Molecular Determination of Tumor Necrosis Factor-alpha, Interleukin-8, Interleukin-10, and C-X-C Chemokine Receptor-2 Genetic Variations and their Association with Disease Susceptibility and Mortality in COVID-19 Patients.

Background: Altered cytokine levels have been associated with poor outcomes among COVID-19 patients. TNF-α, IL-8 and IL-10 are key cytokines in COVID-19 pathogenesis, and CXCR-2 is a major chemokine receptor involved in inflammatory response. Polymorphisms in the genes of these proteins are proposed to influence disease outcomes. In this study, we aimed to find out the association of genetic polymorphisms in TNF-α, IL-8, IL-10 and CXCR-2 genes with susceptibility to and mortality of COVID-19. Methods: The present case-control study was conducted on 230 subjects, among whom 115 were clinically diagnosed and RT-PCR-confirmed COVID-19 patients and 115 healthy control subjects. The polymorphisms in TNFα -308 G>A (rs1800629), IL-8 -251T>A (rs4073), CXCR2 +785 C>T (rs2230054) genes were detected by ARMS -PCR assay whereas for IL-10 (-1082 G>A), rs1800896 G>A allele-specific PCR assay was used and their association with COVID-19 susceptibility and mortality was estimated by multivariate analysis. The results were analyzed for risk of infection and mortality through different inheritance models. Results: Frequencies of TNF-α rs1800629 GA, AA, IL-8 rs4073 TA, AA, IL-10 (-1082 G>A), rs1800896 GA and GG, and CXCR2 rs2230054 CT genotypes were significantly higher in COVID-19 patients compared to the control group (p < 0.05). Furthermore, COVID-19 patients had a higher frequency of the polymorphic A allele of TNF-α, the A allele of IL-8, the G allele of IL-10, and the T allele of CXCR2. The risk of susceptibility to COVID-19 was significantly associated with TNF-α rs1800629 GA, GA+AA genotypes and the A allele, IL-8 rs4073 TA, AA genotypes and A allele, IL-10 rs1800872 GA and CC genotypes and C allele, and CXCR2 rs2230054 CT and CT+CC genotypes. TNF-α-GA and AA genotypes and A allele, IL-8 TA and AA genotypes and A allele and CXCR-2 CC and CT genotypes have significant associations with mortality risk in COVID-19 patients, while GA and GG genotypes of the IL-10 are shown to confer significant protection against mortality from COVID-19. Conclusion: The findings of this study provide important insights into the COVID-19 disease and susceptibility risk. The polymorphisms in TNFα -308 G>A (rs1800629), IL-8 -251T>A (rs4073), IL-10 (-1082 G>A), rs1800896 and CXCR2 +785 C>T (rs2230054) are associated with the risk of susceptibility to COVID-19 and with mortality in COVID-19 patients. Further studies with larger sample sizes are necessary to confirm our findings.

In Silico Investigation of AKT2 Gene and Protein Abnormalities Reveals Potential Association with Insulin Resistance and Type 2 Diabetes.

Type 2 diabetes (T2D) develops from insulin resistance (IR) and the dysfunction of pancreatic beta cells. The AKT2 protein is very important for the protein signaling pathway, and the non-synonymous SNP (nsSNPs) in AKT2 gene may be associated with T2D. nsSNPs can result in alterations in protein stability, enzymatic activity, or binding specificity. The objective of this study was to investigate the effect of nsSNPs on the AKT2 protein structure and function that may result in the induction of IR and T2D. The study identified 20 variants that were considered to be the most deleterious based on a range of analytical tools included (SIFT, PolyPhen2, Mut-pred, SNAP2, PANTHER, PhD-SNP, SNP&Go, MUpro, Cosurf, and I-Mut). Two mutations, p.A179T and p.L183Q, were selected for further investigation based on their location within the protein as determined by PyMol. The results indicated that mutations, p.A179T and p.L183Q alter the protein stability and functional characteristics, which could potentially affect its function. In order to conduct a more in-depth analysis of these effects, a molecular dynamics simulation was performed for wildtype AKT2 and the two mutants (p.A179T and p.L183Q). The simulation evaluated various parameters, including temperature, pressure, density, RMSD, RMSF, SASA, and Region, over a period of 100 ps. According to the simulation results, the wildtype AKT2 protein demonstrated higher stability in comparison to the mutant variants. The mutations p.A179T and p.L183Q were found to cause a reduction in both protein stability and functionality. These findings underscore the significance of the effects of nsSNPs (mutations p.A179T and p.L183Q) on the structure and function of AKT2 that may lead to IR and T2D. Nevertheless, they require further verifications in future protein functional, protein-protein interaction, and large-scale case-control studies. When verified, these results will help in the identification and stratification of individuals who are at risk of IR and T2D for the purpose of prevention and treatment.

