Porphyromonas and Solobacterium showed significantly higher relative abundances, whereas Haemophilus, Corynebacterium, Cellulosimicrobium and Campylobacter showed lower abundances in OLP patients, as compared with healthy controls. In addition, we explored specific microbial co-occurrence patterns in OLP, and revealed significantly fewer linkers of Streptococcus comprising species in erosive OLP. Furthermore, the disease severity and immune dysregulation were also genus-associated, including with Porphyromonas that correlated to disease scores and salivary levels of interleukin (IL)-17 and IL-23. Overall, this study provides a general description of oral microbiome in OLP, and it will be useful for further investigation of their potential roles in the initiation and immune modulation of OLP.Several studies have explored the origin and development mechanism of oral lichen planus (OLP) with limited attention to the role of bacteria in the progression of this common oral disease. Here we utilized MiSeq sequencing of 16S rRNA gene amplicons to identify complex oral microbiota associated with OLP from saliva samples of two subtypes (reticular and erosive) of OLP patients and healthy controls. Our analyses indicated that the overall structure of the salivary microbiome was not significantly affected by disease status. However, we did observe evident variations in abundance for several taxonomic groups in OLP.  Lichen planus is a common chronic mucocutaneous inflammatory disease affecting skin, oral, genital mucosa, scalp and nails. The prevalence of oral lichen planus (OLP) ranges from 0.1–4% in the general population, and females between 30 and 60 are more vulnerable to this disease+ T helper cells subset-Th17 cells were detected in OLP lesions911121314et al. demonstrated Candida infection in 37–50% of OLP patientsHelicobacter pylori and OLP has been shown but remains controversial1718Staphylococcus haemolyticus and Streptococcus agalactiae in OLP lesionsDespite of numerous literatures appraising the OLP origin and development mechanism, its aetiology remains uncertain, and the pathogenesis of this disease still needs more investigations. Several internal and external factors have been implicated, such as immunodeficiencies and microbial infection6In the present study, we aimed to (1) characterize the overall structure of oral microbiota in saliva of OLP patients; (2) seek for OLP-associated microorganisms; (3) elucidate the changes in microbial interactions of OLP patients; and (4) identify key microbial populations associated with disease and immune status. Our results indicated that even though the microbial community composition has not dramatically altered, some abundant genera still have changed obviously. The interactions of microbial-microbial and microbial-immune might play a key role in OLP pathogenesis directly or indirectly.The demographic and clinical characteristics of all subjects are shown in To investigate the changes in structure and composition of salivary microbial communities of OLP, 55 samples were sequenced with Illumina MiSeq technology. After preprocessing, 2,706,496 high quality sequences with average length of 253 bp were obtained in this study, with an average of 49209 sequences per sample. From these sequences, 13 known phyla and 418 genera were identified, and a total of 8717 OTUs were detected, of which 4006 were singletons (detected in only one sample) and were excluded from further statistical analysis. Good’s estimator of coverage was 99.85%, indicating that the 16S rRNA sequences identified in this study likely represent the majority of bacterial sequences present in saliva samples. The sequencing data for details are available in Principal coordinates analysis (PCoA) plot based on Bray-Curtis distances indicated no obvious separation among healthy subjects, reticular OLP and erosive OLP . The disPorphyromonas was only found in erosive OLP patients, while Leptotrichia was detected in reticular and erosive OLP patients. Although Actinomyces was detected in both reticular OLP and healthy controls, this genus was indeed more abundant in reticular OLP. Additionally, using the BLAST method against the HOMD to assign the sequences to bacterial species, we found that Prevotella melaninogenica (OTU5115) was unique in reticular OLP and Veillonella parvula (OTU8105) dominated in reticular and erosive OLP when compared with healthy controls , Rothia aeria (OTU104) and Streptococcus parasanguinis (OTU8510) were less abundant in erosive OLP, while Prevotella melaninogenica (OTU5115) had significantly higher levels in reticular OLP  in healthy controls, Prevotella melaninogenica (OTU661 co-occurring with 31 OTUs) in reticular OLP, and Solobacterium moorei (OTU771 co-occurring with 24 OTUs) in erosive OLP, respectively. In addition, to further explore the biological significance of co-occurrence relationships of oral microbiota, we identified highly connected microbial clusters of the genus Streptococcus in the network  associated with OTU7975 and OTU7832 respectively. In reticular OLP, there was only one module (module C) of Streptococcus associated with OTU8419, but corresponding nodes did not form any modules in erosive OLP. Also, we found that the OTUs representing the genus Streptococcus had significantly fewer linkers in erosive OLP than those in reticular OLP and healthy controls . This indicated that Streptococcus might have played an important role in the development of OLP.The goal of network inference is to identify combinations of microorganisms demonstrating significant co-occurrence or mutual exclusion patterns across samples and to predict ecological relationships. By analyzing and then visualizing the spatial Pearson’s correlations between the bacterial species detected from saliva samples, decreased network complexity, as measured by average connectivity, could be observed for microbial co-occurrence networks in reticular and erosive OLP groups. For example, we captured 1094 associations among 282 nodes in healthy group, 1185 associations among 335 nodes in reticular OLP group, and 985 associations among 324 nodes in erosive OLP group at the OTU level . In the  network . InteresTreponema and Filifactor were the most positively correlated (ρ = 0.945), whereas Capnocytophage and Streptococcus were the most negatively correlated (ρ = −0.601). But in OLP, the most positively correlated genera were Solobacterium and Peptostreptococcus (ρ = 0.839), while Streptococcus and Prevotella were the most negatively correlated (ρ = −0.540). Besides, Opponent co-occurrence patterns were observed for several microbial genera in diseased and healthy status. For example, Treponema and Actinobacillus were correlated negatively (ρ = −0.249) in healthy group, but their correlation converted to be positive (ρ = 0.564) in OLP group. The same pattern was also observed for the genera Actinomyces and Granulicatella.We further employed the co-occurrence and co-exclusion analysis of the top 30 abundant bacterial genera in detail. Two subtypes of OLP patients were combined into the OLP group for this analysis. Out of 26 abundant genera which were shared between the diseased and healthy groups, we found significant changes in the ecologically competitive interactions of these bacteria . In healTreponema, Porphyromonas and Abiotrophia were positively correlated with clinical scores, whereas Oribacterium was negatively correlated with clinical scores. Second, it was observed that many genera were correlated with immunologic factors involved in the inflammation response in OLP. For example, Treponema, Porphyromonas, Gemella and Parvimonas showed significantly positive correlations with the salivary levels of IL-17, whereas Abiotrophia, Porphyromonas, Paraprevotella and Fusobacterium were found to be positively correlated with the salivary levels of IL-23. Finally, some of these genera associated with clinical data had significant correlations with more than one parameter, such as Porphyromonas, which was correlated positively with the clinical scores and salivary levels of IL-17 and IL-23 at the meantime.The concentrations of IL-17 and IL-23 were measured by means of enzyme-linked immunosorbent assay (ELISA) and the results were shown in The role of the viral infection in OLP has already come into notice. However, there is a paucity of knowledge regarding the role of bacteria in the progression of this common oral disease. In the present study, we described the complexity of microbial communities in OLP, and our results uncovered crucial yet striking salivary microbiomic characteristics associated with disease. The microbial community structure displayed a greater variety and less specificity in OLP patients. And the samples from erosive OLP displayed even higher phylogenetic diversity than healthy controls, as its rarefaction curves did not yet reach a plateau with current sequencing. These results are not in agreement with findings of a recent studyTreponema differed between two subtypes of OLP patients, and these microbial taxa may be associated with disease classification of OLP. Porphyromonas was discovered to be dominant in erosive OLP, which was also considered to be the core genus in periodontitis25Porphyromonas had something to do with OLP state. Our findings supported the results of a previous study, in which higher periodontopathogens’ percentages were detected in OLP groups than those in the non-OLP groupsAmong three groups, five major phyla constituted the predominant salivary microbiome , consistent with previous surveys23Veillonella parvula and Prevotella melaninogenica. Also, higher counts of Veillonella parvula was examined at sites with evidence of OLP lesions using DNA-DNA hybridization in a previous studyPrevotella species, the Gram-negative rod-shaped anaerobes, were also implicated involved in many other oral disease, such as endodontic diseasePrevotella melaninogenica, as the hub species with significantly higher abundance, could be regarded as the core bacteria in reticular OLP. Findings reported here underlined the potential role of this reticular OLP-associated species played in the disease. While in this study, Haemophilus parainfluenzae, Rothia aeria and Streptococcus parasanguinis were less abundant in erosive OLP compared with healthy controls. Indeed, in a recent study, Said et al.Veillonella and Prevotella and lower proportions of Haemophilus and Streptococcus than those in healthy controls, and they indicated that the dysbiotic genera Veillonella and Prevotella could elicit an inflammatory response in mucosa. Although the abundance of the genera Veillonella and Prevotella were not significantly different between OLP patients and healthy individuals, they might be important for the pathogenesis of OLP at the species level. However, further investigation is required to clarify the role of these dysbiotic oral microbiota in the progression of this disease.Moreover, analysis at the OTU level confirmed the disease-associated species, which increased in relative abundance and prevalence, including Porphyromonas with higher abundance turned to be correlated significantly with Eubacterium, which was also found to be involved in periodontitisStreptococcus comprising species, also supporting the imbalance of oral microbiota in OLP. Equally, the decrease of Streptococcaceae comprising species associated with oral health was observed in ulcerated sites of recurrent aphthous stomatitis (RAS) patientsOral microorganisms rarely survive in isolation, but instead coexist in complex ecology with various symbiotic relationships24Porphyromonas positively correlated to three clinical parameters deserves more attractions. As one of the most frequently encountered species in human, Porphyromonas gingivalis has recently been demonstrated on its ability to subvert host immunity and promote dysbiosis of oral microbiotaPorphyromonas gingivalis was also indicated to enhance the Th17 cell responses for the development of atherosclerosisPorphyromonas in erosive OLP, our findings highlight the urgency of the further research on the role of this bacteria played in OLP. Additionally, we postulated that some microbiota colonized in saliva of OLP patients might be associated with the severity of disease. The genera Treponema, Porphyromonas and Abiotrophia showed significantly positive correlations with the clinical scores, suggesting these bacteria might be involved in the exacerbation of OLP and their proportion may signify the degree of severity of this disease. Treponema, Porphyromonas, Parvimonas and