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BackgroundMulti System Inflammatory Syndrome in children (MIS-C) associated with COVID-19 is a recently recognised potentially life-threatening entity. There is limited data on post MIS-C sequelae.Methods21 children fulfilling the WHO criteria for MIS-C were included in our study. Data was collected at baseline and at 12-16 weeks post discharge to look for any persistent sequelae mainly relating to the lungs or heart including coronary arteriesResultsFever was the most common presentation, found in 18 (85.7%) patients. All had marked hyper-inflammatory state. Low ejection fraction (EF) was found in 10 (47.6%), but none had any coronary artery abnormality. All received corticosteroids, while 7 (33.3%) children required additional treatment with intravenous Immunoglobulins. 20 children improved while 1 left against medical advice. At discharge, 3 children had impaired left ventricular function. At median 15 weeks’ follow-up, no persistent complications were found. EF had returned to normal and no coronary artery abnormalities were found during repeat echocardiography. Chest radiographs showed no fibrosis and all biochemical parameters had normalized.ConclusionThe children with MIS-C are extremely sick during the acute stage. Timely and adequate management led to full recovery without any sequelae at a median follow-up of 15 weeks.

Background: Kawasaki disease (KD) is a medium-vessel vasculitis having coronary predilection, usually affecting under-5 children presenting as acute febrile illness. Despite being a disease with long-term grievous outcome, only few published literature are available from India, even lesser from its eastern region. Methods: From January 2016 to December 2020, 30 case records of children with a discharge diagnosis of KD were enrolled in this study. Demographic profile, clinical manifestations, laboratory data, echocardiographic findings, and treatment done were extracted from the case records. Laboratory investigations were done at admission and repeated after 24 h of intravenous immunoglobulin administration as per the American Heart Association Guidelines 2004. Echocardiography was carried out at diagnosis, at 2 weeks, and at 6 weeks. Results: Out of 30 children diagnosed with KD, majority belong to 1–5 years of age group (72%) with male predominance. Complete KD was seen in 77% of children. The most common presentation was fever >5 days (100%) followed by oral changes in 26 (87%), conjunctivitis in 25 (83%), extremity changes in 23 (77%), and rash in 21 (70%) children. Desquamation of perineum and reactivation of bacillus Calmette–Guérin scar were seen in 10%. No children with complete KD and three children with incomplete KD developed coronary artery Abnormalities (CAA). Conclusion: Infants with incomplete KD have a higher incidence of CAA. Aggressive management results in better outcome.

Atypical hemolytic–uremic syndrome (aHUS) is a form of thrombotic microangiopathy that occurs due to dysregulation of alternate pathway of complement system, which progressively causes systemic complications, end-stage renal disease, and death. As prognosis is poor compared to typical hemolytic–uremic syndrome, early diagnosis and treatment is crucial for favorable outcome. We came across seven patients of aHUS in our pediatric intensive care unit in the last 5 years. Plasma exchange (PE) along with immunosupressives was used for treatment. First child who did not receive PE died. Rest six patients underwent PE and attained hematological remission; however, one later on progressed to chronic kidney disease and expired. All others are on regular follow-up and doing well. A high index of suspicion is required to diagnose aHUS. Early PE can give a better prognosis.

Significance Chronic neurodegeneration, a major cause of the long-term disabilities that afflict survivors of traumatic brain injury (TBI), is linked to an increased risk for late-life neurodegenerative disorders, including Alzheimer’s disease, Parkinson’s disease, vascular dementia, and chronic traumatic encephalopathy. Here, we report on the restoration of blood–brain barrier (BBB) structure and function by P7C3-A20 when administered 12 mo after TBI. This pharmacotherapy was associated with cessation of chronic neurodegeneration and recovery of normal cognitive function, benefits that persisted long after treatment cessation. Pharmacologic renewal of BBB integrity may thus provide a new treatment option for patients who have suffered a remote TBI, or other neurological conditions associated with BBB deterioration., Chronic neurodegeneration in survivors of traumatic brain injury (TBI) is a major cause of morbidity, with no effective therapies to mitigate this progressive and debilitating form of nerve cell death. Here, we report that pharmacologic restoration of the blood–brain barrier (BBB), 12 mo after murine TBI, is associated with arrested axonal neurodegeneration and cognitive recovery, benefits that persisted for months after treatment cessation. Recovery was achieved by 30 d of once-daily administration of P7C3-A20, a compound that stabilizes cellular energy levels. Four months after P7C3-A20, electron microscopy revealed full repair of TBI-induced breaks in cortical and hippocampal BBB endothelium. Immunohistochemical staining identified additional benefits of P7C3-A20, including restoration of normal BBB endothelium length, increased brain capillary pericyte density, increased expression of BBB tight junction proteins, reduced brain infiltration of immunoglobulin, and attenuated neuroinflammation. These changes were accompanied by cessation of TBI-induced chronic axonal degeneration. Specificity for P7C3-A20 action on the endothelium was confirmed by protection of cultured human brain microvascular endothelial cells from hydrogen peroxide-induced cell death, as well as preservation of BBB integrity in mice after exposure to toxic levels of lipopolysaccharide. P7C3-A20 also protected mice from BBB degradation after acute TBI. Collectively, our results provide insights into the pathophysiologic mechanisms behind chronic neurodegeneration after TBI, along with a putative treatment strategy. Because TBI increases the risks of other forms of neurodegeneration involving BBB deterioration (e.g., Alzheimer’s disease, Parkinson’s disease, vascular dementia, chronic traumatic encephalopathy), P7C3-A20 may have widespread clinical utility in the setting of neurodegenerative conditions.