Surgical Candidacy for Management of Laryngomalacia: A Proposed Scoring System.

Introduction: Identifying surgical candidacy for the management of laryngomalacia is a challenge. Objective: To develop a simple scoring system for surgical candidacy in laryngomalacia. Methods: Eighteen years retrospective observational study of children with laryngomalacia (LM) clinically categorized into mild, moderate and severe LM and were analyzed for surgical candidacy. Results: There were 113 children (age ranging from 5 days to 14 months), 44% being mild, 30% moderate and 26% severe LM. None in mild, 32% in moderate, and all in severe LM had surgical intervention. Presence of stridor on feeding or crying and isolated type 1 or type 2 LM on laryngoscopy were significant indicators for conservative treatment (p-< 0.0001). Moderate failure to thrive, retraction at rest/sleep, with low oxygen saturation while feeding/at rest were significantly higher in both moderate and severe groups with laryngoscopic evidence of combined type 1 and 2 in moderate LM (p < 00,001). Aspiration pneumonia, hospitalization, pectus and mean pulmonary arterial pressure of more than 25 mmHg with laryngoscopic findings of all three combined types were significantly higher in severe LM (p < 0.0001).A simple scoring system was then developed and it revealed that a score of 10 or more required surgical intervention. Conclusion and clinical significance: A clinical scoring system is being reported for the first time in medical literature to identify 'the difficult to treat' subset within moderate laryngomalacia category simplifying decision making in its management for otolaryngologists and pediatricians as well as a referral criterion for pediatric otolaryngologists' services.

An Assessment of the Utility of Serum Fructosamine in the Diagnosis and Monitoring of Diabetes Mellitus.

BACKGROUND: Fructosamine (FA) has gained importance as a new biomarker for hyperglycemia in the past decade and may be of indispensable use in certain conditions where hemoglobin A1c (HbA1c) falls short of utility such as disorders of red blood cells, patients with rapid glycemic excursions requiring more short-term monitoring, pregnancy, chronic kidney disease, etc. Methods: The present study was a hospital-based observational cross-sectional study conducted in the Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), Nagpur, India. Serum HbA1c, FA, albumin-corrected fructosamine (AlbF), total protein-corrected FA (PrF), hemoglobin (Hb), and hematocrit (Hct) were estimated in 32 controls (Group I) and 32 cases of diabetes mellitus (DM) (Group II). The clinical data and lab results were presented as mean±SD/±standard error (SE) of the mean. Student's t-test and ANOVA were used to compare various parameters between the groups. Pearson correlation analysis was performed to assess the correlation between different diagnostic parameters. The receiver operating characteristic (ROC) curve was plotted to assess the diagnostic significance and cut-off value for FA, AlbF, and PrF. RESULTS: The controls and cases were matched for age and gender distribution. Serum HbA1c (p<0.0001), serum FA (p<0.0001), fasting blood sugar (p=0.001), postprandial blood sugar (p<0.0001), random blood sugar (p=0.001), hematocrit (p=0.002), AlbF (p<0.0001), and PrF (p<0.0001) were found to be significantly higher in known diabetic subjects compared to controls. The case group was further subdivided into pre-diabetic and diabetic groups. On correlation analysis of HbA1c with various parameters, a moderate correlation of HbA1c was noted with FA (r=0.522, p<0.0001) and AlbF (r=0.375, p=0.002) in all subjects. Additionally, a moderate correlation of FA (r=0.479, p=0.033), AlbF (r=0.444, p=0.050), and PrF (r=0.441, p=0.065) with HbA1c was also found in subjects with diabetic range glycemia. No such correlation was noted in the pre-diabetic group. No significant correlation was noted between FA and its corrected values in any range of glycemia. None of the parameters assessing glycemia were found to be significantly affected by hemoglobin status. On ROC curve analysis, HbA1c was found to be the best parameter (area under the curve (AUC) =83%, p<0.0001) followed by AlbF (AUC= 80.5%, p<0.0001) and uncorrected FA (AUC=80.5%, p<0.0001) to diagnose DM. CONCLUSION: Serum FA should be considered a valid diagnostic biomarker and of indispensable use in special populations where HbA1c falls short of utility such as patients with red blood cell disorders or those showing rapid glycemic excursions such as those on corticosteroid therapy or insulin therapy, etc. It exhibits additional advantages over HbA1c with respect to lower reagent cost and easy automation on any conventional laboratory instruments based on simple colorimetry.

