Antinuclear Antibody Profiling in Patients of a Tertiary Care Centre in Central India.

The incidence of autoimmune disorders that includes the connective tissue diseases has seen a rise in India in recent times. Antinuclear antibodies, the telltale sign of systemic autoimmune response, thus can be used as a screening tool and also to support the diagnosis of systemic autoimmune disease. The present retrospective cross- sectional analysis aimed to study the antinuclear antibodies profile (patterns and specific antibody reactivity) amongst suspected cases of auto-immune disorders at a tertiary care teaching hospital. The study retrieved and reviewed reports of 644 patients sent for ANA testing by indirect immunofluorescence assay over a period of 1 year by different specialty departments. Positive samples were further processed for anti-ds-DNA antibody and antibodies to extractable nuclear antigen. Data collected was statistically analysed. ANA pattern positivity was observed in 31% of cases and a positive antibody reactivity was seen in 66% of them. Female predominance (82%) was noted in both pattern positivity and antibody reactivity. High levels of pattern positivity and antibody reactivity was found in the young adults (45.9%). Amongst the ANA patterns, the nuclear homogenous pattern was found the commonest. The common antibodies associated with this pattern were anti-dsDNA and U1 Sm/RNP antibodies. A stronger fluorescence intensity on initial screening showed a higher confirmation rate for specific antibodies on immunoassay. High occurrence of positive ANA patterns in autoimmune disorders suggests its utilization as a screening tool for them and would also play an adjuvant to the diagnosis. Early knowledge about future autoimmunity will earn better prognostic achievements through better treatment interventions.

Sickle Cell Trait Presenting as Chronic Calcific Pancreatitis with Pseudocyst- A Case Report.

Sickle cell disease is known to cause acute pancreatitis either due to gall stones obstructing the pancreatic duct or by vaso-occlusive mechanism. However chronic pancreatitis is a very rare complication in sickle cell anemia. We report a case of sickle cell trait presenting with chronic pancreatitis with pseudo cyst. USG abdomen and CT abdomen confirmed the diagnosis of chronic calcific pancreatitis with pseudocyst. Etiological work up for other causes did not reveal anything except sickle cell trait. This case represents a rare association between chronic calcific pancreatitis and sickle cell trait.

Primary Antiphospholipid Syndrome Associated with Autoantibodies Against Golgi Bodies: A Case Report of a 32-Year-Old Female.

Antiphospholipid syndrome is an autoantibody mediated disorder characterised by thrombotic manifestations and/or obstetric morbidity. The autoantibodies are directed against phospholipid binding plasma proteins. Amongst the clinical features abdominal presentation is an unusual feature in this syndrome. We present the case report of a 32-year female whose complaints was abdominal pain for one week and no history of previous foetal loss, who responded well to warfarin and has not developed systemic lupus erythematous even after follow up. Characteristic of this patient is the appearance of auto antibodies against Golgi bodies. Although antinuclear antibodies are seen in patients of rheumatic disease like systemic lupus erythematous, its presence in individuals with unusual presentation of antiphospholipid syndrome may facilitate in diagnosis.

Congruity in Quality Indicators and Laboratory Performance.

Proficiency of laboratory services is the mainstay in clinical medicine in providing error free diagnostic results. The efficiency of the laboratory needs to be evaluated as per standard international criterion. The quality indicators of the different phases of total testing process are considered the fundamental measurable tool for evaluation of laboratory performance. In order to optimize the laboratory's proficiency and accreditate it as per international standard in our newly established lab, the study was conducted to evaluate the frequency of errors incurred by laboratory and nonlaboratory professionals during the whole testing process. Retrospective analysis was done for data received from April 2016 to Dec 2016 in our lab. Total number of samples received was 61,674, out of which 43200 samples could be analyzed for quality indicators. Total numbers of tests processed in these samples were 172,800. In the study samples, 26.5% errors were due to pre-analytical factors whereas 9.4% of errors were contributed by analytical phase and 18% by post-analytical phase. Inappropriateness of test requisition was observed to be the major attributing determinant for pre-analytical errors. Instrumentation efficiency in form of frequent breakdown (~7%), greatly affected the proficiency of analytical phase in our lab. 12% of post-analytical errors were ascribed by excessive turn-around-time. However, timeliness of critical value call out and reporting for STAT samples revealed high proficiency up to 97%. High error rates were observed in pre-pre- and pre-analytical phases that also accorded for high error frequency in post analytical phase. This emphasizes urgent need to formulate guidelines for processing all steps of total testing process and initiate strategic measures for reducing risk of errors and increasing patient safety.

