At the end of the eleven-year conflict in Sierra Leone, a wide range of policies were implemented to address both demand- and supply-side constraints within the healthcare system, which had collapsed during the conflict. This study examines the extent to which households’ exposure to financial risks associated with seeking healthcare evolved in post-conflict Sierra Leone.This study uses the 2003 and 2011 cross-sections of the Sierra Leone Integrated Household Survey to examine changes in catastrophic health expenditure between 2003 and 2011. An Oaxaca-Blinder decomposition approach is used to quantify the extent to which changes in catastrophic health expenditure are attributable to changes in the distribution of determinants  and to changes in the impact of these determinants on the probability of incurring catastrophic health expenditure (coefficient effect).The incidence of catastrophic health expenditure decreased significantly by 18% from approximately 50% in 2003 t0 32% in 2011. The decomposition analysis shows that this decrease represents net effects attributable to the distributional and coefficient effects of three determinants of catastrophic health expenditure – ill-health, the region in which households reside and the type of health facility used. A decrease in the incidence of ill-health and changes in the regional location of households contributed to a decrease in catastrophic health expenditure. The distributional effect of health facility types observed as an increase in the use of public health facilities, and a decrease in the use of services in facilities owned by non-governmental organizations (NGOs) also contributed to a decrease in the incidence of catastrophic health expenditure. However, the coefficient effect of public health facilities and NGO-owned facilities suggests that substantial exposure to financial risk remained for households utilizing both types of health facilities in 2011.The findings support the need to continue expanding current demand-side policies in Sierra Leone to reduce the financial risk of exposure to ill health. Direct payment made by patients at the point of care (out-of-pocket payment) is the major source of financing health care in low income countries . In addiIn recent years, the emphasis has shifted from simply estimating the extent of catastrophic health expenditure within populations to understanding socioeconomic factors that explain variations in households’ exposure to healthcare financial risks. Studies using household-level data have consistently shown strong correlations between a range of household socio-demographic characteristics and the incidence of catastrophic health expenditure. For example, the incidence of catastrophic health expenditure is correlated with health care need – households with a larger proportion of elderly members or children under the age of five are more likely to incur catastrophic health expenditure –12. HousThese studies provide insights into factors that protect households or increase their vulnerability to catastrophic health expenditure and are useful in informing targeted policy decisions. Therefore, in addition to estimating trends in the incidence of catastrophic health expenditure, this paper quantifies the extent to which temporal changes in the determinants of catastrophic health expenditure, including household characteristics and healthcare-seeking behaviours, contributed to changes in the incidence of catastrophic health expenditure in Sierra Leone. Temporal changes in the incidence of catastrophic health expenditure can occur either through a change in the distribution of determinants or through a change in the impact of these determinants. The Oaxaca-Blinder decomposition approach , 19 is aHistorically, Sierra Leone has ranked as one of the poorest countries in the world, an economic situation made worse by the eleven-year 1991–2002) conflict. The conflict was characterized by extensive destruction of life and property, a contraction of the economy and the collapse of public infrastructure across the country. Approximately 2 million people were displaced during the conflict and at the end of the conflict in 2002, an estimated 50,000 people had been killed out of a population of 4.4 million 02 confli.Although Sierra Leone still lags behind other sub-Saharan African countries in terms of its health and economic indicators , some prIn the aftermath of the conflict, several policies were implemented to strengthen the health system. In order to increase responsiveness to local needs in pre-conflict marginalized areas, a Local Government Act was passed in 2004 to decentralize some central government functions  to local councils , 26. ThiIn 2006, a cost-recovery scheme was reintroduced following a failed attempt to fully implement one in 2002 . The aimIn 2010, the Government of Sierra Leone launched a Free Healthcare Initiative (FHCI) aimed at increasing access to health services among vulnerable populations . Under tThese reforms are likely to have important implications for households’ healthcare-seeking behaviours and exposure to healthcare financial risks. For example, the reintroduction of the cost-recovery scheme and user charges in public health facilities can have an effect on households through two possible routes. First, the fall in the use of public healthcare services by some users as a result of higher charges , 31 reduTherefore this paper offers an advantage over previous studies by providing insights not only into the association between household characteristics and the incidence of catastrophic health expenditure but also into the extent to which wider healthcare sector reforms, implemented during the study period and affecting household healthcare-seeking behaviours, may have changed households’ susceptibility to healthcare financial risks.The rest of the paper is organized as follows: section 2 outlines the SLIHS, study variables and the Oaxaca-Blinder decomposition approach; the results are presented and discussed in sections 3 and 4 respectively; and the final section concludes the paper.The SLIHS is a cross-sectional national representative sample of households in Sierra Leone conducted in 2003 and 2011 to track changes in household living standards and wellbeing. In both surveys, households were selected using comparable sampling strategies. Sierra Leone is divided into four provinces and 14 districts. Each district is in turn sub-divided into lower administrative units: first into local councils (19), then chiefdoms (149) and sections (1322). Each section is further sub-divided into area units known as enumeration areas (EAs), which formed the primary sampling unit of the SLIHS. Households were selected from a sample of EAs in a two stage-sampling process: first, EAs were stratified into rural and urban areas and a random sample of EAs selected to ensure a representative number of households from urban and rural areas. A random sample of households was then drawn from selected EAs.In 2011, approximately 6800 households were selected. However, due to financial and human resource constraints, a significantly smaller sample of households (approximately 3700) was interviewed in the 2003 survey.In both years, data on health expenditure were collected using a health questionnaire completed by heads of households on behalf of all household members. The health questionnaire collected data on the use of in- and outpatient health services from both formal and informal service providers as well as expenditure incurred in using these services. Heads of households were asked to report on the use of healthcare services by any household member in a two- or four-week periodIn both years, detailed information was collected on a wide range of food and non-food goods purchased by households. In 2003, household heads were asked to report on food and frequently purchased non-food goods bought in the 3–12 months period prior to the interview date. To limit recall bias, a different approach was adopted in 2011 to collect data for food and frequently purchased non-food goods. Each household received five visits at regular intervals over a one-month period. The monthly food and non-food expenditure for 2011 was estimated as the sum of all purchases made over the entire month.For infrequently purchased non-food goods and services, expenditure data were collected over a 12-month recall period in both 2003 and 2011  using purchasing power parity exchange ratesCatastrophic health expenditure is defined as health expenditure exceeding 10% of household total expenditure. First, health expenditure share of total expenditure is estimated for each household and then a binary variable is generated which equals one, when health expenditure share of total expenditure exceeds 10%.This study makes use of variables shown in previous studies to explain variations in households’ exposure to healthcare financial risks. These include household size, proportion of household members below the age of 5 years and above the age of 65 years, household location  and proportion of unemployed adult household members. Head of household characteristics include age (and age squared), gender, marital status , religion  and education (no education to junior secondary education/some senior secondary education and above).In addition to household demographic characteristics, this study uses other previously identified determinants of catastrophic health expenditure. These include the proportion of household members reporting ill health as well as the proportion of household members utilising inpatient and outpatient healthcare services. To investigate differential effects of health facility-type, health service utilisation is disaggregated by type of service. This is identified using a combination of two variables available in SLIHS – the owner of the healthcare facility visited (public or private) and the health worker type consulted. For outpatient health care use, this includes use of informal healthcare services  = 0 , the change in the incidence of catastrophic health expenditure,where The decomposition of approach , 33:3\dowhere ion term , this stSecond, a detailed decomposition is performed to identify the contribution of each covariate to k represents the kth covariate and where d effect , 35.oaxaca’, specifying a linear probability model [oaxaca’ can support a probit or logit model specification, the linear probability model is used given that ‘oaxaca’ applies the decomposition to the linear predictions from the model [oaxaca’ with a logit or probit model specification will not be expressed on a probability scale