Purvi Purohit

Diseases and Molecular Diagnostics: A Step Closer to Precision Medicine

The current advent of molecular technologies together with a multidisciplinary interplay of several fields led to the development of genomics, which concentrates on the detection of pathogenic events at the genome level. The structural and functional genomics approaches have now pinpointed the technical challenge in the exploration of disease-related genes and the recognition of their structural alterations or elucidation of gene function. Various promising technologies and diagnostic applications of structural genomics are currently preparing a large database of disease-genes, genetic alterations etc., by mutation scanning and DNA chip technology. Further the functional genomics also exploring the expression genetics (hybridization-, PCR- and sequence-based technologies), two-hybrid technology, next generation sequencing with Bioinformatics and computational biology. Advances in microarray “chip” technology as microarrays have allowed the parallel analysis of gene expression patterns of thousands of genes simultaneously. Sequence information collected from the genomes of many individuals is leading to the rapid discovery of single nucleotide polymorphisms or SNPs. Further advances of genetic engineering have also revolutionized immunoassay biotechnology via engineering of antibody-encoding genes and the phage display technology. The Biotechnology plays an important role in the development of diagnostic assays in response to an outbreak or critical disease response need. However, there is also need to pinpoint various obstacles and issues related to the commercialization and widespread dispersal of genetic knowledge derived from the exploitation of the biotechnology industry and the development and marketing of diagnostic services. Implementation of genetic criteria for patient selection and individual assessment of the risks and benefits of treatment emerges as a major challenge to the pharmaceutical industry. Thus this field is revolutionizing current era and further it may open new vistas in the field of disease management.

Clinical and molecular aspects of lead toxicity: An update

Abstract Lead toxicity is a major public health issue in developed and developing countries. Both acute and chronic lead exposure has the potential to cause many deleterious systematic effects including hypertension, frank anemia, cognitive deficits, infertility, immune imbalances, delayed skeletal and deciduous dental development, vitamin D deficiency, and gastrointestinal effects. The underlying mechanisms for all these systemic effects have not been elucidated completely. However, the most plausible cause is free radical damage. In addition to this, lead being a divalent cation can surrogate for calcium at multiple levels affecting various cell signaling pathways. The molecular basis of lead exposure resulting in various systemic effects is being extensively explored. The reports include single nucleotide polymorphisms, epigenetic modifications in susceptible individuals, and the most recent reports also feature regulatory RNA molecules – miRNAs. However, many genetic targets are identified, but their possible mechanisms are still an area to be explored. Additional studies are needed in different population groups to validate the existing findings, as well as to find newer targets that may help in better understanding the molecular mechanisms contributing to lead toxicity. Furthermore, newer strategies for lead risk assessment becomes necessary as the previously recognized “safe” level of lead is also being found to be associated with negative health outcomes.

Evaluation of Antioxidant Status, High Sensitivity C-Reactive Protein, and Insulin Resistance in Male Chronic Opiate Users without Comorbidities

Background: There is a paucity of data on frequency of metabolic syndrome (MS), insulin resistance (IR), and oxidative stress in Indian opiate users without comorbidities. Objectives: To determine the influence of opiate use on frequency of MS, homeostasis model assessment for IR (HOMA-IR), high-sensitivity C-reactive protein (hs-CRP), and oxidative stress in opiate-dependent male patients without comorbidities. Methods: Participants (n = 120) were grouped as controls (Group I), pure opiate dependents (Group II), opiate + tobacco dependents (Group III), and tobacco dependents (Group IV) with a minimum of 1-year dependence participated in the study. Participants were evaluated for anthropometric parameters, blood pressure (BP), fasting blood sugar, insulin, HOMA-IR, lipid profile, hs-CRP, and total antioxidant status (TAS). Frequency of MS was determined based on modified Adult Treatment Panel-III. The data were analyzed using one-way ANOVA, multiple regression by SPSS 21. Results: Frequency of MS in opiate dependents was higher than control. There was a significant difference in serum insulin, HOMA-IR, and TAS levels of the study groups. Multiple regression analysis showed dependence years, body mass index, waist-hip ratio, systolic blood pressure, diastolic blood pressure (DBP), HOMA-IR, and hs-CRP to be significant independent predictors of TAS in Group II and III patients with MS after adjusting for age and education years. TAS and DBP significantly predicted hs-CRP after adjusting for age and education years in Group II and III patients with MS. No such relation was seen in Group I and IV. Conclusions: Chronic opiate-dependent males without comorbidity are a unique group that shows low-grade inflammation, oxidative stress, and prevalence of MS predisposing them to future risk of cardiovascular diseases.