Biochemical aspects of effects of mesenchymal stem cell treatment in chronic wounds progressive healing.

Chronic wounds are a persistent burden for medical professionals. Despite developments and advancements in treatment, these wounds do not heal completely. Mesenchymal stem cells (MSCs) are the epicenter of regenerative medicine that have shown promising results in chronic wound regeneration. Autologous peripheral blood-derived MSCs (PB-MSCs) are comparatively new in wound healing treatment, bone-marrow-derived MSCs (BM-MSCs), and adipose-derived stem cells (ADSCs) are commonly being practiced. In the present study, PB-MSCs treatment was given to chronic wound patients. Various biochemical parameters like random blood glucose, serum urea, serum creatinine, bilirubin (total and direct), Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), total protein, albumin levels, and association of other factors/conditions such as age, sex, addiction of drug/alcohol were also evaluated/compared with complete and without complete healing. The wound area of the ulcer was found to be significantly reduced and the wound was healthier after the treatment. These biochemical parameters could be certainly utilized as biomarkers to anticipate the risk of chronic wounds. These findings may contribute to the development of better wound care treatment strategies and drug discovery in the field of regenerative medicine.

Association of COVID-19 Real-Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Cycle Threshold Value With Surrogate Markers of Disease Severity.

Introduction The cycle threshold (Ct) value in real-time reverse transcription-polymerase chain reaction (RT-PCR) serves as a criterion to diagnose coronavirus disease 2019 (COVID-19) and is inversely proportional to viral load. Levels of inflammatory markers such as aspartate aminotransferase (AST), ferritin, D-dimer, high sensitivity C-reactive protein (hs-CRP), and lactate dehydrogenase (LDH) are used as quantitative measures of COVID-19 severity. We examined the association between these markers and Ct values. Methodology This retrospective data analysis included 400 patients with positive RT-PCR results for COVID-19 who were admitted to a tertiary care hospital. Clinical and biochemical data were accessed from the hospital information management system. Associations of clinical parameters and markers of disease severity (e.g., polymorph, AST, hs-CRP, D-dimer, LDH, and ferritin levels) with Ct values were assessed. Observations LDH, ferritin, D-dimer, and hs-CRP were found to be significantly higher in moderate and severe groups than in the mild COVID-19 group. AST, ferritin, and hs-CRP levels were also significantly higher in severe COVID-19 subjects, compared to moderate COVID-19 subjects. Ct values for the E (envelop) gene and ORF (open reading frame) 1b gene were found to be significantly higher in those with severe COVID-19. Polymorph counts in subjects with Ct values of 25 or higher were significantly increased, compared to those with Ct values under 30. LDH, D-dimer, and hs-CRP levels in subjects with Ct values over 30 were significantly lower than for those with Ct values under 30. Ferritin was the best independent predictor of non-survival in study subjects, with an area under the curve (AUC) of 85.5% (95% confidence interval = 73.2-95.9). The Ct value for the E gene had an AUC of 75.1%, and the ORF1b gene had an AUC of 64.5%. However, no significant correlation was detected between any parameter and Ct value. Conclusion Polymorph, LDH, ferritin, D-dimer, and hs-CRP levels were significantly elevated in subjects with low E gene Ct values. Also, these subjects were at risk of severe disease and fatality. Ct values for the E gene thus could serve as an early indicator for patients at risk of severe disease and death.

