, photoprotection, camouflage, and visual communication -4, and cy system ,6. Squamy system -9. The sDespite such a high potential for complexity and diversity, lizards and snakes remain relatively under-represented in evolutionary developmental studies in general , and in Phelsuma. More specifically, we show that 1) the reflectivity of the white belly, and of the dorsal pigmentary red spots and stripes, is increased by underlying, structurally disorganized broadband reflectors, whereas 2) the hue of the vivid dorsolateral background coloration is generated by structural, highly ordered narrowband reflectors, and is modulated both by the photonic crystal geometry and by a layer of yellow pigments. Most importantly, we show that these interactions require precise colocalization of red and yellow pigment cells with iridophores characterized by different organizations of nanocrystals.Here, we used an integrated approach that combines histology, Raman and UV spectroscopy, mass spectrometry (MS), optics, and numerical simulations to investigate the morphological basis and the physical mechanisms generating variation in color traits in five representative tropical day gecko species of the genus Phelsuma provide excellent models to study the morphological basis of color-pattern variation. Most of the approximately 45 currently recognized species [Phelsuma grandis; Figure Phelsuma klemmeri, Phelsuma quadriocellata and Phelsuma lineata, have lateral black/brown spots or stripes, and a few, such as P. klemmeri lack red markings and exhibit a blue/brown background  both the red and yellow pigments, found respectively in red-skin erythrophores and green-skin xanthophores  are determined by some other factors, such as the pH of the cellular environment of the pigment (as is the case for plant anthocyanins [2) to, the skin of P. quadriocellata or P. lineata, respectively  or the ocyanins ). SupporPhelsuma geckos contains iridophores, with guanine nanocrystals, either well-organized in parallel layers , or highly disorganized   to infinity (when A is high and y0 is close to zero in a perfectly ordered system). Table A/y0 ratio than white or red skin. This indicates that the ordered nanocrystals generate an intense narrowband structural blue or green color through a multilayer interference mechanism. This result is consistent with the pigment-removal experiments  varies both among and within individuals  Figure d. Using s Figure b discussls Table .Phelsuma, we applied mechanical or osmotic pressure to modify the distance between the guanine nanocrystal layers and therefore induce changes in structural coloration [To further test experimentally if the multilayer is responsible for the structural color component of blue/green skin in loration . We founPhelsuma individuals studied here, are shown  colors predicted by the model to color pictures of all samples after pigment removal  and high (ngdg) index layers are close, making the multilayer a narrowband optimal reflector as a result of the collaborative effects of crystal and spacing interferences. However, the iridescence in Phelsuma skin is much weaker than that predicted by the model , whereas green skin is produced, depending on the species and the individual, either by structural green or by the interaction of structural blue with yellow pigments (xanthophores). Dorsal marks are formed by red erythrophores, the reflectivity of which is enhanced by iridophores with disorganized crystals . In addition, the hue produced by erythrophore and xanthophore pigments is pH-dependent or redox-dependent in some species. Most importantly, we show that the color patterns of Phelsuma always require precise colocalization of different sets of interacting pigmentary and structural elements. For example, yellow and red chromatophores are associated with iridophores with ordered and disordered nanocrystals, respectively. This indicates the need to identify the developmental mechanisms responsible for the superposition of specific chromatophore types, opening up new perspectives for Phelsuma lizards as model organisms in evolutionary developmental biology.Extensive variation in background coloration, and in dorsal and lateral color patterns, is present within and among species of the genus a Figure . Our ana, through electron-microscopy and spectroscopy analyses). Hence, exploring the genetic and developmental bases of phenotypic variation in ecologically important color traits in squamate reptiles will require integration of the structural and pigmentary color components and their interactions. The fact that red/yellow coloration in Phelsuma is based on pteridines, the synthesis of which is controlled by enzymes that have been well studied in model organisms [Phelsuma.We also show that the exact combination of parameters producing a given color is difficult to predict without experimental estimation of parameter values  and performed