Comparative secretome study of brown adipocytes and the role of ITIH4 in adipose biology

Adipose tissues in mammals can be broadly classified into two main types: white and brown adipose tissue. Although both are defined as adipose tissues, they differ drastically in their function. The main function of white adipose tissues (WAT) is the storage of fat. Unlike its white counterpart, brown adipose tissue (BAT) specializes in burning fat via thermogenesis and is known to play an important role in non-shivering thermogenesis especially in hibernating animals and newborn babies. Recent evidence of functional BAT in adult humans and its ameliorating effect on metabolic disorders has brought BAT under the spotlight for treatment of metabolic diseases like obesity and type 2 diabetes mellitus. WAT also acts as an endocrine organ by secreting signaling molecules called adipokines such as leptin and adiponectin. Adipokines constitute the secretome of WAT and not only play an important role in WAT function but also affect whole-body energy homeostasis. Various studies have investigated the role of adipose tissue secretome in metabolic disorders like obesity and insulin resistance. The WAT secretome has also been extensively characterized in various settings such as in whole WAT, mature white adipocyte etc. However, the BAT secretome and its adipokines (‘batokines’) have not yet been investigated. Thus, the main aim of this dissertation work was a comparative study of the white and brown adipocyte secretomes using a combination of Click-iT® AHA labeling and pulsed-SILAC (stable isotope labeling by amino-acid in cell culture). In total 1013 proteins were detected and a subset of these proteins was selected based on their secretion with norepinephrine stimulation. An in vitro assay was developed and optimized to test their putative effect on insulin secretion. In addition, one of the secretome candidates, inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) was investigated as a potential batokine and BAT activity marker. Although, the serum levels of ITIH4 did not correlate with BAT activity under cold stimulation, its expression was found to increase with adipogenesis and browning of white adipocytes. Using in vitro knockdown studies, a reduction in differentiation was observed which was characterized by reduction in mature adipocyte functions such as lipolysis, lipid and intracellular triglyceride storage, glucose uptake and lipogenesis. Therefore, rather than being a batokine, ITIH4 was shown to be important for adipogenesis and adipocyte biology. In summary, this dissertation sheds light on BAT secreted proteins and also introduces a new player in field of adipogenesis, both of which might have a significant impact in BAT biology and in the treatment of metabolic disorders like obesity

Perception of the healthcare professionals towards the current trauma and emergency care system in Kabul, Afghanistan: A mixed method study

Background: Trauma and injury contribute to 11% of the all-cause mortality in Afghanistan. The study aimed to explore the perceptions of the healthcare providers (pre and in-hospital), hospital managers and policy makers of the public and private health sectors to identify the challenges in the provision of an effective trauma care in Kabul, Afghanistan.Methods: A concurrent mixed method design was used, including key-informant interviews (healthcare providers, hospital managers and policy makers) of the trauma care system (N = 18) and simultaneous structured emergency care system assessment questionnaire (N = 35) from July 15 to September 25, 2019. Interviews were analyzed using content analysis approach and structured questionnaire data were descriptively analyzed.Results: Four themes were identified that describe the challenges: 1) pre-hospital care, 2) cohesive trauma management system, 3) physical and human resources and 4) stewardship. Some key challenges were found related to scene and transportation care, in-hospital care and emergency preparedness within the wider trauma care system. Less than 25% of the population is covered by the pre-hospital ambulance system (n = 23, 65.7%) and there is no communication process between health care facilities to facilitate transfer (n = 28, 80%). Less than 25% of patients with an injury requiring emergent surgery have access to surgical care in a staffed operating theatre within 2 h of injury (n = 19, 54.2%) and there is no regular assessment of the ability of the emergency care system to mobilize resources (human and physical) to respond to disasters, and other large-scale emergencies (n = 28, 80%).Conclusion: This study highlighted major challenges in the delivery of trauma care services across Kabul, Afghanistan. Systematic improvement in the workforce training, structural organization of the trauma care system and implementing externally validated clinical guidelines for trauma management could possibly enhance the functions of the existing trauma care services. However, an integrated state-run trauma care system will address the current burden of traumatic injury more effectively within the wider healthcare system of Afghanistan

Helmet wearing saves the cost of motorcycle head injuries: A case study from Karachi, Pakistan