Fusobacterium have recently been reported to be the core microbiota in periodontitisTreponema and Porphyromonas in erosive OLP compared to those in reticular OLP and healthy individuals also supported their association with disease severity. While the genus Abiotrophia, which is often associated with culture-negative infective endocarditisThere is an increasing body of evidence suggesting that perturbations to the structure of complex commensal communities can modulate innate and adaptive immune response and lead to the development of immune mediated diseases, with inflammation arising upon the reduction of the proportion of symbiont microbiota or the increase in the number of pathogenic microbiota3639In conclusion, we preliminarily identified the microbial taxa associated with OLP and delineated different features of salivary microbiome between OLP and healthy individuals, providing a comprehensive cross-sectional description of oral microbiota in OLP. Our study afforded deep insight into the dysbiosis in the microbial community of OLP, suggesting a potential microbial role in progression of this disease. Also, Our findings could provide the basis for further exploring the bacteria suspected of being associated with OLP in the future studies.Subjects with reticular OLP (n = 19) and erosive OLP (n = 18) aged between 22–62 years were selected during follow-up clinical examinations from the West China Hospital of Stomatology, Sichuan University, and the OLP patients were diagnosed in accordance with the clinical classification and definition of OLP by the World Health OrganizationAccording to the standard techniques described by NavazeshThe levels of IL-17 and IL-23 in saliva were detected as described previously−1, 1.5 h) treatment step. Quality of the extracts was evaluated by the absorbance ratios at A260/A280 and A260/A230 using spectrophotometry . Only DNA samples with A260/A280 >1.7 and A260/A230 >1.8 were used in downstream experiments. The final concentrations of DNA were quantified using the Pico-Green kit . DNA samples were frozen at −20 °C until further analysis.Genomic DNA was extracted from saliva samples using the QIAamp DNA micro Kit  according to the manufacturer’s instructions with minor modifications by adding a lysozyme  and R806 (primer seq) primers targeting V4 hypervariable regions and then sequenced with the Illumina MiSeq technology at the Institute for Environmental Genomics, University of Oklahoma . The MiSeq 500 cycles 250 × 2 V2 kit was used for sequencing 16S rRNA V4 on MiSeq machine . Data preprocessing and OTU clustering was performed as described previouslyThe sequence data were further analyzed with the following methods: (A) Principal coordinate analysis (PCoA) was used to determine the degree of dissimilarity between pairs of bacterial communities using Bray-Curtis distance method. Three different nonparametric analyses for multivariate data implemented by the “vegan” package in R were also used for community functional structure comparison, including multiresponse permutation procedure (MRPP), nonparametric multivariate analysis of variance (adonis) using distance matrices and analysis of similarities (ANOSIM). (B) Alpha-diversity indices were estimated by the Shannon diversity index, the number of observed OTUs, and Chao1 species richness estimates. Rarefaction curves were applied to calculate the species richness based on the bacterial OTUs and were then compared among three groups by the mothur packaget-test was used to compare microbial diversity and relative abundances of each bacterial taxon. Pearson correlations were used for the analyses of co-occurrence and bacterial associations with the clinical data.Student’s How to cite this article: Wang, K. et al. Preliminary analysis of salivary microbiome and their potential roles in oral lichen planus. Sci. Rep.6, 22943; doi: 10.1038/srep22943 (2016).


Currently available in vitro data on the role of MSX2 as a transcription factor and its links to other players in ameloblast gene regulation are considered. MSX2 modulations are relevant to the interplay between developmental, hormonal and environmental pathways and in vivo investigations, notably in the rodent incisor, have provided insight into dental physiology. Indeed, in vivo models are particularly promising for investigating enamel formation and MSX2 function in ameloblast cell fate. MSX2 may be central to the temporal-spatial restriction of enamel protein production by the dental epithelium and thus regulation of enamel quality  under physiological and pathological conditions. Studies on MSX2 show that amelogenesis is not an isolated process but is part of the more general physiology of coordinated dental-bone complex growth.While many effectors have been identified in enamel matrix and cells via genetic studies, physiological networks underlying their expression levels and thus the natural spectrum of enamel thickness and degree of mineralization are now just emerging. Several transcription factors are candidates for enamel gene expression regulation and thus the control of enamel quality. Some of these factors, such as MSX2, are mainly confined to the dental epithelium. MSX2 homeoprotein controls several stages of the ameloblast life cycle. This chapter introduces MSX2 and its target genes in the ameloblast and provides an overview of knowledge regarding its effects  Msx2 is a member of the family of divergent homeobox-containing genes homologous to the Drosophila Muscle Segment Homeobox (msh) gene. Evolution including the duplication of the ancestral msh gene, has led to three different genes in mice and two in humans. Homeobox-containing genes share a well-conserved sequence of 183 bp coding for a helix-loop-helix motif of 64 amino acids  responsive element , also determinant of cell proliferation  cells and down-regulated when ameloblast cells overtly differentiate  as well as in dental and periodontal mesenchyme  from data obtained in vitro. For example, in the periodontal ligament cells, MSX2 prevents osteo-differentiation in vitro , the main osteoclastic-differentiating factor  inhibitor gene p21, and Shh are not modified  thanks to establishment of cell models that help to decipher molecular mechanisms  mice are a unique model for investigating MSX2 dose effects. Ameloblasts survive and differentiate appropriately in Msx2+/− mice (unlike those in Msx2−/− mice) and the amounts of Msx2 mRNA are half those in wild-type mice. Enamel gene studies have revealed a specific overabundance of amelogenin mRNA in Msx2+/− mice. The enamel phenotype in Msx2+/− mice included increased thickness and, more specifically, a thicker outer prismatic enamel layer and larger prism diameter  and result in hypoplastic enamel without visible prismatic structures  and the National Research Agency (ANR): OSTEODIVERSITY (SVSE 1-2012).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.


This quasi-experimental study integrated family planning, including the Lactational Amenorrhea Method, into community-based maternal and newborn health care and encouraged transition to other modern methods after 6 months to increase birth-to-pregnancy intervals. Community-based distribution of pills, condoms, and injectables, and referral for clinical methods, was added to meet women's demand. This quasi-experimental study integrated family planning, including the Lactational Amenorrhea Method, into community-based maternal and newborn health care and encouraged transition to other modern methods after 6 months to increase birth-to-pregnancy intervals. Community-based distribution of pills, condoms, and injectables, and referral for clinical methods, was added to meet women's demand. Short birth intervals are associated with increased risk of adverse maternal and neonatal health (MNH) outcomes. Improving postpartum contraceptive use is an important programmatic strategy to improve the health and well-being of women, newborns, and children. This article documents the intervention package and evaluation design of a study conducted in a rural district of Bangladesh to evaluate the effects of an integrated, community-based MNH and postpartum family planning program on contraceptive use and birth-interval lengths.The study integrated family planning counseling within 5 community health worker (CHW)-household visits to pregnant and postpartum women, while a community mobilizer (CM) led community meetings on the importance of postpartum family planning and pregnancy spacing for maternal and child health. The CM and the CHWs emphasized 3 messages: (1) Use of the Lactational Amenorrhea Method (LAM) during the first 6 months postpartum and transition to another modern contraceptive method; (2) Exclusive, rather than fully or nearly fully, breastfeeding to support LAM effectiveness and good infant breastfeeding practices; (3) Use of a modern contraceptive method after a live birth for at least 24 months before attempting another pregnancy  to support improved infant health and nutrition. CHWs provided only family planning counseling in the original study design, but we later added community-based distribution of methods, and referrals for clinical methods, to meet women's demand.Using a quasi-experimental design, and relying primarily on pre/post-household surveys, we selected pregnant women from 4 unions to receive the intervention  and pregnant women from 4 other unions  to serve as the comparison group. Enrollment occurred between 2007 and 2009, and data collection ended in January 2013.Formative research showed that women and their family members generally did not perceive birth spacing as a priority, and most recently delivered women were not using contraception. At baseline, women in the intervention and comparison groups were similar in terms of age, husband's education, religion, and parity. CHWs visited over 90% of women in both intervention and comparison groups during pregnancy and the first 3 months postpartum.This article provides helpful intervention-design details for program managers intending to add postpartum family planning services to community-based MNH programs. Outcomes of the intervention will be reported in a future paper. Preliminary findings indicate that the package of 5 CHW visits was feasible and did not compromise worker performance. Adding doorstep delivery of contraceptives to the intervention package may enhance impact. The Population Council first launched a global postpartum family planning program in 1966,Many family planning programs historically have focused on improving access to family planning and on changing people's attitudes about ideal family size and reducing the TFR, rather than on promoting longer birth intervals. In low-income countries, approximately 57% of second and higher-order births occur at intervals shorter than 3 years, and, in some countries, these conditions have not changed in 20 years.Consistent observational data from many settings suggest that birth-to-pregnancy intervals shorter than 24 months  are associated with increased risk of poor maternal, perinatal, and neonatal health outcomes, including increased risk of stillbirth, prematurity, low birth weight, and neonatal mortality.Short birth intervals are associated with poor maternal and newborn health outcomes.Policymakers have called for better integration of family planning services with maternal and child health programs,A 2012 Cochrane review did not find any integrated family planning and maternal, newborn, and child health program in a developing country that had operationalized the growing evidence on the role of family planning in improving child and maternal health .In this article, we describe the intervention package and evaluation design of an operations research study that is nearing completion, called the Healthy Fertility Study (HFS). The study integrated postpartum family planning education and services within an existing community-based maternal and neonatal health (MNH) program, called “Projahnmo” , to improve contraceptive knowledge and practices and birth spacing, focusing specifically on using the Lactational Amenorrhea Method (LAM) during the first 6 months postpartum and transitioning to another modern method afterward. Projahnmo uses female community health workers (CHWs) to deliver MNH services through home visits, complemented by community mobilization activities. The Projahnmo program reduced neonatal mortality by 34% in its first 30 months of implementation in Sylhet district, located in the Sylhet division of rural, northeastern Bangladesh.Many postpartum women have an