MOLECULAR CLONING AND ANALYSIS OF THE PROMOTER FOR RAT HEPATIC NEUTRAL CHOLESTEROL ESTER HYDROLASE

Neutral cholesterol ester hydrolase (CEH) is a key enzyme in regulating hepatic free cholesterol. Using the CEH specific cDNA sequence in the 5\u27-untranslated region as a primer, 1.3 kb of sequence upstream of the ATG initiation codon was amplified and cloned. Primer extension analysis with total RNA from rat primary hepatocytes identified a transcription initiation site, 60 bases upstream from the initiation codon. No typical TATA-box sequences were found upstream from the transcription start site. However, a consensus GC-box, which can bind the positive transcription factor SP1, was found 35 bases upstream from the transcription start site. In addition the promoter also contained several hormone responsive half elements, sterol response elements, ubiquitous transcription factor binding sites and liver specific elements. To determine the promoter activity of the rat CEH gene, the 1.3 kb of 5’-flanking region was fused to a luciferase reporter gene. Smaller 5’-deletion constructs were obtained by generation of unidirectional nested deletion breakpoints in the full-length construct with Exonuclease III. Basal promoter activity, as well as transcriptional regulation by hormones, signal transduction pathways and agents perturbing cholesterol metabolism were studied in human hepatoblastoma HepG2 cells and cultured primary rat hepatocytes by transient transfection assays of the promoter activity of the deletion constructs. Functional glucocorticoid response elements, phorbol ester responsive sequences and sterol responsive sequences were mapped with both the culture systems. Results indicate that the first 599 base pairs upstream of the initiation codon and the region between nucleotides -1317 and -1190 regulate the effects of various physiological stimuli. The effects of various stimuli used in this study were similar in the two cell lines. The rat CEH gene appears to be finely regulated by distinct signals converging to consensus promoter regulatory sequences

The changing trends and tendencies in maternal mortality: a compare and contrast spanning two decades

Background: Though the maternal mortality ratio has been on the constant decline over two decades we shall strive forward to prevent all preventable maternal deaths and we have a long way to reach the SDG 3.1 goal. So, it becomes imperative to further reduce the mortality rates by continuously reviewing and upgrading our health policies. The aim of the study was to compare the trends in maternal mortality in 2002-2011 to the current predicament in 2017- 2019 in our hospital.Methods: Data was collected for all maternal deaths in the Institute of obstetrics and gynaecology, Egmore, Chennai- a centre of excellence for maternal care in South India, in terms of age at time of death, parity, period of death, mode of delivery, the cause of death and the admission to delivery interval and compared between the time periods of 2002-2011 and 2017-2019 to identify the changing patterns.Results: The maternal mortality ratio has consistently declined over the years from 220 in 2002 to 110 in 2019. The age group of 20-29 years and primigravidae continue to amount for most mortality. The postpartum period remains the most susceptible period. There is a significant increase in the number of caesarean deaths (64% from 46%). Significant transition in the cause of death from haemorrhage (20% to 7%) to hypertensive disease of pregnancy (23% to 35%) as leading cause of maternal mortality has been witnessed.Conclusions: Significant difference in the trends over two decades in maternal mortality lies in the fact that the leading cause of maternal mortality has changed from haemorrhage to hypertensive diseases and sepsis and they have continued to occupy the top spots for the past decade. Preventive measures are the solution to reduce maternal mortality due to either of the causes

Bisphosphonates-what’s new?

Bisphosphonates are chemical-related to pyrophosphate. The oxygen atom in pyrophosphate is substituted by a carbon atom in these compounds, resulting in a P-C-P bond. They are potent antiresorptive medicines because they have a strong inhibitory effect on osteoclasts. It lowers fracture risk by reducing bone turnover, increasing bone mineral density, and decreasing fracture risk in the lumbar spine and hip. Bisphosphonates are strongly attracted to bone surfaces, where they accumulate, primarily at remodeling sites. They are rarely associated with systemic side effects due to their selectivity in action. Irritation of the upper gastrointestinal tract is the most common side effect. A strong third-generation bisphosphonate, zoledronate, is currently approved to treat postmenopausal-induced osteoporosis. This review mainly focuses on the mechanism of action, therapeutic uses, and its adverse effects