in accordance with Swiss law.P. klemmeri (n = 1), P. quadriocellata (n = 1), P. lineata (n = 2), P. laticauda (n = 1), and P. grandis (n = 5) were obtained from the pet market or bred in our laboratory. On the phylogenetic tree of the genus Phelsuma[Phelsuma geckos exhibit a vivid dorsolateral coloration, with a background typically of one or more colors . Almost all species have distinct dorsal marks consisting of red to brown spots and stripes of various shapes. In addition, some species have black/brown lateral stripes or spots.Adult  Phelsuma,20, we mmeri n = , P. quad Phelsuma-50): 1) 2 in size were fixed overnight at 4°C in 2% glutaraldehyde and 4% paraformaldehyde in 0.05 mol/L sodium cacodylate buffer (pH 7.4), rinsed with 0.1 mol/L cold sodium cacodylate, and fixed for 1.5 hours on ice in the same buffer with the addition of 1% osmium tetraoxide. Samples were then rinsed in water, stained with 1% uranyl acetate for 2 hours, gradually dehydrated in ethanol, rinsed three times in propylene oxide, incubated overnight in 1:1 propylene oxide/resin, and finally embedded in epoxy resin (Epon). Semi-thin (1 to 2 μm) and ultra-thin (80 to 90 nm) cross-sections were cut with a diamond knife on a Leica UCT microtome. Semi-thin sections were examined under a light microscope. Ultra-thin sections were placed on formavar-coated grids, and viewed with a Tecnai™ G2 Sphera (FEI) TEM at 120 kV to visualize intact guanine crystals, because post-staining inevitably results in loss of crystals [Skin samples were placed in Ringer’s solution for microscopy observation. Cross-sections of 14 to 16 μm were prepared from skin embedded in optimum cutting temperature compound on a Leica  CM1850 cryostat. For TEM, skin pieces of approximately 1 cmcrystals . The griFor each individual, 10 to 20 TEM images of stained sections of iridophores of white, red, and green skin were collected at low  magnification, and 20 to 30 images of unstained sections were collected at high  magnification for green/blue skin. To analyze the size and orientation of crystals relative to skin surface, white ‘holes’  on low magnification images were fitted with ellipses (in JMicroVision ), whose The Gaussian distribution of crystal orientation was calculated as:yo is the intersection of the curve with the y-axis ; A is the amplitude of the Gaussian curve; θ is the angle of the smallest axis of the ellipse relative to the normal to the skin surface; θc is the central value of the distribution, and is equal to 0; and w, the standard deviation, is related to the FWHM of the Gaussian curve, defined aswhere A/y0 ratio varies from zero (when A = 0 in a purely random system) to infinity (when A is high and y0 is close to zero in a perfectly ordered system) as schematically illustrated  by analysis of their UV and high-resolution MS spectra  was seen, but the color balance was rendered correctly. During the mechanical pressure and dehydration experiments, skin color was measured in situ, and compared with simulations in which the distance between the crystals was the only adjusted parameter.For extended UV reflectivity measurements (300 to 800 nm), a probe  was connected to the scanning Xenon source of a spectroscopic Woollam ellipsometer with silicon diode as detector. Lock-in detection was used for ambient light noise reduction. For monitoring color change in real time  showing the effect of dehydration (in air) and rehydration on the structural color generated by iridophores. Note that the yellow pigments had been removed before the experiment.Click here for fileAngular dependence. Angular dependence of the multilayer reflectivity in green skin of Phelsuma grandis.Click here for file


Few studies have investigated the association between objectively measured traffic noise and health-related quality of life. However, as traffic noise has been associated with both cardiovascular disease and diabetes, and health-issues including sleeping problems, annoyance, and stress, it seems plausible that traffic noise is associated with health-related quality of life.Between 1999 and 2002, a cohort of 38,964 Danes filled in the short form-36 (SF-36) questionnaire. Residential exposure to road traffic and railway noise was calculated for all historical addresses for 10 years preceding the SF-36, using the Nordic prediction method. Associations between noise exposure and SF-36 summary scales and the eight sub-scales were calculated using general linear models, adjusted for age, sex, socioeconomic status, and lifestyle.Models adjusted for age, sex and socioeconomic factors showed that a 10 dB higher road traffic noise 1 year preceding SF-36 assessment was associated with a 0.14 lower mental component summary (MCS) score  -0.26, -0.01). However, further adjustment for lifestyle factors  attenuated the association: ). Exposure to more than 55 dB of railway noise in the same time period was borderline significantly