but in terms of log odds (in the case of a logit model) or z-scores (in the case probit models), resulting in difficulties interpreting the results of the decomposition analysis.The decomposition analysis is performed using the STATA user-written command, ‘ty model . While ‘he model . This meTable Positive improvements are also observed with health indicators. The proportion of household members reporting ill health decreased significantly by approximately 21 percentage points points. In terms of household healthcare-seeking behaviours, the proportion of household members utilising formal public outpatient healthcare services remained largely unchanged. However, a higher proportion of household members utilised informal healthcare services (an increase of approximately 1 percentage points) while utilisation of formal private as well as NGO/missionary healthcare services decreased by approximately 0.4 and 1 percentage points, respectively.A shift is observed in the distribution of households across Sierra Leone’s four regions Table . The proTable Compared to households located in the Western region, households in other regions were more likely to experience financial risks associated with seeking healthcare. For example, the probability of incurring catastrophic health expenditure for households living in the Eastern region is approximately 7 percentage points higher than households in the Western region.The pooled cross-sections, while providing useful insights into the determinants of catastrophic health expenditure, mask important changes that occurred over time and the resulting implications for households’ exposure to financial risks. A comparison of coefficient estimates between 2003 and 2011 shows that the magnitude and direction of effect vary between the two years . Similarly, the use of any healthcare facility is associated with a higher probability of incurring catastrophic health expenditure. However in 2011, the magnitude of this effect is higher for formal public outpatient healthcare services by approximately 0.3 percentage points.The Oaxaca-Blinder decomposition analysis Table  quantifiTotal distributional effect is largely driven by changes in the mean distributionTotal coefficient effect is largely driven by changes in the impact of ill health; however, regional variations in the incidence of catastrophic health expenditure as well as changes in the impact of public healthcare service-use have counteracting effects Fig. .Fig. 1SiThe robustness of these findings is tested using different thresholds for defining catastrophic health expenditure Table . The CHEFollowing the end of the brutal civil conflict in Sierra Leone, the health sector underwent a series of reforms aimed at strengthening both demand- and supply-side functions of the health system. These include the devolution of administrative responsibilities and funds to local councils in 2004; the reintroduction of the cost-recovery scheme and user fees in public health facilities in 2006; the Free Health Care Initiative (FHCI) in 2010 and the accompanying human resources for health (HRH) reformsand widespread reconstruction and refurbishment of public healthcare facilities nationwide –29. CoveChanges in both the distribution and coefficient effect of ill health between 2003 and 2011 made the largest contribution to the CHE gap. Between 2003 and 2011, the proportion of household members reporting ill health decreased significantly, contributing to a reduction in the incidence of catastrophic health expenditure. Similarly, the impact of ill health on the likelihood of incurring catastrophic health expenditure reduced significantly between the two study years. In addition to severe disruptions to livelihood, conflicts have been linked to outbreaks of diseases, particularly infectious disease due to disruptions in safe drinking water sources and sanitation . The conThe distributional effect of household regional location provides further indication of general improvements in household economies in the post-conflict period. The distributional effect of regional location, capturing the shift in populations from the least conflict-affected regions (the Western and Southern regions) to the worst affected regions (the Northern and Eastern regions) is indicative of regional economic growth as peace and security returned , 39. ThiBy contrast the regional coefficient effect, capturing regional variations in exposure to financial risk suggests that these positive changes may not have been felt equally across all regions. Compared to households living in the Western region, those in other regions were less likely to incur catastrophic health expenditure in 2003 but more likely in 2011. Changing population distribution and accessibility of health infrastructure is likely to explain this effect. For example, only 16 health facilities were functioning at the height of the conflict in 1996, with the majority of these located in the Western Region . TherefoThe distributional and coefficient effect of facility-type provides interesting insights into the evolution of health service provision in the post-conflict era. A statistically significant decrease in the use of private facilities including NGO and missionary-owned facilities is observed between 2003 and 2011 while an increase is observed in the use of public health facilities . In the aftermath of the conflict, NGOs and private actors played an important role in the provision of healthcare , fillingThe decrease in the use of formal private and NGO healthcare services significantly reduced households’ exposure to financial risk. However, those who continued to use NGO-owned health facilities faced a higher risk of incurring catastrophic health expenditure in 2011 (captured in the coefficient effect). An increase in prices of healthcare services between 2003 and 2011 is likely to explain the observed coefficient effect of NGO- and privately-owned healthcare facilities. NGO-owned facilities in Sierra Leone largely operate on a similar basis to privately owned facilities, charging patients a fee-for-service at the point of care , 46. AltThe distributional effect of public health services suggests an increase in households’ exposure to the financial risk of seeking healthcare. While this effect was not found to be statistically significant, the coefficient effect provides more insight into the implication of public health facility-use on households’ exposure to financial risk. In both study years, the risk of incurring catastrophic health expenditure increased with the proportion of household members using public health services. However, this effect is significantly higher in 2011 suggesting higher financial risks faced by households. This effect may partly be explained by health financing reforms including the reintroduction of the cost recovery scheme and user fees in public health facilities in 2006 which would have resulted in higher out-of-pocket health expenditure for those utilizing these services in 2011 , 31. FurThese findings are in part, sensitive to the catastrophic health expenditure threshold. The CHE gap varies between 11 and 23%, depending on the threshold used for defining catastrophic health expenditure. Nevertheless, the proportionate contribution of the total coefficient and distributional effects to the CHE gap at each threshold are comparable. The detailed distributional effects appear robust to thresholds, while the coefficient effect varies in some cases. For example, the coefficient effect of ill health and public health facility-use is significant only at lower thresholds suggesting equal effects of these determinants in both years at higher thresholds.The robustness of our findings could be affected by the extent of measurement errors in household expenditure data. The precision of household expenditure estimates from survey data is subject to recall periods over which data are collected and the differences in how goods were itemised between the two study years . Given tFurthermore, differences in the recall periods over which health expenditure and other household expenditure were collected may have affected our estimates of the incidence of catastrophic health expenditure. Households are likely to recall and accurately report health expenditure occurring over a shorter recall period , 54. On The findings of this study illustrate an important limitation of this measure of catastrophic health expenditure. Catastrophic health expenditure is conditional on healthcare use and captures the financial risks associated with accessing care. This means that the measure of catastrophic health expenditure defined in this study captures only financial risks for those who seek health care but fails to account for those who do not seek health care when ill due to an inability to pay. Although we find some evidence that regional socioeconomic inequality may have accounted for higher catastrophic health expenditure in 2011, estimates of the concentration index in 2011 suggests that in the total population, catastrophic health expenditure was concentrated amongst those who potentially have the ability to pay for health care when needed. The situations where health care is not sought due to an inability to pay or when health care services are not available may be equally impoverishing – for example an unresolved health problem could prevent adults in a household from working, thus compromising household living standards. The catastrophic health expenditure measure defined in this study does not detect these and is an inevitably partial measure to understanding the links between health, health seeking behavior and impoverishment.The findings of this study suggest that while efforts have been made to address supply-side constraints to accessing healthcare in the years following the conflict in Sierra Leone , financiWhile rebuilding and refurbishing destroyed health infrastructure (supply-side initiatives) is a crucial element to improving access to quality healthcare services in the aftermath of conflict, for policy-makers, this study highlights the importance of complementary demand-side mechanisms aimed at protecting households, at a time when households are themselves recovering from severe losses. These demand-side mechanisms including health insurance schemes and partial or full health care fee waivers  have been shown to reduce out-of-pocket payments amongst the most vulnerable populations in other settings . Althoug