RBD mutations at the residues K417, E484, N501 reduced immunoreactivity with antisera from vaccinated and COVID-19 recovered patients.

Introduction: It is unclear whether induced spike protein-specific antibodies due to infections with SARS-CoV-2 or to the prototypic Wuhan isolate-based vaccination can immune-react with the emerging variants of SARS-CoV-2. Aim/objectives: The main objective of the study was to measure the immunoreactivity of induced antibodies postvaccination with Covishield™ (ChAdOx1 nCoV-19 coronavirus vaccines) or infections with SARS-CoV-2 by using selected peptides of the spike protein of wild type and variants of SARS-CoV-2. Methodology: Thirty patients who had recovered from SARS-CoV-2 infections and 30 individuals vaccinated with both doses of Covishield™ were recruited for the study. Venous blood samples (5 mL) were collected at a single time point from patients within 3-4 weeks of recovery from SARS-CoV-2 infections or receiving both doses of Covishield™ vaccines. The serum levels of total immunoglobulin were measured in both study groups. A total of 12 peptides of 10 to 24 amino acids length spanning to the receptor-binding domain (RBD) of wild type of SARS-CoV-2 and their variants were synthesized. The serum levels of immune-reactive antibodies were measured using these peptides. Results: The serum levels of total antibodies were found to be significantly (p<0.001) higher in the vaccinated individuals as compared to COVID-19 recovered patients. Our study reported that the mutations in the RBD at the residues K417, E484, and N501 have been associated with reduced immunoreactivity with anti-sera of vaccinated people and COVID-19 recovered patients. Conclusion: The amino acid substitutions at the RBD of SARS-CoV-2 have been associated with a higher potential to escape the humoral immune response.

Processing Validation Metrics of Syva Enzyme-Multiplied Immunoassay Technique (EMIT) Methotrexate Assay for Beckman Coulter System.

BACKGROUND: High-dose methotrexate (HDMTX), defined as a dose greater than 500 mg/m2, is used to treat a variety of cancers; and though safe, it can cause major toxicity. Syva enzyme-multiplied immunoassay technique (EMIT) methotrexate (MTX) assay (Gurgaon, India: Siemens Healthcare Diagnostics Ltd.) uses a homogeneous enzyme immunoassay method. Low-end precision performances are very important for laboratory methods, especially when their results have clinical significance at these levels. METHODOLOGY: A total of 25 replicates (five replicates per run, for five runs) were analyzed for profiling. Precision, accuracy, linearity, limit of blank, limit of detection, and limit of quantification were determined using existing guidelines. Imprecision profile and limit of quantitation (LoQ) at 10% were determined by fitting data with hyperbolic regression. RESULTS: The coefficient of variation percentage (CV%) for low, mid, and high-level internal quality control (IQC) was 1.25%, 3.45%, and 1.55%, respectively. Similarly, estimated bias was -4.58%, -3.54%, -7.21% for each level. The assay linearity was maintained from a range of 0.041-1.993 mmol/L with an R2 of 0.959. The limit of detection was estimated to be 0.07 mmol/L. CONCLUSION: Syva EMIT MTX assay can be precisely and accurately used to measure low levels of serum methotrexate at levels lower than claimed by the manufacturer, aiding in the monitoring of toxicity in patients.

An Approach to Re-evaluate the Reference Cutoff of the Parameters of Newborn Screening: An Observational Study.