Background: Nosocomial infection, due to Candida, contracted in the pediatric intensive care unit (PICU), is emerging as a significant healthcare challenge. The incidence of non-albicans Candida as a cause of candidemia is on the rise, unlike in previous decades. Materials and Methods: All the cases of candidemia confirmed by culture, admitted to the PICU during the study period of January 2017 to December 2019, were retrospectively studied. The prevalence, speciation, sensitivity pattern and risk factors of candidemia mortality were recorded and analyzed. Results: There were 1034 admissions to the PICU in the study period, of which 926 blood samples were sent for culture and sensitivity. A total of 31 Candida non-albicans and five Candida albican species were isolated. C. tropicalis was the most common type (44.4%) of Candida species found, followed by C. glabrata (16.7%), C. parapsilosis (16.7%) and C. krusei (5.6%). The sensitivity of all Candida isolates to Amphotericin B, Clotrimazole, Voriconazole, Itraconazole, Ketoconazole, Nystatin, and Fluconazole was 94.4%, 91.7%, 88.9%, 86.1%, 77.8%, 52.8%, and 38.9% respectively. The use of a central venous catheter was a statistically significant contributor to mortality due to candidemia. Conclusion: Non-albican Candida species are the predominant cause of candidemia this study. They are associated with higher fatality rates. Sensitivity of the Candida spp. was more common to Amphotericin-B than azoles.

M2 macrophages are associated with deposition of interstitial collagen and other extracellular matrix proteins during the course wound healing and also inflammatory response to biomaterials. Developing advanced biomaterials to promote the M2 subtype may be an effective way to improve tissue reinforcement surgery outcomes. In this study, the effect of genipin, a naturally derived crosslinking agent, on M0 → M2-polarization was investigated. Genipin was introduced either indirectly by seeding cells on aligned collagen biotextiles that are crosslinked by the agent or in soluble form by direct addition to the culture medium. Cellular elongation effects on macrophage polarization induced by the collagen biotextile were also investigated as a potential inducer of macrophage polarization. M0 and M2 macrophages demonstrated significant elongation on the surface of aligned collagen threads, while cells of the M1 subtype-maintained a round phenotype. M0 → M2 polarization, as reflected by arginase and Ym-1 production, was observed on collagen threads only when the threads were crosslinked by genipin, implicating genipin as a more potent inducer of the regenerative phenotype compared to cytoskeletal elongation. The addition of genipin to the culture medium directly also drove the emergence of pro-regenerative phenotype as measured by the markers (arginase and Ym-1) and through the activation of the pSTAT6-PPAR-gamma pathway. This study indicates that genipin-crosslinked collagen biotextiles can be used as a delivery platform to promote regenerative response after biomaterial implantation. Statement of significance The immune response is one of the key determinants of tissue repair and regeneration rate, and outcome. The M2 macrophage subtype is known to resolve the inflammatory response and support tissue repair by producing pro-regenerative factors. Therefore, a biomaterial that promotes M2 sub-type can be a viable strategy to enhance tissue regeneration. In this study, we investigated genipin-crosslinked electrochemically aligned collagen biotextiles for their capacity to induce pro-regenerative polarization of M0 macrophages. The results demonstrated that genipin, rather than matrix-induced cellular elongation, was responsible for M0 → M2 polarization in the absence of other bioinductive factors and maintaining the M2 polarized status of macrophages. Furthermore, we identified that genipin polarizes the M2 macrophage phenotype via activation of the pSTAT6-PPAR-gamma pathway.