associated with lower MCS. The physical component summary was not associated with traffic noise.The present study suggests a weak association between traffic noise exposure and the mental health component score of SF-36, which may operate through lifestyle. The magnitude of effect was, however, not clinically relevant. Exposure to traffic noise has been associated with a number of illnesses, including cardiovascular disease,2, and dQuality of life is a broad, multidimensional concept, and epidemiologic studies therefore often focus specifically on health-related quality of life (HRQoL), which is a well-accepted construct defined to capture the effect of factors that affect health—both physically and mentally. It is aFew studies have investigated effects of different types of traffic noise on quality of life. In general, airport noise has been found more annoying, and with stronger effects on sleep, than road and railway noise. For airThe above-mentioned studies are relatively small, with the largest cohort including 6,533 participants. Also, mThe study is based on the prospective Diet, Cancer and Health cohort, which has been described in detail previously. BrieflyAt baseline, participants filled in a food frequency questionnaire and a lifestyle questionnaire, and anthropometric measures were collected by trained personnel. In 1999–2002, participants received a follow-up questionnaire regarding diet, lifestyle and HRQoL, and gave self-reported anthropometric data. In total, 45,271 persons  filled in this second questionnaire, and were available for the present study. Reasons for non-participation were death (14.6%), emigration (3.8%), and no reply to the questionnaire (81.7%).Den Videnskabsetiske komite for Københavns og Frederiksberg Kommuner”) Approval no.: (KF) 01–345/93. All participants provided written informed consent, and the study was conducted according to the Helsinki Declaration.The study was approved by the local ethical committees of Copenhagen and Frederiksberg Municipalities (in Danish: “http://www.soundplan.dk/), which implements the joint Nordic prediction method for road traffic noise[Residential address history was collected for all cohort members from 1988 and until follow-up using the Danish civil registration system. Road traffic noise exposure was calculated using SoundPLAN  at the most exposed facade of the dwelling at each address for the day , evening  and night , and was expressed as Lden . A 5 and 10 dB penalty was applied to evening and night respectively. All values < 42 dB were set to 42 dB, as done in previous studies[No information was available on noise barriers or road surface. Road traffic noise was calculated as the equivalent continuous A-weighted sound pressure level . The input variables for the noise model were: point for noise estimation , railway links  and building polygons for all Danish buildings. All noise barriers along the railway are included in the model. Railway traffic noise was expressed as LFor the assessment of both road and railway traffic noise the terrain was assumed flat, a reasonable assumption in Denmark. Urban areas, roads, and areas with water were assumed to be hard surfaces, whereas all other areas were assumed acoustically porous.The follow-up questionnaire included the SF-36 questionnaire in Danish. This contains 36 items, addressing eight dimensions of health: physical functioning, role limitations due to physical health problems, bodily pain, general health, vitality, social functioning, role limitations due to emotional problems, and mental health. Each dimension is scored ranging from 0  to 100 . From these dimensions, two summary scales; physical component summary (PCS) and mental component summary (MCS), can be derived using algorithms specified by the developers. These summary scales are standardized to have a mean of 50 and standard deviation of 10, with a score above 50 representing an above-average health and a score below 50 representing a below-average health. ParticiHRQoL is a multi-dimensional concept that includes the physical, psychological, and social functioning associated with an illness or its treatment. It is oa priori based on existing literature and biological plausibility. The included confounders were measured at time of SF-36. Sex, age, smoking status , gram alcohol/day (continuous), and self-measured waist circumference (continuous) were collected as questionnaire data. Education , cohabiting status , and disposable income  was collected by linking the personal identification number of each participant to the nationwide register Statistics Denmark, which include yearly information since 1980 from the taxation authorities, the register for Education statistics, the Register for Unemployment, and a Company Register for all companies with more than one employee. Finally; models of road traffic noise were adjusted for railway noise and vice versa.The selection of