Zinc is an essential mineral of which its functions have potential implications on exercise performance and beneficial adaptations of physical activity. While the effects of aerobic exercise on zinc metabolism acutely have been well described, the effect of long-term exercise training on zinc status remains unclear. The present review aims to determine the effects of exercise training on markers of zinc status in an apparently healthy adult population. We conducted a systematic literature search on PubMed, Scopus, SPORTDiscus and Cochrane Library from inception to 28 January 2016 to identify interventional or cohort studies that investigated the effects of exercise training on indices of zinc status. Pairwise comparisons of mean differences in within-group change were calculated and summarised visually in forest plots. Six studies satisfied the inclusion criteria for the systematic review, of which 5 studies included data on changes in serum zinc concentrations and 3 studies provided changes in dietary zinc intake. Two comparisons showed significantly higher increase of serum zinc concentrations in the exercise group compared to control, while one comparison reported significantly lower change in serum zinc for the exercising group. The exercise groups consumed significantly higher dietary zinc compared to controls in two comparisons. The present review revealed an incomplete evidence base in evaluating the effect of long-term exercise training on markers of zinc status. Further well-designed investigations are required to elucidate the relationship for establishment of dietary recommendation in populations who are continuing exercise interventions. Zinc is an essential trace element with numerous metabolic functions . While zExercise and physical activity have been the cornerstone of lifestyle recommendations for the healthy population and those with chronic diseases ,11. The A number of cross-sectional studies have investigated the association between exercise/physical activity and zinc status, in particular the zinc status in athletic and control populations. While some studies showed lower serum zinc concentration in athletes ,17, otheWe have previously reported significant acute fluctuations in serum zinc concentrations as a result of a single bout of aerobic exercise ,21. The th of January 2016. The search terms included: zinc, exercis*, athlet*, physical activity, train*. Related terms and MeSH terms were used where appropriate. Searches were limited to human subjects and the English language. http://www.crd.york.ac.uk/PROSPERO/ (CRD42015026336).A literature search of PubMed, Scopus, SPORTDiscus and the Cochrane Library electronic databases were conducted from inception to the 28Interventional and cohort studies that were published in peer-reviewed journals and examined the effects of exercise training on zinc status were included. Exercise training was defined as an intervention, over a defined period of time, which involves more than one bout of continuous physical activity . Males aTwo investigators extracted published data from all included studies independently, and any differences were resolved by discussion. Data extracted included descriptive information, such as the study authors, year of publication, country where the study was conducted, the number, sex and other characteristics of the control and study populations. The duration, intensity and mode of exercise performed in the study were detailed in the data extraction process. Primary outcomes extracted were plasma/serum zinc concentration and dietary zinc intake, before and following exercise training. Plasma and serum concentrations were grouped to represent systemic zinc concentration; in this report the term “serum” will be used to represent both serum and/or plasma. Other potential markers of zinc status, including zinc concentrations in urine and erythrocyte, and activity or concentration of Cu-Zn superoxide dismutase were extracted as secondary outcomes.low, unclear and high. Studies with exercise group matched with control group in age, sex and body composition represent a low risk of bias in recruitment. Valid measurement of serum zinc was determined by the use of trace element free collection tubes, appropriate method of analysis i.e. atomic absorption spectroscopy or inductively coupled plasma mass spectroscopy, and timing of blood collection. Similarly, low risk of bias for the valid measurement of dietary zinc was achieved by the use of appropriate method of collection , reporting of energy or macronutrient intake and consideration of supplemental intake. Data on risk of bias in individual studies were entered into Review Manager 5.3 [All included studies were assessed for risk of biases that are applicable for non-randomised interventional studies as recommended by the Grading of Recommendations Assessment, Development and Evaluation (GRADE) guidelines . Two indager 5.3 .Changes in the primary outcome measures of serum zinc concentration and dietary zinc intake were calculated by subtracting final from baseline values, with calculated estimation of SD of change using an assumed correlation of 0.7 between the two values . The difThe electronic database search identified a total of 4919 citations following removal of duplicates. After initial screening of titles and abstracts, 4802 citations were excluded as they were irrelevant to the current review. Of the remaining 117 full texts assessed for eligibility, six studies met the inclusion criteria. The characteristics of studies included in the current review are described in et al.’s study. A small percentage of participants displayed low serum zinc concentration in another study [Five studies –32 provier study , with 5%et al. [Three studies ,30,31 pret al.  reportedResults for other zinc outcomes are presented in low risks of bias for recruitment with control group matched in age, sex and BMI with the exercise group [low or unclear risks of bias for valid measurement of serum zinc, with the exception of one study scoring high risk of bias, where non-fasting blood samples were collected for the analysis of serum zinc [low risk of bias for valid measurement of dietary zinc. Most of the studies reported follow up data for all participants thereby scoring low risk of bias for incomplete accounting of participants. The majority of the studies reported all relevant outcomes with the exception of one study that failed to report measured dietary intake data [se group ,28,30–32ake data  and anotThe current review of interventional and cohort studies has identified incomplete evidence for the effects of exercise training on zinc status in an apparently healthy adult population. In total, six studies satisfied the inclusion criteria for the present analysis. The limited evidence suggests that exercise is associated with higher dietary zinc intake and erythrocyte SOD activity; while minimal differences were observed in serum zinc concentrations. We have revealed distinct gaps in the current literature regarding the effects of exercise training on zinc status with implications on establishing dietary zinc requirements in populations that exercise regularly.The studies included in the current review consisted of exercise interventions that are dissimilar in mode, duration and intensity. Three out of six studies included exercise that is predominately aerobic in nature, for example running and swimming ,29,31. ABased on six comparisons for exercise-induced changes in serum zinc concentration, the estimated mean difference of change for two comparisons indicated that exercise induces significant increase in serum zinc; while one comparison suggested that serum zinc changes are lower in the exercise group compared to control. The current evidence suggests equivocal changes in serum zinc concentration as a result of exercise interventions. The limitations of serum zinc concentration as a marker of zinc status in humans are well described ; changesThe most commonly measured zinc outcome, other than serum zinc concentration, was erythrocyte Cu, Zn-SOD activity. SOD plays a key role in antioxidative activity by providing catalytic function for the disassociation of the radical oxygen species, superoxide, into oxygen or hydrogen peroxide. In erythrocytes where oxygen exchange occurs with haemoglobin, significant intracellular oxidative stress is induced by exercise acutely through increases in oxygen consumption . As a reExercise also impacts on immunity, both acutely and chronically. Following a bout of prolonged high intensity exercise, acute decreases in multiple components of immune function are evident including the reductions in B lymphocyte production of immunoglobulins and antigen-presenting capacity . Whilst On the basis of four comparisons, dietary zinc intake appears to be higher in exercise groups compared to control in two comparisons; therefore, we deemed the effects of exercise training on dietary zinc intake to be equivocal. In a study by Lukaski et al., while dietary zinc intake increased in male swimmers over the competition period, zinc density (mg/MJ) remained unchanged . This suAll included studies within this review were low in number of participants and therefore contributed to the limited power of the comparisons in determining the effect of exercise training on markers of zinc status. Further, four out of six included studies recruited men only; similar level of sex bias were noted previously , highligThis review is the first to determine the effects of exercise training on markers of zinc status in an adult population. The current report extends our knowledge of the acute effects of exercise on markers of zinc metabolism ,21. In tThe present review revealed an incomplete evidence base in evaluating the effect of long term exercise training on markers of zinc status. Further, the strength of the presented evidence is limited by the majority of study designs reported being non-randomised or cohort studies. The limited evidence suggests that dietary zinc intake may increase in those who are physically active, by homeostatic adjustments for increases in energy expenditure. Additional studies, with different study populations such as those participating in moderate levels of physical activity, are required to extend the evidence to the general population. Future evaluation on the effects of zinc intake on physical performance will present implications for clinical sports nutrition practice. Exercise training was found to be associated with increases in SOD activity as part of the adaptations of exercise, consistent with the current literature . As the S1 Table(DOCX)Click here for additional data file.S1 File(DOC)Click here for additional data file.S2 File(XLSX)Click here for additional data file.


