PAI-1 Study in Thalassemia Major Patients Receiving Multiple Blood Transfusion.

Thalassemia is a congenital hemolytic disease which is treated by repeated blood transfusion. Chronic iron overload is currently considered to be the primary cause of mortality in ß-thalassemia, mainly due to the induction of left-sided cardiac failure. Iron overload results from a number of mechanisms associated with the disease itself. In addition to chronic iron overload thalassemic patients are more prone for procoagulant status which in turn lead to clinical thrombotic events. The hypercoagulable state in thalassemia is due to multiple elements, a combination of which is often the drive behind a clinical thromboembolic events. PAI-1 study was done in thalassemia major patients receiving multiple blood transfusion as a marker for procoagulant status. Total of 30 thalassemic patients on repeated blood transfusion was included in the study and total of 30 healthy age and sex matched controls were included in the study. It was also found that there was significant differences between cases and controls. The mean level of PAI 1 in controls was 3047 ± 414 pg/ml, the value in cases was 3683 ± 358 pg/ml. The level was significantly increased (p < 0.05) in the cases compared to controls. PAI-1 levels were also compared with the total number of blood transfusion which correlates well.

Comparative study of serum amylase and lipase in acute pancreatitis patients.

To establish utility of single enzymatic marker for the diagnosis of acute pancreatitis. This is a cohort study. Tertiary care centre proven cases of acute pancreatitis (n = 50) admitted in surgery ward between December 2011 and May 2013 were included in the study. Serum amylase and lipase were performed along with many analytes. All relevant data including serum lab values and imaging were collected. All 50 patients included in the study had raised serum lipase, 42 patients had both amylase and lipase raised, 8 patients had amylase normal but lipase raised. In smaller hospitals where limited lab and radiological facilities are available, estimation of serum lipase will be a better choice over serum amylase in diagnosis of acute pancreatitis.

Correlation of Microalbuminuria with Estimated GFR (eGFR) by Cockcroft-Gault and MDRD Formula in Type 2 Diabetics and Hypertensives.

Increase in urine albumin excretion rate (AER) precede a fall in glomerular filtration rate in patients developing diabetic chronic kidney disease (CKD). Our results have shown that 7 (50 %) of diabetic and hypertensive individuals with decreased GFR do not have increased AER. In this cross-sectional study, we measured AER of 75 patients with type 2 diabetes and hypertension by immunoturbidimetric method. We correlated the results with eGFR values obtained by Cockcroft-Gault and MDRD method. The method used was not a compensated method. We measured serum creatinine by modified Jaffe's kinetic method in autoanalyzer XL-600. Analysis of data showed positive correlation between eGFR and microalbuminuria by both the methods with eGFR <60 mL/min/1.73 m(2). Pearson's correlation co-efficient (r) was 0.9 (p = 0.0001) by Cockcroft-Gault formula and 0.69 (p = 0.0063) by MDRD formula. Our results concluded that there was positive correlation between AER and eGFR <60 mL/min/1.73 m(2). We have recognized that these two parameters provide a complimentary benefit in management of cases with CKD.

SARS-CoV-2 infecting endothelial cells, biochemical alterations, autopsy findings and outcomes in COVID-19, suggest role of hypoxia-inducible factor-1.

Researchers around the world have experienced the dual nature of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), 'tragically lethal in some people while surprisingly benign in others'. There have been congregating studies of the novel coronavirus disease (COVID-19), a disease that mainly attacks the lungs but also has mystifying effects on the heart, kidneys and brain. Researchers are also gathering information to ascertain why people are dying of COVID-19, whether it is solely a respiratory disorder, a coagulation disorder or multi-organ failure. Alterations in laboratory parameters like lactate, ferritin and albumin have been established as risk factors and are associated with outcomes, yet none have not been sub stantiated with a scientific biochemical rationale. SARSCoV-2 affects the alveolar type II epithelial cells which significantly disturbs its surfactant homeostasis, deprives Na,K-ATPase of ATP, thereby disturbing the alveolar lining fluid which then gradually decreases the alveolar gaseous exchange initiating the intracellular hypoxic conditions. This activates AMP-activated kinase, which further inhibits Na,K-ATPase, which can progressively cause respiratory distress syndrome. The virus may infect endothelial cell (EC) which, being less energetic, cannot withstand the huge energy requirement towards viral replication. There - fore glycolysis, the prime energy generating pathway, must be mandatorily upregulated. This can be achieved by Hypoxia-inducible factor-1 (HIF-1). However, HIF-1 also activates transcription of von Willebrand factor, plasminogen activator inhibitor-1, and suppresses the release of thrombomodulin. This in turn sets off the coagulation cascade that can lead to in-situ pulmonary thrombosis and micro clots. The proposed HIF-1 hypothesis justifies various features, biochemical alteration, laboratory as well as autopsy findings such as respiratory distress syndrome, increased blood ferritin and lactate levels, hypoalbuminemia, endothelial invasion, in-situ pulmonary thrombosis and micro clots, and multi-organ failure in COVID-19.