unmet need for family planning.We chose to conduct the study in Sylhet division because it had the highest neonatal, infant, and under-5 child mortality rates among the 7 divisions of Bangladesh.TFR of 3.7 compared with 2.7 for Bangladesh overallUnmet need for contraception of 26% compared with 17% for Bangladesh overall57% of second and higher-order births occurred less than 36 months after the preceding birth compared with 37% for Bangladesh overallNonetheless, the presence of government-run health and family planning facilities and exposure to family planning messages in the media in Sylhet division are comparable with other parts of Bangladesh. However, women's access to clinical family planning services in Sylhet district is severely constrained. One-quarter of the public-sector positions slated for provision of clinical family planning services are unfilled . When clinical family planning services are available, they are offered only at specific times during the week. Nongovernmental and private-sector providers also largely lack clinical skills in family planning. Equally important, in Sylhet, cultural norms restrict women from leaving their homes without a chaperone.We conducted formative research between January 2006 and June 2006 to inform the intervention design for postpartum family planning service delivery. Formative research activities included:80 unstructured, in-depth interviews with household members  to elicit information on birth spacing, timing of first birth, contraceptive practices, and other issues related to healthy fertility40 semi-structured interviews with mothers who practice and who do not practice birth spacing and their mothers-in-law, using free-listing exercises in which participants listed the benefits of birth spacing and risks associated with short birth intervals6 focus groups with mapping exercises among recently delivered women (3) and older women (3) to explore community perceptions about birth spacing, as well as to compile a list of health care providers who offered family planning servicesA household survey with 1,612 systematically sampled, recently delivered women to gather data on household sociodemographic factors, contraceptive knowledge and practices, and access to and use of health services31 semi-structured interviews with health care providers and 4 focus group discussions with community opinion leaders, including religious leaders, members of Union Councils , and village government leaders, to obtain their opinions on healthy timing and spacing of pregnanciesInterviewers transcribed and coded the interview and focus group discussions, and the research team reviewed the transcripts and codes to identify themes and relevant findings. When religious leaders understood that family planning was being integrated to support women's and children's health, they stated that they had no objection to this activity.All 40 of the mothers and mothers-in-law that participated in the free-listing exercises were able to list at least 4 risks to either the mother or child associated with closely spaced births. Similarly, 38 of the 40 participants were able to list at least 3 benefits associated with a healthy birth-spacing interval, including physical, emotional , and economic benefits.Nearly 78% of recently delivered women said that they desired a birth-spacing interval of 3 years or longer. During in-depth interviews, some men stated an economic interest in spacing births, and some were also aware of the physical toll that closely spaced births would have on the health of the mother.Respondents generally did not perceive birth spacing as a high priority relative to other family needs. Mothers-in-law considered birth spacing of secondary importance to the provision of grandchildren, which they saw as a necessary and critical obligation of their daughters-in-law. Mothers themselves also said that birth spacing was not a very serious issue for them. Having many children within short intervals is common and perceived as normal in their communities; as a result, the women had not questioned the practice previously. Many of the husbands interviewed considered birth spacing as more important to their wives than to them.Community members could identify benefits of longer birth intervals, but birth spacing generally was not a high priority for them.Despite recognition of the risks of closely spaced births and the benefits of longer intervals, nearly 75% of the recently delivered women (who were not sterilized) were not using any contraceptive method to prevent pregnancy.Among health care providers, family planning service delivery reportedly focused more on limiting family size than on advocating birth spacing. Knowledge of LAM, including its objectives and criteria, was limited. . Each CHW serves a population of about 4,000, or 4 villages.The CHWs are young women from the local community with grade 10 education. All CHWs in both intervention and comparison areas received 21 days of basic MNH training, including skills development for behavior change communication, clinical assessment of neonates, and hands-on clinical training under supervision in a tertiary care hospital and in households. CHWs in the intervention area received an additional 3 days of training about healthy timing and spacing of pregnancy and postpartum family planning, including LAM, and 4.5 days of training about contraceptive methods and logistics management, according to protocols from the government of Bangladesh.In the existing Projahnmo MNH program, CHWs visited pregnant and postpartum women 6 times during the antenatal and postpartum period. They also made 1 visit every 2 months within their catchment area to identify pregnant women.In the intervention area, the HFS integrated postpartum family planning counseling within 3 of the 6 existing MNH visits: at 30–32 weeks of pregnancy, 6 days postpartum, and once between days 29–35 days postpartum. The study also added 2 more visits, at 2–3 months and 4–5 months postpartum , to educWe designed 3 family planning interventions specifically to support improvements in newborn and child health. The purpose was to ensure that adding family planning activities to the existing MNH services did not undermine program achievements to date in newborn health outcomes.Use of LAM during the first 6 months postpartum to prevent high-risk pregnancies. LAM can be considered a maternal and child survival intervention because its use prevents high-risk pregnancy during the critical 6-month period after a birth. A recent meta-analysis showed that infants conceived during the first 6 months after a live birth are at elevated risk of multiple adverse outcomes.Use of LAM to increase the duration of exclusive breastfeeding and improve breastfeeding practices. LAM use is associated with improved exclusive breastfeeding practices. At the time of the HFS design, the median duration of exclusive breastfeeding in Sylhet division was less than 1 month (0.7).fully or nearly fully breastfeeding, the HFS team decided to communicate the breastfeeding criterion of LAM as exclusive breastfeeding, to prevent the possibility of undermining exclusive breastfeeding practices promoted by the ongoing MNH program. The team hoped to replicate results achieved in Jordan, where one study found that use of LAM promoted good exclusive breastfeeding practices.Use of other modern methods to achieve a 24-month birth-to-pregnancy interval, which is associated with reduced risk of under-5 mortality and improved maternal and child nutrition status. Nearly half of all contraceptive users in Bangladesh stop using their method within 12 months of starting.LAM is often referred to as a “gateway” method of family planning, because many LAM users eventually accept another modern method.supplementary material):After conducting a desk review of existing information, education, and communication (IEC) materials in Bangladesh from NGOs and the government, we adapted materials on postpartum care from the Bangladesh Rural Health Service Delivery Program and developed, field tested, and refined into the local language 3 new leaflets on exclusive breastfeeding and birth spacing, LAM, and return to fertility after delivery  CHWs distributed pills and condoms during their routine pregnancy surveillance visits every 2 months.Community-based distribution of contraceptives was added to the study design to meet women's demand.active referrals for clinical contraceptive methods—that is, CHWs accompanied women desiring clinical methods to the facility.Beginning in March 2011, CHWs also started providing progestin-only injectable contraceptives to women enrolled in the HFS. Women must receive the injection every 3 months, but they have a window period and can receive the injection up to 2 weeks early or up to 4 weeks late  We intended to enroll women over a 7-month period , but we received additional funding that allowed us to increase the study area to 8 unions, thus expanding the enrollment period through July 14, 2009 .Using a quasi-experimental design, and relying primarily on pre/post-household surveys among study participants, we purposefully selected 4 unions to receive the intervention and 4 other unions to serve as the comparison group, receiving standard maternal and newborn care promotion only.The study unions were purposively selected to:Minimize contamination between intervention and comparison groups and with other ongoing nongovernmental organization (NGO) programsUpazila (sub-district) Health Complex , as this would represent a large difference in availability of health servicesExclude unions having an Include unions with functional government facilities and workers designated for providing family planning servicesThe study was approved by the Johns Hopkins University (JHU) Institutional Review Board and the Bangladesh national ethics committee of Bangladesh Medical Research Council and is registered as a Clinical Trial (Identifier: NCT01702402).Following recommendations from the 2005 WHO Technical Consultation on Birth Spacing,Although birth-to-birth intervals can be measured more accurately than birth-to-pregnancy intervals, from a programmatic perspective, it is easier to communicate messages about birth-to-pregnancy intervals, because women and their partners think in terms of the time from the birth of one child to the conception of the next child. Thus, the household survey questionnaires refer to birth spacing in terms of birth-to-pregnancy intervals. However, birth-to-birth interval is the study's primary outcome measure.To calculate sample size, we hypothesized that the proportion of women having another birth within 33 months of the last birth will be 12% in the intervention area and 16% in the comparison area—a 25% difference. To measure a 25% decrease in the proportion of women with a birth interval shorter than 33 months with 80% power and 5% significance level required a sample size of 1,181 pregnant or postpartum women per study arm. Taking into account a design effect of 1.5, we estimated we would need to increase the sample size to 1,772 women per study arm. We assumed a 20% loss to follow-up (10% per year), which further increased the sample size to 2,215 per arm . This sample size is sufficient to measure hypothesized changes in contraceptive knowledge and use and birth-to-birth intervals of 33 months. The total number of women enrolled was 4,570, slightly higher than anticipated as a result of ongoing field operations in several locations simultaneously . The final study cohort was comprised of 4,504 women  for whom data on pregnancy outcomes were available.A team of data collectors not involved in implementing the intervention conducted 8 data collection visits for each study participant:1 visit during the antenatal period completed before weeks 30–32 of pregnancy (considered the baseline)7 follow-up visits during the postpartum period In addition to the independent data collectors, CHWs also collected data used to evaluate the intervention, including date of birth of newborns, birth weight, and neonatal outcomes. A separate team of data collectors conducted verbal autopsies of all reported infant deaths occurring in both study arms.P = .10).Women in the intervention and comparison groups were similar in terms of age, husband's education, religion, and parity . Women'sP<.05). Still, the mean number of CHW visits was higher in the intervention than in the control group . This was not unexpected since CHWs provided contraceptive methods to women in the intervention group, which probably required