Derangement in systemic metabolic homeostasis is tightly associated with widespread activation of resident and circulating immune cells, a phenomenon known as ‘metaflammation’. Numerous studies have explored the role of tissue resident and circulating macrophages in contributing to metaflammation, obesity, and their sequelae; however, there is a dearth of information regarding targetable transcriptional regulators of the genesis and persistence of metabolic disease. Here, we identify myeloid Krüppel-like factor 2 (KLF2) as a novel regulator of metabolic disease. Previous reports demonstrate that KLF2 serves as a critical regulator of myeloid cell quiescence and is downregulated in numerous acute and chronic inflammatory states. Specifically in the context of chronic metaflammation, we note that KLF2 expression is decreased in circulating immune cells of obese patients and in adipose tissue macrophages of high fat diet (HFD) fed mice, which is consistent with the hypothesis that KLF2 regulates metaflammation. To explore this further, we utilized mice with myeloid cell-specific deletion of KLF2 (K2KO) which exhibit accelerated obesity and insulin resistance. K2KO mice have widespread central (i.e. CNS) and peripheral metaflammation both in the basal and HFD-stimulated states. To discern whether the effect of myeloid deletion of KLF2 on metabolism is due to deletion in microglia in the feeding centers of the hypothalamus or in peripheral immune cells, bone marrow chimeras with head shielding were created. 50% reconstitution of circulating immune cells with K2KO cells in wildtype (WT) mice was sufficient to maintain the metabolic disease phenotype, while mice with K2KO microglia + WT circulating cells had only slightly improved outcomes compared to K2KO mice. Conversely, ablation of microglia in K2KO mice using PLX5622 formulated in HFD also successfully attenuated the aberrant feeding behavior, weight gain, and glucose dyshomeostasis seen in K2KO mice. Together, these data demonstrate a role for loss of KLF2 in hematopoietic and CNS resident cells in causing metabolic disease. Given that myeloid KLF2 expression decreases under metabolic stress in WT mice and humans, we sought to explore whether maintenance of KLF2 expression in these cells would be protective against diet-induced metabolic disease. Indeed, mice with myeloid-specific overexpression of KLF2 demonstrated a markedly improved metabolic phenotype when challenged with HFD, providing evidence that targeting KLF2 expression in myeloid cells may prove to be a therapeutic option against metaflammation.

Metabolic diseases and their serious sequelae such as non-alcoholic fatty liver disease (NAFLD) pose a substantial clinical burden. It is now well recognized that skeletal muscle is a major site for the metabolism of all major macronutrients, and derangements in these muscle processes significantly contribute to metabolic disease. Studies over the last 15 years have identified the transcription factor Krüppel-like factor 15 (KLF15) as an important regulator and effector of metabolic processes across various tissues, and furthermore, genome-wide studies have identified human KLF15 variants with increased body mass index and diabetes. Given the importance of skeletal muscle in maintaining metabolic homeostasis, we generated a skeletal muscle specific KLF15 knockout (K15-SKO) mouse to study the role of skeletal muscle KLF15 in regulating systemic metabolism. We found that this animal is prone to developing obesity and insulin resistance at baseline, a phenotype that is greatly exacerbated in response to high fat diet (HFD). Strikingly, K15-SKO mice show a propensity toward developing NAFLD, as demonstrated by increased micro- and macrovesicular steatosis, hepatocellular ballooning, increased hepatic fatty acid and triglyceride deposition, and elevated Cd36 expression. A potential cause of NAFLD is the accumulation of excess lipids and lipid intermediates due to defects in the lipid flux pathway in extrahepatic tissues. Indeed, we see defects in the expression of genes involved in the carnitine shuttle and a paucity of long-chain acylcarnitines in K15-SKO skeletal muscle. Furthermore, RNA sequencing of skeletal muscle from K15-SKO mice shows downregulation in a number of pathways involved in lipid handling. This indicates that KLF15 serves as a novel extrahepatic molecular regulator of hepatic health. It has been previously shown that a diet rich in short-chain fatty acids (SCFA) can bypass defects in lipid handling and ultimately improve metabolic health. To explore this therapeutic avenue, we gave K15-SKO mice either normal chow (NC) or a SCFA-rich diet for 7 weeks. We observed decreased weight gain and improved glucose homeostasis in SCFA-rich diet fed mice. In addition to being a preventative strategy, SCFA-rich diets may also serve as a potential therapy to rescue from metabolic disease. To this end, we gave K15-SKO mice HFD for 5 weeks followed by 7 weeks of either NC or SCFA-rich diet. We observed that providing SCFAs can improve metabolic health and ameliorate the phenotype seen due to defects in skeletal muscle lipid handling: mice given SCFA-rich diet following HFD had significantly decreased weight gain and improved insulin sensitivity. These studies demonstrate that skeletal muscle KLF15 serves as an important regulator of lipid flux and hepatic health, and that SCFA-rich diets are a promising candidate for metabolic disease resultant of impaired lipid handling.

Drought adversely affects crop production across the globe. The root system immensely contributes to water management and the adaptability of plants to drought stress. In this study, drought-induced phenotypic and transcriptomic responses of two contrasting chickpea (Cicer arietinum L.) genotypes were compared at the vegetative, reproductive transition, and reproductive stages. At the vegetative stage, drought-tolerant genotype maintained higher root biomass, length, and surface area under drought stress as compared to sensitive genotype. However, at the reproductive stage, root length and surface area of tolerant genotype was lower but displayed higher root diameter than sensitive genotype. The shoot biomass of tolerant genotype was overall higher than the sensitive genotype under drought stress. RNA-seq analysis identified genotype- and developmental-stage specific differentially expressed genes (DEGs) in response to drought stress. At the vegetative stage, a total of 2161 and 1873 DEGs, and at reproductive stage 4109 and 3772 DEGs, were identified in the tolerant and sensitive genotypes, respectively. Gene ontology (GO) analysis revealed enrichment of biological categories related to cellular process, metabolic process, response to stimulus, response to abiotic stress, and response to hormones. Interestingly, the expression of stress-responsive transcription factors, kinases, ROS signaling and scavenging, transporters, root nodulation, and oxylipin biosynthesis genes were robustly upregulated in the tolerant genotype, possibly contributing to drought adaptation. Furthermore, activation/repression of hormone signaling and biosynthesis genes was observed. Overall, this study sheds new insights on drought tolerance mechanisms operating in roots with broader implications for chickpea improvement.