Purpose: To estimate the difference in healthcare cost of head injuries among motorcycle helmet users and non-users.Methods: Motorcycle crash victims with head injuries that were brought to a public, tertiary care emergency room in Karachi were studied through a descriptive cross-sectional design. A standard questionnaire was used to collect data on demographics, injury pattern, helmeting practice, length of hospital stay, out-of-pocket payments (OOPs), and healthcare service utilization at the facility to estimate total healthcare and other costs applying micro-costing methods during the hospitalization period.Results: A total of 323 motorcyclists involved in crash were brought to a public tertiary care ER, 112 patients had head injuries and were enrolled in the study. The helmeted motorcyclists had a significantly lower median total healthcare cost of PKR 10,796 ($69) [IQR 9851 ($63)-PKR 12,581 ($80)] compared to higher cost of PKR 12,113 ($77) [IQR 10,431 ($66)-50,545 ($322)] (p value = 0.046) in non-helmeted. Helmet users expended significantly less cost on laboratory tests, PKR 365 ($2) [IQR 365 ($2)-548 ($3)] compared to PKR 3650 ($23) [IQR 365 ($2)-5840 ($37)] (p value =0.027) among non-users. Furthermore, cost of radiological investigations was also low among helmeted patients compared to non-helmeted ones, median PKR 4096 ($26) [IQR 3166 ($20)-5678 ($36)] vs 4750 ($30) [3166 ($20)-11,358 ($72)] (p value =0.049). The out-of-pocket payments (OOPs) for healthcare services were lower among helmet users as compared to non-users, with cost of PKR 17,750 ($113) [IQR 16,650 ($106)-18,000 ($115)] vs PKR 19,800 ($126) [IQR 12,300 ($78)-30,900 ($197)] (p value =0.03), respectively.Conclusion: The result of this study demonstrates that helmet use among motorcyclists significantly reduced healthcare costs and healthcare resource utilizations during hospitalization for head injuries in Pakistan. Thus, it is important to implement strict helmet wearing laws to decrease head injuries and the cost burden on the healthcare facility and patients

Public exposure from I-131 hospitalized isolated patients in NIMRA Jamshoro Pakistan

Purpose/ Background: To treat the cancerous tissues the unsealed radioisotopes are being in clinical practice since 7 decades. From these unsealed sources, I-131 is the choice of treatment for the treating thyroid cancers. Orally administered I-131 patients are to be kept isolated in hospital for some period until captured activity in the body reaches to national and international limits for the avoidance of unacceptably high radiation exposures to patients' family members and the general public to keep ALARA (as low as reasonably achievable) principal in mind. The main rationale of this study was to calculate the exposure/dose of the general public. Material and Methods: This work presents the exposure rate and dose to the general public in the corridor and the non-radioactive patients admitted in adjacent room from I-131 administered isolated patients treated at NIMRA (Nuclear Institute of Medicine and Radiotherapy) Jamshoro Pakistan. In this study exposure from 23 thyroid cancer patients treated with different activities of I-131 (50 to 200 mCi) from January 2011 to December 2012 were included. Results: The average exposure rate in the corridor was about 5.17 µSv/hr (2.14 µSv/hr to 8.15 µSv/hr) and the cumulative dose to nonradioactive patients residing in an adjacent room was 0.647 mSv (0.192 mSv to 1.664 mSv). Conclusion: This study concludes that the exposure rate to the general public especially the admitted non-radioactive patient in the adjacent room is almost within the limits of 1 mSv as specified in national and international standards.-----------------------------------------Cite this article as: Memon SA, Laghari NA, Qureshi ST, Ahmad A, Khan AA, Hussain MM. Public exposure from I-131 hospitalized isolated patients in NIMRA Jamshoro Pakistan. Int J Cancer Ther Oncol 2014; 2(2):020214. DOI: 10.14319/ijcto.0202.1

Drug safety Africa: An overview of safety pharmacology & toxicology in South Africa.