additional CHW visits for resupply of methods.There was no difference in coverage of women by CHWs in the intervention versus control groups . During This article describes the design of an operations research study that integrates postpartum family planning education and services within an existing community-based maternal and newborn care program in a low-resource setting.The HFS adds to the literature on postpartum family planning. In developed countries, promotion of postpartum family planning is a standard component of obstetric care.Results of the study described in this article are currently being analyzed and will be published in a future paper.The HFS intends to fill a gap in the literature on postpartum family planning by providing guidance about a scaled-up package of integrated community-based care, including about the appropriate number and timing of CHW household visits. Our postpartum family planning intervention package included 1 antenatal visit and 4 postpartum visits during the first 5 months postpartum, plus 1 visit every 2 months for pregnancy surveillance and contraceptive distribution.One antenatal and 3 postpartum visits are feasible and sustainable in many community-based MNH programs, including in Bangladesh. For example, the Maternal and Child Health Integrated Program (MCHIP) in Bangladesh funded by the U.S. Agency for International Development (known as the MaMoni project) is implementing an MNH program in 2 districts that involves 3 postpartum visits with integrated family planning counseling and services. Although the 5 CHW visits in the HFS intervention package may be considered intensive, preliminary results indicate that the large majority of women received the full number of scheduled visits and that the schedule did not compromise worker performance. Based on programmatic experience, the HFS team concluded that fewer visits would lead to gaps in continuation of breastfeeding and the transition from LAM to other modern contraceptive methods.Although making 5 CHW visits may be considered intensive, preliminary results indicate it was feasible.Doorstep delivery of contraceptives was not originally part of the HFS intervention package. The government of Bangladesh had moved away from community-based distribution of contraceptives toward clinic-based provision of family planning services.The HFS builds on important lessons from the Bangladesh Matlab Family Planning–Health Services ProjectThe HFS relies on service delivery through locally recruited CHWs with a grade 10 education. In contrast, the Navrongo project achieved results by using more highly educated health care professionals—nurses—a cadre not widely available in many developing countries.The HFS targets family planning services to pregnant and postpartum women—an underserved group of women with demonstrated high unmet need for family planning—not to all women in the community, as with the MatlabThe Matlab project demonstrated progress in improving contraceptive continuation rates,Matlab made progress in helping women to space their pregnancies, yet currently in Sylhet, 47% of second-order and higher births are spaced less than 36 months apart, and for the 15–19 age group nationally, this percentage rises to 80%.Navrongo demonstrated reductions in child mortality,The HFS design adds postpartum family planning education and services into an ongoing and successful maternal and newborn care program, whereas with the Matlab project, maternal and child health services were gradually added to the responsibilities of the family planning field worker.The findings from the HFS trial are anticipated to help practitioners provide women and infants in low-resource settings with an integrated, community-based continuum of care.


To investigate the effects of different intensities of acute exercise on insulin sensitivity and protein kinase B/Akt activity in skeletal muscle of obese mice.Swiss mice were randomly divided into four groups, and fed either a standard diet (control group) or high fat diet (obese sedentary group and obese exercise group 1 and 2) for 12 weeks. Two different exercise protocols were used: swimming for 1 hour with or without an overload of 5% body weight. The insulin tolerance test was performed to estimate whole-body sensitivity. Western blot technique was used to determine protein levels of protein kinase B/Akt and phosphorylation by protein Kinase B/Akt in mice skeletal muscle.A single bout of exercise inhibited the high fat diet-induced insulin resistance. There was increase in phosphorylation by protein kinase B/Akt serine, improve in insulin signaling and reduce of fasting glucose in mice that swam for 1 hour without overload and mice that swan for 1 hour with overload of 5%. However, no significant differences were seen between exercised groups.Regardless of intensity, aerobic exercise was able to improve insulin sensitivity and phosphorylation by protein kinase B/Ak, and proved to be a good form of treatment and prevention of type 2 diabetes. The major effect of exercise is the increase in intracellular proteins expression of insulin signaling pathway especially the glucose transport in skeletal muscle.-3Physical exercise increases insulin sensitivity independently of body mass reduction and changes in body composition.Obese individuals and resistant to insulin have as common characteristics the problem in insulin signaling due to subclinical inflammatory process. This process shows a decrease in action of this hormone in target cells and is influenced by the presence of fasting hyperglycemia and hyperinsulinemia. Such metabolic changes are associated with a decrease in glucose uptake by skeletal muscle in response to insulin.i.e., the serine/threonine protein kinase B/Akt, is emphasized. Protein Akt, widely studied in the last years, is an important physiological regulator that controls several cellular functions such as growth, survival, proliferation and metabolism.Among mechanisms involved in insulin signaling, the distal protein by molecular pathway of this hormone, . A previous study showed that Akt1 knockout mice have small body size and increased apoptosis rates than their wild type mice. That situation revealed the importance of this isoform for cellular survival. Akt2 knockout mice developed characteristics of type 2 diabetes mellitus that were crucial for maintenance of glycemia. Animals with protein Akt3 absence had an impairment in brain development.-8Protein Akt has three isoforms , and each isoform exerts a specific function inside the cell The IR activation results in phosphorylation on tyrosine of several substrates, including insulin receptor substrates 1 and 2 (IRS-1 and IRS-2). After phosphorylation on tyrosine the IRS-1 and IRS-2 can bind and activate phosphatidylinositol 3-kinase (PI3-kinase). This protein triggers phosphorylation of membrane phosphoinositides and the recruiting of Akt close to the plasmatic membrane. The phosphoinositides promote and activate phosphorylation of phosphoinositidedependent kinase (PDK). Finally, PDK activates the Akt when phosphorylating it in its catalytic domain (Thr308). The activated Akt protein has the ability to phosphorylate and activate several metabolic targets. The Akt, for example, stimulates the glucose uptake, synthesis of glycogenic and proteins in skeletal muscle.,10-12Intracellular insulin signaling begin with its linking with a specific receptor of the membrane, a heterotetrameric protein with kinase activity, composed by two alpha subunits and two beta subunits named insulin receptor (IR).,12The glucose uptake stimulated by Akt in skeletal muscle involves phosphorylation and activation of Akt substrate, which is a protein of molecular weight of 160 kDa, named AS160. When phosphorylated in threonine 642 this protein is dissociated from vesicles that contain glucose transporters type 4 (GLUT-4), enables the translocation and exocytosis of these vesicles into the membrane, and increases GLUT-4 expression which consequently increases glucose uptake.diabetes mellitus. Studies on cells in vitro, experimental animal model of diabetes and with human patients showed that a single bout of exercise increased the insulin sensibility. This effect was associated with higher phosphorylation (activation) of Akt and its substrates in skeletal muscle. As a consequence, we observed an increase of expression and translocation of GLUT-4 transporter and glucose uptake in skeletal muscle during and after physical exercise.-17Currently physical exercise is considered one of the most important strategies for prevention and treatment of insulin resistance and type 2 It is clear in the literature that physical exercise promotes significant improvement on insulin signaling in skeletal muscle and on insulin sensibility, however, further investigation should be done to identify physiological and molecular effects of physical exercises of different intensities in organisms with insulin resistance and diabetes.This study investigated the effects of different intensities of acute exercise on insulin sensibility and Akt phosphorylation in skeletal muscle of obese mice.Universidade Estadual de Campinas (UNICAMP), protocol nº 2805-1. The experiments were conducted in the first semester 2013. The sample included 24, 6-weeks-old, Swiss mice bred at the Multidisciplinary Center for Biological Research at UNICAMP. Animals were housed in single cages and had free access to water and two types of diet: standard rodent chow (C) or hyper fat diet (HFD) during experimental period. Details of diets are given in All experiments followed care principles and procedures for the use of experimental animals and were approved by ethical committee of the The acute exercise protocol consisted in animals swimming, where animals swan in group of six at a cylindrical tank of 45 cm in length and 60cm depth, with water temperature of 32± 1°C. Animals in group OE-1, exercised in single bout for 1 hour without additional overload, and animals in group OE-2 exercised for the same time with overload of 5% of body weight attached to animals' tail. Before the beginning of bout of exercises, to acclimatize to water, animals were placed in the swimming tank with water up to chest level for three consecutive days. The other groups of animals were submitted to the same procedure to simulate the stress in the water received by group OE-1 and OE-2. All experiments were conducted 24 hours after the exercise bout and with previous fasting for 8 hours.At the end of experimental period, animals total body mass , glucose and insulin blood levels were assessed. In addition, animals of each experimental group were submitted to insulin tolerance test (ITT).Plasmatic glucose dosage was conducted using the enzymatic colorimetric of glucose oxidase. Plasmatic insulin samples were evaluated by the ELISA method.1/2. The glucose t1/2 was measure from analysis curve of minimum squares of glucose concentration during the linear decay phase.ITT was conducted 24 hours after the bout of swimming exercise. Animals had been fasting for 8 hours before blood sample was collection to verify the glycemia at time 0 of the test. Subsequently, insulin (2U/Kg of body mass) was injected intraperitoneally and then blood samples were collected from the animals' tail at 5, 10, 15, 20, 25 and 30 minutes to determine the blood glucose value using a portable blood glucose meter . Glucose decay constant rate (Kitt) was measured using the formula 0.693/t-6 mol/l) was injected. We collected samples of gastrocnemius muscles (mixed portion) after 90 seconds of insulin that were homogenized in immunoprecipitation buffer containing 1% Triton X 100M Tris (pH 7.4), 100mM of sodium pyrophosphates, 100mM of sodium fluoride, 10mM of EDTA, 10mM of sodium vanadate, 2mM of PMSF and 0.1mg/mL of aprotinin at 4°C. The homogenized was centrifuged at 11,000 rpm for 30 minutes. The protein concentration was determined in supernatant solution by Bradford method. Subsequently, a total extract determination was conducted and an immunoprecipitation assay with specific antibodies. At the end of the experimental procedures, the epididymal adipose tissue was removed for assessment of its total mass using an analytical balance.Mice were anesthetized using intraperitoneal injection of ketamin/diazepam (50/5mg/kg). The level of anesthesia was monitored by the corneal and pedal reflexes. Animals' abdominal cavity was opened and after the portal vein was identified, 0.2mL of saline solution or insulin . Separated proteins in SDS-PAGE were transferred to nitrocellulose