Markers for differentiation of tubercular pleural effusion from non-tubercular effusion.

BACKGROUND: The limitations of the conventional methods for diagnosing tuberculosis (TB) have spurred multi-faceted research activities throughout the world. This study aims to explore the levels of adenosine deaminase (ADA) and interleukins in pleural effusion of tuberculous, malignant, and miscellaneous origin for differential diagnosis of tubercular and non-tubercular effusion. METHOD: Adenosine deaminase was estimated by kinetic method employing xanthine oxidase while interleukins were measured using commercially available ELISA kits in pleural fluids of tubercular and non-tubercular origin. RESULTS: Pleural fluids INF-γ, sIL-2R, TNF-α and ADA were significantly higher in TB group (n = 48) as compared to the non-TB group (n = 33) (mean ± SD: INF-γ; 1,958.7 ± 896.5 pg/mL vs 356.9 ± 733.6 pg/mL, sIL-2R; 6,101 ± 1,753.8 pg/mL vs 3,166 ± 2,611.1 ± pg/mL, TNF-α; 195.5 ± 292.1 pg/mL vs 59.7 ± 128.9 pg/mL, ADA; 123.6 ± 81.8 IU/L vs 48 ± 48.5 IU/L, P < 0.01). CONCLUSION: INF-(is more sensitive and specific than ADA for the diagnosis of TB and should be added to the armamentarium of the diagnostic workup of pleural fluids for timely and accurate diagnosis of TB and differentiation of tubercular pleural effusion from non-tubercular effusion.

Analysis of short-term variation and long-term drift during reagent kit lot change in an NABL accredited clinical biochemistry laboratory.

BACKGROUND: Kit lot change in clinical biochemistry labs leads to variations in patient results. This study planned to identify variations during 60 reagent lot changes in our laboratory during the period from June 2018 to May 2019. METHODS: A statistical analysis was performed to identify the difference between patient samples results variations and QC results. The long term drift was analyzed using a regression test. RESULTS: There was a significant difference between the patient and QC results in 16.7% of reagent lot changes. Moreover, the extent of variation in QC results was 3.3%. No long-term drift was seen in three analytes which were studied using regression analysis. CONCLUSIONS: Our results showed that, during reagent kit lot change, along with QC material, the patient samples should also be run in order to identify the variation. However, this practice is presently ignored by most of the laboratories. There was no accumulated effect in our laboratory due to reagent kit lot change.

Olfactory Dysfunction Among Asymptomatic Patients with SARS CoV2 Infection: A Case-Control Study.

Olfactory dysfunction (hyposmia, anosmia) is a well-recognized symptom in patients with coronavirus disease-19 (COVID-19). Studies of olfactory dysfunction in asymptomatic patients have not been reported. We conducted a study looking for the presence of olfactory dysfunction with an objective assessment tool in asymptomatic Covid 19 and compared it with patients with mild COVID-19 and age-matched controls. We recruited 57 male patients each of Mild COVID-19, asymptomatic Covid 19, and healthy controls for the study. All participants underwent evaluation of smell threshold by Butanol Threshold test (BTT) and ability to distinguish common odors by Smell identification test. The scores of each test were recorded on a numerical scale. The participants in all three arms were matched for age, history of smoking, and pre-existing medical conditions. The mean scores of the Butanol Threshold test in Mild COVID-19, asymptomatic Covid 19 and controls were 2.95 ± 2.25 (0-7.5), 3.42 ± 2.23 (0-7.5), and 4.82 ± 1.86 (0-8), respectively. A one-way ANOVA showed a significant difference between groups (df 2, MS 53.78, F 11.94, p < 0.005). Intergroup differences using the student T-test showed significantly low BTT scores in Mild COVID-19 (p < 0.005) and asymptomatic (p < 0.005) as compared to control. BTT scores could not distinguish between asymptomatic patients and control. The smell threshold was impaired in asymptomatic Covid 19 and Mild COVID-19. Butanol Threshold Test score could not differentiate between asymptomatic Covid 19 and controls.

