Neurological Manifestations of Glutamic Acid Decarboxylase Autoimmunity in Indian Patients.

Objective: To study the neurological manifestations of glutamic acid decarboxylase (GAD 65) autoimmunity in Indian patients. Methods: Retrospective study conducted in a tertiary care referral hospital in South India. Patients who tested positive for GAD 65 antibodies from February 2013 to July 2019 were included. Results: We identified 922 patients who underwent GAD 65 testing, of which 81 tested positive (8.78%) [mean age 55.42 years (SD 17.39, range 9-86 years, median age 57 years)]. Males (n = 47) outnumbered the females (n = 34). All the GAD values measured were <5000 IU/ml. There were 34 cases (42%) of atypical parkinsonism (16/34, 47% fulfilled the diagnostic criteria for autoimmune atypical parkinsonism) in our series forming the most common group with GAD 65 positivity, followed by autoimmune encephalitis (8 cases, 9.88%). Men were more affected with atypical parkinsonism (22/34; 64.70%), stiff person syndrome (2/3; 66.66%), and neuropathy (4/7; 57.1%) while women were more with autoimmune encephalitis (6/8; 75%). Eighteen (22.6%) had underlying autoimmunity (three had type 1 diabetes mellitus). Six (7.4%) had underlying neoplasm. Thirty-three out of 43 patients responded to immunotherapy (76.74%). Five had spontaneous improvement. Conclusion: Glutamic acid decarboxylase65 antibody values were much lower in our study population. Male-dominant autoimmunity was seen unlike that in Western literature. The most striking was the high preponderance of atypical parkinsonism in GAD 65-positive patients. We also found that GAD 65 positivity is a useful marker for a positive response to immunotherapy in suspected autoimmune neurological syndromes irrespective of their titers.

The utility of serum amylase as a prognostic marker in multiple myeloma.

Background: Ectopic production of amylase by tumor cells is known since 1951. Elevated amylase in multiple myeloma (MM) was first described in 1988. It has been postulated that translocation of chromosome 1, where amylase gene is situated, is responsible for ectopic production from the malignant plasma cells. Anecdotal reports have shown hyperamylasemia in MM to be associated with extensive bone disease, rapid progression, and shorter survival. Serum amylase estimation is a ubiquitous test. This prospective study was conducted to ascertain the degree of elevated amylase, its clinical utility, and implications in MM patients. Materials and Methods: In an 18-month period, all consenting patients with newly diagnosed or relapsed MM were tested for serum amylase levels. The study excluded patients with elevated lipase, abnormal creatinine clearance, and evidence of intestinal obstruction or perforation. Patients with amylase value >100 U/L were designated to have "elevated amylase level" for the purpose of this study. Results: We enrolled 58 patients with MM, of which 29.3% (n = 17) were found to have elevated serum amylase levels. The median age of patients with elevated amylase was 65 years. The male-to-female ratio was 1.9:1. There was no statistical association between age, gender, type of heavy chain class, light chain, or high-risk cytogenetics. Among patients with the International Staging System (ISS), Stages I, II, and III, 20.8% (n = 5), 31.3% (n = 5), and 41.2% (n = 7) were noted to have elevated amylase levels. A statistically significant association was noted between the presence of extramedullary disease (EMD) and elevated amylase level (P = 0.028). Higher mortality (29.4% versus 17%) and shorter mean survival of (30.2 ± 3.3 months versus 51.7 + 4.9 months) were recorded in patients with elevated amylase levels in comparison to those with normal levels. Conclusions: Elevated serum amylase level in MM is indicative of advanced ISS stage, the presence of EMD, higher risk of mortality, and shorter survival. Serum amylase can be used as a cost-effective tool in myeloma management.

Determination of sensitivities and specificities of cerebrospinal fluid free light chains to diagnose multiple sclerosis- a multicentric case-control study.