Evaluating Ischemia-Modified Albumin as an Early Biomarker for Hypertensive Disorders During Pregnancy: A Case-Control Study.

Background Ischemia-modified albumin (IMA) is looked upon as a newer marker of myocardial ischemia. There is a paucity of literature however with regard to studies correlating levels of IMA in patients with hypertensive disorders of pregnancy. The present study therefore aimed at estimating the levels of IMA in patients with gestational hypertension and assessing its utility in predicting hypertensive disorders of pregnancy. Methods The present study was a hospital-based case-control study conducted in the Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), Nagpur. IMA was estimated in 30 controls (Group I) and 20 cases of gestational hypertension (Group II) using a spectrophotometric assay detecting free unbound Cobalt left behind. The clinical data and lab results were presented as mean ± SD. Student's t-test was applied and Pearson's correlation coefficient was calculated. A value of p < 0.05 was taken as statistically significant. The ROC (Receiver Operator Characteristic) curve was used to establish the cut-off of serum IMA levels in pregnancy-induced hypertension (PIH). Results There was no significant difference in age and period of gestation (POG) at the time of sample collection between the groups. There was a significant difference in the systolic and diastolic blood pressures (BPs) of both groups. The mean level of serum IMA was significantly higher in cases of gestational hypertension (0.88 ± 0.14 absorbance units {ABSU}) as compared to controls (0.69 ± 0.08 ABSU) (p<0.001). On correlation analysis, the systolic and diastolic BPs were found to be highly positively correlated with serum IMA levels (p<0.001). ROC curve analysis suggested that at a cut-off of 0.73 ABSU, IMA has 85% sensitivity and 80% specificity for predicting gestational hypertension. Conclusion Statistically significant results of serum IMA levels obtained in gestational hypertension which falls on the lesser severe spectrum of the disease imply that serum IMA can be used for early diagnosis of gestational hypertension and impending Pre-eclampsia (PE) and Eclampsia.

Evaluation of promoter hypermethylation of tumor suppressor gene BRCA1 in epithelial ovarian cancer.

Context: Epithelial ovarian cancer (EOC) is a serious gynecological issue worldwide and its late detection is the major encumbrance in treatment procedures. Hypermethylation-mediated BRCA1 gene silencing results in failure of the repair system of damaged DNA playing an important role in ovarian carcinogenesis. BRCA1 gene hypermethylation can serve as a safe and highly specific clinical marker for EOC. Aims: The present study was conducted to evaluate the promoter hypermethylation of BRCA1 gene in EOC patients. Settings and Design: This hospital-based case-control study carried out in the tertiary care hospital in New Delhi. Subjects and Methods: Promoter hypermethylation of BRCA1 gene was examined in 30 EOC diagnosed untreated cases confirmed by histopathological examinations and compared with 30 normal healthy controls matched for age using methylation specific-polymerase chain reaction. Results: We found significantly higher BRCA1 promoter hypermethylation in the serum of EOC cases as compared to controls with P < 0.0001. BRCA1 gene methylation was found to have 70% sensitivity for the diagnosis of EOC with 100% specificity. A significant difference was observed in the range of CA125 levels, B12 and Folate levels between EOC cases and controls. Conclusions: We conclude that BRCA1 gene is significantly hypermethylated in EOC patients and thus can prove to be a noninvasive diagnostic tool. Our results provide prefatory evidence that epithelial ovarian epigenome can be influenced by dietary nutrients.