Vitamin C: Current Concepts in Human Physiology

ca. 200 words; this text will present the book in all promotional forms (e.g. flyers). Please describe the book in straightforward and consumer-friendly terms. [Vitamin C is synthesized by almost all animals. However, for humans, it is a vitamin that needs constant replenishment in the diet. While its role as an anti-oxidant and for preventing scurvy have been known for a long time, novel functions and unrecognized associations continue to be identified for this enigmatic molecule. In the past decade, new details have emerged regarding differences in its uptake by oral and intravenous modes. While vitamin C deficiency remains largely unknown and poorly addressed in many segments of the population, novel pharmacological roles for high-dose, intravenous vitamin C in many disease states have now been postulated and investigated. This has shifted its role in health and disease from the long-perceived notion as merely a vitamin and an anti-oxidant to a pleiotropic molecule with a broad anti-inflammatory, epigenetic, and anti-cancer profile. This Special Issue comprises original research papers and reviews on vitamin C metabolism and function that relate to the following topics: understanding its role in the modulation of inflammation and immunity, therapeutic applications and safety of pharmacological ascorbate in disease, and the emerging role of vitamin C as a pleiotropic modulator of critical care illness and cancer.

Intravenous Vitamin C Administered as Adjunctive Therapy for Recurrent Acute Respiratory Distress Syndrome

This case report summarizes the first use of intravenous vitamin C employed as an adjunctive interventional agent in the therapy of recurrent acute respiratory distress syndrome (ARDS). The two episodes of ARDS occurred in a young female patient with Cronkhite-Canada syndrome, a rare, sporadically occurring, noninherited disorder that is characterized by extensive gastrointestinal polyposis and malabsorption. Prior to the episodes of sepsis, the patient was receiving nutrition via chronic hyperalimentation administered through a long-standing central venous catheter. The patient became recurrently septic with Gram positive cocci which led to two instances of ARDS. This report describes the broad-based general critical care of a septic patient with acute respiratory failure that includes fluid resuscitation, broad-spectrum antibiotics, and vasopressor support. Intravenous vitamin C infused at 50 mg per kilogram body weight every 6 hours for 96 hours was incorporated as an adjunctive agent in the care of this patient. Vitamin C when used as a parenteral agent in high doses acts “pleiotropically” to attenuate proinflammatory mediator expression, to improve alveolar fluid clearance, and to act as an antioxidant

Self-Care Behaviours and Glycemic Control among Adults with Type 2 Diabetes

Aim: To explore self-care behaviours and glycemic control among adults with type 2 diabetes. Design: A descriptive cross-sectional design was used for the study. Methods: The revised summary of diabetes mellitus questionnaire was used to collect data and glycated haemaglobin. A random sample comprising of 350 Omani adults with type 2 diabetes were selected between January-June 2010. Structural equation modelling and ANOVA were used for analysis. Results/Findings: One-third of the adults with type 2 diabetes followed diet, foot care and medications (on an average of 3 days/7 days) compared to half percentage of them adhering to foot care. 27% of the total variance in self-car activities was accounted by diet, 32% by exercise and 17% by medications. Blood glucose monitoring, foot care, and smoking and HbA1c accounted for 60%, 78%, and 51% variances. The standardized path coefficients of diet, exercise, smoking, foot care, blood sugar monitoring and medications had a significantly positive influence on self-care behaviours. Conclusion: There were inadequate self-care behaviours among the majority of adults with type 2 diabetes with poor glycaemic control. Nurses should use the self-care management model when designing tailored educational interventions to enhance glycemic control

Ascorbate-dependent vasopressor synthesis: a rationale for vitamin C administration in severe sepsis and septic shock?

Severe systemic inflammatory response to infection results in severe sepsis and septic shock, which are the leading causes of death in critically ill patients. Septic shock is characterised by refractory hypotension and is typically managed by fluid resuscitation and administration of catecholamine vasopressors such as norepinephrine. Vasopressin can also be administered to raise mean arterial pressure or decrease the norepinephrine dose. Endogenous norepinephrine and vasopressin are synthesised by the copper-containing enzymes dopamine β-hydroxylase and peptidylglycine α-amidating monooxygenase, respectively. Both of these enzymes require ascorbate as a cofactor for optimal activity. Patients with severe sepsis present with hypovitaminosis C, and pre-clinical and clinical studies have indicated that administration of high-dose ascorbate decreases the levels of pro-inflammatory biomarkers, attenuates organ dysfunction and improves haemodynamic parameters. It is conceivable that administration of ascorbate to septic patients with hypovitaminosis C could improve endogenous vasopressor synthesis and thus ameliorate the requirement for exogenously administered vasopressors. Ascorbate-dependent vasopressor synthesis represents a currently underexplored biochemical mechanism by which ascorbate could act as an adjuvant therapy for severe sepsis and septic shock