The establishment of agricultural matrices generally involves deforestation, which leads to fragmentation of the remaining forest. This fragmentation can affect forest dynamics both positively and negatively. Since most animal species are affected, certain groups can be used to measure the impact of such fragmentation. This study aimed to measure the impacts of agricultural crops (matrices) on ant communities of adjacent lower montane Atlantic rainforest fragments. We sampled nine forest fragments at locations surrounded by different agricultural matrices, namely: coffee (3 replicates); sugarcane (3); and pasture (3). At each site we installed pitfall traps along a 500 m transect from the interior of the matrix to the interior of the fragment . Each transect was partitioned into four categories: interior of the matrix; edge of the matrix; edge of the fragment; and interior of the fragment. For each sample site, we measured ant species richness and ant community composition within each transect category. Ant richness and composition differed between fragments and matrices. Each sample location had a specific composition of ants, probably because of the influence of the nature and management of the agricultural matrices. Species composition in the coffee matrix had the highest similarity to its corresponding fragment. The variability in species composition within forest fragments surrounded by pasture was greatest when compared with forest fragments surrounded by sugarcane or, to a lesser extent, coffee. Functional guild composition differed between locations, but the most representative guild was ‘generalist’ both in the agricultural matrices and forest fragments. Our results are important for understanding how agricultural matrices act on ant communities, and also, how these isolated forest fragments could act as an island of biodiversity in an ‘ocean of crops’. In many parts of the world, agricultural practices are the main causes of deforestation and forest fragmentation . FragmenThe impact of fragmentation on forest species emerges from two components: the reduction of area (habitat loss per se) and the isolation between the remaining habitat fragments ,4. Both The nature of the surrounding agricultural matrices affects forest species dynamics  at localForest fragmentation also generates a transitional area (edge) between the matrix and the forest areas. The abiotic characteristics of this zone are different and could in turn generate several “edge effects” ,25. ThesIn general, to measure the effects of forest fragmentation a variety of organisms can be used ,32 and tAmong the arthropods, ants are a group that are commonly used as bioindicators , especiaThe focus of the current study was to elucidate how ant alpha-diversity (i.e. species richness), species composition and functional guild composition of ground ants in fragments of lower montane Atlantic rainforest in southeastern Brazil are influenced by different types of agriculture in order to better understand how the nature of forest fragmentation influences such communities. We also considered whether all matrices of a particular crop influence the ant fauna to the same degree. We expected that agricultural matrices would have fewer species of ants than forest fragments and that the similarities in species composition among the same agricultural matrices would be greater than among the remaining forest fragments. We also expected that the predominant functional guilds in the forest fragments would be predators, while in the agricultural matrices they would be generalists.The work was conducted in three regions, Alfenas, Areado and Serrania, which comprise the Alfenas microregion in southern Minas Gerais State, Brazil  . The AlfCoffea arabica), pasture (Brachiaria sp.) and sugarcane (Saccharum officinarum). The fragments studied were all preserved remnants of lower montane Atlantic rainforest )  . The posmunities . The nMDmunities . The strur model . In the accard]) .2 = 41.928; df = 11; p < 0.001). There was a difference between the interior of agricultural matrix of pasture and sugarcane , and pasture and coffee . In addition, there were differences between interior of forest fragments of pasture and sugarcane , and interior of forest fragment of coffee and interior of forest fragment of sugarcane . Likewise, we found a difference between the interior of the agricultural matrix of coffee and interior of the forest fragment embedded in coffee . Ant species richness among the points fluctuated and then, generally, stabilized in the forest fragment . The similarity analysis showed that the major similarity was among matrices of the same type. The pasture matrices were more similar among themselves than other matrices or forest fragments.We found a difference in ant species richness among the transect categories  than in the forest fragments (48.83%). More representatives from the predator guild were recorded in the forest fragments (32.03%) than in the matrices (16.67%). Arboreal ants were more abundant in the pastures (2.89%). However, we did not find arboreal ants in the forest fragments surrounded by pasture. Army ants were more abundant in forest fragments surrounded by pastures (1.5%) and were not found in pastures. We recorded more Attina guild species in the forest fragments than in the matrices . There wAccording to our results, the agricultural matrices that we studied exert an influence on the embedded forest fragments and this affects ant community richness and composition. The results of the similarity analyses and the differences in the proportion of the functional guilds on each site support this statement.Pheidole was the most represented genus in our samples, being found in all locations. This representativeness could be due to it being a hyperdiverse, cosmopolitan genus of generalist ants which can use several types of resources [Atta (26.13%). Ants of this genus build giant nests [Ant species richness varied among the locations. esources ,38. On tnt nests  with up nt nests ; we beliWe found difference between ant richness and composition in the sugarcane crops and pasture. The lower richness in the pastures can be explained by the environmental factors that act direct on the pasture, such as sun exposition and winds. The habitat exerts a pressure on the ant community and that pressure can shape the richness and composition in those locations . FurtherThere were no differences in ant richness between locations . However, the communities were different among the sampled sites. Some species are very flexible and can survive in the homogeneous conditions provided in this agricultural matrix . Other rThese results for richness could be related to the turnover of ant species in the assemblages . SpeciesThe similarity between the coffee matrices and the embedded forest fragments, albeit not extremely high, could be indicating that the coffee matrices can provide similar habitat features for ant species to those of forest fragments. The coffee matrices are characterized by having arboreal, and perennial vegetation, and this may confer permeability to the matrix, facilitating species movement . This peOn the other hand, the ant composition diverged between the edge and the interior of forest fragments surrounded by pasture matrices. As mentioned before, the composition and the vegetation type of the matrix can directly influence the forest fragment ,68. The Forest fragments surrounded by pasture are more influenced by the matrix compared to the other locations ,69. In sRegarding the proportion of the functional guilds, the high number of generalist species in the agricultural matrices indicates that these species can survive and even dominate in homogeneous habitats ,71, whilIn forest fragments, however, there were more predator species than agricultural matrices. We expected more predator species in heterogeneous environments . PredatoWe add adjacent vegetation type to Lawton's hypothesis  in whichThe findings of our work supplement those from similar studies elsewhere in the world –19 and pS1 FigLines in red are agricultural matrices and the green lines are the forest fragments.(DOCX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.