BACKGROUND: CSF free light chains help diagnose multiple sclerosis, but no data is available on the Asian population. Our objective was to study the diagnostic utility of CSF free light chains for diagnosing multiple sclerosis in Indian patients. METHODS: Prospective multicentric case-control study. Cases included those who were tested for oligoclonal bands and fulfilled the modified McDonald criteria 2017 for multiple sclerosis and clinically isolated syndromes. Those tested for oligoclonal bands (OCB) but with other diagnoses- inflammatory and non-inflammatory were included as controls. Clinical details were collected from electronic medical records. CSF and serum kappa and lambda free light chains were measured, apart from oligoclonal bands, immunoglobulin, and albumin in paired serum and CSF samples. RESULTS: There were 70 patients (31 cases and 39 controls). The mean age was 43.41(SD 16.073) years, and 43(61.4%) were females. CSF kappa showed highest specificity 97.4%, at a cut off 2.06 mg/L (sensitivity 71%) and highest sensitivity 90.3%, at a cut off 0.47 mg/L (specificity 79.5%). Best balance of sensitivity and specificity for CSF kappa was seen at a cut-off of ≥ 0.63 mg/L {sensitivity 87·1 (CI - 70.17-96.37), and specificity 87·18 (CI -72.57-95.70)}. The ratio of Kappa/lambda showed highest specificity of 100%(similar to OCB) with a sensitivity of 71% at a cut off of 1.72. The ratio of sum of kappa and lambda light chains, and Qalb (∑CSF FLC/Qalb), showed the highest specificity (94.87%)among the blood brain barrier corrected ratios. CONCLUSION: This study showed that the diagnostic utility of CSF kappa was comparable to OCB to diagnose multiple sclerosis in sensitivity, but not specificity, so can be a screening test before testing for OCB in our population.

High Prevalence of Plasminogen Activator Inhibitor-1 4G/5G Polymorphism among Patients with Venous Thromboembolism in Kerala, India.

Venous thromboembolism (VTE) is a multifactorial clotting disorder in which inherited and environmental factors synergistically contribute to its pathogenesis. The aim of this case-control study was to analyze the prevalence of hereditary thrombophilic risk factors, provoking and non-provoking environmental risk factors in patients with VTE from Kerala, India. We have observed a low prevalence of factor V Leiden (7%), prothrombin G20210A (2%), and prothrombin G20030A (2%) mutations and a high prevalence of plasminogen activator inhibitor-1 (PAI-1) 4G/5G (52%), PAI-1 4G/4G (24%) genotypes in the VTE patients (n = 147). Deficiency of anticoagulants, antithrombin (3.4%), and protein C (4.1%) was relatively low. None of the risk factors were observed in 17% of the patients. Majority of VTE patients were younger than 50 years with a median age of 43 years. In conclusion, our results indicate a high prevalence of PAI-1 4G/5G polymorphism among the VTE patients which is in concordance with previous studies in the Asian population. The PAI-1 4G/5G polymorphism could be a potential biomarker for assessing VTE risk, particularly among the Indian population.

Role of fibroblast growth factor-23 (FGF-23) in the prediction of postoperative atrial fibrillation after coronary artery bypass surgery.

Introduction: Fibroblast growth factor-23 (FGF-23) is a bone-derived hormone which had shown a significant association with the occurrence of atrial fibrillation (AF) in patients with chronic kidney disease. We hypothesized that FGF-23 could be a very useful predictive biomarker for atrial remodeling and in turn for postoperative atrial fibrillation (POAF) after coronary artery bypass grafting (CABG). We also looked at the correlation of cardiac remodeling in right atrial biopsy and its correlation with POAF and with the FGF-23 level. Methods: This study was a single-center cross-sectional observational study. All the patients who were planned for CABG with no prior history of AF were included in the study. All the patients were tested for preoperative serum FGF-23 level. During CABG, biopsy specimen of the right atrial appendage was sent for histopathology evaluation. All patients were monitored for POAF until discharge from the hospital. Results: This study involved 60 patients who underwent elective CABG. Multivariate analysis revealed a significant association between preoperative FGF-23 levels with the occurrence of POAF (p < 0.001). The area under the curve of FGF-23 was 0.894 with a sensitivity of 92.3% and specificity of 87.23%. The cut-off value for serum FGF-23 levels was found to be 6.50 ng/ml. Analysis of biopsy specimens revealed that the presence of hypertrophic myocytes had a significant association with the occurrence of POAF/POAE and with FGF-23. Conclusion: Preoperative FGF-23 levels can predict the incidence of POAF in post-CABG patients. Histopathologically, the presence of hypertrophic myocytes correlated well with the occurrence of POAF.