covariates was done Furthermore, we used information on Charlson Comorbidity Index in the stratified analyses , calculax exposure was calculated with the Danish AirGIS dispersion modeling system for the same years as exposure to traffic noise, for all addresses where each individual had lived, as previously described in details[x did only result in minor changes in the estimates, and it was therefore not included in the final model.NOn details. AdjustmLinear regression models were used to examine the association between residential traffic noise and PCS, MCS and the eight sub-scales. To test the robustness of our models, we calculated three different versions, with increasing level of adjustment: model 1 with adjustment for age and sex; model 2, as model 1 plus adjustment for socioeconomic factors and railway noise; and model 3 as model 2 plus individual lifestyle factors.All continuous variables were evaluated by investigating linearity both graphically and by linear spline models. We found significant deviation from linearity for age, alcohol and waist circumference, and these were therefore included in the model as splines with boundaries at 60 years (age), 2 g/day , and 78 centimeters (waist circumference).For train noise, we calculated both linear estimates and categorical analyses with three categories . This allowed us to examine the estimate for the large proportion of the study population, which had no train noise exposure (80.5%). The use of 55 dB as cut-point was chosen based on consistent use in existing literature.Effect modification by sex, age, education, Charlson Comorbidity Index  and railway noise exposure was examined by including an interaction parameter in the models, and tested by the Wald test.P-values < 0.05 were considered statistically significant. Results for railway and road traffic noise are reported as changes in the dependent variable per 10 dB increase in noise exposure, with corresponding 95% confidence intervals (CI). Results for railway noise are also reported as categorical results, to allow comparison between those with no exposure and groups of exposed.All analyses were performed using SAS 9.3 , except for the graphical representation of the association between traffic noise and PCS/MCS score, which was produced using restricted cubic splines in the rms library of the R software (version 2.13.1).In total, 45,271 persons, who filled in the follow-up questionnaire, were available for the present study. Of these, 384 persons were excluded, because they had a diagnosis of cancer before baseline, 5,662 were excluded due to missing information on either exposure variables, outcome variables or included covariates, and 261 were excluded because they were exposed to airfield/air-traffic noise. This rendered an analytical cohort of 38,964 persons with complete information on all included covariates.The Pearson correlation coefficient between road traffic noise 1 and 10 years preceding HRQoL was 0.91. The crude analyses (Model 1) showed statistically significant associations between road traffic noise and both PCS and MCS, suggesting that traffic noise had a negative effect on HRQoL . The models adjusted for socioeconomic factors (Model 2) showed an association between road traffic noise and MCS with a 0.14 lower MCS score  per 10 dB higher road traffic noise 1 year preceding SF-36. The exposure-response curve found no specific window of effect, and showed that the magnitude of effect of traffic noise on HRQoL was not of clinical relevance—especially not after adjustment for individual lifestyle factors , Fig. 1.General health ), and Mental health ) per 10 dB higher road traffic noise 1 year preceding SF-36. In contrast, in the fully adjusted model, there was a positive association with Role—Physical: 0.13 , whereas the association between road traffic noise and Mental Health remained negative: -0.18  per 10 dB higher road traffic noise. The association with General Health was attenuated after adjustment for lifestyle , suggesting that railway noise was associated with a poorer HRQoL. However, in the continuous analyses, this was non-significant: -0.08  per 10 dB .Mental health subscale both before and after adjustment for lifestyle covariates.The present study investigated the association between residential noise exposure and HRQoL, measured by the SF-36. There was no association with the physical (PCS) or mental (MCS) component summaries in the fully adjusted models, but a significant, negative association with MCS in models adjusted for socioeconomic factors only. There was a suggestion of a negative association between train noise and MCS in the categorical analyses. There was a significant, inverse association between road traffic noise and the When interpreting the clinical relevance of SF-36 results, differences of 2–3 points on the PCS scale, 3 points on the MCS and 5–10 points on the sub-scales are considered to be