KRAS and BRAF mutations is a crucial step for the correct therapeuticapproach and predicting the epidermal growth factor receptor (EGFR)-targeted therapyresistance of colorectal carcinomas. The concomitant KRAS and BRAF mutations occurrarely in the colorectal cancers (CRCs) with the prevalence of less than 0.001% of the cases.In patients with KRAS-mutant tumors, BRAF mutations should not regularly be testedunless the patient is participating in a clinical trial enriching for the presence of KRAS orBRAF-mutated tumor. The current report demonstrates a case with advanced adenocarcinomaof the colon showing the coexistence of KRAS and BRAF mutations and may haveprofound clinical implications for disease progression and therapeutic responses. The detection of  KRAS gene between the primary and metastatic tumors  is one of the most common cancers, especially in the developed countries and its worldwide mortality rates exceed 700,000 deaths per year. CRC usually causes by the mutations in the epithelial cells of the gastrointestinal surfaces resulting in hyperactivity of the signaling pathways and finally transforms these cells into the adenomatous polyps. Accumulation of the inherited or acquired mutations transits the adenomatous polyps to malignancy. The genetic mutations and morphological changes of this progression path, have been well described in the previous studies ,2. Signic tumors -5. KRAS protein is a key factor in the regulation of cell mitosis. KRAS activates its primary downstream target protein named BRAF, a serine- threonine protein kinase that mediates the KRAS signal toward the downstream effectors, Mitogen- activated protein (MAP) kinase extracellular signal-regulated kinase (MEK) and the extracellular signal-regulated kinase (ERK)  . In normelopment . Mutatioreported ,9. KRAS abnormal activity in cancer cells invasion has been well described by Liao et al.  expression level, and may promote cancer aggressiveness. Also, the elevated level of mutated KRAS enhances the carcinoembryonic antigen (CEA) level and blocks the monolayer formation of colon epithelial cells  signaling pathways play an important role in the colorectal pathogenesis and are associated with the primary resistance to the EGFR inhibitors  and BRAF (exon 15) by using polymerase chain reaction (PCR) and direct sequencing. PCR cycling conditions were performed according to the manufacturer’s instruction. The ABI 3130 genetic analyzer  and the Big Dye Terminator  were used for the sequencing reactions of this study. After obtaining informed consent from patient, the EGFR immunohistochemical expression profile was investigated by anti-EGFR monoclonal antibody according to the manufacturer’s descriptions. The paraffin-embedded tissue sections were collected on microscopic slides. Tumor and tumor-free areas of the samples were identified within 15 μm thick deparaffinized, lightly hematoxylin-counterstained sections and subjected to the pathological studies. Each microdissected area was applied to the DNA extraction followed by the mutational analysis of To investigate microsatellite instability (MSI), the DNA samples from the blood and microdissected tumoral areas of the patient were examined by using the BAT26, BAT25, NR24, NR21 and NR27 mononucleotide microsatellite markers. These markers are known to be the most sensitive markers of MSI status and widely uses for identification of the CRC cases with the concomitant mismatch repair (MMR) defects. KRAS and BRAF mutational status analysis was performed on the representative lesion of the colon, using the PCR-sequencing method and MSI analysis. The identified mutations in KRAS and BRAF genes were a mutation in codon 12 of exon 2 and a missense nucleotide base change in codon 600 of exon 15 (GTG to GAG), respectively. However, MSI was not detected. KRAS mutations in primary colorectal tumors play a prognostic role as a predictor of resistance to the anti-EGFR antibodies (KRAS mutational analysis of the primary tumor usually match with the metastases in only 5 to 10% of the cases, the KRAS mutational status is heterogeneous between the primary tumors and the metastases (KRAS and BRAF mutations in the same colorectal tumor has been reported in few studies. These studies have also reported the correlation of the concomitant KRAS and BRAF mutations with the clinical and morphological characteristics (CRC is estimated as the third commonly diagnosed type of cancer and the fourth common cause of the cancer mortality after the lung, stomach and liver cancers, in both males and females. Considering the invention of new therapeutics including Irinotecan and Oxaliplatin in addition to the development of targeted therapies such as Cetuximab and Bevacizumab, the survival rates of diagnosed patients have been significantly improved. It has been shown in several experiments that the tibodies ,18,19. Atastases -32. Thistastases ,34. Moreeristics ,26-28. KRAS and BRAF mutant tumors have a different biology and natural history than singly KRAS or BRAF mutant tumors, or which of the two mutations play the dominant role in driving the tumor proliferation. It has been noticed that the proportion of the concomitant mutations is associated with the degree of transmural invasion of the tumor; moreover, 2.8 and 9.4% of concomitant mutations occur in the T2 and T4 tumors, respectively, suggesting the activation role of both genes in the tumor progression (KRAS and BRAF mutant tumors have different mutation results and probably depict the different over-activated signaling cascades (KRAS and BRAF mutations is associated with more aggressive clinical manifestations. Because of the rarity of this observation, it is not clear whether the concomitant gression . Moleculcascades . Currentcascades . ConsideKRAS mutation status and microsatellite instability analysis are the only well-explained prognostic biomarkers as the negative predictors of the therapy efficiency in colorectal cancer. On the other hand, concomitant KRAS and BRAF-mutated colorectal tumors are relatively rare, so that the routine analysis of BRAF mutations in tumors with the KRAS mutations is not recommended. The case shows, that the concomitant KRAS and BRAF mutations is associated with more severe disease. This may provide clinical implications for cancer progression and management. Our case study suggests that the analysis of BRAF mutation, especially in KRAS-positive tumors would be highly advisable. In addition, the future clinical trials related to colorectal cancer, should specifically consider the eligibility of patients with concomitant KRAS and BRAF-mutated tumors. Currently,


Trichoderma. In general, almost half of the variations of transcriptome and protein level are due to translational control. Thus, studies are required to integrate raw information from different “omics” approaches for accurate depiction of translational response of BCAs in interaction with plants and plant pathogens. The studies on translational status of only active mRNAs bridging with proteome data will help in accurate characterization of only a subset of mRNAs actively engaged in translation. This review highlights the associated bottlenecks and use of state-of-the-art procedures in addressing the gap to accelerate future accomplishment of biocontrol mechanisms.Genome-wide studies of transcripts expression help in systematic monitoring of genes and allow targeting of candidate genes for future research. In contrast to relatively stable genomic data, the expression of genes is dynamic and regulated both at time and space level at different level in. The variation in the rate of translation is specific for each protein. Both the inherent nature of an mRNA molecule to be translated and the external environmental stimuli can affect the efficiency of the translation process. In biocontrol agents (BCAs), the molecular response at translational level may represents noise-like response of absolute transcript level and an adaptive response to physiological and pathological situations representing subset of mRNAs population actively translated in a cell. The molecular responses of biocontrol are complex and involve multistage regulation of number of genes. The use of high-throughput techniques has led to rapid increase in volume of transcriptomics data of  Trichoderma is a cosmopolitan and cardinal representative soil microflora of various climatic conditions . Genome sequencing and its annotation in mycoparasitic species have depicted genome sizes of 38.8 and 36.1 Mb for T. viride and of T. atroviride for biocontrol strains, respectively, compared to 34 Mb of T. reesei an industrial strain. Annotation of complete genome depicted a gene pool of 11,800 genes for T. atroviride and 12,400 genes for T. viride, compared to 9,143 genes in saprophytic strain T. reesei. The abundance of gene pool in mycoparasitic strains of Trichoderma genome . Recent studies showed that the gene expression of mycoparasitic T. harzianum and T. atroviride strains changes not only to plant-pathogenic fungi . The interaction of Trichoderma with host plants reprograms not only the gene expression of biocontrol strains but also of its associated host plant . For example, strains of Trichoderma are explored for growth promotion and boosting immune responses, root development, and activation of seed germination or amelioration of abiotic stresses  transcripts were found significantly differentially expressed in mycoparasitic interactions. The differentially expressed transcripts were also investigated during pathogenic attack on Phytophthora capsici, Botrytis cinerea, and R. solani . A comparative account of genome revealed presence of seven chromosomes in industrial strain T. reesei  during the early interaction of T. atroviride with B. cinerea and R. solani were identified .Studies involving translational regulation of gene expression are emerging as a prominent tool for the understanding the