Risk Factors associated with COVID-19 Patients in India: A Single Center Retrospective Cohort Study.

BACKGROUND AND OBJECTIVES: The coronavirus disease 2019 (COVID-19) outbreak has caused a worldwide pandemic, resulting in >3.8 million deaths. Our aim is to identify the risk factors associated with in-hospital mortality using survival analysis considering the characteristics and outcomes of COVID-19 patients admitted to a dedicated tertiary-care hospital in Mumbai, India. MATERIALS AND METHODS: In a retrospective cohort study, 565 patients admitted from 28th March 2020 to 30th June 2020 were enrolled, and a follow-up was conducted till August 2020. To investigate the impact of COVID-19, survival analysis was performed using the Kaplan-Meier method. Potential risk factors associated with mortality were analyzed using logistic regression models for multivariate analysis and the Cox proportional hazards model for estimating hazard ratios (HRs). RESULTS: From the 565 positive COVID-19 cases, 49 patients died (8.7%) and 516 (91.3%) were discharged. Overall, 119 patients (20%) required intensive care unit (ICU) admission, of which 70 (58%) patients survived. The Kaplan-Meier survival curve showed a significant association of COVID-19 infection with age (≥60; p = 0.008), hypertension (p = 0.03), dialysis (p = 0.0001), lung commodities (p = 0.01), breathlessness (p = 0.0001), severe disease upon high-resolution computed tomography (HRCT) analysis (p = 0.0001), ICU admission (p = 0.0001), and low lymphocyte count at admission (p = 0.0001). Additionally, patients receiving tocilizumab (p = 0.0001) and deprived of hydroxychloroquine (HCQ) + azithromycin (azee) (p = 0.0001) were estimated at a high risk of mortality. INTERPRETATION AND CONCLUSION: Coronavirus disease 2019 (COVID-19) increased the risk of mortality in patients with increased age, comorbidities, and severe symptoms upon treatment with an immunosuppressant (tocilizumab). However, patients treated with HCQ + azee showed favorable results due to their antiviral effects in vitro.

A 30-day Survival and Safety of Percutaneous Tracheostomy in Moderate-to-severe COVID-19 Pneumonia Patients: A Single-center Experience.

Aims and objectives: In coronavirus disease-2019 (COVID-19) pneumonia, guidelines on timing and method of tracheostomy are evolving. The aim of the study was to analyze the outcomes of moderate-to-severe COVID-19 pneumonia patients who required tracheostomy and the safety with regard to the risk of transmission to the healthcare workers. Materials and methods: We retrospectively analyzed 30-day survival outcome of a total of 70 moderate-to-severe COVID-19 pneumonia patients on a ventilator, wherein tracheostomy was performed only in 28 (tracheostomy group), and the remaining were with endotracheal intubation beyond 7 days (non-tracheostomy group). Besides demographics, comorbidities and clinical data including 30-day survival and complications of tracheostomy were analyzed in both groups with respect to the timing of tracheostomy from the day of intubation. Healthcare workers were monitored for COVID-19 symptoms by carrying out periodical COVID tests. Results: The 30-day survival of the tracheostomy group was 75% as compared to 26.2% of the non-tracheostomy group. The majority of the patients (71.4%) had severe disease with PaO2/FiO2 (P/F ratio) <100. The first wave showed an 80% (4/5) while the second wave 100% (8/8) thirty days survival in the tracheostomy group performed before 13 days. All patients during the second wave underwent tracheostomy before 13 days with a median of 12th day from the day of intubation. These tracheostomies were performed percutaneously at the bedside, without any major complications and no transmission of disease to healthcare workers. Conclusion: Early percutaneous tracheostomy within 13 days of intubation demonstrated a good 30-day survival rate in severe COVID-19 pneumonia patients. How to cite this article: Shah M, Bhatuka N, Shalia K, Patel M. A 30-day Survival and Safety of Percutaneous Tracheostomy in Moderate-to-severe COVID-19 Pneumonia Patients: A Single-center Experience. Indian J Crit Care Med 2022;26(10):1120-1125.