Pertussis is a serious and life-threatening infection of infancy. Recurrent apnea with a paroxysm of cough is the early clue for its diagnosis. The rate of pertussis increased worldwide with the occurrence of regular outbreaks globally, including India. The resurgence of pertussis is multifactorial, and it includes antigenic shifts in bacteria, decreasing vaccine immunity, reduced duration of protection by acellular pertussis vaccine, and improved method of surveillance and diagnosis. Family members, especially mothers and siblings, are an important source of pertussis transmission to vulnerable infants. Maternal vaccination for pertussis during pregnancy should be done as many cases of infantile pertussis was found before primary immunization. A 45 days infants admitted to the pediatric intensive care unit with recurrent apnea and bradycardia, require prolonged mechanical ventilation with intense cardiorespiratory monitoring. Real-time polymerase chain reaction of the nasopharyngeal swab for pertussis was positive. Successfully discharged after a long course of hospitalization.

Background: Capsule endoscopy (CE) has an established role in evaluating obscure gastrointestinal bleeding (OGIB). The aim was to know the diagnostic yield of CE and spectrum of OGIB. Materials and Methods: In this retrospective study, we evaluated all the patients with obscure gastrointestinal bleed using MiroCam capsule endoscope (IntroMedic, Seoul, Korea) between February 2014 and March 2018. Clinical data, ancillary investigations, and response to specific treatment were considered to confirm CE findings. Results: Out of 102 patients included in the study (mean age 54.5 ± 16.1 years, male: female ratio = 1.83:1) OGIB‑overt and OGIB‑occult was present in 46 and 56 patients, respectively. Diagnostic yield of CE was similar in both the groups (overt‑37/46, 80.4% versus occult‑37/56, 66.5%) (P ≥ 0.05), although there was trend to find more lesions in overt group. Overall positive diagnostic yield was 72.5%. Lesions detected were vascular malformations in 21 (20.5%), nonsteroidal anti‑inflammatory drug enteropathy in 13 (12.7%), small bowel ulcerations in 27 (26.4%), which were further divided into three subgroups (a) nonspecific ulcerations 11 (10.7%), (b) tubercular ulcer with/without stricture in 7 (6.8%) and (c) serpiginous ulcers and fissuring with cobble‑stone appearance suggestive of Crohn’s disease in 9 (8.8%), portal hypertensive enteropathy in 5 (4.9%), worm infestation (hookworms in 3, roundworms 1) in 4 (3.9%), and small bowel tumour in 1 (0.98%) patient. Overall, 56.7% patients were having definitive (P2) lesions (Saurin classification). Two patients had retention of capsule, but none developed intestinal obstruction. Capsule was removed with surgical intervention. Conclusion: CE has high diagnostic yield, relative safety and tolerability, and it is an important diagnostic tool for OGIB. Small bowel tuberculosis, Crohn’s disease and Worm infestation continue to be commonly recognized causes of OGIB in developing countries like India.

Infected walled-off necrosis (WON) is a well-known complication of acute necrotizing pancreatitis, with higher mortality and morbidity. An infected or symptomatic WON requires drainage. Occasionally, WON may spontaneously fistulize into the gastrointestinal lumen or may rupture into the peritoneum. We describe a case of spontaneous rupture of WON in the transverse colon with uncomplicated spontaneous resolution, which is an extremely rare event.

Adipose tissue stores energy in the form of triglycerides. The ability to regulate triglyceride synthesis and breakdown based on nutrient status (e.g., fed versus fasted) is critical for physiological homeostasis and dysregulation of this process can contribute to metabolic disease. Whereas much is known about hormonal control of this cycle, transcriptional regulation is not well understood. Here, we show that the transcription factor Kruppel-like factor 15 (KLF15) is critical for the control of adipocyte lipid turnover. Mice lacking Klf15 in adipose tissue (AK15KO) display decreased adiposity and are protected from diet-induced obesity. Mechanistic studies suggest that adipose KLF15 regulates key genes of triglyceride synthesis and inhibits lipolytic action, thereby promoting lipid storage in an insulin-dependent manner. Finally, AK15KO mice demonstrate accelerated lipolysis and altered systemic energetics (e.g., locomotion, ketogenesis) during fasting conditions. Our study identifies adipose KLF15 as an essential regulator of adipocyte lipid metabolism and systemic energy balance.