regulation of protein abundance in adaptive responses of the host . In the Figure 3). Polysomal profiling discovered in 1960s involves the separation of actively translated mRNAs bound by several ribosomes from free RNA by sucrose gradient centrifugation and then mRNAs can be coupled to northern blot or RT-qPCR or cDNA microarrays, or RNA-seq on a global level  polysomal profiling, (b) ribosomal profiling, and (c) ribosome affinity purification (RAP) .The post-transcriptional events such as translation regulation and protein stability are the principle causes of weak correlations and variations in proteomic, transcriptomic, and genomic data. The associated errors in transcriptome analysis are subjected to arise from the suppression by microarrays which can further impede the identification of active candidate transcripts. On the other side, methods opted for protein staining, limitations associated in visualizing low-abundant and co-migrating proteins seriously hampers proteomic based study. The recent developments in proteomics methods such as use of mass spectrometric (MS) and liquid chromatography (LC) techniques have made quantitative proteomic profiling, currently a driving force for identification of proteins. The highly stable and reproducible performance of mass spectrometers such as Q Exactive hybrid quadrupole-Orbitrap mass spectrometer MS and Triple TOF 5600 MS is capable of identification of both proteomics  and charTrichoderma spp. has accelerated our research on understanding of the behavior of different species of this genus and how the information on their gene pool determines their capabilities and limitations. The genomes of Trichoderma which is known to contain thousands of genes encoding different glycosyl hydrolases, secondary metabolites, antibiotics, lectins with insecticidal properties, and transporters with potential in bioremediation involved in antibiotics biosynthesis, and several other candidate genes , translatome level (ribosome loaded mRNAs) and the proteome is experimentally feasible in a high-throughput way, the integration of these omic technologies is still far away. Systematic global analyses aims at integrating transcriptome, translatome, and proteome level can provide accurate view of widespread adaptive mechanisms of interaction between Trichoderma–plant–pathogen.The genomic comparison of mycoparasitic species of Trichoderma arsenal involved in its success as BCAs as well as industrial sectors. In such instances, the integration of the translatome using ribosomal profiling and coupling it with proteomic approaches such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) for both extracellular and intracellular proteins offers a lot of scope for accurate characterization of active molecular components involved in biocontrol and then subsequently their utilization of various applications.In future, integrated efforts will help us to better understand, identify, and then explore the molecular behavior of Trichoderma–plant–plant pathogens in three way interaction will play vital role in accurate characterization of transcripts responsible for cosmopolitan nature of Trichoderma and then targeting the promising one for agricultural based applications. The latest advancements and complete genome sequencing have already provided a platform of gene pool. Further integration with latest functional techniques such as translatome will lead another step close to identification of targets in the form of active transcripts involved in a complex interaction of plant–BCA–plant pathogens.A comparative multiomic coupled insights of VS and RS prepared the manuscript. PS and AG edited the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In 2014, over 600 million adults worldwide were obese. Obesity increases the risk of developing major risk factors for chronic kidney disease (CKD), like diabetes and hypertension, and it has a direct impact on the development of CKD and end-stage renal disease (ESRD). The good news is that obesity is largely preventable. Education and awareness of the risks of obesity and a healthy lifestyle, including proper nutrition and exercise, can dramatically help in preventing obesity and kidney disease. This article reviews the association of obesity with kidney disease on the occasion of the 2017 World Kidney Day.Numerous studies have shown an association between measures of obesity and both the development and the progression of CKD. In general, the associations between obesity and poorer renal outcomes persist even after adjustments for possible mediators of obesity’s cardiovascular and metabolic effects, suggesting that obesity may affect kidney function through mechanisms in part unrelated to these complications. The deleterious effect of obesity on the kidneys extends to other complications such as nephrolithiasis and kidney malignancies.1The exact mechanisms whereby obesity may worsen or cause CKD remain unclear. Some of the deleterious renal consequences of obesity may be mediated by comorbid conditions such as diabetes mellitus or hypertension, but there are also effects of adiposity which impact the kidneys directly via production of (among others) adiponectin, leptin and resistin . These i2 The incidence of the so-called obesity-related glomerulopathy has increased 10-fold between 1986 and 2000.These various effects result in pathologic changes in the kidneys including ectopic lipid accumulation and increased deposition of renal sinus fat, glomerular hypertension and increased glomerular permeability, and ultimately the development of glomerulomegaly, and focal or segmental glomerulosclerosis .2 The in3Obesity is associated with a number of risk factors contributing to nephrolithiasis, such as lower urine pH and increased urinary oxalate, uric acid, sodium and phosphate excretion. The insulin resistance characteristic of obesity may also predispose to nephrolithiasis through its impact on tubular Na–H exchanger and ammoniagenesis, and the promotion of an acidic milieu.4The putative mechanisms behind the increased risk of kidney cancers observed in obese individuals include insulin resistance, chronic hyperinsulinemia and increased production of insulin-like growth factor 1, which may exert stimulating effects on the growth of various types of tumour cells. More recently, the endocrine functions of adipose tissue, its effects on immunity and the generation of an inflammatory milieu with complex effects on cancers have emerged as additional explanations.5 It is possible that the seemingly protective effect of a high body mass index (BMI) is the result of the imperfection of BMI as a measure of obesity. However, there is also evidence to suggest that higher adiposity, especially subcutaneous  fat, may also be associated with better outcomes in ESRD patients. Such benefits may be present in patients who have very low short-term life expectancy, such as most ESRD patients, including among others benefits from better nutritional status.In a seemingly counterintuitive manner, obesity has been consistently associated with lower mortality rates in patients with CKD and ESRD.6 which may serve as a platform to improve the prevention of obesity-related CKD. Campaigns aiming at reducing the obesity burden are now at centre stage worldwide and are strongly recommended by the WHO, and it is expected that these campaigns will reduce the incidence of obesity-related complications, including CKD.In the United States, Healthy People 2020, a programme that sets 10-year health targets for health promotion and prevention goals, focuses on both CKD and obesity. A successful surveillance system for CKD has already been implemented in some places such as the United Kingdom,7 In a recent meta-analysis collating experimental studies in obese CKD patients, interventions aimed at reducing body weight showed coherent reductions in blood pressure, glomerular hyperfiltration and proteinuria.8 Bariatric surgical intervention has been suggested for selected CKD and ESRD patients.Obesity-related goals in obese CKD patients remain vaguely formulated, largely because of the paucity of high-level evidence intervention studies to modify obesity in CKD patients. In overweight or obese diabetic patients, a lifestyle intervention including caloric restriction and increased physical activity compared with a standard follow-up reduced the risk for incident CKD by 30%.Globally, these experimental findings provide a proof of concept for the usefulness of weight reduction and angiotensin converting enzyme (ACE) inhibition interventions in the treatment of CKD in the obese. Studies showing a survival benefit of increased BMI in CKD patients, however, remain to be explained. These findings limit our ability to make strong recommendations about the usefulness and the safety of weight reduction among individuals with more advanced stages of CKD. Lifestyle recommendations to reduce body weight in obese people at risk for CKD and in those with early CKD appear justified, particularly recommendations for the control of diabetes and hypertension.The worldwide epidemic of obesity affects the earth’s population in many ways. Diseases of the kidneys, including CKD, nephrolithiasis and kidney cancers are among the more insidious effects of obesity, but which nonetheless have wide ranging deleterious consequences, ultimately leading to significant excess morbidity and mortality and excess costs to individuals and the entire society. Population-wide interventions to control obesity could have beneficial effects in preventing the development or delaying the progression of CKD. It is incumbent upon the entire health care community to devise long-ranging strategies towards improving the understanding of the links between obesity and kidney diseases, and to determine optimal strategies to stem the tide. The 2017 World Kidney Day is an important opportunity to increase education and awareness to that end.