Metabolic Syndrome in Hypothyroidism Leading to Type 2 Diabetes Mellitus: A Cross-sectional Study of Western Rajasthan

Aim: We aimed to diagnose latent diabetic hypothyroid patients presenting with symptoms of metabolic syndrome (MS) based on the Adult Treatment Panel-III (ATP-III) guidelines. Background: Type 2 diabetes mellitus (DM) coexisting with thyroid disorders is difficult to manage. With an ever-increasing incidence of both these disorders and an increasing risk of secondary complications due to their coexistence, newer correlative studies are needed for the early diagnosis of these diseases. Subjects and Methods: The present study was conducted on 100 healthy controls and 150 newly diagnosed hypothyroid patients. The patients were selected based on symptomatology and thyroid function tests. They were then analyzed for body mass index (BMI), blood pressure, fasting blood sugar (FBS), fasting serum insulin, homeostatic model assessment-insulin resistance (HOMA-IR), lipid profile, and apolipoprotein B (apo-B) and apolipoprotein (apo-A 1 ). Statistical Analysis: Analysis was done using the Students t test and Spearmans coefficient of correlation. Results: For hypothyroid patients who presented with raised BMI, diastolic hypertension and dyslipidemia were further investigated for underlying latent diabetes. Of the total hypothyroid patients, 53.3% had raised FBS, 48% had diastolic hypertension, 86.6% had hypertriglyceridemia and 66.67% patients fulfilled three conditions for MS as per the ATP-III guidelines. There was highly significant correlation of serum insulin and HOMA-IR with lipid fractions and cardiovascular disease (CVD) risk ratios (total cholesterol/high-density lipoprotein cholesterol) and apo-B/apo-A 1 in hypothyroid patients. Conclusion: All hypothyroid patients should be closely watched for presence of DM and MS for prevention of atherogenic dyslipidemia, which may lead to CVDs. The estimation of serum insulin, apo-A 1 and apo-B, along with the traditional lipid profile may be useful in such patients.

A correlation study of CVD risk factors in Type 2 diabetics of Western Rajasthan

The aim was to evaluate the CVD risk in type 2 diabetes mellitus subjects on diagnosis using the traditional lipid profile and apo-proteins B and A1 in correlation with serum insulin and IR. The study was conducted on 280 subjects (100 Controls and 180 type 2 DM subjects) who first reported to our OPDs. We analysed FBG, serum Insulin, HOMA – IR, Lipid profile and apo –B and A1. There was a female preponderance, obesity, hyperinsulinemia, insulin resistance (IR) and diabetic dyslipidaemia (hypertriglyceridemia and reduced HDLc). Correlative analysis of BMI showed significant correlation with SBP, lipid profile (except HDLc) and apo –B. Similarly correlative analysis of serum insulin and HOMA –IR showed significant correlation with FBG, lipid profile (except HDLc with HOMA –IR which was Non – significant) and apo –B. CVD risk ratios T. Chol./HDLc and apo –B/apo –A1 showed a significant association with serum insulin and HOMA – IR. Type 2 diabetics from Western Rajasthan, especially those with high BMI, are at increased risk of complications due to atherosclerosis. The positive association of the two CVD risk ratios with serum insulin and IR showed that type 2 diabetics’ risk of developing CVD increases with rise in serum insulin and IR.

Genetic Engineering: Towards Gene Therapy and Molecular Medicine

Abstract Gene therapy, or the use of genetic entities after manipulation for disease management, is derived from advances in genetics, molecular biology, clinical medicine, and human genomics. Molecular medicine, the application of molecular biological techniques to disease treatment and diagnosis, is derived from the development of human organ transplantation, pharmacotherapy, and elucidation of the human genome. The promise of gene therapy, permanent reversal or amelioration of disease symptoms without dependence on a long-lasting intake of drugs, has come within reach because of these conceptual and technical advances in molecular biology. The incidents came at a time when technical advances in the manipulation of DNA had led to widespread testing of gene-based therapies. Gene therapy is generally categorized as either in vivo, in which the gene is delivered directly into recipient cells in the site of target, or ex vivo, in which the gene of interest is inserted in vitro into a targeted cell population (usually stem cells or fibroblasts) and the cells are delivered to the desired site in vivo. These two gene delivery strategies are usually termed “in vivo gene delivery” and “cell-mediated gene delivery,” respectively. Since the beginning of the first gene therapy (clinical trial), i.e., approximately 25 years ago, the field of human gene therapy has gone through numerous successions of ups and downs. Gene therapy has made a profound impact not only in the treatment of genetic disease such as sickle cell anemia as single gene disorder but also open new vistas in neurological diseases, cardiovascular diseases, and cancer; hence, we have started looking at human diseases as a whole.