IL-2 and IL-1ß Patient Immune Responses Are Critical Factors in SARS-CoV-2 Infection Outcomes.

BACKGROUND: Immune dysregulation has been linked to morbidity and mortality in COVID-19 patients. Understanding the immunology of COVID-19 is critical for developing effective therapies, diagnostics, and prophylactic strategies to control the disease. AIM: The aim of this study was to correlate cytokine and chemokine serum levels with COVID-19 disease severity and mortality. SUBJECTS AND METHODS: A total of 60 hospitalized patients from the Tabuk region of Saudi Arabia with confirmed COVID-19 were included in the study. At hospital admission, the IL-1 ß, IL-2, IL-8, IL-10, LT-B4, and CCL-2 serum levels were measured. The cytokine levels in COVID-19 patients were compared to the levels in 30 healthy matched control subjects. RESULTS: The IL-1 ß, IL-2, LTB-4, CCL-2, and IL-8 levels (but not IL-10) were significantly higher in all COVID-19 patients (47 survivors and 13 non-survivors) compared with the levels in the healthy control group. In the non-survivor COVID-19 patients, patients' age, D-dimer, and creatinine kinase were significantly higher, and IL-1 ß, IL-2, and IL-8 were significantly lower compared with the levels in the survivors. CONCLUSION: Mortality rates in COVID-19 patients are associated with increased age and a failure to mount an effective immune response rather than developing a cytokine storm. These results warrant the personalized treatment of COVID-19 patients based on cytokine profiling.

Neutrophil-to-Lymphocyte Ratio as a Predictor of Disease Severity and Mortality in Coronavirus Disease 2019: Prospective Study From Central India.

BACKGROUND: Clinical presentation of coronavirus disease 2019 (COVID-19) varies from an asymptomatic state to severe disease characterized by acute respiratory distress syndrome, respiratory failure, thrombosis, and multi-organ dysfunction syndrome. The neutrophil-to-lymphocyte ratio (NLR) has been reviewed as one of the laboratory factors that have been proposed to predict the severity of disease and mortality in COVID-19 pandemic. AIM AND OBJECTIVES: To evaluate the association between NLR and the disease severity and mortality in COVID-19. MATERIALS AND METHODS: After approval from Institutional Ethics Committee, this prospective cohort study was carried out in a tertiary-care teaching medical institute of Central India. COVID-19 patients of the age group 18 years and above admitted during the study period were included. Cases were categorized into four groups as asymptomatic (Group A), mild (Group B), moderate (Group C), and severe (Group D) based on clinical symptoms, respiratory rate, oxygen saturation, and chest imaging. NLR was calculated by doing a complete blood count at the time of hospitalization by the Mindray BC-6000 auto hematology analyzer. The outcome of the disease was classified as recovery and death during hospitalization. Receiver operating characteristic (ROC) curve analysis was used to assess the ability of NLR at admission to predict severe COVID-19 or mortality. Ordinal regression analysis was used to assess the impact of NLR on disease severity and mortality. RESULTS: Mean NLR was significantly higher in the severe COVID-19 group as compared to the mild/moderate group and in deceased as compared to discharged cases. ROC curve analysis revealed NLR to be an excellent predictor of disease severity as well as a prognostic parameter for risk of death. NLR was found to be a significant independent positive predictor for contracting the severe disease (Odd's ratio 1.396, 95% CI=1.112-1.753, p=0.004) and mortality (Odd's ratio 1.276, 95% CI=1.085-1.499, p=0.003). CONCLUSION: High NLR was significantly associated with the disease severity and mortality in COVID-19.

Acute Cholecystitis Leading to Elevated High-Sensitive Cardiac Troponin I in a Young Female Without Any Cardiac Ailment: A Rare Case Presentation.