in BIO-RAD media transfer device. Nitrocellulose membrane was incubated overnight using a specific antibody. The linking of antibody to non-specific proteins was minimized by pre-incubation of nitrocellulose membrane with blocking buffer  for 1.5 hour. The identifying method used was the chemiluminescence in which proteins of interest are detected by incubation of membranes with primary antibodies and subsequently with conjugated-secondary antibodies with horseradish peroxidase (HRP). Immunoreactive bands were detected by the chemiluminescence and densitometry determined by uptake system and imaging analysis.Antibodies used for immunoblot experiment were anti-Akt and anti-phosphoserine Akt (Ser473) .t test was used for non-paired data to compare the two groups. When necessary the variance analysis was used with the Bonferroni post hoc test for multiple comparisons of means. The significance level of p<0.05 was adopted.Results were expressed as mean±standard error of the mean. The Student's Fasting glycemia values found in animals of groups OB, OE-1 and OE-2 were significantly higher than the control group. Mice that exercised from OE-1 and OE-2 had satisfactorily higher hexose than the group OB. But no statistical difference was found among groups submitted to difference intensities of acute exercise.Insulinemia values were significantly higher in mice of group OB, OE-1 and OE-3 than in the control group. This result states that acute exercise did not change insulin values in animals.We verified that phosphorylation of Akt in skeletal muscle increased in all experimental groups  when stimulated by insulin compared to control group that received saline (C1) . In OB mWe verified that sedentary mice fed with high fat diet had lower rate of glucose uptake than mice in the control group . However, mice that performed acute exercise had an increase in glucose uptake rate in TTI  compared with sedentary obese animals, but these values did not differ from the control group. Such results showed that a single bout of exercise can increase glucose uptake in obese mice .,3,20 This study did not find difference in total body mass and epididymal fat content between experimental groups. This result was expected once a single bout of exercise is not enough to produce changes in body composition of mice. However, the protocol for acute swimming exercise was able to increase insulin sensibility and phosphorylation of Akt in obese animals. This protein constitutes a key for insulin signaling pathway.Studies have shown that insulin sensibility increases with practice of acute or chronic physical exercise, even when there is no reduction of body fat mass. verified that sensibility and insulin signaling (by IRS/PI3-kinase/Akt) in skeletal muscle and epydidymal fat tissue of lean mice increased after six weeks of swimming training with attached overload corresponding to 5% of animal body mass. This fact showed that exercise is efficient to increase the insulin response in peripheral tissues sensitive to insulin, such as skeletal muscle and fat of healthy animals.Luciano et al. Recent studies had observed that some types of fatty acids from diet, especially the saturated fatty acids, are capable to linking and activating proteins of membrane named TollLikeReceptor 4 (TLR4). When activated, these receptors trigger an inflammatory response and this activation promotes negative consequences for insulin signaling pathway.,22 This occurs because TLR4 has the ability to active proteins serine-kinase IKKβ (Ikappakinase beta) and JNK , known as pro-inflammatory action. In this sense, these proteins promote phosphorylation in serine 307 of insulin receptor and insulin receptor subtracts 1 and 2 (IRS1 and 2) decreasing the insulin signal in different tissues.,22However, in obesity there is an increase in production and release of pro-inflammatory cytokines, from the fat tissue infiltrated with macrophages, along with increase of free fatty acids from lipolysis and intake of saturated fat-rich diet that cause damages to insulin signaling and hyperglycemia.B/NFB pathway. When this protein is activated, the IKKβ promotes phosphorylation of IB that provokes dissociation of IB/NFB complex. Because it is a transcription factor, when it free inside the cell, the NFB translocate to the nucleus and promotes transcription of new inflammatory cytokines, such as tumor necrosis factor alpha (TNF-α), protein phosphatases as the protein phosphatases 1B (PTP1B), and inducible nitric oxide synthase (iNOS) with recognized negative effect on molecular insulin signal. For this reason, an increase of inflammatory subclinical feature is associated with obesity and consequently with the insulin resistance state in obesity.,22In addition, the TLR4 receptor triggers activation of IKK/I Oliveira et al. observed that suppression of inflammatory pathway lasted for 24 and 36 hours after the last bout of exercise. These authors used a protocol of swimming exercise for rodents, the similar model used in our study. However, Oliveira et al sample was composed by Wistar rats that, although develop obesity and insulin resistance, do not always present hyperglycemia as Swiss mice.In this inflammatory feature associated with obesity and insulin resistance, researchers of several parts of the world have been investigating the role of physical exercise to restore insulin sensibility, reduce activating of TLR4, serine-kinase proteins JNK and IKKβ in obese animals. who observed increase of sensibility to insulin in obese rats submitted to acute exercise protocol. Although, these author's study used a protocol with high volume of exercises with bouts for 3 hours each and intervals of 45 minutes. Besides that, no additional overload was used during swimming exercise so that these exercises could be considered of low intensity.These findings are consistent with Ropelle et al. compared two physical exercise protocols of different volume and intensity and observed that the protocol of high volume of exercise  and the protocol of low volume of exercise  were efficient to restore insulin sensibility, suppress protein expression and phosphorylation of serine kinase, JNK and IKKβ, and increase protein phosphorylation of insulin signaling pathway . Therefore, these authors showed that shortterm exercise can promote similar results of those found with long-term exercises. Although these results are of great importance to the literature because low volume of exercise is more easily tolerable by obese, especially for the difficult caused by obesity in performing of exercises that require body mass movement. Unfortunately, these authors did not check differences in responses of different intensity exercises at low volume protocols.Da Silva et al.In our study, the obese Swiss mice induced by high fat diet were insulin resistant and had low phosphorylation of Akt protein than lean animals from the control group. A different result was seen in animals submitted to acute exercise protocol for 1 hour, without or with additional weight of 5% of body mass, that were more sensible to insulin and had an increase in phosphorylation of Akt than obese mice that not exercised. However, independently of physical exercise protocol these variables did not present difference. This result indicates that obese animals that exercised without additional weight attached to their body had similar benefit as those that exercised with more intensity . These results have a great relevance for practical application of physical exercise protocols in obese population considering that low intensity and volume aerobic exercises are more easily tolerable by this public, mainly in the beginning of practice of exercise.2peak, done in stationary bicycle, was enough to increase sensibility to insulin in those individuals. However, after seven bouts of exercise with the same volume and intensity of acute bout of exercise, researchers observed increases in glucose uptake by the hyperinsulinemic-euglycemic clamp test and expression of type 4 glucose transporter (GLUT4) in obese patients with type 2 diabetes. However, when they analyzed proteins of insulin signaling they did not observe differences between basal status and after physical exercise. This phenomenon could be explained because exercise can stimulate independent pathways of insulin to increase glucose uptake as the pathway of the kinase protein activated by AMP (AMPK).,16 The AMPK is activated in situations of energetic deficit and increased GLUT4 translocation for muscle cell membrane. In addition, because physical exercise can achieve glucose uptake in other insulinsensitive tissues, such as fat tissue.O'Gorman et al. when studied the increase of induced glucose uptake by exercise in obese individuals with type 2 diabetes reported that an acute bout of exercise for 1 hour to 75% of VO submitted animals to physical training for seven weeks and they observed higher response of signaling pathway of insulin in adipocytes with increasing in glucose uptake and phosphorylation in tyrosine of insulin receptor subtracts 1 and 2 (IRS-1 and IRS-2), high association of IRS-1 with PI3-Kinase and, as a consequence, increase Akt protein phosphorylation. Besides, physical exercise, as previous reported, stimulates glucose uptake by AMPK pathway favoring the entrance of these hexoses in contracted muscle,,17 even when the signaling pathway of insulin is impaired.In our study we observed that acute exercise could increase insulin sensibility in obese animals. This change occurred at least in parts for the increase of Akt protein phosphorylation in skeletal muscle of exercised obese rodents. However, exercise has shown to be able to increase glucose uptake in response to insulin also in fat tissue. Peres et al.Although in our study did not evaluate this aspects, it is possible that they collaborate for the increase of total body insulin sensibility observed insulin tolerance test in obese animals submitted to acute exercise and, for this reason, which had contributed for homeostasis of glucose in these animals. These mechanism needs to be further investigate in different physical exercise protocols. reported that eight weeks of aerobic exercises for 45 minutes and 70% VO2peak of intensity were efficient to increase glucose uptake in both obese volunteers and obese with diabetes. Physical training also enabled an increase in GLUT4 expression of Akt protein in the two groups, however, this increase in Akt expression was not followed by increase in phosphorylation neither by changes in other proteins of insulin signaling pathway. Hence, the importance of further studies with acute and chronic exercise protocols in obese and obese with diabetes population is clear, particularly to understand better the effects of physical exercise on insulin signaling pathway.Other study composed by obese individuals and obese with type 2 diabetes by Christ-Roberts et al.,22,23 As previously presented, some pro-inflammatory proteins with increased activity in obesity are able to impair the insulin signaling pathway, acting especially on proximal proteins of insulin signaling pathway. Among these proteins, the kinase serine JNK and IKK could phosphorylate in serine 307 the receptor and insulin receptor subtracts  and impair the insulin signal, which results in low Akt phosphorylation. It is well know that exercise reduces the activity of these proteins and restores insulin action.,22,23In addition, the increase of Akt phosphorylation with exercise could be a result of an improvement in inflammatory process observe by previous studies in the literature.,28 A study conducted by Matos et al. showed that physical exercise could decrease TRB3 expression and restore the Akt phosphorylation in skeletal muscle in obese animals. This is another molecular mechanism that physical exercise can increase Akt phosphorylation. Although our study did not analyze the TRB3 expression, there is the possibility that both exercises protocols had reduced TRB3 expression and, contributed with increase in Akt phosphorylation after an acute bout of physical exercise.The most recently discovery, the TRB3 protein  has its expression increased in obesity and it has the ability to physically bind to Akt, therefore avoiding Akt activation and consequently impairing the biologic effects of this activity such as the GLUT4 translocation to the muscle cell membrane.Our findings showed that physical exercise protocols of low intensity and moderate intensity are different strategies to restore insulin sensibility efficiently in obese organisms.