This case report describes the endodontic treatment of an idiopathic perforated internal root resorption. A 24-year-old male Malay patient presented with internal root resorption of two of his anterior teeth. The medical history was non-contributory and he had no history of traumatic injury or orthodontic treatment. Cone-beam computed tomography (CBCT) determined the nature, location and severity of the resorptive lesion. Non-surgical root canal treatment of tooth #22 and combined non-surgical and surgical approach for tooth #11 were carried out using mineral trioxide aggregate (MTA) as the filling material. The clinical and radiographic examination three years after completion of treatment revealed evidences of periapical healing. The appropriate diagnosis and the treatment of internal root resorption allowed good healing of these lesions and maintained the tooth in function for as long as possible. Internal root resorption (IRR) is an inflammatory condition that results in progressive destruction of intra radicular dentin along the middle and apical thirds of the canal walls due to osteoclastic action , 2. It iIRR could be asymptomatic and may present on routine radiographic examinations . PatientDiagnostic accuracy of internal root resorption based on conventional radiographic examination is limited by the fact that the produced images only provide a two-dimensional representation of three-dimensional object. Radiographic imaging is unable to reveal the location and nature of the resorptive defect as well as the thickness of the remaining root canal dentine . FurtherThis case report describes the successful management of an idiopathic perforated IRR associated with two anterior teeth.A healthy 24-year-old male Malay patient was referred by his dentist to the endodontic specialist clinic in 2011 for the management of teeth #11 and 22. The patient reported an acute episode of throbbing pain approximately two weeks prior to his referral. He was taking a course of antibiotic prescribed by the dentist. The medical history was non-contributory. He had no history of traumatic injury or orthodontic treatmentIn the clinical examination, teeth #11 and 22 had discoloured mesial composite restorations with intact margins. The probing depth was within normal limit for both teeth. Tooth #11 was tender on palpation and percussion elicited a different sound. The tooth was not mobile. Soft swelling of buccal tissues was noted at apical area of tooth #11 and it was also tender to palpation. A periapical radiographic examination revealed a large, well demarcated oval radiolucency at the middle third of root canal of tooth #11 with the remaining small portion of the apex . Tooth #22, was not tender to percussion and palpation nor mobile. Pulp sensibility test  was performed and gave a positive reading at 20. Radiographic examination revealed a radiolucent area at the cervical third of the root canal and the canal could not be followed through the lesion . The defTo assess the nature, location and severity of the resorptive lesion, limited CBCT 3D  of the region associated with tooth #11 was taken after verbal consent obtained. Following analysis of the axial, sagittal and coronal slices, it was evident that the internal root resorption of tooth #11 was extensive and had perforated at the middle third of canal in the palatal  and bucc1) emergency treatment of tooth #11 as it was symptomatic, 2) non-surgical root canal treatment of tooth #22 to arrest the progressive resorption and 3) combined non-surgical and surgical approach for tooth #11. The patient was informed of the clinical findings, and various treatment options were discussed. The potential technical difficulties, in particular, the problem of effective sealing of the resorptive defect and the poor prognosis of the endodontic treatment for both teeth were explained to the patient. The agreed treatment plan was; At the first visit, the root canal of tooth #11 was accessed and there was pus and blood present in the canal. Canal was cleaned and dressed with calcium hydroxide and access cavity was filled with temporary cement .Treatment was then carried out for tooth #22. The root canal was accessed under surgical microscope  and there were lots of tissue in the resorptive cavity and excessive bleeding present in the canal. It was not possible to negotiate the canal beyond the resorption even after two visits. Mineral trioxide aggregate (MTA)  was placed in the resorptive cavity up to the orifice of the canal.Patient was scheduled for the management of tooth #11. However, he was out of the city and came back to see us a year later in 2012. Upon review, the tooth #22 was asymptomatic and periapical radiography showed absence of periradicular radiolucency . Root caThe patient was reviewed again recently three years after treatment completion. Patient was asymptomatic and clinically, the teeth and the surrounding tissues were healthy . The perwait and see approach is not appropriate [Management of IRR is an endodontic challenge especially if the resorptive lesion is extensive and perforating. A correct diagnosis is important as the management of IRR is different from ERR. Once the diagnosis and prognosis of IRR have been established, root canal treatment is the treatment of choice . The ratropriate . The aimropriate , disinferopriate . Howeverropriate . Sodium ropriate . Radiographically, internal resorption may be described as symmetrical or eccentric lesion with sharp, smooth and clearly defined margin, with a uniform density of radiolucency, and the outline of pulp chamber or root canal could not be followed through the lesion . In contSeveral case reports and studies have confirmed the usefulness of CBCT in diagnosing and managing resorptive lesions , 14. AltInformation obtained from CBCT scan also influenced the method of root canal filling used. Internal root resorption defects can be difficult to obturate adequately. In this case, there would be a problem of significant extrusion of root filling material through the perforation if flowable gutta percha was used. MTA was chosen instead because of its superior sealing properties, bacteriostatic effects, biocompatibility and its ability to set in presence of blood . SurgicaThe prognosis of conventional root canal treatment for non-perforated IRR is high which was in accordance to previously reported study . HoweverThis case report was intended to share information on management of idiopathic internal root resorption, the use of CBCT in decision-making process and MTA as the material of choice to treat root perforation. Diagnosing the types of root resorption lesion is a challenge as the clinical and radiographic presentation might be similar. CBCT has been very beneficial in the assessment and management of perforating internal root resorption. In internal root resorption, when endodontic treatment is adequate, lesion can be halted and evidence of bony healing can be seen radiographically.


With wildlife tourism rapidly expanding, it is critical to understand its effects on the species it targets. Accelerometer loggers show that white shark activity increases when they are interacting with cage-diving operators. The study highlights the importance of considering bioenergetics when evaluating the effects of wildlife tourism on wildlife. Carcharodon carcharias). High frequency three-axis acceleration loggers were deployed on ten white sharks for a total of ~9 days. A combination of multivariate and univariate analysis revealed that the increased number of strong accelerations and vertical movements when sharks are interacting with cage-diving operators result in an overall dynamic body acceleration (ODBA) ~61% higher compared with other times when sharks are present in the area where cage-diving occurs. Since ODBA is considered a proxy of metabolic rate, interacting with cage-divers is probably more costly than are normal behaviours of white sharks at the Neptune Islands. However, the overall impact of cage-diving might be small if interactions with individual sharks are infrequent. This study suggests wildlife tourism changes the instantaneous activity levels of white sharks, and calls for an understanding of the frequency of shark-tourism interactions to appreciate the net impact of ecotourism on this species’ fitness.Anthropogenic activities are dramatically changing marine ecosystems. Wildlife tourism is one of the fastest growing sectors of the tourism industry and has the potential to modify the natural environment and behaviour of the species it targets. Here, we used a novel method to assess the effects of wildlife tourism on the activity of white sharks ( Understanding how species respond to human-induced changes has become an important research pursuit . In receResearch on the effects of tourism on elasmobranchs is on the rise, with previous studies investigating impacts of provisioning on elasmobranch physiology e.g. , changesCarcharodon carcharias) is a large marine apex predator with a global distribution, occurring in temperate, sub-tropical and tropical waters (Triaenodon obesus) for tourism increases their daily energy expenditure by elevating activity levels during periods when they normally rest  products to attract sharks present in the area to the vessels. Tethered baits of tuna sections or gills and entrails of up to several kilograms are also used to improve client experience by keeping sharks within visual range of divers in the cage. The third operator does not use berley or tethered bait but uses sound transmitted from an underwater speaker to attract sharks.The Neptune Island group is located near the approach to Spencer Gulf, about 30 km from the South Australian mainland Fig. . While tWhite sharks were equipped with an ‘accelerometer package’ that included several devices to record the activity of white sharks and ensure the recovery of the device  during AA plastic cable connected to a time-scheduled release mechanism  bound the package to a fin-clamp . AccelerFor each shark, the total deployment period was divided into shorter periods, which were assigned one of five contexts based on time of the day, the presence/absence of the shark and cage-diving operators, and the behaviour of the shark Table . The preSwim speeds (m/s) were estimated from logger propeller rotation values, using a relationship between rotation and speed determined in a flow-tank. Attachment angle  were used to determine the effects of contexts on each of these influential metrics, with Interaction as the base level and being compared against the other contexts. The inclusion of individual shark as a random effect enabled the analysis to account for the lack of independence in behaviour within each identified shark. The most appropriate statistical family, error distribution, and validity of the model were determined through an examination of the distribution of the response variable, a visual inspection of the residuals for the saturated models, and an ANOVA test between the fitted and residual values of the model. Modelling was undertaken using the ‘glmmPQL’ function of the MASS R package and accounting for serial autocorrelation. Model fitness was assessed based on R squared values following The accelerometer package was deployed on ten sharks  between 2.9 and 4.3 m total length for a total of 211.5 h  = 0.0002), with Interaction being significantly different to all other contexts, but no other contexts being different to each other , where white sharks associating with cage-diving vessels switch from patrolling to area restricted searching (ODBA increased from ~0.034 when sharks were at the Neptune Islands (i.e. ‘Absent’ and Present contexts) and at ‘Night’ to ~0.055 during Interactions, representing a 61% increase. Considering that several studies have validated ODBA as a reliable proxy for metabolic rate , this inWhile ODBA was significantly higher during Interactions, swim speed and tailbeat frequency were similar between contexts. The number of bursts and change in manoeuvring during Interactions might have not been accompanied by measurable changes in mean speed or tailbeat frequency. Tailbeat frequency was also based on dominant frequency from Fast Fourier Transformation, which is best estimated during regular swimming. Extensive turns and body movement such as during Interactions might affect the ability to identify dominant frequency. Swim speed was not estimated for two sharks because of the deployment angle of the accelerometer package, and there were a few instances when the swim speed stopped recording during strong acceleration. These might explain the discrepancy between swim speed, tailbeat frequency and ODBA, and result in ODBA being a better proxy for activity in this study than swim speed or tailbeat frequency.T. obesus (T. obesus is for a species which typically spends a large portion of the day resting on the seabed (Although interacting with the cage-diving industry increases ODBA while at the Neptune Islands, the amount of time that white sharks interact with operators is variable (e seabed . These sAlthough sharks are enticed to the cage-diving vessels with baits, industry regulations do not allow operators to feed white sharks and strict limits on the amount of bait and berley are now in place in South Australia and at other white shark cage-diving locations . Energy The white shark cage-diving industry in South Australia is managed using an adaptive management framework based on the residency of white sharks at the Neptune Islands, which is estimated annually . The curThe present study provides evidence of the effect of wildlife tourism on the activity of a marine apex predator and potential implications for its daily energy budget, and ultimately improves our understanding of behavioural responses to anthropogenic influences. Future research should quantify the amount of time white sharks interact with cage-diving operators and estimate its effect on white sharks in relation to their daily energy budget. Estimation and comparison of the energy obtained from natural prey vs. bait would also facilitate a better understanding of the effect of the cage-diving industry on the energy budget of white sharks (Supplementary DataClick here for additional data file.