Clinical Profile of Patients with Severe Acute Respiratory Syndrome Coronavirus 2 Infection Developing Pulmonary Barotrauma on Mechanical Ventilation.

Background: There is limited information on clinical profile and outcomes of patients on mechanical ventilation (MV) who developed pulmonary barotrauma (PBT) in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Patients and methods: In a retrospective observational study, all SARS-CoV-2 pneumonia patients admitted from March 28, 2020, to August 31, 2020, at Sir HN Reliance Foundation Hospital and Research Center and Seven Hills Hospital (Reliance Facility), Mumbai, India, of 18 years and above on MV and developed PBT, were included. Results: A total of 14 SARS-CoV-2 patients of 45 on MV (31.0%) developed PBT of 1,029 hospitalized. All patients were male and divided as per admission into PaO2/FiO2 (P/F) ≤100 (median 80) and P/F >100 (median 222) group. Pneumothorax developed in seven and six cases of P/F ≤100 and P/F >100 groups, respectively. Three patients in each group developed subcutaneous emphysema, while four developed pneumomediastinum in P/F >100 group. Twelve patients (7, P/F ≤100, and 5, P/F >100) were on invasive, while two (P/F >100) were on noninvasive MV. The mean P/F on the day of PBT was reduced by 27.5 and 65.3%, while peak inspiratory pressure was elevated with a median of 36 and 28 cm H2O in P/F ≤100 and P/F >100 groups, respectively. The median highest tidal volume (420 mL), positive-end expiratory pressure (8 vs 6 cm H2O) on the day of PBT, and length of hospital stay (11 vs 25 days) did not differ between two groups. Survival was 28.6% (4/14). Conclusion: SARS-CoV-2 patients requiring MV with PBT had poor outcomes. Clinicians should be vigilant about the diagnosis of PBT. How to cite this article: Kargirwar KV, Rathod D, Kumar V, Patel M, Shah M, Choudhury H, et al. Clinical Profile of Patients with Severe Acute Respiratory Syndrome Coronavirus 2 Infection Developing Pulmonary Barotrauma on Mechanical Ventilation. Indian J Crit Care Med 2022;26(5):613-618.

Gene expression of klotho & antioxidative enzymes in peripheral blood mononuclear cells of essential hypertension patients in Indian population.

BACKGROUND & OBJECTIVES: Oxidative stress is known to have a causal role in hypertension. Klotho has emerged as a novel anti-aging molecule to inhibit oxidative stress at cellular level. This study aimed at evaluating the gene expression of klotho and antioxidative enzymes, manganese superoxide dismutase (Mn-SOD) and catalase, in peripheral blood mononuclear cells of essential hypertensive patients as compared to normotensive healthy controls. METHODS: Ninety-nine newly diagnosed hypertensives and 103 age- and BMI-matched controls were recruited. The participants were non-diabetic and not on any medication. Soluble α-klotho levels were detected using enzyme-linked immunosorbent assay. Gene expression was evaluated by quantitative real-time polymerase chain reaction. RESULTS: Soluble α-klotho levels were significantly lower (27%, P=0.001) in patients as compared to controls. The trend remained same when compared against 44 out of 103 controls considered for gene expression analysis. Relative gene expression of klotho and catalase were 3-fold and 1.25-fold lower in patients as compared to controls, respectively. ΔCt value-based gene expression were also significantly lower for both genes (P=0.001). A decreasing but non-significant trend was observed for Mn-SOD gene expression. ΔCt value-based gene expression of catalase positively correlated with that of Mn-SOD in patient (rs=0.448) and control (rs=0.547) groups (P<0.001). In patients, the gene expression of Klotho positively correlated with that of catalase (rs=0.498, P=0.001), but not Mn-SOD (rs=0.155, P=0.126). INTERPRETATION & CONCLUSIONS: In the present study on newly diagnosed hypertensives, klotho and catalase gene expression were found to be significantly lower as compared to controls, indicating the role of oxidative stress in this patient group. In addition, a significant correlation between Klotho and catalase gene expression suggests a role for klotho in essential hypertension with respect to antioxidant defence.