Association of Follicular Fluid Advanced Glycation End-Products with Oocyte Response and Clinical Pregnancy in Assisted Reproduction Cycles.

CONTEXT: Advanced glycation end-products (AGEs) are toxic metabolic end-products of lipids, nucleic acids, and proteins. Their accumulation in the ovaries can alter the follicular microenvironment and affect stimulation response. AIMS: We aimed to study the association of AGEs in follicular fluid (FF AGE) with oocyte response and clinical pregnancy in ART-Assisted Reproductive Technology cycles. SETTINGS AND DESIGN: This prospective study involved 84 individuals undergoing ART. METHODS AND MATERIAL: FF was collected during oocyte retrieval, and the level of AGEs was measured by enzyme-linked immunosorbent assay. Oocyte response was grouped as below target (<7 MII oocytes) or above target response (≥7 MII). STATISTICAL ANALYSIS USED: The association of FF AGE with the oocyte response and clinical pregnancy rate was analyzed by Mann-Whitney U-test. The strength of association of FF AGE with the outcome variables was analyzed with receiver operating characteristic (ROC) curve. RESULTS: The median FF AGE was 17.6 (8.5) µg/ml. It was significantly higher in the below target than the normal ovarian response group (18.5 [17.8] vs. 16.3 [7.8] µg/ml, P = 0.046). Similarly, it was significantly higher in those who did not conceive (19.9 [7.3] vs. 13.5 [5.9] µg/ml, P < 0.001). The cutoff of FF AGE obtained by ROC curve analysis was 16.5 µg/ml above which there were significantly lower oocyte response and clinical pregnancy. CONCLUSIONS: Elevated FF AGE can be a significant negative predictor of clinical pregnancy and ovarian response to stimulation in ART cycles. The FF AGE level above the cutoff value of 16.5 µg/ml was associated with significantly lower oocyte response and clinical pregnancy.

Somatic mutation detection efficiency in EGFR: a comparison between high resolution melting analysis and Sanger sequencing.

BACKGROUND: High resolution melting curve analysis is a cost-effective rapid screening method for detection of somatic gene mutation. The performance characteristics of this technique has been explored previously, however, analytical parameters such as limit of detection of mutant allele fraction and total concentration of DNA, have not been addressed. The current study focuses on comparing the mutation detection efficiency of High-Resolution Melt Analysis (HRM) with Sanger Sequencing in somatic mutations of the EGFR gene in non-small cell lung cancer. METHODS: The minor allele fraction of somatic mutations was titrated against total DNA concentration using Sanger sequencing and HRM to determine the limit of detection. The mutant and wildtype allele fractions were validated by multiplex allele-specific real-time PCR. Somatic mutation detection efficiency, for exons 19 & 21 of the EGFR gene, was compared in 116 formalin fixed paraffin embedded tumor tissues, after screening 275 tumor tissues by Sanger sequencing. RESULTS: The limit of detection of minor allele fraction of exon 19 mutation was 1% with sequencing, and 0.25% with HRM, whereas for exon 21 mutation, 0.25% MAF was detected using both methods. Multiplex allele-specific real-time PCR revealed that the wildtype DNA did not impede the amplification of mutant allele in mixed DNA assays. All mutation positive samples detected by Sanger sequencing, were also detected by HRM. About 28% cases in exon 19 and 40% in exon 21, detected as mutated in HRM, were not detected by sequencing. Overall, sensitivity and specificity of HRM were found to be 100 and 67% respectively, and the negative predictive value was 100%, while positive predictive value was 80%. CONCLUSION: The comparative series study suggests that HRM is a modest initial screening test for somatic mutation detection of EGFR, which must further be confirmed by Sanger sequencing. With the modification of annealing temperature of initial PCR, the limit of detection of Sanger sequencing can be improved.

Possible Effects of Perchlorate Contamination of Drinking Water on Thyroid Health.