Image Analysis as a Tool for Estimation of Red Peppers’ Color

Red pepper is an excellent source of natural red color. The estimation of color is of great importance, as it is being used for grading red peppers to suit various applications in the food industry. A novel method for quick and easy determination of the color of red peppers by analyzing the color images obtained from a flatbed scanner was developed and validated. ImageJ software was used for measuring the RGB values of the images and the RGB values thus obtained were converted into the industrially accepted American Spice Trade Association (ASTA) color values in red peppers by using an empirical formula. The results were compared with color values obtained through ASTA chemical analytical method. The developed image analysis method was used for the analysis of red peppers with color values ranging from 15 to 154 ASTA units. The image analysis method showed good agreement with the chemical method for color values ranging from 40 to 140 ASTA units. The new method is also fast and easy to adopt and is deemed useful in the field, of processing and storage facilities, where access to sophisticated instrumentation for color estimation and color stability studies is limited.</p

Large-eddy simulations of a turbulent jet impinging on a vibrating heated wall

High-resolution large-eddy simulations (LES) are performed for an incompressible turbulent circular jet impinging upon a vibrating heated wall supplied with a constant heat flux. The present work serves to understand the flow dynamics and thermal characteristics of a turbulent jet under highly dynamic flow and geometric conditions. The baseline circular vibrating-wall jet impingement configuration undergoes a forced vibration in the wall-normal direction at the frequency, f = 100 Hz. The jet Reynolds number is = 23,000 and the nozzle-exit is at y/D = 2 where the wall vibrates between 0 and 0.5D with amplitude of vibration, A = 0.25D. The configuration is assembled through validation of sub-systems, in particular the method for generating the turbulent jet inflow and the baseline circular jet impingement configuration. Both time-mean and phase-averaged results are presented. The mean radial velocity increases upon positive displacement of the wall and decreases upon negative displacement but this correlation changes with increased radial distance from the stagnation point. Vortical structures are shown to play a major role in convective heat transfer even under the vibrating conditions of the impingement wall. Periodic shifts in the secondary Nusselt number peak are observed that depend upon the travelling eddy location and strength of large-eddy structures. Enhancement in heat transfer is seen in the stagnation region but this beneficial effect of vibration on heat transfer is confined to the impingement region, r/D < 1.5

A Preliminary Investigation towards the Risk Stratification of Allogeneic Stem Cell Recipients with Respect to the Potential for Development of GVHD via Their Pre-Transplant Plasma Lipid and Metabolic Signature

The clinical outcome of allogeneic hematopoietic stem cell transplantation (SCT) may be influenced by the metabolic status of the recipient following conditioning, which in turn may enable risk stratification with respect to the development of transplant-associated complications such as graft vs. host disease (GVHD). To better understand the impact of the metabolic profile of transplant recipients on post-transplant alloreactivity, we investigated the metabolic signature of 14 patients undergoing myeloablative conditioning followed by either human leukocyte antigen (HLA)-matched related or unrelated donor SCT, or autologous SCT. Blood samples were taken following conditioning and prior to transplant on day 0 and the plasma was comprehensively characterized with respect to its lipidome and metabolome via liquid chromatography/mass spectrometry (LCMS) and gas chromatography/mass spectrometry (GCMS). A pro-inflammatory metabolic profile was observed in patients who eventually developed GVHD. Five potential pre-transplant biomarkers, 2-aminobutyric acid, 1-monopalmitin, diacylglycerols (DG 38:5, DG 38:6), and fatty acid FA 20:1 demonstrated high sensitivity and specificity towards predicting post-transplant GVHD. The resulting predictive model demonstrated an estimated predictive accuracy of risk stratification of 100%, with area under the curve of the ROC of 0.995. The likelihood ratio of 1-monopalmitin (infinity), DG 38:5 (6.0), and DG 38:6 (6.0) also demonstrated that a patient with a positive test result for these biomarkers following conditioning and prior to transplant will be at risk of developing GVHD. Collectively, the data suggest the possibility that pre-transplant metabolic signature may be used for risk stratification of SCT recipients with respect to development of alloreactivity