Over the course of a 24-h day, demand on the heart rises and falls with the sleep/wake cycles of the organism. Cardiac metabolism oscillates appropriately, with the relative contributions of major energy sources changing in a circadian fashion. The cardiac peripheral clock is hypothesized to drive many of these changes, yet the precise mechanisms linking the cardiac clock to metabolism remain a source of intense investigation. Here we summarize the current understanding of circadian alterations in cardiac metabolism and physiology, with an emphasis on novel findings from unbiased transcriptomic studies. Additionally, we describe progress in elucidating the links between the cardiac peripheral clock outputs and cardiac metabolism, as well as their implications for cardiac physiology.

The texture evolution during hot compression of extruded ZK60A-T5 magnesium alloy plate loaded along the extrusion direction (ED) and the normal direction (ND) has been examined with the help of pole figures obtained on specimens deformed in the ranges of 200 &deg, C to 500 &deg, C and 0.0003 s&minus, 1 to 10 s&minus, 1. The results are interpreted in terms of the operating slip systems and mechanisms identified based on processing maps developed for the above two initial specimen orientations. The processing map for the initial ED orientation exhibited three domains. In Domains 1 and 3, first-order pyramidal slip {10 1 &macr, l} &lt, 11 2 &macr, 0&gt, occurs, while in Domain 2, second-order pyramidal slip {11 2 &macr, 2} &lt, 3&gt, occurs. The pole figures obtained on specimens deformed in Domains 1 and 3 are strikingly similar, indicating that the operating slip system controls the texture evolution. Compression in Domains 1 and 3 nearly randomizes the intense basal texture in the as-received specimens, while a new texture is generated in Domain 2 with basal poles at 45&deg, to ND or transverse direction (TD). This new texture will promote basal slip when loaded in a transverse direction. When loaded in the normal direction (ND), the processing map exhibited four domains. In Domains 1 and 4, {10 1 &macr, slip occurs, while {11 2 &macr, slip occurs in Domains 2 and 3. The pole figures obtained from specimens deformed in Domains 1 and 4 have similar features, while those deformed in Domains 2 and 3 exhibited similar features to one another, confirming that the operating slip systems control the texture development since they are the same in each pair. The compression along ND produces strong basal textures with the basal planes normal to the ED. The texture gets intensified with increased temperature of deformation and causes strong anisotropy in mechanical properties.

Chronic glucocorticoid therapy has serious side effects, including diabetes and fatty liver. However, the molecular mechanisms responsible for steroid-induced diabetes remain largely enigmatic. Here, we show that hepatic Krüppel-like factor 9 (Klf9) gene expression is induced by dexamethasone and fasting. The overexpression of Klf9 in primary hepatocytes strongly stimulated Pgc1a gene expression through direct binding to its promoter, thereby activating the gluconeogenic program. However, Klf9 mutation abolished the stimulatory effect of dexamethasone on cellular glucose output. Adenovirus-mediated overexpression of KLF9 in the mouse liver markedly increased blood glucose levels and impaired glucose tolerance. Conversely, both global Klf9-mutant mice and liver-specific Klf9-deleted mice displayed fasting hypoglycemia. Moreover, the knockdown of Klf9 in the liver in diabetic mouse models, including ob/ob and db/db mice, markedly lowered fasting blood glucose levels. Notably, hepatic Klf9 deficiency in mice alleviated hyperglycemia induced by chronic dexamethasone treatment. These results suggest a critical role for KLF9 in the regulation of hepatic glucose metabolism and identify hepatic induction of KLF9 as a mechanism underlying glucocorticoid therapy–induced diabetes.

Skeletal muscle lipid metabolism significantly modulates systemic metabolic status; unsurprisingly, aberrant skeletal muscle nutrient handling is closely associated with metabolic diseases. The mechanisms underlying skeletal muscle regulation of lipid metabolism remain to be fully elucidated. Previous studies have shown that mice deficient in nuclear receptor PPARδ suffer from deranged skeletal muscle lipid handling; a similar phenotype is observed in mice deficient in the transcription factor Krüppel-like factor 15 (KLF15). Preliminary data support the existence of KLF15-PPARδ cooperativity in regulating skeletal muscle lipid metabolism, and thus we sought to 1) define the metabolic role of skeletal muscle specific KLF15; and 2) elucidate the molecular basis of KLF15-PPARδ interaction and impact on skeletal muscle lipid metabolism. We generated a skeletal muscle specific KLF15 knockout (K15-SKO) mouse and characterized the metabolic phenotype of this animal. K15-SKO mice have increased body weight and fat mass, elevated circulating free-fatty acids and triglyceride, insulin insensitivity, and glucose intolerance compared to controls. Importantly, K15-SKO mice demonstrated decreased skeletal muscle expression of a number of lipid flux genes, many of which are targets of PPARδ. To determine the necessity of KLF15 for PPARδ-mediated gene expression, we depleted KLF15 in C2C12 cells and looked at a number of PPARδ targets. The expression levels of Fatp1, Cpt1b, and Scl25a20 were attenuated - this response was unchanged in the presence or absence of GW501516 (GW), a PPARδ agonist. We used Seahorse cell metabolism analyzer to assess fatty acid oxidation and observed that knockdown of KLF15 reduces GW induction of palmitate oxygen consumption rates. We conducted co-transfection studies and determined that KLF15 and PPARδ act synergistically on the Fatp1 promoter. Co-immunoprecipitation studies confirmed physical interaction between KLF15 and PPARδ. Finally, control and K15-SKO mice were gavaged with GW for 10 days, and K15-SKO animals demonstrated attenuated induction of a number of PPARδ targets in skeletal muscle. Taken together, these data suggest that skeletal muscle specific KLF15 is critical in the regulation of lipid handling and necessary for optimal PPARδ-mediated regulation of skeletal muscle lipid metabolism. References: 1. Manickam, R., Wahli, W. Roles of peroxisome proliferator-activated receptor β/δ in skeletal muscle physiology. Biochimie136 (2017). 2. Haldar, S.M., et al. Kruppel-like factor 15 regulates skeletal muscle lipid flux and exercise adaptation. Proc Natl Acad Sci U S A109 (2012). 3. Tanaka, T., et al. Activation of peroxisome proliferator-activated receptor delta induces fatty acid beta-oxidation in skeletal muscle and attenuates metabolic syndrome. Proc Natl Acad Sci U S A100 (2003). Funding sources: NCATS, T32; NHLBI, T32; NIDDK, R01 Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. Abstracts presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.