Identification of a single nucleotide polymorphism indicative of high risk in acute myocardial infarction.

Background & objectives: Acute myocardial infarction (AMI) is a major health concern in India. The aim of the study was to identify single nucleotide polymorphisms (SNPs) associated with AMI in patients using dedicated chip and validating the identified SNPs on custom-designed chips using high-throughput microarray analysis. Methods: In pilot phase, 48 AMI patients and 48 healthy controls were screened for SNPs using human CVD55K BeadChip with 48,472 SNP probes on Illumina high-throughput microarray platform. The identified SNPs were validated by genotyping additional 160 patients and 179 controls using custom-made Illumina VeraCode GoldenGate Genotyping Assay. Analysis was carried out using PLINK software. Results: From the pilot phase, 98 SNPs present on 94 genes were identified with increased risk of AMI (odds ratio of 1.84-8.85, P=0.04861-0.003337). Five of these SNPs demonstrated association with AMI in the validation phase (P=0.05). Among these, one SNP rs9978223 on interferon gamma receptor 2 [IFNGR2, interferon (IFN)-gamma transducer 1] gene showed a significant association (P=0.00021) with AMI below Bonferroni corrected P value (P=0.00061). IFNGR2 is the second subunit of the receptor for IFN-gamma, an important cytokine in inflammatory reactions. Interpretation & conclusions: The study identified an SNP rs9978223 on IFNGR2 gene, associated with increased risk in AMI patient from India.

Peripheral Blood Mononuclear Cell ABCA1 Transcripts and Protein Expression in Acute Myocardial Infarction.

BACKGROUND: ATP binding cassette transporter-A1 (ABCA1) facilitates the formation of high density lipoprotein (HDL). HDL due to its anti-atherosclerotic, anti-inflammatory and anti-thrombotic activities provides protection against atherothrombosis or myocardial infarction (MI). The aim was to investigate the role of peripheral blood mononuclear cell (PBMNC) ABCA1 expression in MI. METHODS: The participants comprised 29 males with acute MI (AMI) and 20 healthy controls. AMI patients were normotensive, not on statins, with triglycerides < 200mg/dl and categorized into AMI with type 2 diabetes (T2DM) (N = 12) and without T2DM (N = 17). The PBMNC ABCA1 mRNA transcripts were analysed by quantitative real-time polymerase chain reaction (qRTPCR) and protein by enzyme linked immunosorbent assay (ELISA). RESULTS: PBMNC ABCA1 mRNA transcript and protein levels were not significantly different in AMI patients or when sub-grouped into with/without T2DM, as compared to controls. ABCA1 protein correlated positively with HDL-cholesterol (r = 0.655, p = 0.021) in AMI patients with T2DM and negatively with age (r = - 0.525, p = 0.031) in AMI patients without T2DM and it was more strongly associated in latter group with smoking and alcohol habit. CONCLUSION: In the present study, the effects of metabolites of diabetes and ischemia were observed on PBMNC ABCA1 protein and thus on HDL-C in AMI patients. Further influence of risk factors such as smoking and alcohol consumption observed in the present study can be evaluated in larger sample size. The control of these cardiovascular associated risk factors may increase stability of PBMNC ABCA1 protein and thus HDL-C levels.

Polymorphisms of MDR1, CYP2C19 and P2Y12 genes in Indian population: effects on clopidogrel response.