BACKGROUND: Perchlorate is an anion that occurs as a contaminant in groundwater. It originates from the improper disposal of ammonium perchlorate, a component of rocket fuel. The objective of this study was to explore whether the exposure to perchlorate in drinking water had an impact on the thyroid function of the population residing near an ammonium perchlorate plant in Kerala. Methodology. Using an ecological study design, we compared the serum levels of thyroid-stimulating hormone, thyroxine, and thyroid peroxidase antibodies among a representative sample of 289 study subjects from the area surrounding the ammonium perchlorate enrichment plant to 281 study subjects in a control area. RESULTS: The perchlorate concentration in the groundwater varied from 1600 ppb to 57,000 ppb in the 10 samples from the contaminated area and was below 24 ppb in all locations in the control area. No significant differences were found in the mean serum TSH concentration and mean T4 levels between the subjects from the contaminated area and the control area. On regression analysis, perchlorate contamination was not found to be a significant predictor of TSH. CONCLUSION: This study did not find any significant association between perchlorate in drinking water and changes in thyroid hormone levels. Our findings indicate the need for further investigation of this hypothesis using urinary perchlorate as a measure of individual exposure.

Effect of Non-Surgical Periodontal Therapy on the Serum Sialic Acid Levels in Diabetic Patients with Periodontitis.

Sialic acid (SA), a family of acetylated derivatives of neuraminic acid, an acute phase reactant by itself. It usually occurs as a terminal component at the non-reducing end of carbohydrate chains of glycoproteins and glycolipids. SA participates in multiple physiological functions, such as cell-to-cell interactions, cell migration and proliferation. Diabetes mellitus (DM) is a chronic metabolic disorder characterized by rise in blood glucose level. Periodontitis is a chronic inflammatory disease of the periodontal tissue, leading to destruction of bone surrounding the tooth and ultimately tooth loss. There is a two way relationship between diabetes mellitus and periodontitis. Periodontitis is the sixth complication of diabetes along with retinopathy, nephropathy, neuropathy, macrovascular disease, and altered wound healing. Inflammatory mediators like interleukin-6 and tumor necrosis factor-alpha produced during periodontal inflammation can interfere with the actions of insulin receptors and worsen the glycemic control of diabetic patients. Periodontitis is a major cause of tooth loss, affecting over 300 million people and bacteria associated with periodontitis are also linked with systemic problems like endocarditis, atherosclerosis. Recent work has highlighted a major role for the host sugar sialic acid in the biofilm physiology and host-pathogen interactions of T. forsithya, a key periodontal pathogen. There exists a need for a biomarker, for early detection of disease evolution and more robust therapy efficacy measurements. Serum sialic acids were estimated in Indian population by diphenylamine method and Thiobarbituric acid method. The average values were 68 ± 2.6 mg percent by DPA method and 56 ± 5 mg percent by TBA (thiobarbituric acid assay) method. Age and sex showed no influence on serum sialic acid level. Objectives of the present study was to compare (TSSA) level in healthy subjects, subjects with (CMP) with and without (NIDDM) and its effect on non-surgical periodontal therapy. In the present study, the participants were divided into three groups: Group A, B and C. Group A consists of systemically healthy subjects, Group B consists of subjects with (CMP) while Group C consists of subjects with (CMP) with (NIDDM) and results of this study indicated that, at baseline, there were significant differences between Group A, B and Group C with respect to all the clinical parameters, including (GI), (OHI-S), (PPD), (CAL), (TSSA) and (HbA1c) levels. Thus (TSSA) level could be considered as novel biomarker in the progression of periodontal disease and diabetic status. Periodontitis could be considered as a potential, modifiable, and independent risk factor for the development of diabetes. Early detection of elevated (TSSA) level may help in interpreting the progression of periodontitis, risk of development of diabetes mellitus in future and also to prevent complications.

Diagnostic Value and Prognostic Use of Presepsin Versus Procalcitonin in Sepsis.