DM is an autosomal dominant disorder whose multisystemic features originate form nucleotide expansions: (CTG)n in the dystrophy myotonic protein kinase (DMPK) gene on chromosome 19q13 in DM type 1 (DM1), or (CCTG)n in intron 1 of the CNBP gene  on chromosome 3q21 in DM type 2 (DM2). Sarcopenia is an age-related condition characterized by the decline of muscle mass, strength, and function, whose causes are still poorly known and probably manifold . Interestingly, skeletal muscles in both DM and sarcopenia show myofiber atrophy, fiber size variability, and centrally located nuclei, as well as a reduced satellite cells’ effectiveness. Based on ex vivo and in vitro studies, we have demonstrated that both myofibers and satellite cells of DM and sarcopenic muscles exhibit a massive nuclear rearrangement of the structural and molecular factors responsible for pre-mRNA transcription and maturation; the impairment in the pre-mRNA post-transcriptional pathways would thus account for the aging-reminiscent muscle phenotype of DM patients suggesting that the skeletal muscle wasting observed in DM and sarcopenia may result from similar cellular mechanisms.In recent years, histochemistry at light and electron microscopy has increasingly been applied to investigate basic mechanisms of skeletal muscle diseases; in particular, the study n nucleotide sequences in the 3′ untranslated region (3′-UTR) of the DMPK gene, located on chromosome 19 (ch19q13)  gene    undergo extensive modifications before becoming mature mRNAs to be exported to the cytoplasm. This processing occurs in the spliceosome, i.e., the molecular complex composed of five small nuclear ribonucleoproteins (snRNPs)  and many non-snRNP splicing factors, as well as by a large number of regulating molecules  undergo accumulation and altered intranuclear distribution in both DM1 and DM2  from aged subjects  and cytoplasmic modifications ; in addition, these myoblasts have dramatically reduced myogenic capability giving rise to structurally and functionally defective myotubes .Abnormal intranuclear distribution of splicing factors has been described in satellite cells of aged muscles, suggesting that RNA pathways undergo alterations also in these quiescent cells, possibly hampering their response to muscle damage  to transcriptionally inactive domains as much as it occurs in the skeletal muscle nuclei from sarcopenic individuals. Moreover, combined fluorescence and immunoelectron microscopy conclusively demonstrated that, in DM skeletal muscle, nuclear foci sequester not only MBNL1 but also two major classes of splicing factors – snRNPs and hnRNPs – which are essential for the early processing phases of pre-mRNAs (Malatesta et al., It has repeatedly been demonstrated that the proper location and composition of the RNP-containing nuclear domains is an essential pre-requisite for transcription and pre-mRNA processing to correctly take place (Mahadevan et al., Ex vivo and in vitro studies demonstrated that both myofibers and satellite cells of DM and sarcopenic muscles exhibit a massive nuclear reorganization of the RNP-containing domains where the molecular factors responsible for pre-mRNA transcription and maturation do localize; we hypothesize that the impairment in the RNA post-transcriptional pathways may account for the aging-reminiscent muscle phenotype of DM patients suggesting that the skeletal muscle wasting observed in DM and sarcopenia may result from similar cellular mechanisms.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.


Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) which are frequently found infecting cassava, one of sub-Saharan Africa’s most important staple food crops. Each year these viruses cause losses of up to $100 million USD and can leave entire families without their primary food source, for an entire year. Twelve new whole genomes, including seven of CBSV and five of UCBSV were uncovered in this research, doubling the genomic sequences available in the public domain for these viruses. These new sequences disprove the assumption that the viruses are limited by agro-ecological zones, show that current diagnostic primers are insufficient to provide confident diagnosis of these viruses and give rise to the possibility that there may be as many as four distinct species of virus. Utilizing NGS sequencing technologies and proper phylogenetic practices will rapidly increase the solution to sustainable cassava production.Cassava brown streak disease is caused by two devastating viruses,  Manihot esculenta Crantz) productivity in sub-Saharan Africa. CBSD is caused by two distinct species of single-stranded RNA (ssRNA) viruses, Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), belonging to the genus Ipomovirus of the family Potyviridae [Bemisia tabaci Gennadius in the field [Nicotiana benthamiana) or mechanically under laboratory conditions [Cassava brown streak disease (CBSD) presents the most formidable threat to cassava  developing molecular diagnostic tools for early detection of CBSD-associated virus isolates, 2) generating information on genetic variability, 3) understanding the evolutionary forces acting on the virus genes and 4) clarifying the taxonomy of the virus or viruses associated with CBSD. However, progress has been hampered by conventional methods such as Real-Time PCR that may be too specific to a particular species or even strain of a virus . With thPhylogenetic relationships among the available CBSV whole genome sequences have shown a high level of intra-and inter-genus diversity , 7. HoweThe ideal phylogenetic tree construction methods are the maximum-likelihood and Bayesian methods, which are model-based and use statistics that best describe the data. Maximum-likelihood gives the most likely tree and takes into consideration the rate of change in every single sequence in the alignment (among site rate variation) i.e rate of change in the alignment and branch lengths. A Bayesian approach on the other hand provides many trees by sampling of the tree spaces using Markov Chain Monte Carlo sampling  and provIn this study, we have combined NGS data and the southern hemisphere’s most powerful supercomputer, Magnus, to resolve the phylogenetic relationships of 26 whole genomes of CBSV, including 12 new isolates from Tanzania, Serengeti (in the Lake Zone of Tanzania), Tanga (in the East Coast Zone), Nyasa (in the Southern Zone) and the Mafia district on the island off the Tanzanian mainland. The analysis was performed on 384 cores of the Supercomputer Magnus and provides greater insight into the genomic diversity of CBSV and UCBSV present in sub-Saharan Africa than ever before. The addition of supercomputing applications has allowed for exciting advances in phylogenetic and species delimitation analyses that give us greater confidence in detailing, for the first time, the presence of further closely related species of viruses in the complex mix that makes up CBSD.Cassava fields (3–6 months old) in the major cassava growing zones of Tanzania  were inspected for CBSD symptoms . A totalRNA was extracted from approximately 100mg of cassava leaf using the CTAB (cetyltrimethyl ammonium bromide) , 20. TheTMPlant according to the manufacturer’s instructions . Briefly, after rRNA depletion and RNA fragmentation, first and second strand cDNA was synthesized, adapters were ligated to the 5′ and 3′ ends of the fragments and the fragments enriched by PCR. cDNA libraries final size and concentration of each library was estimated using a Bioanalyzer  and the Qubit , respectively. Ten nM library pools were prepared by mixing the libraries to achieve an equal molar concentration of each. Libraries were normalized, pooled and sequenced using a 2×300 -cycle PE V3 Illumina kit. Paired end reads were generated using the Illumina MiSeq System at the Biosciences Eastern and Central Africa–International Livestock research Institute (BecA-ILRI) Hub in Nairobi, Kenya.Total RNA extracts that presented 260/280 and 260/230 purity indices equal to or greater than 2.0 and integral RNA in electrophoresis and Bioanalyzer measurements (RIN>8) were selected. The cDNA libraries were prepared from 1 μg of total RNA using the IlluminaTruSeq Stranded Total RNA Sample Preparation kit with Ribo-Zerode novo assembly function of CLCGW with automatic word size, automatic bubble size, minimum contig length 500, mismatch cost two, insertion cost three, deletion cost three, length fraction 0.5 and similarity fraction 0.9. Contigs were sorted by length and the longest subjected to a BLAST search (blastn and blastx) [For each sample, reads were first trimmed using CLC Genomics Workbench 6.5 (CLCGW) (CLC Bio) with the quality scores limit set to 0.01, maximum number of ambiguities to two and removing any reads with <30 nucleotides (nt). Contigs were assembled using the  blastx) . In addi blastx)  (Biomatt blastx) . Open re blastx) , 24, andTwelve CBSV (5) and UCBSV (7) whole genomes were downloaded from GenBank and imported into Geneious, and the Mauve plugin was used to align these with the 12 new whole genome sequences. Nucleotide alignments were translated into protein using the MAFFT translate align option in Geneious and then visually inspected.P value cutoff of 0.05. A recombination pattern was considered if detected by four or more of these programs, and anything less than four programs were not considered a valid recombination event [The RDP4 package  was usedon event , 33, 34.Bayesian analyses were conducted using ExaBayes version 1.4.1;  and wereThe tip to root approach of species delimitation  was emplBased on the analysis of whole genome virus sequences, both CBSV and UCBSV are widely distributed in Tanzania. In coastal areas, CBSV and UCBSV occurred widely. Three CBSV isolates  were identified in the Tanga district from unnamed local CBSD-affected cassava plants. One UCBSV isolate (TZ: Tan–23) was found to occur in the same district. One CBSV isolate (TZ: MAF–49) and two UCBSV isolates (TZ: MAF–51 and 58) were identified from cassava cultivar Kilembe in Mafia district, an island located in the Indian Ocean off the Tanzanian mainland. In the Lake zone in northwestern Tanzania, in Serengeti district of the Mara region, two CBSV isolates (TZ: Ser–5 and 6) were identified from Rumala, a local cassava cultivar. Another isolate UCBSV (TZ: Ser–6) was found in a co-infection with a CBSV isolate (TZ: Ser–6) in the same plant. In southern Tanzania, we found two CBSV isolates (TZ: Nya–36 and 38) from CBSD-affected cassava cultivar ‘Nkangawandu’ in the Nyasa district, on the shore of the Lake Nyasa near Mbamba Bay in the Ruvuma region. These findings represent the first report of CBSD occurrence in the district. The geographical locations from where the CBSV and UCBSV were collected, symptom severity score distribution and the identity of the genetic group of the isolates