DNA methylation is an epigenetic mark that controls gene expression in response to internal and environmental cues. In this study, we sought to understand the role of DNA methylation in response to desiccation and salinity stresses in three rice cultivars (IR64, stress-sensitive; Nagina 22, drought-tolerant and Pokkali, salinity-tolerant) via bisulphite sequencing. We identified DNA methylation patterns in different genomic/genic regions and analysed their correlation with gene expression. Methylation in CG context within gene body and methylation in CHH context in distal promoter regions were positively correlated with gene expression. However, methylation in other sequence contexts and genic regions was negatively correlated with gene expression. DNA methylation was found to be most dynamic in CHH context under stress condition(s) in the rice cultivars. The expression profiles of genes involved inde-novomethylation were correlated with methylation dynamics. Hypomethylation in Nagina 22 and hypermethylation in Pokkali in response to desiccation and salinity stress, respectively, were correlated with higher expression of abiotic stress response related genes. Our results suggest an important role of DNA methylation in abiotic stress responses in rice in cultivar-specific manner. This study provides useful resource of DNA methylomes that can be integrated with other data to understand abiotic stress response in rice.HighlightBisulphite sequencing revealed single base resolution DNA methylation, and cultivar-specific differential methylation patterns and correlation with gene expression that control desiccation and salinity stress response in the rice cultivars.

Dermatopathy as an initial manifestation of cystic fibrosis (CF) in the newborn period is unusual. The eruption is usually first noted in the perineum, typically appears in infancy from age 2 weeks to 15 months after birth. It subsequently spreads to the extremities and trunk. We report a two months old male baby who presented with failure to thrive, hypoproteinaemia, anaemia and a cutaneous eruption resembling acrodermatitis enteropathica. Oral zinc supplementation resulted in temporary resolution of the dermatitis. A further work-up revealed the diagnosis of cystic fibrosis.The rash was responsive to nutritional and pancreatic enzyme supplementation.

Background Limb ischemia resulting from peripheral vascular disease is a common cause of morbidity. Vessel occlusion limits blood flow, creating a hypoxic environment that damages distal tissue, requiring therapeutic revascularization. Hypoxia‐inducible factors ( HIF s) are key transcriptional regulators of hypoxic vascular responses, including angiogenesis and arteriogenesis. Despite vascular smooth muscle cells’ ( VSMC s’) importance in vessel integrity, little is known about their functional responses to hypoxia in peripheral vascular disease. This study investigated the role of VSMC HIF in mediating peripheral ischemic responses. Methods and Results We used Arnt SMKO mice with smooth muscle–specific deletion of aryl hydrocarbon receptor nuclear translocator (ARNT, HIF ‐1β), required for HIF transcriptional activity, in a femoral artery ligation model of peripheral vascular disease. Arnt SMKO mice exhibit impaired perfusion recovery despite normal collateral vessel dilation and angiogenic capillary responses. Decreased blood flow manifests in extensive tissue damage and hypoxia in ligated limbs of Arnt SMKO mice. Furthermore, loss of aryl hydrocarbon receptor nuclear translocator changes the proliferation, migration, and transcriptional profile of cultured VSMC s. Arnt SMKO mice display disrupted VSMC morphologic features and wrapping around arterioles and increased vascular permeability linked to decreased local blood flow. Conclusions Our data demonstrate that traditional vascular remodeling responses are insufficient to provide robust peripheral tissue reperfusion in Arnt SMKO mice. In all, this study highlights HIF responses to hypoxia in arteriole VSMC s critical for the phenotypic and functional stability of vessels that aid in the recovery of blood flow in ischemic peripheral tissues.