Background Unless a cutoff level of the parameters of newborn screening (NBS) is defined, a screening test's results would end in high recall rates and apprehensive parents. The study aimed to establish a cutoff level of the healthy term newborns. Materials and methods The study was a retrospective observational data analysis on a cohort of 1158 term newborns who underwent NBS in our institute. The percentile distribution of the NBS parameters was computed and the 99th percentile value was considered the new cutoff. For lower values, such as neonatal glucose 6-phosphate dehydrogenase (nG6PD) and neonatal biotinidase (nBIOT), low percentile values were regarded as new cutoff value. Results Neonatal thyroid stimulating hormone (nTSH), nG6PD, neonatal immunoreactive trypsinogen (nIRT), and nBIOT showed a wide variation in the distribution. Most newborns had neonatal galactose (nGAL), nIRT, and nBIOT values above the median. The 99th percentile value of nTSH was 14.5 mIU/L, and that of neonatal 17-hydroxyprogesterone (n17-OHP) was 43.7 nmol/L. The 1.0th percentile value for nG6PD was decreased to 2.18 IU/gHb. The new cutoff values for nBIOT, nIRT, neonatal phenylketonuria (nPKU) and nGAL were 48.59 U, 95.3 µg/L, 2.3 mg/dL and 15.9 mg/dL. The mean and median nTSH values did not significantly differ (p=0.99) in the first five days of birth. On the contrary, the study population depicted considerably raised levels of n17-OHP on day 3, followed by a sharp decrease (p=0.029). Similarly, nIRT displayed significant differences in the first five days (p=0.017). Conclusion Using the 99th percentile values of the NBS parameters as the new cutoff levels might be beneficial in terms of the recall rates and cost burden.

Association of Methylenetetrahydrofolate Reductase Gene Polymorphism in Mothers With Adverse Clinical Outcomes in Neonates.

BACKGROUND: The presence of polymorphic methylenetetrahydrofolate reductase (MTHFR) in mothers poses a risk for numerous detrimental outcomes in neonates. The present study investigated the association of maternal MTHFR A1298C and C677T single nucleotide polymorphisms (SNPs) with the clinical outcomes in their neonates. MATERIALS AND METHODS: The cross-sectional study included 60 mothers and their neonates. Blood samples from mothers were analyzed for MTHFR A1298C and C677T SNP genotyping by real-time polymerase chain reaction. Clinical details of mothers and neonates were documented. Study groups were stratified based on wild, heterozygous, and mutant genotypes for the respective polymorphisms observed in mothers. Multinomial regression was applied for the association, followed by gene model formulation to estimate the impact of the genetic variants on the outcomes. RESULTS: The frequency percentages of mutant CC1298 and TT677 genotypes were 25% and 8.06%, respectively, and the mutant allele frequencies (MAF) were 42.5% and 22.5%. Percentages of adverse outcomes such as intrauterine growth restriction, sepsis, anomalies, and mortality were higher in neonates born to mothers with homozygous mutant genotypes. Maternal C677T MTHFR SNPs revealed a significant association with neonatal anomalies (p = 0.001). The multiplicative risk model depicted OR (95% CI) for CT vs. CC+TT as 3.0 (95% CI: 0.66-13.7), and for TT vs. CT+CC was 15 (95% CI: 2.01-112.12). The C677T SNP in mothers predicted a dominant model for neonatal death (OR (95% CI): 5.84 (0.57-60.03), p = 0.15), whereas the A1298C reported recessive model for 1298CC mothers (OR (95% CI): 11 (1.05-115.5), p = 0.02). Both the genotypes assumed a recessive model for adverse neonatal outcomes: OR (95%CI) for CC vs. AA+AC was 3.2 (0.79-12.9, p = 0.1), and for TT vs. CC+CT was 5.48 (0.57-175.7, p = 0.2). The risk for sepsis in neonates was nearly six times higher in those born from mothers with homozygous CC1298 and TT677 than in the wild and heterozygous variants. CONCLUSION: Mothers with C677T and A1298C SNPs are highly susceptible to adverse outcomes in their neonates. Hence, screening the SNPs during the antenatal period can purposefully serve as a better predictive marker, following which proper clinical management could be planned.

Altered T-cell profile in sickle cell disease.

Background: Impaired immune status due to altered T-cell response in sickle cell disease (SCD) might provide substantial insight into immune activity in SCD patients. Materials & methods: A total of 30 healthy control, 20 SCD patients in a crisis state and 38 SCD patients in a steady state were evaluated for T-cell subsets. Results: A significant decrease in CD8+ (p = 0.012) and CD8+45RA-197+ (p = 0.015) T-cells were observed among SCD patients. Naive T-cells (45RA+197+; p < 0.01) were elevated and effector (RA-197-) and central memory (RA-197+) T-cells were grossly reduced in the crisis state. Negative regression of naive T-cells with CD8+57+ affirmed immune inactivation. The predictor score reflected 100% sensitivity for predicting the crisis state (area under the curve = 0.851; p < 0.001). Conclusion: Monitoring naive T-cells with predictive scores can help assess the early shift from a steady state to a crisis state.