Cardiac troponin I (cTnI) is regarded as a gold standard investigation for the diagnosis of acute myocardial infarction (AMI). However, cTnI may be elevated in certain non-AMI cardiac conditions and even in certain noncardiac conditions. We report a case of a young female presenting with symptoms suggestive of acute cholecystitis with elevated high-sensitive cardiac troponin I (hs-cTnI). The patient developed acute chest pain during the hospital stay. On evaluation, quantitative assay for hs-cTnI was found to be elevated; however, other markers of cardiac damage such as creatinine kinase-MB (CK-MB), qualitative cTnI by card test, and even echocardiogram (ECG) were found to be negative. As the patient was a young female with no significant history of coronary diseases, the spurious elevation of hs-TnI due to a noncardiac ailment was suspected. The patient was managed with minimal cardiological management till AMI was excluded. The hs-cTnI levels returned to normal post-cholecystectomy. A patient presenting with symptoms suggestive of cholecystitis and elevated hs-cTnI must be carefully evaluated before resorting to any invasive management for AMI. In most cases, hs-cTnI will return to normal post-cholecystectomy.

Molecular Detection of Streptococcus mutans and Streptococcus sobrinus in Dental Plaque Samples in Children Aged Six to Nine Years.

Context Dental caries is a widespread threat, usually in children, although it has been observed at other stages of life. Various pieces of literature have confirmed the prevalence of S treptococcus mutans and S treptococcus sobrinus in the progression of the disease. However, establishing procedures to detect these species remains a challenge, posing a barrier to treatment plans. Aim The aim of this study is to detect the species in dental plaque samples from children aged six to nine years by polymerase chain reaction (PCR) and correlate their prevalence in various dentitions. Material and Methods This is an observational analytical cross-sectional study conducted in a tertiary care dental hospital. After sample isolation, microbiological processing was performed, genomic DNA was isolated, and PCR run was performed using specific primers to detect the species. SPSS for Windows Version 17 (IBM Corp., Armonk, NY) and Microsoft Excel (Microsoft Corporation, Redmond, WA, USA) were used to perform statistical analysis. A p-value of <0.05 was considered statistically significant. Results The technique could identify S. Mutans and S. Sobrinus in a short turnaround time. The frequency of S. mutans and S. sobrinus infections was higher in individuals with dental caries. Conclusions Molecular detection via PCR is a reliable, economical, and less time-consuming method for detecting S. mutans and S. sobrinus in dental plaque samples.

The COVID TIDE Approach: A Protocol for Safe Tracheostomy Practice in COVID Patients.

We all are aware of COVID 19 pandemic. As the numbers are increasing, the critical care demand is also increasing. Tracheostomy is one of the commonest procedures which has been performed on COVID positive ventilated patients. It is important to understand and follow the utmost safe practices for the patient and the health care workers for such aerosol generating procedures. The aim of this study is to identify the lacunae in tracheostomy practices during this COVID times and suggest a systematic approach for the safe practices. An online questionnaire survey-based study was performed in September 2020. The target population was practicing otolaryngologists of India with various years of experience. The aim of the study was to evaluate the lacunae in tracheostomy safe practices and to create a systematic approach for the safety of health care workers. Data compilation and analysis was done by using Microsoft Excel. A systematic COVID TIDE tracheostomy safe practices approach was designed after reviewing various tracheostomy guidelines and recommendations. Total 114 otolaryngologists responded with a complete survey report. 72.2% responders were not up to date with their knowledge of tracheostomy safe practices. 79.8% were not performing this procedure in a negative pressure room. 15.8% were not aware of the personal protective equipment level they are using. Only 56.1% survey responders were holding the ventilation before tracheal incision. Overall, 94.7% responders were keen to know about the safe approach of tracheostomy in COVID positive patients. Tracheostomy is an aerosol generating procedure, lacunae in the knowledge can cause major risk to health care professionals. Finally, in such crises, consideration should be taken for simulation exercises, dedicated airway teams and a systematic COVID TIDE approach to improve the safety of the staff and patients. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12070-021-02370-w.