minimally important clinical differences. The effThe results of the present study add complexity to the findings of previous studies on objectively measured road and railway traffic noise and HRQoL. However, some of the contradicting results may be explained by the modest size of the previous studies, rendering the statistical power rather low. Also, different methods were used to estimate noise, which do not allow direct comparison of results. Different estimations include vehicle traffic density as a proxy for noise exposure,21, stanThe present study design allowed estimation of associations across different exposure windows in time. When comparing the results of the two time-periods included; 1 and 10 years before SF-36, the association between road traffic noise exposure and HRQoL seemed stronger over the longer exposure period, especially in model 1  and 2 , which may suggest that longer term exposure is important in relation to HRQoL. However, the fact that the difference between estimates is largest in Model 1, may also suggest that the long-time exposure information is more confounded than the short-time exposure information. More studies with long-time exposure information are needed in order to investigate this in detail.Mental health with increasing traffic noise exposure adds strength to the similar findings in four smaller, cross-sectional studies, which all found an inverse association between road traffic noise and mental health[Our finding of a lower score on the subscale al health,29–31. Wal health,33. A UKal health. In contal health.Role physical which contributes to the overall positive association. This suggests that participants with a high noise exposure have fewer problems with their job or other daily activities because of their physical health[The non-significant, positive association between traffic noise exposure and PCS in the fully adjusted models of our study is unexpected. When examining the subscales individually, it is the scale al health. Given tIn general, the direction of the association between road traffic noise and HRQoL differs markedly between model 2 and model 3 in our study. In model 3 we further adjust for lifestyle factors: smoking, alcohol and waist circumference. Only few studies have previously investigated the association between traffic noise and HRQoL, with no strict strategy of which covariates to include. It is possible, that part of the effect of traffic noise on HRQoL actually operates through pathways where traffic noise affects lifestyle, and through this also HRQoL. E.g., a Dutch study found a positive association between traffic noise and smoking, and a SIn order to investigate this further, we tried adding the individual lifestyle factors  to model 2 one at a time, in order to clarify if a specific variable caused the shift in estimates from model 2 to model 3. There was no clear indication that this difference was attributable to one specific lifestyle factor, although adding waist circumference did show the strongest effect on the estimates. This is in line with the above-mentioned study finding an association between noise and obesity, and stuThe present study includes objective noise assessment only. It has been proposed, that the subjective assessment of noise moderates the association between noise exposure and health as well as quality of life,38. NoisThe strengths of the present study include the study size, which increases the statistical power. Furthermore, we had detailed address-histories for all participants, allowing collection of virtually complete exposure data for different exposure time windows. The Nordic Prediction Model used to calculate exposure is well-known and regularly used in studies of exposure and health. The outcome, HRQoL, was assessed using the well-validated SF-36 questionnaire; one of the most widely used measures of HRQoL. In the present study, we focused on the two component summary scores. This minimized multiple testing. We did, however, also investigate the eight sub-scales, in order to focus on effects on quality of life, which only register on selected sub-scales. A limitation of the study is that the norm-based scoring of the SF-36 data is based on an American standard population, and may thus differ from the Danish population; this miIn conclusion, the present study suggests an association between road traffic and railway noise exposure and the mental component summary of SF-36, which may operate through individual lifestyle factors. Despite being significant, however, the association is small and does not seem clinically relevant. There was no association between traffic noise and the physical component summary. Furthermore, the study contributes to a general discussion on the pathways through which the effects of traffic noise operate on HRQoL and encourages careful consideration on which confounders to include in future studies of the association.