The sickle-shaped hemoglobin in sickle cell disease (SCD) patients are known to cause frequent episodes of blockage in small vessels. Repeated episodes of blockage result in tissue injury and create a state of chronic inflammation. In response, a series of inflammatory reactions initiate such that the immune response in these patients is quite altered. To understand these changes, this study was conducted to observe alterations in T-cell subtypes and gain substantial insight into immune activity in SCD patients. A total of 30 healthy control, 20 SCD patients in a crisis state and 38 SCD patients in a steady state were evaluated for T-cell subsets. The SCD patient experienced a gross decrease in T-cells with killing ability and memorizing ability for immune responses. The SCD patients in crisis state reported a significant increase in inactivated T-cells but the levels of activated T-cells that can defend and memorize the immune response were quite low. The finding suggested that this group of SCD patients had compromised immune activation that hindered the activation and differentiation of inactivated T-cells to their effector and memory cells. An equation was derived considering all the parameters that were significantly altered to derive a predictive score that showed 100% sensitivity for predicting a crisis state. Hence, it is proposed that monitoring the inactivated T-cell population or predictive score might help clinicians to assess clinical severity at an early stage and initiate appropriate preventive measures.

Rhinosporidiosis- Epidemiological, Clinicoradiological, Immunological Profile.

Introduction: Rhinosporidiosis is an enigmatic disease with many unsolved queries right from taxonomy to treatment. This study has been done to understand the disease characteristics with a peek into the lesser known immunological aspects of it by studying the changes in levels of certain primarily cell-mediated immunity (CMI)-specific cytokines in rhinosporidiosis patients. Materials and Methods: A prospective observational study was performed. Detailed epidemiological and clinicoradiological assessment was done along with selected inflammatory and immunological markers. The tests for immunological parameters were done by ELISA and CLIA and data were compiled and analyzed using appropriate statistics. Results: Disease showed male predominance and all patients gave a universal pond bathing history. Majority patients had O+ve blood group. Right side was affected most with nasal obstruction being commonest symptom. Nasal cavity was involved in majority of cases with inferior turbinate and meatus being sites of maximum occurrence and attachment. Nasopharynx, oropharynx were other involved sites. Extra-nasal sites included skin and parotid gland. Endoscopic and CECT findings were similar and confirmed intraoperatively. Tests for inflammatory markers showed no significant change in patients. Immunological markers -IL-6, TNF-beta- levels showed significant increase though no such increase was found with IFN-gamma levels. Conclusion: Rhinosporidiosis has a definite epidemiological and clinical-radiological profile. A clear association with exposure to contaminated water is present which could not be further associated with disease duration or recurrence. The immunological profile needs to be further investigated upon since it remains quite elusive.

Serological Evaluation of Antibody Titers After Vaccination Against COVID-19 in 18-44-Year-Old Individuals at a Tertiary Care Center.

Background The evaluation of the effectiveness of the vaccines (ChAdOx1-nCOV; Covishield and BBV-152; Covaxin) against coronavirus disease 2019 (COVID-19) is necessary to assess their efficacy. Because most antibodies that neutralize the coronavirus are directed against the receptor binding domain within the spike protein of the virus, these antibodies serve as markers for viral neutralizers and, in turn, for vaccine response. The present study aimed to evaluate the anti-neutralizing antibody (receptor binding domain (RBD)) and immunoglobulin G2 (IgG2) titers following the completion of the vaccination schedule (both vaccines) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Methodology In this longitudinal prospective study, conducted in a tertiary care center, 30 sequentially (two doses) vaccinated study participants between the ages of 18 and 44 years were sampled for estimation of anti-RBD antibody titer and IgG2. All statistical analysis was done using SPSS version 20 (IBM Corp., Armonk, NY, USA). P-values less than 0.05 were considered significant. Results There was a statistically significant increase in the neutralizing antibody titer after one month of the second dose (z = -4.597, p < 0.001), while a significant decrease was seen in the IgG2 levels (z = -3.075, p = 0.002). The results showed a significant neutralizing effect of the vaccines being used, with Covishield being more effective than Covaxin. The levels of neutralizing antibodies were independent of all demographic variables such as age, sex, and body mass index. Conclusions This study evaluating the efficacy of the two vaccines, namely, Covishield and Covaxin, is the first of its kind in the state of Chhattisgarh. The results of this study are similar to previous studies conducted in India and outside India, concluding that Covishield is a more effective vaccine.