AIMS/OBJECTIVE: Influence of genetic variations on the response of clopidogrel, an antiplatelet drug is implicated. In the present study, the prevalence of single nucleotide polymorphisms of MDR1 (C3435T), CYP2C19 [CYP2C19*2 CYP2C19*3, CYP2C19*17] and P2Y12 (i-T744C) in Indian population and their effects on clopidogrel response was analyzed. METHODS AND RESULTS: To analyze the prevalence of polymorphisms, 102 healthy individuals were recruited. Clopidogrel response was assessed by ADP induced platelet aggregation in clopidogrel naïve acute myocardial infarction (AMI) patients (n = 26) screened from 100 AMI cases, before loading dose of 300 mg, at 24 h before next dose and 6 days after on 75 mg per day and platelet aggregation inhibition (PAI) was calculated between these time intervals. Genotyping was carried out by PCR-based restriction enzyme digestion method for C3435T of MDR1 and i-T744C of P2Y12, by multiplex PCR for CYP2C19*2 (G681A) and CYP2C19*3 (G636A) and by nested PCR for CYP2C19*17 (C806T). The effect of the above mentioned genetic variations on PAI was analyzed. Variant allele of CYP2C19*3 was not observed while the prevalence of 3435T of MDR1 (0.524), CYP2C19*2 (681A, 0.352); i-744C of P2Y12 (0.088), as well as wild type allele CYP2C19*17 (C806, 0.897) associated with decrease clopidogrel response were observed. Trend toward poor response to clopidogrel was observed at 24 h with the variant genotypes of CYP2C19*2 and i-T744C of P2Y12 as compared to wild type. CONCLUSION: The present study did show a trend toward impaired response of clopidogrel to inhibit platelet aggregation with variant genotypes of CYP2C19*2 and iT744C of P2Y12 compared to respective wild type genotype at 24 h.

Levels of cathepsins in acute myocardial infarction.

AIMS/OBJECTIVE: Over expression of matrix degrading enzymes have been implicated in plaque destabilisation and rupture. Cathepsins associated with extracellular matrix breakdown make them intriguing suspects. The aim of the study was to analyse peripheral levels of cathepsin B and cathepsin K and their inhibitor cystatin C during acute myocardial infarction (AMI). MATERIALS AND METHODS: Study population included AMI patients at acute event (AMI group, n=48), stable angina patients (stable angina group n = 17), and healthy individuals (Control group, n=31). Cathepsin B, cathepsin K, cystatin C, and matrix metalloproteinases (MMP)-9 were analysed by enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Cathepsin B (45.9%) and cathepsin K (92.31%) at acute event of myocardial infarction (AMI group) increased (P=0.001) while cystatin C decreased marginally (12.5%) as compared to controls. Stable angina group, demonstrated only marginal reduction in all the parameters studied as compared to controls. CONCLUSION: Cathepsin B and cathepsin K can be further evaluated as biomarkers in identifying high-risk individuals for AMI.

Prevalence of VKORC1 and CYP2C9 gene polymorphisms in Indian population and its effect on warfarin response.

OBJECTIVES: The clinical effectiveness of Warfarin is established. Patients require different warfarin dosages to achieve the target therapeutic anticoagulation. The variability of Warfarin dosage is largely genetically determined, and it can be partly explained by the C1173T and G-1639A polymorphisms of vitamin K epoxide reductase complex subunit 1 (VKORC1) which is its target and *2 and *3 allele of Cytochrome P-450 (CYP) 2C9 [CYP2C9] enzyme which metabolizes to its inactive form. Aim of the present study was to determine the prevalence of these variant alleles known to influence the warfarin dose and correlate genotypes with the average INR as well as mean dose of Warfarin required to maintain INR, in the Indian population. METHODS: Study population included 100 healthy individuals and 83 patients operated for Aortic or Mitral Valve replacement and prescribed warfarin thereafter. Of these 83 patients records of INR for the period of six months and mean maintenance dose (stable therapeutic dose) of warfarin required to maintain INR were available for 26 patients. For the remaining patients, apart from their demographic data only maintenance dose was available. Genotyping of above mentioned polymorphisms was carried out by using PCR-based restriction digestion method. RESULTS: Although less as compared to wild type alleles, the variant alleles of CYP2C9*2 and *3 as well as of VKORC1 polymorphisms (C1173T and G-1639A) were observed in our study population. Mean maintenance dose (mg/day) of Warfarin was in the decreasing order of patients as compared to the wild type genotypes for all above mentioned polymorphisms. The decrease in the dose was in the order of heterozygotes for CYP2C9*2 to CYP2C9*3 to C1173T and G-1639A of VKORC1 (P<0.001). There was significant correlation (r=0.51, P<0.001) observed between the dose estimated by pharmacogenetic algorithm of Sconce et al (2005) and actual stable therapeutic dose. INR was high for mutant variants (3.8 to 4) after first dose suggesting that they require decreased mean daily dose of Warfarin. CONCLUSION: In the present study the effect of CYP2C9*2, *3, and VKORC1 (C1173T and G-1639A) genotypes on warfarin dose was observed. However, the genotyping has not been incorporated into daily practice. Perhaps more practical approach would be for clinicians to take genotype information into consideration along with other factors when dosing warfarin.