This meeting report is based on presentations given at the first Drug Safety Africa Meeting in Potchefstroom, South Africa from November 20-22, 2018 at the North-West University campus. There were 134 attendees (including 26 speakers and 34 students) from the pharmaceutical industry, academia, regulatory agencies as well as 6 exhibitors. These meeting proceedings are designed to inform the content that was presented in terms of Safety Pharmacology (SP) and Toxicology methods and models that are used by the pharmaceutical industry to characterize the safety profile of novel small chemical or biological molecules. The first part of this report includes an overview of the core battery studies defined by cardiovascular, central nervous system (CNS) and respiratory studies. Approaches to evaluating drug effects on the renal and gastrointestinal systems and murine phenotyping were also discussed. Subsequently, toxicological approaches were presented including standard strategies and options for early identification and characterization of risks associated with a novel therapeutic, the types of toxicology studies conducted and relevance to risk assessment supporting first-in-human (FIH) clinical trials and target organ toxicity. Biopharmaceutical development and principles of immunotoxicology were discussed as well as emerging technologies. An additional poster session was held that included 18 posters on advanced studies and topics by South African researchers, postgraduate students and postdoctoral fellows

HAND2 is a novel obesity-linked adipogenic transcription factor regulated by glucocorticoid signalling

Aims/hypothesis Adipocytes are critical cornerstones of energy metabolism. While obesity-induced adipocyte dysfunction is associated with insulin resistance and systemic metabolic disturbances, adipogenesis, the formation of new adipocytes and healthy adipose tissue expansion are associated with metabolic benefits. Understanding the molecular mechanisms governing adipogenesis is of great clinical potential to efficiently restore metabolic health in obesity. Here we investigate the role of heart and neural crest derivatives-expressed 2 (HAND2) in adipogenesis.MethodsHuman white adipose tissue (WAT) was collected from two cross-sectional studies of 318 and 96 individuals. In vitro, for mechanistic experiments we used primary adipocytes from humans and mice as well as human multipotent adipose-derived stem (hMADS) cells. Gene silencing was performed using siRNA or genetic inactivation in primary adipocytes from loxP and or tamoxifen-inducible Cre-ERT2 mouse models with Cre-encoding mRNA or tamoxifen, respectively. Adipogenesis and adipocyte metabolism were measured by Oil Red O staining, quantitative PCR (qPCR), microarray, glucose uptake assay, western blot and lipolysis assay. A combinatorial RNA sequencing (RNAseq) and ChIP qPCR approach was used to identify target genes regulated by HAND2. In vivo, we created a conditional adipocyte Hand2 deletion mouse model using Cre under control of the Adipoq promoter (Hand2AdipoqCre) and performed a large panel of metabolic tests.Results We found that HAND2 is an obesity-linked white adipocyte transcription factor regulated by glucocorticoids that was necessary but insufficient for adipocyte differentiation in vitro. In a large cohort of humans, WAT HAND2 expression was correlated to BMI. The HAND2 gene was enriched in white adipocytes compared with brown, induced early in differentiation and responded to dexamethasone (DEX), a typical glucocorticoid receptor (GR, encoded by NR3C1) agonist. Silencing of NR3C1 in hMADS cells or deletion of GR in a transgenic conditional mouse model results in diminished HAND2 expression, establishing that adipocyte HAND2 is regulated by glucocorticoids via GR in vitro and in vivo. Furthermore, we identified gene clusters indirectly regulated by the GR-HAND2 pathway. Interestingly, silencing of HAND2 impaired adipocyte differentiation in hMADS and primary mouse adipocytes. However, a conditional adipocyte Hand2 deletion mouse model using Cre under control of the Adipoq promoter did not mirror these effects on adipose tissue differentiation, indicating that HAND2 was required at stages prior to Adipoq expression.Conclusions/interpretation In summary, our study identifies HAND2 as a novel obesity-linked adipocyte transcription factor, highlighting new mechanisms of GR-dependent adipogenesis in humans and mice.Data availability Array data have been submitted to the GEO database at NCBI (GSE148699).</p

“Takotsubo cardiomyopathy presenting with QT prolongation: an atypical presentation.”