Effect of the Angiotensin-Converting Enzyme (ACE) (I/D) Polymorphism in COVID-19 Patients and Their Healthy Contacts.

Introduction The quest to understand the pathophysiology behind the deleterious effects of the coronavirus disease 2019 (COVID-19) outbreak took a turn when involvement of the angiotensin converting enzyme (ACE) receptors in different organs, especially the lungs, could explain all the clinical manifestations and adverse events in patients. The I/D polymorphism in the ACE gene, having been attributed in various studies, was also seen to have an effect in this pandemic. Present study aimed to analyze the effect of this I/D mutation in COVID-19 patients and in their healthy contacts. Methods Patients with past history of COVID-19 infection and their healthy contacts were enrolled in the study after obtaining ethical clearance and informed consent. The polymorphism was studied by real-time polymerase chain reaction (PCR). Data was analyzed in SPSS version 20 (IBM Corp., Armonk, NY, USA). p value less than 0.05 was taken as significant. Results The allelic distribution followed the Hardy-Weinberg equilibrium, with the wild 'D' allele being dominant in the population. Between the case and controls, the mutant 'I' allele was observed more in the controls, and the association was statistically significant. Conclusion From the results of the present study, it could be concluded that while the wild 'D' allele led to higher chances of being affected with COVID-19, the polymorphism to 'I' allele was relatively protective in nature.

A pilot study to evaluate the changes in venous blood gas parameters and hypoxia biomarkers in health care workers using different kinds of masks.

Background: The study is aimed to investigate the metabolic alterations and changes in biochemical parameters associated with extended mask. Methods: It was a prospective comparative study conducted on 129 participants comprised of 37 healthy controls and 92 health care workers using different kind of masks like, cloth mask, surgical masks and N95-FFR/PPE. Two samples on day-1 and day-10 were collected for analysis of blood gas parameters, serum hypoxia-inducible factor-α (HIF-α), and erythropoietin (EPO). Results: Oxygen saturation percentage (sO2) of 72.68 (P = 0.033) was significantly low, whereas, Na+ (P = 0.05) and Ca2+ (P < 0.001) were raised in exposed individuals than the healthy controls. The serum HIF-α level of 3.26 ng/mL, was considerable higher in the exposed individuals than controls (P = 0.001). pO2 and sO2 were the lowest and HIF-α and EPO were raised in N95-FFR/PPE of all mask users (P < 0.01). A significant difference was evidenced for pCO2, pH, Na+, Ca2+, and EPO in the exposed group. A positive correlation between the duration of mask use (in hours) with HIF-α (r = 0.247, P = 0.005) and Ca2+ (r = 0.306, P < 0.001) was observed. The major complaints in N95-FFR/PPE users were headache (15.2%) and polydipsia (33.3%). Conclusion: The study findings depicted a significant metabolic alterations in PPE/N95 users which could be due to chronic hypoxic exposure of the tissues.

Machine learning algorithm to evaluate risk factors of diabetic foot ulcers and its severity.

Early identification of the risk factors associated with development of diabetic foot ulcer (DFU) can be facilitated using machine learning techniques. The aim of this study is to find out the association of various clinical and biochemical risk factors with DFU and develop a prediction model using different machine learning algorithms. Eighty each of type 2 diabetes mellitus (T2DM) with DFU and (T2DM) without DFU were enrolled for this observational study. Clinical and laboratory data were analysed using different machine learning algorithms: Support vector machines (SVM-Poly K), Naive Bayes (NB), K-nearest neighbour (KNN), random forest (RF) and three ensemble learners: Stacking C, Bagging and AdaBoost for constructing prediction models for discriminating between the two groups (stage I classification) and ulcer type classification (stage II classification). Ensemble learning performed better than individual classifiers in terms of various performance evaluation metrics. New risk factors like ApoA1 and IL-10 for development of DFU in diabetes mellitus were identified. IL-10 along with uric acid could discriminate the grades of ulcers according to its severity. Decision fusion strategy using Stacking C algorithm resulted in enhanced prediction accuracy for both the stages of classification which can be used as a complementary method for computational screening for DFU and its subtypes. Current methodology for T2DM with DFU/T2DM without DFU and ulcer type classification.