are shown in Foliar CBSD symptom types and severity varied from mild to severe on the plants that were surveyed. Field infected cassava plants collected in Mafia district from which the CBSV isolate TZ:MAF–49 and the UCBSV isolates TZ:MAF–51 and 58 were found displayed pronounced varying patterns of chlorosis, which appeared as feathery patterns with chlorotic blotches along the margins of secondary veins, tertiary veins, and main veins . In addiIn the Tanga district, cassava plants infected by UCBSV isolates TZ:Tan-19-1 and 2, TZ:Tan–26 and UCBSV isolates TZ:Tan–23 displayed severe yellow chlorosis on leaves  . Die-bacde novo assembly of the reads for each individual sequence, also using CLCGW, the number of contigs produced ranged from 224 to 7,411. The contig of interest lengths were 5,339 to 9,074 with average coverage of 39 to 721 times and the numbers of reads mapped to each contig were 2,027 to 48,557. After mapping to a reference genome in Geneious, the lengths of the consensus sequences were 8,549 to 10,044 with average coverage of 40 to 673 times with the numbers of reads mapped to the reference sequence ranging from 2,701 to 49,252. Final overall sequence lengths consisted of a consensus between the de novo contig of interest and the mapped consensus sequence and were 8,945 to 9,070 nt. Eight of the samples yielded one sequence of interest each and two others yielded two sequences of interest each, with a total of 12 new sequences, five of UCBSV and seven of CBSV. All sequences were submitted to GenBank with the accession numbers KR108828-KR108839. Raw reads forming part of the assembled genomes can be made available to individuals upon request.Ten samples were sequenced on an Illumina MiSeq, which produced numbers of raw reads ranging from 2,071,164 to 5,719,724 for each sample. Raw reads can be accessed from the European Nucleotide Archive accession number: PRJEB10634. After trimming for quality using CLCGW, these numbers were reduced to 2,025,314 to 5,582,134 . FollowiWhen the complete coding regions of the 12 new sequences, along with those retrieved from GenBank were analyzed, eight firm recombination events were identified amongst the CBSV sequences , Fig 4A Bayesian phylogenetic analysis clearly separated CBSV and UCBSV into 2 groups, indicating that they are distinct species . The anaCBSV and UCBSV are supported as distinct species as shown in In the present study, we report the occurrence of 12 new whole genome sequences for the devastating cassava viruses (five new isolates of CBSV and seven UCBSV) from East Africa. We have utilized existing data, NGS technology and Supercomputing to identify new viruses and scrutinize the distribution of CBSV and UCBSV in Tanzania. In doing so, we have found that both viruses are widely distributed in different agro-ecologies  in Tanzania. These findings are contrary to previously published reports that CBSV occurrence is limited to low and medium altitude areas below 1000 m above sea level, while UCBSV occurred in the highland areas (>1000 m above sea level.) of East Africa . We founIn the field, cassava plants affected by CBSD displayed a range of symptoms, depending on the cultivar and agro-ecology. In this study, CBSV and UCBSV were associated with varied foliar symptoms partly because the CBSD resistance and or tolerance levels of the sampled farmers' cassava cultivars could not be established. Our study therefore confirms earlier findings that there is no clear correlation between cassava brown streak virus species and CBSD symptom types and variability .Cassava Brown Streak Virus and Ugandan Cassava Brown Streak Virus are the two recognized viral species associated with the devastation of cassava crops in East Africa in recent years [http://www.ictvonline.org/proposals/2010.001aP.A.v2.Ipomovirus-Sp.pdf). Species delimitation has moved beyond relying on sequence percentage similarity and more robust techniques are needed to distinguish these species. The species delimitation metrics utilized in our study indicates support for these two species but also an additional three species within the UCBSV clade. The new species we have identified are labeled C, D and E . While it is clear that UCBSV is a separate species to CBSV, it is not found exclusively in Uganda, nor did it originate there. Therefore, referring to the species as 'Uganda' has led many to believe that this species has only recently spread or been moved (through use of infected cuttings) to the wider geographical scope it is found in Kenya, Rwanda, Tanzania including mafia and Zanzibar Islands in the Indian Ocean. It would be inconceivable that through recent human effort, UCBSV-infected materials were moved to these islands in the Indian Ocean, which are many miles away from Uganda. Therefore, great care must be taken when naming the clades, groups and species associated with a particular set of sequences and their phylogenetic analysis. It has been common practice to name these according to things such as host, geographic location or country of origin, however this is problematic. It is becoming clear that the use of a numbered, lettered or latinised numeral system may be a useful tool particularly regarding species delimitation and strain differentiation of some plant viruses [The International Committee on Taxonomy of Viruses (ICTV) has the mandate to name new viruses. One of the essential principles of virus nomenclature aims to avoid or reject the use of names, which might cause error or confusion . One cas viruses , 48. Thi viruses , 48, 49.Unbiased species delimitation is a crucial first step in identifying out-breaking viral strains. If traditional species aren’t questioned or evaluated, many new species will go undetected, possibly leading to increased damage due to lack of adequate diagnostics. The use of the Magnus supercomputer made these analyses possible, and without it the 7-hour runtime required to generate one phylogenetic tree would balloon out to approximately 609 hours on a modern laptop or PC. The analyses has allowed us to determine that new diagnostic primers for the individual species should be redesigned for the CI region of the genome–As in previous studies, we found no evidence of recombination between CBSV and UCBSV, although there is evidence for homologous recombination . As moreBemisia tabaci, that are currently being discovered [A way forward in identifying plant viruses affecting cassava should include the rigor that has been established in the current study, NGS technologies coupled with proper phylogenetics and species delimitation metrics are necessary for identifying cassava plant viruses. The data presented shows there is a need to reevaluate the current diagnostic primers (coat protein and 3’ UTR) for CBSV/UCBSV as they are not detecting the diversity, this is key to ensuring clean seeds and cuttings are moved throughout the region. In addition, it will be crucial as more viruses are discovered to map the transmission efficiency of these viruses with the new species of the vector, scovered , 55. Int


Hormones produced by the adrenal glands and adipose tissues have important roles in normal physiology and are altered in many disease states. Obesity is associated with changes in adrenal function, including increase in adrenal medullary catecholamine output, alterations of the hypothalamic-pituitary-adrenal (HPA) axis, elevations in circulating aldosterone together with changes in adipose tissue glucocorticoid metabolism, and enhanced adipocyte mineralocorticoid receptor activity. It is unknown whether these changes in adrenal endocrine function are in part responsible for the pathogenesis of obesity and related comorbidities or represent an adaptive response. In turn, adipose tissue hormones or “adipokines” have direct effects on the adrenal glands and interact with adrenal hormones at several levels. Here we review the emerging evidence supporting the existence of “cross talk” between the adrenal gland and adipose tissue, focusing on the relevance and roles of their respective hormones in health and disease states including obesity, metabolic syndrome, and primary disorders of the adrenals. The significance of the adrenal gland as an endocrine organ and the roles of adrenal hormones in physiology and disease have been recognized well over a century , 2. In fThe adrenal gland is composed of an outer cortex producing steroid hormones including mineralocorticoids, glucocorticoids, and sex steroids and an inner medulla of neuroectodermal origin producing catecholamine hormones. Each of these hormones acts on adipose tissue, which is readily apparent from the well-described clinical manifestations of primary adrenal endocrinopathies such as Cushing syndrome, Addison disease, and pheochromocytoma.Adipose tissues are a heterogeneous group of tissues classified based on histology and function as white adipose tissue (WAT) and brown adipose tissue (BAT). Adipose tissues can be further categorized by anatomical location as visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) . While rα (TNF-α), C-reactive protein (CRP), serum amyloid A (SAA), and interleukin-6 (IL-6) which are secreted not only from adipocytes, but also from myeloid-derived cells such as macrophages located in the adipose tissue stroma [The adipose tissues are a source of several hormones termed “adipokines” or “adipocytokines” including adipocyte-derived factors such as leptin, adiponectin, resistin, and proinflammatory cytokines such as tumor necrosis factor-e stroma , 8. While stroma –11.The growing body of knowledge describing two-way interactions between adrenal hormones and the “endocrine adipocyte” has resulted in the proposal of the existence of an “adipose-adrenal axis” . In thisAldosterone is the primary mineralocorticoid hormone produced by the adrenal glands in humans and exerts its effects via the mineralocorticoid receptor (MR). While the well-described “classical” effects of aldosterone on transepithelial Na transport are mediated by MR present in epithelial cells, MR has been shown to be present in a number of other cell types including adipocytes .