Incidence of microalbuminuria in hypertensive patients.

The prevalence of microalbuminuria was assessed in 174 albustix negative hypertensive patients by estimating albumin in the morning random urine samples by immunoturbidimetric method within four hours of voiding of urine. The urine samples were not stored and collected without any preservatives. The urinary albumin was calculated in terms of ratio with respect to urinary creatinine and expressed as albumin creatinine ratio (mg/g). Out of 174 albustix negative hypertensives, 58 (33.3%) patients were found to have microalbuminuria. The prevalence of microalbuminuria in males and females was found to be 34% and 30.7% respectively. No correlation was found between the Body Mass Index (BMI) and albumin excretion (r(2) = 0.0271) and between duration of hypertension and urinary albumin excretion (r(2) = 0.0042). Prevalence of microalbuminuria in nonsmokers and non-alcoholic hypertensives was 20%. The prevalence in alcoholics, smokers and both smokers and alcoholics was found to be 35%, 42% and 41% respectively. The high prevalence of microalbuminuria than the various reported studies on the subject demands establishment of a screening programme for microalbuminuria, implementation of specific intervention methods and education of hypertensive patients about the consequences of smoking and alcohol on possible involvement of renal system.

A kinetic method amenable to automation for ceruloplasmin estimation with inexpensive and stable reagents.

OBJECTIVES: To develop a kinetic, inexpensive, automatable assay for ceruloplasmin as ferroxidase. DESIGN AND METHODS: Dithiothreitol has been used to stabilize the substrate and quinolones to complex the ferric. RESULTS: Mean ferroxidase activity in healthy adults, Wilson's disease and pregnancy was 787.29, 178.5 and 1828.09 IU/L, respectively. Correlation coefficient of ferroxidase vis-à-vis copper, ferroxidase by ferrozine and ceruloplasmin were 0.90, 0.94 and 0.92, respectively. CONCLUSIONS: Norfloxacin method is a simple, inexpensive and automatable kinetic assay.

Serum ferroxidase albumin ratio as a marken in pulmonary tuberculosis.

Serum ferroxidase and albumin levels were determined in 98 patients of tubercuiosis, of whom 49 were freshly diagnosed, sputum positive (group-I) & 49 were completely treated patients (group-II). Forty nine age and sex matched healthy individuals were taken as controls. Mean±SD of serum ferroxidase and albumin levels in controls, group-I and group-II was found to be 864.35±106.35 IU/L & 3.91±0.234 g/dL, 1603.76±222.65 IU/L & 3.24±0.518 g/dL and 1001.78±201.63 IU/L & 3.82±0.43 g/dL, respectively. Serum ferroxidase in group I was significantly higher as compared to controls and group-II (p<0.01). The decreased levels of serum albumin in group I, as compared to control and group-II was statistically significant (p<0.01). Serum ferroxidase: albumin ratio (Ferroxidase in International Unit per gram of albumin) in group I (50.47±10.36 IU/g) was significantly higher than controls (22.22±3.3 IU/g), (p<0.001) while in group II it was significantly lower (26.72±7.18 IU/g, p<0.001) than group-I and close to control values. Serum ferroxidase: albumin ratio (IU/g) can therefore be incorporated as a surrogate marker to assist in diagnosis and prognosis of pulmonary tuberculosis.

Urinary N-acetyl beta glucosaminidase and gamma glutamyl transferase as early markers of diabetic nephropathy.