BACKGROUND: Sepsis is a medical problem beyond belief despite the use of modern antibiotics and frequently updated guidelines on resuscitation therapies. It is at this crucial juncture where inflammatory biomarkers have been providing an incredible and easy way to detect sepsis. Various biomarkers have been used over the past decades in medicine to evaluate sepsis, each one better than the previous and having its own pros and cons. In this study, we assess the role of presepsin, which is a CD-14 polypeptide, and procalcitonin which has for some time been the inflammatory marker of choice in sepsis. This is the first study of presepsin as a sepsis biomarker in Indian adults. METHODS: A prospective observational study was conducted in 48 patients who were diagnosed to have sepsis either on admission to the hospital or during their stay in hospital according to the American College of Chest Physicians/Society of Critical Care Medicine (ACCP/SCCM) guidelines during the period from October 2015 to January 2017, after fulfilling all inclusion and exclusion criteria. Efficacy of both inflammatory markers was studied from blood drawn from the patient at the same time, with the same prick. RESULTS: A total of 48 patients were included in this study. The superiority of presepsin over procalcitonin was evident with presepsin having a sensitivity of 46.2 and specificity of 100 and procalcitonin having a sensitivity of 46.2 and specificity of 31.8. The P value of the presepsin results was significant at <0.001. Along with it, presepsin also proved to be a very reliable marker for 28-day mortality with all 12 patients in the presepsin positive group expiring (P value: <0.001). Receiver operating characteristic (ROC) curve was plotted to try and define an optimal normal value for presepsin in an Indian population and the value calculated was 93.71 with a sensitivity of 65.4 and a specificity of 68.2. CONCLUSION: This study shows the superiority of presepsin over procalcitonin as it has much better specificity and a similar sensitivity than procalcitonin and is a better indicator of 28-day mortality. The new cut off that we have postulated here for presepsin improves the efficiency of the inflammatory marker by increasing its sensitivity at the cost of decreasing its specificity slightly.

Association of stress, salivary cortisol level, and periodontitis among the inmates of a central prison in Kerala.

BACKGROUND: The aim of this cross-sectional study was to evaluate the association between stress, salivary cortisol, and periodontitis among the inmates of the central prison. MATERIALS AND METHODS: Seventy inmates were grouped depending on their pocket depth into Group A (pocket depth >4 mm and <6 mm), Group B (at least four sites with pocket depth ≥6 mm), and Group C (pocket depth ≤3 mm). The clinical parameters such as the oral hygiene index-simplified, gingival index, pocket depth, and the clinical attachment levels (CALs) were recorded. Stress was measured using the Depression, Anxiety, and Stress Scale along with prison time served. Saliva samples were collected, and cortisol levels were determined using electrochemiluminescence assay. Chi-square test was used for finding the association between the clinical parameters. The correlation between clinical parameters, stress, salivary cortisol levels, and time served was done using Pearson's rank correlation coefficient. RESULTS: The CALs, the stress score and the salivary cortisol levels were significantly higher in Group B (P < 0.001). Pearson's correlation showed a positive correlation between stress, cortisol level, and pocket depth. A positive correlation which was statistically significant was obtained between salivary cortisol level and prison time served by the inmates. CONCLUSION: Within the limits of this study, it can be concluded that there is a positive relation between stress and periodontal disease. The study suggests that salivary cortisol level can be used as a marker to assess stress.

A randomized study of coconut oil versus sunflower oil on cardiovascular risk factors in patients with stable coronary heart disease.

BACKGROUND AND RATIONALE: Coronary artery disease (CAD) and its pathological atherosclerotic process are closely related to lipids. Lipids levels are in turn influenced by dietary oils and fats. Saturated fatty acids increase the risk for atherosclerosis by increasing the cholesterol level. This study was conducted to investigate the impact of cooking oil media (coconut oil and sunflower oil) on lipid profile, antioxidant mechanism, and endothelial function in patients with established CAD. DESIGN AND METHODS: In a single center randomized study in India, patients with stable CAD on standard medical care were assigned to receive coconut oil (Group I) or sunflower oil (Group II) as cooking media for 2 years. Anthropometric measurements, serum, lipids, Lipoprotein a, apo B/A-1 ratio, antioxidants, flow-mediated vasodilation, and cardiovascular events were assessed at 3 months, 6 months, 1 year, and 2 years. RESULTS: Hundred patients in each arm completed 2 years with 98% follow-up. There was no statistically significant difference in the anthropometric, biochemical, vascular function, and in cardiovascular events after 2 years. CONCLUSION: Coconut oil even though rich in saturated fatty acids in comparison to sunflower oil when used as cooking oil media over a period of 2 years did not change the lipid-related cardiovascular risk factors and events in those receiving standard medical care.