Association of Inflammatory and Liver Markers with Cardiometabolic Risk Factors in Patients with Depression

Metabolic syndrome (MS) is found to be more prevalent in patients with psychiatric disorders including depression. This study aimed to assess the association of inflammatory and liver markers with cardiometabolic risk factors in patients with Depressive disorders. Prevalence of MS by using Modified NCEP ATP-III Criteria and liver enzymes and CRP were assessed in 382 patients with depressive disorders. MS prevalence was 27.7% and lower HDL level was the commonest metabolic abnormality. ALT, GGT, and CRP levels were positively correlated with weight and BMI. ALT, GGT, and CRP levels were significantly greater in patients with abnormal waist circumference, triglyceride levels and raised blood pressure, compared to patients with normal indices. Such association was not found with abnormal HDL cholesterol and hyperglycemia. Levels of GGT and CRP were significantly greater in patients with MS compared to patients without MS and CRP was significant predictor for MS. To conclude, one-fourth of depressed patients had MS. MS and metabolic abnormalities were associated with inflammatory marker and liver enzymes. Patients with depression should be regularly evaluated for cardiovascular risk factors, liver enzymes, and inflammatory markers.

Effect of Severity of Opiate Use on Cardiometabolic Profile of Chronic Opiate Dependents of Western Rajasthan

Lack of cardiometabolic profile data based on severity of opiate dependence for opiate abusers. The study aimed to evaluate the effect severity of opiate abuse on the cardiometabolic profile of male opiate abusers without co-morbidities. The study included 30 healthy controls (HCs), 90 prospective chronic opiate (opium and heroin) abusers, with and without co-dependence of smoking and tobacco-chewing. The subjects were categorized based on severity of opiate dependence questionnaire (SODQ) and metabolic syndrome (MS) based on NCEP ATP-III criteria and fasting blood samples analyzed for sugar, insulin, insulin resistance (IR), lipid profile, Hs-CRP and total antioxidant capacity (TAC). There was higher prevalence of MS in opiate abusers as compared to HCs. Majority of the patients fell in grade 2 and 3 of severity. There was significant difference across groups for WHR (p < 0.001), SBP (p < 0.03), FBS (p < 0.001), insulin (p < 0.02), IR (p < 0.03) and TAC (p < 0.01). Multiple regression analysis of SODQ grades 2 and 3 independently predicted TAC by Hs-CRP (p = 0.032 and 0.042). There was a significant correlation of TAC with serum insulin, IR and Hs-CRP in SODQ grade 2 and serum insulin and Hs-CRP in SODQ grade 3. Chronic opiate abuse is not benign and predisposes abusers to cardiometabolic risk with increasing severity of dependence, owing to oxidative stress and chronic low-grade inflammation.

Hypocalcemia in an 11 Year Old Child: A Difficult Case to Treat

AbstractHypocalcemia is a laboratory and clinical abnormality that is observed especially in neonates and paediatric patients. Laboratory hypocalcaemia is often asymptomatic but it can manifest as central nervous system irritability, paraesthesia, tetany (i.e. contraction of hands, arms, feet, larynx, bronchioles), seizures, and even psychiatric changes in children. Cardiac function may also be impaired because of poor muscle contractility. We report a unique case of an eleven year old male child who presented with chronic kidney disease associated with severe hypocalcemia, tonic-clonic seizures, hypovitaminosis D but normal electroencephalogram and electrocardiography. The child required prolonged intravenous calcium gluconate therapy to correct his ionised calcium levels.

Metabolic syndrome in pediatric age - A group requiring intensive review

Metabolic syndrome or MS as it is called is a compilation of most of the non-communicable disease components which is a life style disease with strong genetic predisposition, culminating in a morbid adulthood. The increasing number of subjects under the umbrella of MS is alarming. Diabetes mellitus, hypertension is the end result of this insulin resistance as it is sometimes called. Apart from genes, it also has strong inflammatory components that are responsible for early onset of MS. MS is marked by a gamut of inflammatory and non-inflammatory markers. If diagnosed early by screening the population, the percentage of florid cases of this debilitating syndrome can be brought down. Active intervention and constant monitoring of affected or predisposed young subjects will curtail the progress of MS and prevent full blown diabetes and hypertension, thus reducing cardiovascular complications, other morbidity and early mortality. Prevention is always better than cure.