Albumin and enzymes-N-acetyl-beta-glucosaminidase (NAG) and gamma glutamyl transferase (GGT) were estimated in the morning random urine samples of 196 albustix negative diabetic patients to evaluate the clinical utility of these urinary enzymes as early markers of diabetic nephropathy. Albumin was estimated by immunoturbidimetric method and enzymes by linetic essay within six hours of voiding of urine. The urinary albumin and urinary enzyme concentration was calculated in terms of ratio with respect to urinary creatinine. Correlation coefficient (r) bewween urinary albumin and urinary enzymes in normoalbuminuric, microalbuminuric and overall diabetic cases was 0.23, 0.32 and 0.40 respectively for NAG, and 0.08, 0.06 and 0.18 respectively for GGT. NAG excretion was found increased in 34%, 63.7% and 49.5% of normoalbuminuric, microalbuminuric and overall diabetic cases respectively while GGT in 6.4%, 24.5% and 15.8%. The correlation coefficient between urinary albumin and NAG in normoalbuminuric, microalbuminuric, and overall diabetic patients with increased NAG excretion was found only 0.31, 0.27 and 0.35 respectively. No correlation was found between duration of diabetes and enzyme excretion. The study suggests that urinary NAG or GGT or both together do not have any clinical significance as an early marker of diabetic nephropathy.

Diagnostic Utility of Biomarkers in COPD.

BACKGROUND: COPD will become the third leading cause of death by 2020. There are many situations in which spirometry, the primary tool for diagnosis of COPD, cannot be performed, and thus, the staging and status of these patients cannot be determined. To date, there is no known biochemical marker used for diagnosing COPD. This study aimed to explore the utility of biomarkers for diagnosis of COPD. METHODS: This was an observational study composed of 96 stable subjects with COPD and 96 subjects with normal lung function. Each group contained an equal number of smokers and nonsmokers. Serum levels of superoxide dismutase 3, glutathione peroxidase, catalase, ceruloplasmin ferroxidase activity, C-reactive protein, and surfactant protein D (SPD) were estimated. Ferroxidase activity was estimated by a kinetic method, whereas the other analytes were measured by enzyme-linked immunosorbent assay. The cutoff value, sensitivity and specificity at the cutoff value, and area under the curve for each analyte were determined from receiver operating characteristic curve. RESULTS: Significantly decreased superoxide dismutase 3 and increased ferroxidase activity, SPD, glutathione peroxidase, and C-reactive protein levels were found in subjects with COPD. For all subjects and nonsmoking subjects with COPD, the area under the curve was highest for ferroxidase activity, followed by glutathione peroxidase, SPD, and C-reactive protein, with a sensitivity and specificity of > 73%. For smoking subjects with COPD, the area under the curve was highest for SPD, followed by glutathione peroxidase, ferroxidase activity, and C-reactive protein, with a sensitivity and specificity > 67%. Some combinations of markers were found to give either a sensitivity or specificity of > 95%, which can be utilized to rule in and rule out COPD. CONCLUSIONS: Biomarkers can be reliably utilized in the diagnosis of COPD. Of all the markers, SPD appears to be the most promising in smokers, whereas ferroxidase activity shows promise in nonsmokers. To rule out COPD, ferroxidase activity or glutathione peroxidase can be potentially useful, whereas to rule in COPD, ferroxidase activity and glutathione peroxidase appear to be the most promising combination in both nonsmoking and smoking subjects.

METHODS FOR ESTIMATION OF BLOOD GLUCOSE : A COMPARATIVE EVALUATION.

Glucose in blood is the most frequent analyzed parameter in a clinical chemistry laboratory. In Armed Forces Laboratories, copper reduction method (Modified Folin Wu) is commonly used. Here we have compared this method as well as O-Toluidine and GOD-POD method with reference UV-Hexokinase method. Both Modified Folin Wu and O-Toluidine showed upward deviation with substantial imprecision (CV=4.9% - 3.5% and 6% - 5.8% respectively) and inaccuracy (Average deviation = 5.76% and 10.68% respectively). GOD-POD method was found linear (up to 500 mg/dl), with good precision (CV=0.7% to 1.4%) and accuracy (Average deviation= -0.97%). This method is simple, rapid, economical and sensitive, and can be adopted to a routine colorimeter.