A study of apolipoprotein A1(ApoA1) and interleukin-10(IL-10) in diabetes with foot ulcers.

Background: The high morbidity, mortality and associated economic burden have entailed to identifying early biomarker of diabetic foot ulcers (DFU). Pro-inflammatory and anti-inflammatory molecules play a role in the chronic inflammation associated with diabetic foot ulcers (DFU). Aim: This study aims to find the association between ApoA1, IL-10, TNF-α and diabetic foot ulcers, and whether their levels can assess the severity of the disease. Method: Two groups, diabetic mellitus without foot ulcers and diabetes with foot ulcers were recruited for the study. Detailed clinical history was obtained and blood was collected to measure TNF-α , IL-10 and Apo A1. The association between variables was analysed using Pearson correlation test. ROC analysis was used to identify cut-off values of ApoA1, IL-10 and TNF-α in diabetes patients with foot ulcers. Results: The presence of pro-inflammatory parameter, TNF-α , was higher and anti-inflammatory biomarkers, HDL, ApoA1 and IL-10 were lower in patients of DFU than those without foot ulcers (p < 0.001). Increasing age, smoking, retinopathy, eGFR and inflammatory biomarkers like low levels of ApoA1 (p < 0.005) and IL-10 (p < 0.001) significantly contributed to the development of diabetic foot ulcers. ROC curve identified the cut-off for ApoA1 and IL-10 as 89.82mg/dL and 78.80pg/mL respectively. Conclusion: In the light of this study, ApoA1 has the potential to predict DFU. The finding proposes IL-10 (b = -0.37, p < 0.001) could be considered in stratifying DFU as per its severity.

First-Trimester Reference Intervals for Thyroid Function Testing among Women Screened at a Tertiary Care Hospital in India.

Objectives Due to differences in the method of assay and population-specific factors, each laboratory needs to establish its own gestation-specific reference intervals (GRIs) for thyroid hormones. Materials and Methods Three-hundred forty-one women with less than 14 weeks gestation were screened at a tertiary care hospital in Chhattisgarh, India. Serum levels of thyroid-stimulating hormone (TSH), free thyroxine (fT4), and thyroid peroxidase antibody (anti-TPO) were measured using an ADVIA Centaur XP immunoassay. GRIs (2.5th and 97.5th percentiles) were determined for TSH and fT4. TSH and fT4 concentrations were converted to multiples of the median (MoM) values. Effect of maternal age, gestational age, and maternal weight was analyzed. Statistical Analysis Quantitative variables were expressed as means and standard deviations (SD), and qualitative variables were expressed as frequencies and percentages. Normality of the data was checked using the Kolmogorov-Smirnov test. Values that were normally distributed were expressed only as means and SD. Those that were not normally distributed were expressed as medians and interquartile range. For all statistical analysis, p < 0.05 was considered as statistically significant. Results First-trimester GRI was 0.245 to 4.971 mIU/L for TSH, 10.2 to 18.9 pmol/L for fT4, and 27.0 to 56.89 kIU/L for anti-TPO. There was no significant difference in the mean serum TSH ( p = 0.920), fT4 ( p = 0.714), or anti-TPO ( p = 0.754) values among women in 4 to 7th week and 7 to 14th week of gestation. The 1st and 99th centile MoMs were 0.03 and 4.09 for TSH and 0.66 and 1.39 for fT4. There was a significant positive correlation between the maternal weight and TSH MoM values ( p = 0.027, r = 0.120). Conclusion These laboratory- and first-trimester-specific GRI for TSH and fT4 shall help in proper diagnosis and treatment of subclinical thyroid dysfunctions. TSH and fT4 MoM values can be used to indicate high or low values in a quantitative manner independent of the reference ranges and may be used by other laboratories.

Influence of laboratory biomarkers on inflammatory indices for assessing severity progression in COVID-19 cases.