We conducted a study using a case-crossover design to clarify the risk of acute effects of zolpidem and benzodiazepine on all-sites of fractures in the elderly.Case-crossover design.Elderly enrollees (n = 6010) in Taiwan’s National Health Insurance Research Database with zolpidem or benzodiazepine use were analyzed for the risk of developing fractures.After adjusting for medications such as antipsychotics, antidepressants, and diuretics, or comorbidities such as hypertension, osteoarthritis, osteoporosis, rheumatoid arthritis and depression, neither zolpidem nor benzodiazepine was found to be associated with increased risk in all-sites fractures. Subjects without depression were found to have an increased risk of fractures. Diazepam is the only benzodiazepine with increased risk of fractures after adjusting for medications and comorbidities. Hip and spine were particular sites for increased fracture risk, but following adjustment for comorbidities, the associations were found to be insignificant.Neither zolpidem nor benzodiazepine was associated with increased risk of all-site fractures in this case cross-over study after adjusting for medications or comorbidities in elderly individuals with insomnia. Clinicians should balance the benefits and risks for prescribing zolpidem or benzodiazepine in the elderly accordingly. Fractures are a significant and growing health problem for elderly individual, and are associated with increased morbidity and mortality –3. ThougPrevious studies have shown that hypertension , osteoarIn previous studies, zolpidem –21 and bThis study used the Taiwan National Health Insurance Research Database (NHIRD) to determine the association of zolpidem or BZDs and all-sites of fractures in the elderly from 2000 to 2010. As of June 2009, Taiwan’s National Health Insurance (NHI) Program covered 22,928,190 people, exceeding 99% of the population. The NHI also has contracts with 97% of medical providers in Taiwan . The NHIThis research was based on a population-base cohort study. The data were taken from the Longitudinal Health Insurance Database (LHID) for 2005 published by the NHI Bureau. Taiwan’s NHI was launched in March 1995  and coveThe longitudinal database is a subset of the NHIDatabase consisting of 1 million samples from Taiwanese covered by the NHI in 2005 . It contThe present study tracked patients over the age of 65 who were using zolpidem or BDZs. In addition, the study also discussed patients diagnosed with sleep disorders (ICD-9-CM codes 307.41–307.49 and 780.50–780.89) and their impact on fractures. The index date was defined as the date on which subjects were diagnosed with fractures by ICD-9-CM codes for fractures of the hip (820), humerus (812), forearm (813), wrist (814), spine , and any other sites (800–829). Patients who had been previously diagnosed with fractures before the index date were excluded. For patients, who had suffered a stroke and were paralyzed for a period of time, it was impossible to measure the extent of paralysis and the recovery time. Therefore, we excluded subjects with a stroke diagnosis. Regarding diseases, such as dizziness and locomotor problems, it could not be confirmed whether the patient was paralyzed or not. Consequently, we did not take these diseases into account. All study subjects had used hypnotics within six months prior to the index date.Comorbidities or coexisting medications that might interact with the risk of fractures were adjusted for. The confounding factors with potential to change between case and control groups were adjusted in our analysis based on Chen, et al. . These cThe protocol for this study conformed to the Helsinki Declaration, and was approved by the Institutional Review Board of Cathay General Hospital (permission code: CGH-P101089). Personal identifiers from the database were encrypted or otherwise stripped prior to analysis, and the need for written informed consent was waived by the Institutional Review Board.The case-crossover design is widely used to study the acute effects of drug over short durations , 30. EacStatistical Product and Service Solution (SPSS) v19.0 is widely used for analytical computing, data mining, and predictions. The present study used conditional logistic regression analyses odds ratio (OR) and adjusted OR. Within 95% confidence interval p < 0.05 was the level of significance for fracture. Stratified analysis was used to predict changes to risk of fracture under different conditions of BDZ and zolpidem use.p < 0.001), 1.80  after adjusting for use of antidepressants, antipsychotics and diuretics; and 1.49  after adjusting for comorbidities including hypertension, osteoarthritis, osteoporosis, rheumatoid