α, monocyte chemotactic protein-1 (MCP-1), and IL-6 in WAT, while decreasing the thermogenic activity and lowering uncoupling protein 1 (UCP1) transcription of BAT [Activation of MR appears to have important roles in adipose tissue including differentiation of preadipocytes to mature adipocytes and promotion of a proinflammatory state via induction of cytokines including TNF-n of BAT . MR mRNAn of BAT .Since glucocorticoids (cortisol in humans and corticosterone in rodents) and mineralocorticoids  both bind to MR with relatively high affinity, the MR specificity of aldosterone in epithelial cells in humans is due to the intracellular inactivation of cortisol to cortisone (and corticosterone to 11-dehydrocorticosterone in rodents) by the enzyme 11-beta-hydroxysteroid dehydrogenase type 2 (11HSD2) . AdipocyIncreased serum aldosterone levels have been observed with both obesity-associated hypertension and the metabolic syndrome. While several mechanisms may be at play, this finding is most likely due in large part to the upregulation of the renin-angiotensin-aldosterone system (RAAS) in obesity, a proposed mechanism linking hypertension with obesity that is reversed by weight loss .A second, more novel mechanism of increased aldosterone production in obese individuals, exemplifying the concept of adrenal-adipose “cross talk,” is the identification of adipose-derived factors able to directly stimulate adrenal aldosterone production . This fiA third candidate mechanism contributing to our understanding of the proposed role of MR activation in adipose tissues was brought up by a recent report demonstrating both in vitro production and in vivo production of aldosterone by adipocytes, resulting in both autocrine and paracrine regulatory effects on adipocyte function .Clinical implications of these insights into the role of adipocyte MR in obesity include whether MR blockade would be of benefit in treating obesity-associated morbidities such as hypertension, diabetes, and the metabolic syndrome. While studies of MR blockade in obese mice have shown benefits on adipose tissue dysfunction , 20, cliPrimary hyperaldosteronism (PA), known also by its eponymous name Conn syndrome, is a state of continuous aldosterone excess due to autonomous production of aldosterone by an adrenal adenoma or bilateral adrenal zona glomerulosa hyperplasia and thus provides a relevant model for investigating the results of systemic aldosterone excess on adipose tissue.While continuous infusion of aldosterone over 12 days resulted in weight gain in adrenalectomized rats , both crGlucocorticoids have effects on adipose tissue development, metabolism, and adipocyte secretory function. Studies investigating the effects of GC on adipogenesis and adipocyte lipid metabolism provide discordant results, while the anti-inflammatory nature of GR activation in adipocytes is a consistent finding , 28. GluAlterations of circulating cortisol dynamics, mainly as hypothalamic-pituitary-adrenal (HPA) axis dysfunction, as well as local metabolism of glucocorticoids in adipose tissue, have been linked to obesity and the metabolic syndrome.The dramatic changes in fat distribution characterized by central adiposity with wasting of subcutaneous fat depots, typical of Cushing syndrome, have led to the hypothesis that alterations in GC action may play a role in more common forms of visceral obesity. While circulating cortisol levels are not increased in obese individuals, increased net cortisol production occurs locally in the adipose tissue due to the above-described alterations in adipocyte HSD enzymes, and thus obesity has been described as “Cushing syndrome of the omentum”  Figure .An important confounder in interpreting studies of GC action on adipose tissue is the fact that GC effects in adipose tissue are mediated via the MR receptor as well as the GR receptor. Due to the lack of adipocyte activity of 11HSD2, an enzyme which inactivates endogenous GC in mineralocorticoid target tissues, much of the observed effect of naturally occurring GC in adipocytes is due to its effect on MR, rather than GR  Figure .Many studies have suggested increased 11HSD1 activity in obesity , 34. TraWhile studies of the effects of GC on adipogenesis and adipocyte lipid metabolism have yielded inconsistent results, the effects of GC in suppressing inflammatory signaling from adipocytes are remarkably consistent. Experiments of selective GR stimulation with dexamethasone have generally resulted in decreased expression of proinflammatory cytokines by adipocytes, while GR knockdown with siRNA has the opposite effect . InvestiObesity and metabolic syndrome are associated with alterations in the HPA axis including changes in adrenal cortisol production, peripheral cortisol metabolism, and dynamic tests of the HPA axis. Whether these changes play a role in the development and pathophysiology of obesity and related comorbidities or represent an adaptation to the obese state is unknown. Basal levels of ACTH and cortisol appear to be undisturbed in obesity. While 24-hour urine-free cortisol generally has not been found to be increased in obese subjects, one study described significantly elevated night time (7 pm–7 am) urine-free cortisol in women with abdominal obesity . The mosThe adrenal cortex produces androgen hormones in both sexes, including dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEA-S), and androstenedione. In humans DHEA-S circulates at higher concentrations than any other steroid hormone and levels decline substantially with age, paralleling the changes in body composition characteristic of aging . Of partA potential confounder in interpreting these data is the fact that DHEA acts as a precursor hormone and is converted into both testosterone and estrogens, which have independent effects on adipose tissue and body composition . ReceptoThe adrenal medulla functions similarly to a sympathetic ganglion, secreting catecholamine hormones, primarily epinephrine, into the bloodstream under the control of the sympathetic nervous system (SNS). Epinephrine has potent lipolytic effects on adipose tissue  and impaThere is increasing data indicating cross talk between catecholamines and adipokines, with evidence supporting stimulation of adrenal medullary function by leptin, while resistin may suppress medullary catecholamine secretion . Closingα, IL-6, and adiponectin.Obesity is associated with overactivity of the sympathetic nervous system. This SNS “overdrive” has been implicated as a possible contributor to the pathogenesis of obesity-associated morbidities and end-organ damage attributed to the obese state. One theory explains the increased SNS output of obesity as a homeostatic counterregulatory mechanism aimed at redirecting excess energy supply into adrenergic thermogenesis, thus protecting from the harms of further fat storage . AdiposeThe relationship between leptin and catecholamine production may be of particular significance in the discussion of obesity as leptin has been implicated as a direct inducer of increased SNS outflow in obesity . ConversRecent reports have highlighted the importance of catecholamines in regulating BAT in adults. Several observational studies of patients harboring PHEO describe that active BAT, as evidenced by increased uptake on 18-fluorodeoxyglucose-positron emission tomography (FDG-PET) by BAT, is diminished following surgical treatment of pheochromocytoma , 68. IntAdrenal masses are discovered in approximately 4% of CT or MRI studies performed for reasons other than suspicion of adrenal disorder . Such inWhile the primary regulation of adrenal hormone production, classical glucocorticoids and androgens by ACTH, aldosterone by the RAAS, and catecholamines by the SNS, has been well described, it has more recently been proposed that adipokines may have a role in modulating adrenal function. While a comprehensive review of the effects of all adipokines on adrenal function is beyond the scope of this review, we will summarize the effects of two adipocytokines of particular relevance, leptin and adiponectin, as follows.Since its discovery as a WAT-derived circulating satiety factor in 1994 , the effWhile the reciprocal relationship between leptin and sympathetic nervous system activity has been detailed above, leptin has also been shown to exert direct effects on the adrenal medulla. Leptin stimulates secretory activity of porcine chromaffin cells , while bγ) signaling pathways. Adiponectin receptors are present in both human and mouse adrenals [Adiponectin is the most abundant adipokine in the circulation with insulin-sensitizing, antiatherogenic and anti-inflammatory properties mediated mainly by the adenosine monophosphate-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor gamma (PPARadrenals , 82. Pasadrenals . Short tadrenals , 84. In adrenals .The emergence of obesity as a worldwide epidemic has brought the study of adipocyte biology and secretory functions of adipose tissue to the forefront of biomedical and endocrine research. The interaction between adipokines and other endocrine organs is an area of recent intensive investigation. In this regard, the hormones of the adrenal glands are particularly relevant given their well-established effects on lipid metabolism, inflammation, and the “stress” response. Here we have discussed the physiologic and pathophysiologic actions of each of the adrenal hormones in the adipose tissues with focused discussions of the specific alterations observed in obesity and metabolic syndrome as well as mechanistic considerations of their roles in obesity and associated comorbidities. We have also examined the evidence linking adipocyte dysfunction to primary adrenal disorders and elucidated several examples of two-way interactions or cross talk between hormones of the adrenals and adipokines. Further research is needed to improve our collective understanding of the significance of the described changes in adipocyte function in primary adrenal diseases and particularly of the changes in adrenal function resulting from the adipokine milieu characteristic of obesity; a fundamental concept in need of further exploration is whether these are adaptive mechanisms representing a protective response to the obese state or, conversely, contributors or “drivers” in the morbidity associated with excess adiposity. Lastly, we have provided a theoretical basis for potential future pharmacological interventions aimed at adrenal hormone targets in the adipose tissue.


Growing evidence shows that the expression and release of a number of inflammatory cytokines are increased in hypertension ,2 and ar2, Haematoxylin and Eosin-stain), fibrosis  and inflammation parameters .12-week-old male SHR and Wistar rats were infused through osmotic pumps with CNP  or saline (S) for 14 days. Systolic blood pressure  was recorded by tail-cuff method. At the end of the treatment, the left ventricle (LV) was extracted to determine left ventricular mass index , the myocyte area (µmCharacteristic high SBP values in SHR are accompanied by hypertrophy, fibrosis and a higher presence of inflammatory cytokines.Our results show that chronic treatment with CNP attenuates the expression of proinflammatory markers and the early signs of fibrosis in cardiac tissue of hypertensive rats. These effects, combined with the drop in blood pressure we observed, indicate that CNP could possibly have an important pathophysiological and therapeutical role in preventing or even reversing cardiac fibrosis and inflammation accompanying left ventricular remodelling in arterial hypertension.