VEGF Promoter Region 18-bp Insertion-Deletion Polymorphism in Sickle Cell Disease Patients with Microalbuminuria: A Pilot Study.

PURPOSE: Vascular endothelial growth factor (VEGF) is a potent inducer of micro vascular permeability thus leading to nephropathy. Insertion/deletion (I/D) polymorphism of 18 bp at - 2549 position in VEGF gene causes increased transcription leading to increased production of VEGF. Thus, we aimed to associate I/D polymorphism of the 18 bp fragment at - 2549 position of the promoter region of VEGF gene with sickle cell nephropathy (SCN). METHODS: This observational analytical case control study included 30 subjects each of SCN, sickle cell disease (SCD) without nephropathy and the control group. The subjects were assessed for various hematological and biochemical parameters. Further, 18 bp I/D polymorphism of VEGF gene in all three study groups was assessed by polymerase chain reaction followed by electrophoresis and compared. RESULT: Though increased frequency of both DD genotype and D allele was found in SCN compared to SCD and control, only frequency of D allele was found to be significantly higher (p = 0.04). D allele posed marginal risk of microalbuminuria in SCD subjects compared to controls (OR = 2.11) as well as to SCD without MA subjects (OR = 1.84). CONCLUSION: D allele in I/D polymorphism in the promoter region of VEGF gene may be associated with marginal increase in risk of susceptibility to sickle cell nephropathy.

Polymorphism of CYP1A1 gene variants rs4646903 and rs1048943 relation to the incidence of cervical cancer in Chhattisgarh.

Cytochrome P450 CYP1A1 is a phase 1 xenobiotic metabolizing enzyme involved in the metabolism of toxins, endogenous hormones and pharmaceutical drugs. It is therefore possible that polymorphism of CYP1A1 gene producing functional changes in the enzyme may be susceptible factors in cervical carcinogenesis. This study was aimed to look association of CYP1A1m1 (T>C) and m2 (A>G) gene polymorphisms in Chhattisgarh population. In this case-control study, we analyzed leukocyte DNA from a total of 200 subjects form Chhattisgarh (100 cases and 100 controls). All subjects were genotyped for CYP1A1m1 (T>C) and m2 (A>G) using PCR-RFLP with statistical analysis by using SPSS version 16.0 and VassarStats (online). Among the two gene variants rs4646903 (T>C) and rs1048943 (A>G), individuals with AG and GG genotypes of CYP1A1m2 polymorphism have significantly higher and increased risk of cervical cancer (OR=2.0, 95%CI=1.04-3.84, p=0.035; OR=62.9, 95%CI=3.72-1063.83, p=0.004 respectively) and the association of CYP1A1m1 polymorphism did not show any significant relationship with cervical cancer patients (p=0.23). The 'G' allele showed strong association with the disease (p<0.0001). Thus, CYP1A1m2 polymorphism showed an increased risk in the population leading to cervical cancer. Our study suggested that the presence of 'C' allele of rs4646903 (T>C) showed no risk and 'G' allele of rs1048943 (A>G) might be a leading allele to cause increased cervical cancer susceptibility due to significant association of CYP1A1m2 gene polymorphism.

Assessment of Serum Vascular Endothelial Growth Factor Levels in Pregnancy-Induced Hypertension Patients.

Objective. The objective of the study was to assess the serum vascular endothelial growth factor (VEGF) levels in peripheral blood of patients with pregnancy-induced hypertension (PIH) and find association between serum VEGF levels and PIH. Methods. Thirty-five PIH subjects, 35 normal pregnant females, and 20 normal healthy females were included in the study. Detailed history, clinical examination, and relevant biochemical parameters were assessed; serum VEGF levels were estimated using Double-antibody enzyme-linked immunosorbent assay. Results. The study groups were found to be age matched (p = 0.38). VEGF level in the pregnancy-induced hypertensive group (median = 109.19 (3.38 ± 619)) was significantly higher than the normal pregnant (median = 20.82 (1.7-619)) and control (median = 4.92 (1.13-13.07)) group and the difference between these three groups was significant (p < 0.0001). The 3 groups are found to be significantly different in terms of RBS (p = 0.01), urea (p < 0.0001), creatinine (p = 0.0005), AST (p = 0.0032), ALT (p = 0.0007), total protein (p = 0.0004), albumin (p < 0.0001), calcium (p = 0.001), and sodium (p = 0.02), while no statistically significant difference was found between total bilirubin (p = 0.167), direct bilirubin (p = 0.07), uric acid (p = 0.16), and potassium (p = 0.14). Conclusion. Significantly higher levels of serum VEGF were noted in PIH subjects compared to normal pregnant and control subjects.