Cardiac metabolism is highly adaptive in response to changes in substrate availability, as occur during fasting. This metabolic flexibility is essential to the maintenance of contractile function and is under the control of a group of select transcriptional regulators, notably the nuclear receptor family of factors member PPARα. However, the diversity of physiologic and pathologic states through which the heart must sustain function suggests the possible existence of additional transcriptional regulators that play a role in matching cardiac metabolism to energetic demand. Here we show that cardiac KLF15 is required for the normal cardiac response to fasting. Specifically, we find that cardiac function is impaired upon fasting in systemic and cardiac specific Klf15-null mice. Further, cardiac specific Klf15-null mice display a fasting-dependent accumulation of long chain acylcarnitine species along with a decrease in expression of the carnitine translocase Slc25a20. Treatment with a diet high in short chain fatty acids relieves the KLF15-dependent long chain acylcarnitine accumulation and impaired cardiac function in response to fasting. Our observations establish KLF15 as a critical mediator of the cardiac adaptive response to fasting through its regulation of myocardial lipid utilization.

The transcription factor Kruppel-like factor 2 (KLF2) displays anticarcinogenic activities but the mechanism that underlies this activity is unknown. We show here that KLF2 is markedly downregulated in human breast cancers and that its expression positively correlates with breast cancer patient survival. We show further that KLF2 suppresses tumor development by controlling the transcriptional activity of the vitamin A metabolite retinoic acid (RA). RA regulates gene transcription by activating two types of nuclear receptors: RA receptors (RARs), which inhibit tumor development, and peroxisome proliferator-activated receptor β/δ (PPARβ/δ), which promotes tumorigenesis. The partitioning of RA between these receptors is regulated by two carrier proteins: cellular retinoic acid-binding protein 2 (CRABP2), which delivers RA to RARs, and fatty acid-binding protein 5 (FABP5), which shuttles ligands to PPARβ/δ. We show that KLF2 induces the expression of CRABP2 and RARγ and inhibits the expression FABP5 and PPARβ/δ thereby shifting RA signaling from the pro-carcinogenic FABP5/PPARβ/δ to the growth-suppressing CRABP2/RAR path. The data thus reveal that KLF2 suppresses tumor growth by controlling the transcriptional activities of RA.

Maintenance of vascular integrity in the adult animal is needed for survival, and it is critically dependent on the endothelial lining, which controls barrier function, blood fluidity, and flow dynamics. However, nodal regulators that coordinate endothelial identity and function in the adult animal remain poorly characterized. Here, we show that endothelial KLF2 and KLF4 control a large segment of the endothelial transcriptome, thereby affecting virtually all key endothelial functions. Inducible endothelial-specific deletion of Klf2 and/or Klf4 reveals that a single allele of either gene is sufficient for survival, but absence of both (EC-DKO) results in acute death from myocardial infarction, heart failure, and stroke. EC-DKO animals exhibit profound compromise in vascular integrity and profound dysregulation of the coagulation system. Collectively, these studies establish an absolute requirement for KLF2/4 for maintenance of endothelial and vascular integrity in the adult animal.

Mononuclear phagocytes are a heterogeneous family that occupy all tissues and assume numerous roles to support tissue function and systemic homeostasis. Our ability to dissect the roles of individual subsets is limited by a lack of technologies that ablate genefunction within specific mononuclear phagocyte sub-populations. Using Nr4a1-dependent Ly6Clow monocytes, we present a proof-of-principle approach that addresses these limitations. Combining ChIP-seq and molecular approaches we identified a single, conserved, sub-domain within the Nr4a1 enhancer that was essential for Ly6Clow monocyte development. Mice lacking this enhancer lacked Ly6Clow monocytes but retained Nr4a1 gene expression in macrophages during steady state and in response to LPS. Because Nr4a1 regulates inflammatory gene expression and differentiation of Ly6Clow monocytes, decoupling these processes allows Ly6Clow monocytes to be studied independently.

Non-coding RNAs constitute a major portion of the transcriptome in most of eukaryotes. Long non-coding transcripts originating from the DNA segment present between the protein coding genes are termed as long intergenic non-coding RNAs (lincRNAs). Several evidences suggest the role of lincRNAs in regulation of various biological processes. In this study, we identified a total of 2248 lincRNAs in chickpea using RNA-seq data from eight successive stages of flower development and three vegetative tissues via an optimized pipeline. Different characteristic features of lincRNAs were studied and compared with those of predicted mRNAs in chickpea. Further, we utilized a method using network propagation algorithm to reveal the putative function of lincRNAs in plants. In total, at least 79% of the identified chickpea lincRNAs were assigned with a putative function. A comprehensive expression profiling revealed differential expression patterns and tissue specificity of lincRNAs in different stages of flower development in chickpea. In addition, potential lincRNAs-miRNA interactions were explored for the predicted lincRNAs in chickpea. These findings will pave the way for understanding the role of lincRNAs in the regulatory mechanism underlying flower development in chickpea and other legumes.