Decisional Balance and Processes of Change are generally addressed in motivational interventions for the treatment of cannabis use disorders. However, specific aspects of these multifaceted constructs, with greater relevance for severe cannabis users, need to be ascertained to enable better interventions. This study aimed to compare the different facets of decisional balance and processes of change between mild and severe cannabis users in a community-based sample of young undergraduates. Thirty-one severe cannabis users and 31 mild cannabis users, indicated with the Severity of Dependence Scale, were assessed using the Decisional Balance Questionnaire (DBQ) and the Processes of Change Questionnaire (PCQ). We found that severe cannabis users had higher scores in the DBQ dimensions of Utilitarian Gains for the Self, Utilitarian Gains for Significant Others, and Self-approval, as well as in the total subscale of Gains but not Losses. The group of severe cannabis users also had higher scores in the PCQ dimensions of Self-revaluations and Counter-conditioning. Our results pinpoint specific dimensions of Decisional Balance and Processes of Change that are endorsed by severe cannabis users. This knowledge could be applied to inform motivational interventions targeting severe cannabis users. Cannabis is the most commonly used illegal drug in Europe; 14.6% of young adults ages 15 to 34) have used cannabis within the past month  and disadvantages (losses) of using cannabis versus quitting cannabis. Cognitive processes linked to disadvantages of smoking and gains of quitting have shown to shift the decisional balance in favour of quitting as indicated by research in nicotine dependence . HoweverThe Processes of Change represent 10 different behavioural and experiential strategies for changing behaviour. They have been classified in cognitive-affective processes (defined as changes in the way people think and feel about their smoking) versus behavioural processes (defined as coping strategies that are utilized to change cannabis use behavior). Cognitive-affective processes include consciousness raising (CR), Dramatic Relief (DR), Environmental Re-evaluation (ER), Self-revaluation (SR), and Social Liberation (SO). Behavioural processes include Stimulus Control (SC), Counter Conditioning (CC), Reinforcement Management (RM), Self- Liberation (SL), and Helping Relationships (HR). Cognitive-affective processes are more often utilized during early stages of change, fostering awareness and control of thoughts, feelings and health-related goals , 14 and Decisional Balance and Processes of Change facets are typically addressed in some of the most successful treatment interventions for cannabis dependence, such as motivational interventions. However, it would be relevant to determine which of these facets are actually endorsed by severe cannabis users, as the facets that better resonate with their own motivations will be likely to have stronger therapeutic effects. To address this unmet need, this study aims to compare Decisional Balance and Processes of Change in severe versus mild to moderate cannabis users in a community-based sample of young undergraduates. This comparison will identify the specific dimensions of Decisional Balance and Processes of Change that are endorsed by severe cannabis users, which are similar to treatment-seeking populations. Therefore, the results obtained in this community sample can be useful to inform the contents of motivational interventions in clinical samples. We hypothesised that severe cannabis users compared to mild users: 1) would endorse less gains and more losses related to using cannabis; 2) would endorse more behavioral and less cognitive processes of change.The sample comprised community-dwelling young adults between 17 and 45 years of age, recruited from the student population of the University of Granada. Participants were recruited by university faculty during class breaks and were selected using a probabilistic sampling design. In particular, a cluster stratified sample design was adopted. Strata were based on the different university faculties. Cluster samples were extracted such that majors and years of study were represented in proportion to the total number of students in each faculty. Finally all students of the cluster sample were included in the final sample. There were 856 student participants recruited between September 2013 and June 2014. They were informed about the aims of the study and provided signed informed consent. Ethical approval was obtained from the Research Ethics Committee from the University of Granada.All 856 participants completed the Severity of Dependence Scale (SDS) and 115 reported that they were cannabis users. We divided these 115 cannabis users in two groups. The first group comprised cannabis users of moderate severity (n = 84) and the second group comprised severe cannabis users (n = 31) as indicated by the SDS cut-off defined by Cuenca-Royo et al. . In ordeThe protocol used is included in Supporting information  and inclThe severity of cannabis use was measured using the Spanish version of the SDS . The SDSThis instrument measures the intensity of physical addiction to nicotine. We was used the Spanish version . The tesIn the context of cannabis use the Decisional Balance questionnaire evaluates the costs and benefits  of continuing to use cannabis , 28. It This questionnaire evaluates the Process of Change as a personal mechanism that permits progression from one stage to another. It includes 30 items that evaluates 10 different processes using a 5-point Likert scale that measures the frequency of use in the past month (1 = Never to 5 = Repeatedly). This questionnaire was designed to assess the frequency in which participants make use of experiential processes , and behavioral processes  to change cannabis abuse. Internal consistency was established with alpha coefficients ranging from .69 to .92 –34.We conducted two multivariate analyses of variance (MANOVA) using Group as the independent variable (as defined by the SDS cut-offs: Severe versus Mild to moderate users) and the scores of the different dimensions of Decisional Balance (DBQ) and Processes of Change (PCQ) as the two dependent variables. These MANOVAs were followed by univariate between-group ANOVAs on the different dimensions of the DBQ and the PCQ. All analyses were replicated using sex and tobacco dependence indicated by the Fagerström Test as covariates.11,50 = 83.900; p< 0.001; Eta = 0.949). Univariate tests of between-group effects revealed significant effects of Group on the total score of Gains  and in the specific dimensions of Utilitarian Gains for the Self , Utilitarian Gains for Significant Others  and Self-approval . In all cases, the severe group had higher scores than the mild group. There were no significant differences in Losses. Mean, standard deviations and significance are presented in We found a significant effect of Group on Decisional Balance . Univariate tests of between-group effects revealed significant effects of Group on Self-Reevaluation  and Counter-Conditioning . In all cases, the severe group had higher scores than the mild group. Mean, standard deviations and significance are presented in We also found a significant effect of Group on Process of Change ; Utilitarian Gains for Others (my cannabis use bothers other people) and Self approval (I am embarrassed to smoke cannabis) are specifically endorsed. These dimensions have shown to be associated with positive behavioral changes after treatment interventions in alcohol and tobacco dependent users , 17, 18.The Processes of Change findings suggest that severe cannabis users are more likely to use particular behavioral Processes of Change , whereas they did not differ from mild cannabis users with regards to cognitive Processes of Change. These behavioral Change Processes have been shown to be associated with reduction of problematic drug use in clinical samples – 14. TheIn conclusion, our results are preliminary and need to be complemented by direct research on the treatment pathways of motivational interventions in cannabis dependence. Recent meta-analytical studies have examined these mechanisms, but have focused in the interventions format and design, but not specifically in the motivational ingredients , 36. OurS1 Table(DOCX)Click here for additional data file.S1 Data(SAV)Click here for additional data file.