Association of 18bp insertion/deletion polymorphism, at -2549 position of VEGF gene, with diabetic nephropathy in type 2 diabetes mellitus patients of North Indian population.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent multifunctional cytokine which plays a key role in the pathogenesis of diabetic micro-vascular complications. Human VEGF gene is said to be highly polymorphic. Insertion/deletion (I/D) polymorphism of the 18 bp fragment at -2549 position of the promoter region in VEGF gene is said to be of particular interest. The study was aimed to evaluate association of Insertion/deletion (I/D) polymorphism of the 18 bp fragment at -2549 position of the promoter region in VEGF gene, with diabetic nephropathy in type 2 diabetes mellitus. METHODS: This cross sectional study enrolled 40 subjects each of diabetic nephropathy (DN), diabetes mellitus without nephropathy (DM) and normal control subjects. DNA was isolated from peripheral blood leukocytes. Genotyping of the VEGF gene insertion/ deletion (I/D) polymorphism was done by the polymerase chain reaction (PCR) methods. The frequency of VEGF alleles and genotype distribution were compared in diabetic nephropathy, uncomplicated diabetic and control groups. RESULTS: DD genotype and D allele were found to be significantly associated with DN group (p = 0.009 and 0.02 respectively) in comparison to DM group. Also DD genotype conferred significant risk of diabetic nephropathy in DM group (OR = 4.2) (against combined frequency of ID and II genotype) so does D allele 2.09 (against I allele). CONCLUSION: DD genotype and D allele in I/D polymorphism at -2549 position of VEGF gene is associated with increased susceptibility to diabetic nephropathy in north Indian population.

Use of red vacutainer for collection of CSF causes falsely high estimation of proteins by pyrogallol red method.

BACKGROUND: Though plain plastic/glass tubes are recommended for CSF collection, many laboratories use the commercially available red topped evacuated tubes for CSF collection for biochemical analysis. These red vacutainers affect the assay of some serum parameters. AIM: We evaluated the effect of using red vacutainer for CSF collection on estimation of proteins. METHODS: CSF samples of 50 patients were collected in plain plastic containers. One milliliter from these were transferred to red vacutainers and mixed gently. Protein by pyrogallol red method was estimated directly from plastic containers and from the sample that was poured to red vacutainers. We further prepared different concentrations of bovine serum albumin in normal saline and estimated the O.D. on a spectrophotometer and protein levels on a clinical chemistry analyzer, similarly before and after transferring 1 ml to red vacutainer. RESULTS: The CSF protein levels were significantly higher (p=<0.0001) when transferred to a red vacutainer (median: 81.5 and range:32-324 mg/dl) than that estimated directly from a plain plastic container (Median: 50.5 and range: 20-300 mg/dl) and affected the interpretations in 50% cases. The protein levels were 25, 50, 50 and 355% higher when BSA prepared at concentrations of 100, 50, 25 and 10mg/dl respectively were transferred to red vacutainers. But for 750, 1500 and 3000 mg/dl of BSA concentrations, the increase in red vacutainer was 1.4, 2 and 0.7% respectively. CONCLUSION: Collection of CSF in red vacutainer significantly affects CSF protein estimations, probably due to the presence of clot activator. This might alter the interpretation of result and thus management of the subject. So the red vacutainer should not be recommended for CSF collection.