Takotasubo Cardiomyopathy (TCM), also known as broken heart syndrome, is a syndrome of transient and reversible cardiac dysfunction in the absence of obstructive coronary artery disease following an unpleasant emotional event. TCM commonly presents as chest pain mimicking acute coronary syndrome (ACS) in character. The most common electrocardiogram (EKG) findings associated with TCM are ST-segment elevation or T wave inversion. Herein, we present a case of TCM in an individual who presented with a sudden onset chest pressure following a stressful event. She was found to have QT interval prolongation on EKG and elevated troponins. Initially thought to have non-ST elevation myocardial infarction (NSTEMI). Cardiac catheterization did not reveal coronary artery disease. The ventriculography suggested apical ballooning and the diagnosis of TCM with atypical EKG presentation. Our case is unique as we describe an atypical electrocardiographic presentation of TCM. Our case emphasizes that physicians should refrain from prescribing QT-prolonging drugs to patients with TCM, to avert potential QT prolongation and progression to torsade de pointes (TDP)

Road safety hazards for children while commuting to school: Findings from a pilot study in Karachi, Pakistan

Introduction: Road traffic injuries are a leading cause of mortality and morbidity among children. Travelling to and from school is a major risk exposure for children around the globe.Objective: The purpose of this study was to assess road traffic injury hazards for school children during dropp-off or picked-up times.Methods: This observational cross-sectional study included 94 public and private schools in Karachi, Pakistan. A structured observational tool was used to collect data on school demographics, the road traffic environment, infrastructure, injury hazards in vehicles used by school children, and child pedestrian injury risk and road use behaviors.Results: A total of 860 observations of school children, drivers of vehicles transporting children, schools, and vehicles were recorded. Most schools (n = 83, 88%) did not have designated parking spaces around the school; only one public school had a parking area. Only one private school had a zebra crossing around the school premises. Very few schools (n = 13, 14%), mostly private (n = 12) had pedestrian sidewalks. Only 35 (18%) adult motorcyclists, out of 199, were wearing a helmet, and eight (6%), out of 145, car passengers were wearing seatbelts. Compressed natural gas (CNG) cylinders were installed in 83 (35%), out of 235, observed vehicles. The remaining 152 (65%) did not have CNG cylinders or they were not visible to our data collectors. In 55 (23%) observations, bus passengers stepped off the bus in the middle of the road. Most pedestrians (n = 266, 99.5%) did not use a Zebra crossing. More than a quarter (n = 74, 28%) of pedestrians looked left and right before crossing the road.Conclusion: While traveling to school, either by walking or taking vehicular trips, children face many road traffic injury hazards in Karachi. Pedestrians and passengers exhibited risky behaviors while using roads. Further initiatives are advised from a public health viewpoint aiming at minimizing transport-related hazards

Tip-over injuries among children: Data from an urban emergency department of Karachi, Pakistan

Introduction: Most unintentional injuries among children occur in the home environment. Tip-overs, defined as incidents where heavy objects fall on children due to some type of interaction, are one of the reasons for injuries inside the home. This study aims to determine injury patterns and outcomes for child injuries resulting from tip-overs in the home environment as reported in the emergency department.Methods: We performed a retrospective chart review of pediatric (under 18 years) tip-overs injuries occurred in years 2010 to 2015 at the Aga Khan University Hospital. Furthermore, parents of injured children participated in phone interviews to provide information about the injury scene. File review and telephonic interviews were conducted in the year 2015 and 2016.Results: A total of 75 children visited the emergency department with tip-over injuries, out of which 55 (73%) were boys. The majority of incidents (75.5%) happened inside the home, and the most common places were the living room and bedroom (32% and 21% respectively). More than half (53%) of the children were not under adult supervision at the time of the incident and less than half (47%) of the household took safety measures after the incident. Tip-over injuries were common among 3-year-old children with decreasing frequency as children grew older. The most common causes of tip-overs were TV/TV trolley (32%), followed by furniture (28%), and wall and roof (23%). The most common sites of injuries were head (n = 33, 44%) and extremities (n = 33, 44%). A majority of the cases (n = 66, 88%) were admitted to the hospital from the emergency department, under care of both general (n = 51, 68%) and critical care units (n = 15, 20%). More than a quarter (n = 27, 36%) required at least one surgical procedure during their hospital stay. The median length of hospital stay was one days (interquartile range, IQR 1-5 days). There were two cases of mortality (3%).Conclusion: Most tip-over injuries among children were caused by TV, furniture, and TV trolleys. These injuries can be prevented with public education around home safety measures, such as mounting them on the wall