Study of C-reactive protein and myocardial infarction in the Indian population.

To analyse the association of high sensitivity C-reactive (hsCRP) protein levels and -717A/G single nucleotide polymorphism of CRP with acute myocardial infarction (AMI) in the Indian population. Study population included 100 MI cases wherein 32 patients had experienced previous MI (MI-Group-1), 68 MI cases were recruited at presentation (MI-Group-2) and equal number of age and gender matched healthy individuals. hsCRP levels were determined by ELISA and genotyping of -717A/G was carried out by polymerase chain reaction-based restriction digestion method. The -717A/G genotypes did not influence hsCRP level and their distribution did not differ between groups. However, in the present study hsCRP demonstrated significant correlation with BMI in controls of both the genders and with triglycerides in females of AMI at presentation who otherwise are with low risk profile. Identifying traditional risk factors associated with inflammation may help in controlling the acute event.

Platelet polymorphisms: frequency distribution and association with coronary artery disease in an Indian population.

Platelets play a critical role in normal blood hemostasis and thrombus formation in myocardial infarction (MI). Several polymorphisms of genes involved in platelet activation and fibrinolysis have been reported to be associated with MI. The aim of the present study was to determine the frequency distribution and association of polymorphisms in these genes with coronary artery disease (CAD) among Indians. A case-control genetic association study was performed for polymorphisms in platelet glycoprotein receptors (GPIIb/IIIa [HPA1a/1b], GPIb-IX-V [VNTR], and GPIa/IIa [C807T]), fibrinogen ß-chain (BclI), α-chain (Aα312), tissue plasminogen activator (tPA) [I/D] and plasminogen activator inhibitor-I (PAI-1) [4G/5G] in 473 healthy controls and 446 patients with stable and unstable angina. Genotyping was either by PCR-based restriction endonuclease digestion or allele-specific primers. The I allele frequency of the tPA I/D polymorphism was significantly higher in our patients (χ(2)=7.33, P<0.01) and no other polymorphisms varied significantly between patients and controls. Also, none of the polymorphisms seemed to affect the severity of the disease, the only exception being the mutant alleles of ß chain of fibrinogen gene, which were significantly elevated in single vessel disease. This is the first study to evaluate the role of gene polymorphisms in both the thrombotic and fibrinolytic pathway in the Indian population and suggests that tPA I/D polymorphism confers CAD risk in our population.

In search for novel biomarkers of acute coronary syndrome.

Prerequisites for a biomarker to enter clinical routine in acute coronary syndrome (ACS) are several folds. The most important features are that the biomarker can be offered on a routine platform to diagnose and identify high-risk individuals of ACS, provide higher sensitivity and specificity than the ECG in predicting outcome, and should have impact on therapeutic decision making. In recent years, a deeper understanding of the pathobiology of the atherothrombosis as the underlying mechanism of ACS has directed scientific studies towards the evaluation of certain pathogenic components involved in the process as potential biomarkers for the clinical settings of ACS. Under investigations are markers to identify early injury/ischemia, markers for detection of vulnerable plaque, its disruption, of thrombosis and markers for inflammation. The aim of this paper is to review the current contribution of biochemical markers to clinical cardiology and also to discuss some important developments in this field.