Heart-type fatty acid-binding protein (H-FABP) as an early diagnostic biomarker in patients with acute chest pain.

BACKGROUND: Heart-type fatty acid-binding protein (H-FABP) is an emerging biomarker, which was found to be sensitive for the early diagnosis of acute myocardial infarction (AMI). We prospectively investigated the usefulness of H-FABP determination for the evaluation of acute chest pain in patients arriving at the emergency department. METHODS: Fifty-four patients presenting with acute ischemic chest pain were evaluated. H-FABP was estimated at admission using latex-enhanced immunoturbidimetric assay. Serial cardiac troponin I (cTnI), creatinine kinase-MB (CK-MB) determination, ischemia workup with stress testing, and/or coronary angiogram (CAG) were performed according to standard protocols. RESULTS: The sensitivity and specificity of H-FABP was 89.7% and 68%, for cTnI it was 62.1% and 100%, and for CK-MB it was 44.8% and 92%, respectively for diagnosis of AMI. The sensitivity of H-FABP was found to be far superior to initial cTnI and CK-MB, for those seen within 6h (100% vs. 46.1%, 33% respectively). On further evaluation of patients with positive H-FABP and negative cTnI, 71.4% of the patients had significant lesion on CAG, indicating ischemic cause of H-FABP elevation. Six patients with normal cTnI and CK-MB with high H-FABP had ST elevation on subsequent ECGs and were taken for primary angioplasty. CONCLUSION: H-FABP is a highly sensitive biomarker for the early diagnosis of AMI. H-FABP as early marker and cTnI as late marker would be the ideal combination to cover the complete diagnostic window for AMI. Detection of myocardial injury by H-FABP may also be applied in patients with unstable angina. H-FABP can also be used as a marker for early detection of STEMI before the ECG changes become apparent.

Pregnancy Associated Plasma Protein-A (PAPP-A) Levels in Acute Coronary Syndrome: A Case Control Study in a Tertiary Care Centre.

In recent years, an important objective of cardiovascular research has been to find new markers that would improve the risk stratification and diagnosis of patients presenting with symptoms of acute coronary syndrome (ACS). Established biomarkers for diagnosis of ACS includes troponins and creatine kinase MB (CK-MB). Pregnancy associated plasma protein A (PAPP-A) is an emerging marker which has been described as a marker of plaque instability. PAPP-A is a large metalloproteinase involved in insulin-like growth factor signaling and has been shown to be involved in pathological processes like atherosclerosis. Many studies have been published regarding release of PAPP-A in circulation during ACS. The objective of this study was to evaluate the role of PAPP-A as an early marker of ACS by comparing levels of PAPP-A in patients with acute myocardial infarction (AMI) and unstable angina (UA) with asymptomatic controls. The association of PAPP-A with markers of myocardial necrosis and the association of PAPP-A levels to the presence of risk factors for coronary artery disease was also studied. We measured PAPP-A levels in patients with AMI (30), UA (23) and asymptomatic controls (45). PAPP-A was estimated using PAPP-A US (ultra sensitive) ELISA manufactured by DRG (Germany). PAPP-A levels were significantly elevated in patients with AMI and in patients with UA (mean levels 64.26 ± 1.05 and 36.23 ± 1.05 ng/ml respectively; p < 0.001). Mean PAPP-A levels in controls were 10.68 ± 1.04 ng/ml. In UA cases PAPP-A levels were elevated when the troponin I and CK-MB levels were within the normal range. No correlation was observed between PAPP-A with markers of myocardial necrosis. PAPP-A can serve as a useful biomarker in the diagnosis of ACS, especially UA, where cardiac troponin levels and CK-MB levels are not elevated and ECG changes are inconclusive.