arthritis, and depression. Use of other BDZs showed no risk after adjusting for medications or comorbidities.p < 0.05) and, when adjusted for medications, an OR of 1.49 . For spine fractures, zolpidem was associated with a crude OR of 1.38  and, when adjusted for medications, an OR of 1.36 . BDZ was associated with a crude OR of 1.22  and, when adjusted for medications, an OR of 1.20 . However, neither site was related to increased risk of fractures after adjusting for comorbidities.A case-crossover design was used to account for acute drug effects in patients with continued drug use over the study period. In the case-crossover design, the case has its own control, thus limiting bias resulting from gender and age, along with other genetic and physiological factors. Furthermore, it shows the immediate effects within 24 hours, rather than exposure-to-fall time of up to 6 months as in some other studies , 18. ZolSeveral prior case control or case cross-over studies have found that BDZ , 23 and Some previous studies of hypnotics use in elderly patients with insomnia, found an elevated fracture risk among female patients over 60 years-old . HoweverOne study reported that zolpidem use was associated with a greater risk of fractures among elderly patients than alprazolam and lorazepam us, but a similar risk to that associated with diazepam use . Other sSeveral previous studies focused on the impact of hypnotics use on the risk of hip fractures , 21, 45 p < 0.05). A possible explanation for this elevated fracture risk is that the elimination half-life of diazepam is prolonged in the elderly [We also compared use of several of the BDZs most frequently prescribed to elderly people including alprazolam, bromazepam, diazepam, estazolam, fludiazepam, flunitrazepam, lorazepam and midazolam , 46, 47. elderly , suggestPrevious studies found that depression is a risk factor associated with fractures in elderly individuals with osteoporosis , 50. HowSleep disorders have been found to increase the risk of depression , 53, suiPatients with missing data in the LHID 2005 dataset were excluded from consideration, which could introduce minor biases in the analysis. Additionally, this study used claimed data, and we could not observe clinical events such as the extent of symptom, recovery time, actual BDZ and zolpidem dosages or side effects. In addition, before 2000 NHIRD data were coded using A-codes, which cannot be analyzed. For the ICD codes in diagnoses, there must be some individual differences, however, as aforementioned studies have shown, the NHIRD is accurate in diagnoses. Despite these limitations, the LHID provides large samples and sufficient information about disease diagnoses and drug usage, and LHID data have been used for many studies of interactions between drugs and disease or between disease and disease.This case cross-over study revealed that zolpidem or BDZ use is not associated with increased risk of all-site fractures in elderly, but that diazepam use is associated with elevated fracture risk. Increased fracture risk among patients without depression requires further investigation. Hip and spine fractures were a particular concern for elderly users of zolpidem or BDZ hypnotics, although the association was found not to be significant after adjusting for comorbidities.


Canada Foundation for Innovation is also a funder for this study. Please refer to the correct funding and acknowledgement statements below:Funding:JCM was supported by a postdoctoral fellowship from the U.S. National Institutes of Health (5F32AI084427). This work has been funded by grants from ISCIII PS09/00130, Ministerio Economía y Competitividad (MINECO) SAF2012-31166, Dpto. Salud Gobierno Navarra 359/2012, and MinisterioEducación PRX 12/00191 to JG, PS09/01904 to JL, and Canadian Institute of Health Research to RJR. CIBERES is an initiative from ISCIII, Spain. Sequencing was performed at the Pharmaceutical Sciences Sequencing Centre at the University of British Columbia, supported by grants from the CFI (Canada Foundation for Innovation) and the faculty of Pharmaceutical Sciences. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Acknowledgements:We thank Javier Moleres and Antonio López-Gómez for their technical help. We thank Sunita Sinha and Corey Nislow for library preparation and next-generation sequencing, performed at the Pharmaceutical Sciences Sequencing Centre at the University of British Columbia, and supported by grants from the Canada Foundation for Innovation and the faculty of Pharmaceutical Sciences to C. Nislow. We thank Christopher J. Grassa for critical help with the genome assembly pipeline.