The adaptive immune response is tightly regulated by complex signals in dendritic cells (DCs). Although Th2 polarization is dictated by defined functional DC subsets, the molecular factors that govern the amplitude of these responses are not well understood. Krüppel-like factor 2 (KLF2) is a transcription factor that negatively regulates the activation of numerous immune cells in response to stimuli. Here, we demonstrate that suppression of KLF2 in conditioned DCs preferentially amplifies Th2 responses in two model systems, one of which is a prototypical intracellular pathogen and the other an allergen. This elevation in Th2 responses was dependent on contact-mediated Notch signaling in vitro and in vivo. A deficiency of KLF2 increased the expression of Notch ligand Jagged2 via hypoxia-inducible factor 1α (HIF-1α), which led to Th2 amplification. Our results revealed a novel circuit in DCs for Th2 polarization that is governed by KLF2., IMPORTANCE Dendritic cells are the key element that bridges innate and adaptive immunity. A complex and not-well-understood area in dendritic cell biology is the regulatory network that predetermines or moderates their function to shape the adaptive immune response. Our study for the first time demonstrates that KLF2, a transcription factor, conditions dendritic cells to regulate Th2 responses via a Jagged2/Notch axis. Downregulation of KLF2 expression in dendritic cells may provide a beneficial effect for treatment of diseases such as obesity or parasitic infections but may be deleterious in the case of invasion by intracellular pathogens. Strategies to tune KLF2 may be useful for future therapeutic approaches to particular diseases of mankind.

The phenomenon of exercise‐induced hemoptysis is still relatively underrecognised in humans. We report a case of recurrent hemoptysis brought on by vigorous exercise. A 33‐year‐old male presented with several episodes of intermittent fresh small‐volume hemoptysis reproducible on vigorous exercise. There was no other significant medical history other than a past history of testicular tumor, treated with orchidectomy and adjuvant Bleomycin‐based chemotherapy 1 year prior to onset of symptoms. Computed tomography scan showed no major abnormalities other than few small bilateral non‐specific nodules. Computed tomography aortogram and pulmonary angiogram, ventilation/perfusion scan, and echocardiography yielded no significant abnormalities. Infectious, autoimmune disease, coagulopathy, vasculitis, and malignant causes were excluded. Bronchoscopy showed possible endobronchial bleeding. This phenomenon is thought to be due to vulnerability of pulmonary capillaries to stress or mechanical failure during strenuous exercise at high cardiorespiratory workload.

Background— Although metabolic reprogramming is critical in the pathogenesis of heart failure, studies to date have focused principally on fatty acid and glucose metabolism. Contribution of amino acid metabolic regulation in the disease remains understudied. Methods and Results— Transcriptomic and metabolomic analyses were performed in mouse failing heart induced by pressure overload. Suppression of branched-chain amino acid (BCAA) catabolic gene expression along with concomitant tissue accumulation of branched-chain α-keto acids was identified as a significant signature of metabolic reprogramming in mouse failing hearts and validated to be shared in human cardiomyopathy hearts. Molecular and genetic evidence identified the transcription factor Krüppel-like factor 15 as a key upstream regulator of the BCAA catabolic regulation in the heart. Studies using a genetic mouse model revealed that BCAA catabolic defect promoted heart failure associated with induced oxidative stress and metabolic disturbance in response to mechanical overload. Mechanistically, elevated branched-chain α-keto acids directly suppressed respiration and induced superoxide production in isolated mitochondria. Finally, pharmacological enhancement of branched-chain α-keto acid dehydrogenase activity significantly blunted cardiac dysfunction after pressure overload. Conclusions— BCAA catabolic defect is a metabolic hallmark of failing heart resulting from Krüppel-like factor 15–mediated transcriptional reprogramming. BCAA catabolic defect imposes a previously unappreciated significant contribution to heart failure.

Chickpea ranks third among the food legume crops production in the world. However, the genomic resources available for chickpea are still very limited. In the present study, the transcriptome of chickpea was sequenced with short reads on Illumina Genome Analyzer platform. We have assessed the effect of sequence quality, various assembly parameters and assembly programs on the final assembly output. We assembled ∼107million high-quality trimmed reads using Velvet followed by Oases with optimal parameters into a non-redundant set of 53 409 transcripts (≥100 bp), representing about 28 Mb of unique transcriptome sequence. The average length of transcripts was 523 bp and N50 length of 900 bp with coverage of 25.7 rpkm (reads per kilobase per million). At the protein level, a total of 45 636 (85.5%) chickpea transcripts showed significant similarity with unigenes/predicted proteins from other legumes or sequenced plant genomes. Functional categorization revealed the conservation of genes involved in various biological processes in chickpea. In addition, we identified simple sequence repeat motifs in transcripts. The chickpea transcripts set generated here provides a resource for gene discovery and development of functional molecular markers. In addition, the strategy for de novo assembly of transcriptome data presented here will be helpful in other similar transcriptome studies.