Soluble levels of cell adhesion molecules (CAMs) in coronary artery disease.

AIMS: To analyze soluble levels ofcell adhesion molecules (CAM) such as Intercellular CAM (ICAM), vascular CAM (VCAM-1), platelet endothelial CAM (PECAM-1), Endothelial (E)-selectin, and Platelet (P)-selectin in coronary artery disease patients and correlate with degree of severity of the disease. METHODS: Study population included patients who suffered myocardial infarction at presentation (N=49) and those with unstable angina (N=79) and stable angina (N=14). Soluble levels of CAMs were measured by ELISA. RESULTS: At acute event in AMI patients, there was significant rise of soluble (s) E-selectin (4.5 fold, P = 0.001), sVCAM-1 (65.6%, p = 0.001), sPECAM-1 (46.2%, p = 0.02), sP-selectin (42.7%, p = 0.001) and sICAM-1 (20.1%, p = 0.003) as compared to controls. In unstable angina group as compared to AMI there was significant decrease in the levels observed in, sE-selectin (62.7%, p = 0.001), sPECAM-1 (47.5%, p = 0.001) as well as sVCAM-1 (17.9%, p = 0.04) and insignificant decrease with respect to sICAM-1 and no change with respect to sP-selectin levels. Stable angina group as compared to unstable angina group demonstrated no significant difference in sCAMs and the trend with AMI group was similar to that seen between unstable angina and AMI group. Significantly elevated levels of sE-selectin, sVCAM-1 and sPECAM-1 at acute event suggest them to be causal molecules as well as markers of plaque destabilization. Levels of sP-selectin in stable angina were similar to that observed in AMI and unstable angina groups suggesting elevated platelet activation in stable angina as well.

Matrix metalloproteinase-3 (MMP-3) -1612 5A/6A promoter polymorphism in coronary artery disease in Indian population.

Matrix metalloproteinases (MMPs) play important role in the pathogenesis of coronary artery disease (CAD). 5A allele of -1612 5A/6A polymorphism of MMP-3 is associated with two fold higher activity than 6A allele. Present study was designed to analyse the association of this polymorphism with CAD in Indian population. Subjects included in the study were patients with stable angina (n=35), unstable angina (n=53), patients with recent event of myocardial infarction (MI) (MI Group-1, n=56) and patients at presentation of the acute MI (MI Group-2, n=49). Controls were healthy individuals (n=99). Genotyping of MMP-3 5A/6A polymorphism was carried out by PCR-based restriction digestion method. The genotype distribution of patient groups did not deviate from controls. Serum MMP-3 levels were significantly elevated at presentation of the acute MI by 36.8% (P=0.031) as compared to controls and more associated with 6A genotype suggesting discrepancy between in vitro transfection experiment and peripheral MMP-3 levels.

Circulating thrombotic and haemostatic components in patients with coronary artery disease.

The study aimed to analyze the circulating levels of thrombotic and haemostatic components; tissue factor, tissue factor pathway inhibitor, tissue plasminogen activator and plasminogen activator inhibitor-1 in patients with acute myocardial infarction at presentation (Group 1, n=49), unstable angina and Non-ST elevated MI after treatment (Group 2, n=22), stable angina (Group 3, n=18) and healthy individuals (Group 4, n=31). Significant finding was increase in tissue factor not only in Group 1 (2.0 fold, P=0.001), Group 2 (2.2 fold, P=0.015) but also in Group 3 (1.8 fold, P=0.018) as compared to controls. In Group 1 Plasminogen activator inhibitor-1 increased significantly (35.8%, P=0.02). Tissue factor pathway inhibitor and tissue plasminogen activator demonstrated increase in Group 1 of age<40 years while insignificant changes in elder patients. Increased thrombotic and decreased fibrinolytic conditions in acute myocardial infarction patients were observed. Increase TF in stable angina demonstrates procoagulant status in these patients as well.

Role of matrix metalloproteinases in coronary artery disease.

AIM: Matrix metalloproteinases (MMPs) contribute both in the formation as well as in the destabilization of atherosclerotic plaque. In the present study we analyzed circulating levels of MMP-7 that acts on chondroitin sulphate a proteoglycan that is particularly abundant in atherosclerotic plaque and MMP-8 which acts on Type I collagen, the synthesis and degradation of which is important for stability of the plaque and correlate with the degree of severity of coronary artery disease (CAD). METHODS: Circulating levels of MMP-7 and MMP-8 and tissue inhibitors of MMP (TIMP) -1 and TIMP-2 were analysed by Enzyme Linked ImmunoSorbent Assay (ELISA7rpar;, in patients with acute myocardial infarction (AMI) at presentation (N=48), acute coronary syndrome (ACS) group (N=227rpar; (on treatment) and stable angina group (N=17) (on treatment). RESULTS: There was significant rise in MMP-8 (88.23%, P=0.001), in AMI group which decreased in ACS treated group 7lpar;15.9%, non-significant) as compared to controls. There was increasing trend of MMP-7 in AMI and ACS group and strong correlation with hsCRP. MMP-7 predominated in stable angina group. There was significant decrease in TIMP-2 in AMI group and TIMP-1 and TIMP-2 in ACS and stable angina group as compared to controls. CONCLUSION: Significant increase in MMP-8 and decrease in TIMP-2 during acute stage of AMI suggests MMP-8 and TIMP-2 are markers for vulnerable plaque independent of hsCRP for AMI. MMP-7 was found to be elevated in stable angina patients and was correlated with hsCRP at acute phase of AMI suggesting persistent at all stages of CAD.

Circulating levels of cell adhesion molecules in hypertension.

Hypertension causes complications such as coronary atherosclerosis and thrombosis wherein inflammatory factors play significant role. In the present study inflammatory molecules such as cell adhesion molecules (CAMs); endothelial (E)-selectin, platelet (P)-selectin, intercellular CAM-1 (ICAM-1), vascular CAM-1 (VCAM-1) and platelet endothelial CAM-1 (PECAM-1) were analysed in subjects newly diagnosed with hypertension with no secondary cause against normotensive healthy individuals. In each group 57 subjects were recruited and soluble (s) levels of CAMs were analysed by ELISA. As compared to controls median of sE-selectin (49.2%, P=0.001), sP-selectin (54.3%, P=0.001), and sICAM-1 (18.9%, P=0.012) were significantly elevated in hypertensive subjects. Significant negative correlation was observed of sP-selectin (spearman rank correlation coefficient (rs) =-0.345, p=0.027) and sPECAM-1 (rs =-0.446, p=0.003) with age in hypertension group. Hypertension may increase expression of certain CAMs while younger hypertensives in addition are also at increased risk of atherothrombosis.

Bone marrow cells for myocardial repair-a new therapeutic concept.

AIM: To assess the safety and feasibility of transfusing autologous bone marrow stem cells (ABMSC) into the culprit coronary artery after an acute anterior wall myocardial infarction (MI) and further to see the ability of ABMSC to promote improvement in Left Ventricular lsqb;LV] function at follow-up. METHODS: In an ongoing phase I clinical trial, twenty-seven patients of uncomplicated acute anterior wall MI treated as per the current practicing guidelines have been included. Among these, seventeen patients received intra-coronary unfractionated ABMSCs from 77ndash;15 days after acute MI (ABMSC group) and ten patients acted as controls. RESULTS: All the procedures carried out were without any complications. After 6 months, cardiac function analysis of ten patients from the ABMSC group by LV angiography and Cardiac Magnetic Resonance Imaging (MRI) demonstrated a significant rise of 12.74% (p = 0.001) and 7.1% (p = 0.001), respectively in the LV ejection fraction [LVEF]. There was an improvement in the LV systolic function wherein LV end systolic volume (LVESV) decreased significanty to 28.75% (p = 0.010) and 16.49% (p = 0.022) by LV angiography and cardiac MRI, respectively. LV end diastolic volume (LVEDV) decreased marginally by LV angiography (p = 0.548) and by cardiac MRI (p = 0.514). Five patients of the control group by LV angiography demonstrated non-significant rise of 1.0% (p = 0.706) in LVEF, 12.79% (p = 0.332) in LVEDV and 22.56% (p = 0.308) in LVESV. By cardiac MRI controls demonstrated significant rise in EF of 3.2% (p = 0.0367rpar; but non-significant fall of only 2.32% (p = 0.812) in LVEDV and 6.47% (p 7equals; 0.508) in LVESV. CONCLUSION: This study shows that intracoronary infusion of ABMSC is safe and feasible after acute MI and shows a favourable trend towards the improvement of LV function and prevention of ventricular remodeling which determines long-term survival.

Standardization of homocysteine on HPLC using DTT as reductant.

Determination of plasma total homocysteine by high-pressure liquid chromatography (HPLC) usually requires reduction of protein bound or free homocysteine-disulphides into thiols by a reducing agent and the liberated thiols are then derivatized by a fluorescent marker. In this study we have standardized the HPLC method for homocysteine measurement using dithiothreitol (DTT) as reductant.The results of plasma total homocysteine values obtained by HPLC were compared with IMx method. The difference between the two means was statistically insignificant [P=0.616847 (two tail)] Linear regression analysis showed strong correlation between the two methods (r=0.983). Using this method we have analyzed 132 controls and 130 Coronary Heart Disease (CHD) patients for plasma total homocysteine, wherein, the mean plasma total homocysteine levels were 10.51±8.36 and 11.51±10.06 µmol/L respectively. Our research study suggests that DTT method is a simple and inexpensive assay for homocysteine determination in human plasma for research application.

Growth hormone (GH-1) gene deletions in children with isolated growth hormone deficiency (IGHD).

OBJECTIVE: To detect growth hormone GH-1 gene deletions (6.7 kb, 7.6 kb, 7 kb) in familial/nonfamilial isolated growth hormone deficiency (IGHD) and note their clinical and investigative profile. METHODS: Thirty (M16,F14) prepubertal IGHD patients aged 0.25 to 14 y, from 25 families were screened. Duration of growth failure, relevant history, clinical phenotype, and height SDS were recorded. Peak GH response to Clonidine (0.15 mg/m(2)), IGF-1, IGFBP-3 and pituitary/target gland hormones were studied. Genomic DNA of patients and family was analysed by PCR and DNA fragments were visualized on agarose gel electrophoresis. RESULTS: This series was divided into deletion +ve, Group I (n=12,40%) inclusive of six familial/six nonfamilial patients, and deletion -ve Group II (n=18,60%), 5 familial/13 nonfamilial cases; in total 11/30 were familial. Onset of growth failure was earlier in Group I (p<0.001) mean 1.1 vs 4.7 y. Mean height SDS was -7 vs. -4.5 in Groups I/II (p<0.01), age at presentation 5.1 vs 8.6 y. Overhanging forehead, prominent eyes, hypoplastic facies characterized Group I with FBS <50 mg/dl in 50% and very low peak GH <0.04 vs 2.04 ng/ml (p<0.001) in Group II. In both groups IGF-1 and IGFBP3 were low, other hormones were normal and MRI showed hypoplastic adenohypophysis. 40% had GH-1 gene deletion (6.7 kb deletion in 83%, 7.6 kb and a compound heterozygote in 8% each). CONCLUSIONS: In this series of 30 IGHD patients, frequency of GH-1 gene deletions (12/30) was 40%, and 54% among familial patients, and 31% with height SDS>-4. 83% had 6.7 kb deletion. Height SDS>-4, clinical phenotype, peak GH<1 ng/ml and hypoglycemia characterised IGHD Type IA.