Background and Objective: This study explored the role of various laboratory biomarkers on inflammatory indices for predicting disease progression toward severity in COVID-19 patients. Methods: This retrospective study was conducted on 1233 adults confirmed for COVID-19. The participants were grouped undermild, moderate, and severe grade disease. Serum bio-inflammatory index (SBII) and systemic inflammatory index (SII) were calculated and correlated with disease severity. The study variables, including clinical details and laboratory variables, were analyzed for impact on the inflammatory indices and severity status using a sequential multiple regression model to determine the predictors for mortality. Receiver operating characteristics defined the cut-off values for severity. Results: Among the study population, 56.2%, 20.7%, and 23.1% were categorized as mild, moderate, and severe COVID-19 cases. Diabetes with hypertension was the most prevalent comorbid condition. The odds for males to have the severe form of the disease was 1.6 times (95% CI = 1.18-2.18, P = 0.002). The median (inter-quartile-range) of SBII was 549 (387.84-741.34) and SII was 2097.6 (1113.9-4153.73) in severe cases. Serum urea, electrolytes, gamma-glutamyl transferase, red-cell distribution width-to-hematocrit ratio, monocytopenia, and eosinopenia exhibited a significant influence on the SpO2, SBII, and SII. Both SBII (r = -0.582, P < 0.001) and SII (r = -0.52, P < 0.001) strongly correlated inversely with SpO2 values [Figures 3a and 3b]. More than 80% of individuals admitted with severe grade COVID-19 had values of more than 50th percentile of SBII and SII. The sensitivity and specificity of SBII at 343.67 for severity were 81.4% and 70.1%, respectively. SII exhibited 77.2% sensitivity and 70.8% specificity at 998.72. Conclusion: Serial monitoring of the routinely available biomarkers would provide considerable input regarding inflammatory status and severity progression in COVID-19.

Plasma Interleukin-33 Cannot Predict Hip Osteonecrosis in Patients With Sickle Cell Disease: A Case-Control Study.

Background Plasma interleukin-33 (IL-33), a cytokine associated with inflammatory and autoimmune disease, has been described to be significantly raised in osteonecrosis of the femoral head (ONFH) and hence was recommended for use as a marker for ONFH. The concentration of plasma interleukin-33 level has not been estimated in any studies conducted in patients with sickle cell disease (SCD); hence, we investigated the levels of plasma interleukin-33 in patients with sickle cell disease with or without ONFH to assess whether it can be used as a marker for the early detection of ONFH in this disease also. Methods Forty-four consecutive patients with sickle cell disease with osteonecrosis of the femoral head and matched controls without ONFH were evaluated for plasma interleukin-33 levels by enzyme-linked immunosorbent assay (ELISA). All patients were confirmed for sickle cell disease using high-performance liquid chromatography (HPLC). ONFH was diagnosed in patients with sickle cell disease using clinical-radiological findings. Univariate and multivariate analyses were performed using the IL-33 level as the dependent variable. Results Plasma IL-33 levels were comparable in 44 patients with sickle cell disease with osteonecrosis of the femoral head as compared with 24 patients with sickle cell disease without ONFH (2.05 ± 4.57 pg/mL versus 1.50 ± 2.89 pg/mL, p-value = 0.590). There was no significant difference in IL-33 levels in different stages of avascular necrosis (AVN). Conclusions Plasma interleukin-33 levels cannot act as a marker of ONFH as were being considered in idiopathic ONFH or ONFH caused by other causes such as trauma and chronic steroid or alcohol usage.

Early Biochemical Markers in Predicting the Clinical Outcome of COVID-19 Patients Admitted in Tertiary Care Hospital.

Introduction An array of routinely accessible serum biomarkers was assessed to explore their overall impact on severity and mortality in coronavirus disease 2019. Materials and Methods A retrospective analysis of 1,233 adults was conducted. The study groups comprised 127 nonsurvivors and 1,106 survivors. Data for demographic details, clinical presentations, and laboratory reports were recorded from the medical record section. The predictors were analyzed for their influence on mortality. Results The mean (+ standard deviation) age of the patients in the nonsurvivor group was 58.8 (13.8) years. The mean age (56.4 years) was highest in severe grade patients. The odds ratio for death was 2.72 times for patients above the age of 40 years. About 46% of nonsurvivors died within 5 days of admission. Males were found to be more prone to death than females by a factor of 1.36. Serum urea depicted highest sensitivity (85%) for nonsurvival at 52.5 mg/dL. Serum albumin (3.23 g/dL), albumin-to-globulin ratio (0.97), and C-reactive protein-to albumin ratio (CAR) (2.08) showed a sensitivity of more than 70% for mortality outcomes. The high hazard ratio (HR) for deceased patients with hyperkalemia was 2.419 (95% confidence interval [CI] = 1.96-2.99; p < 0.001). The risk for nonsurvival was increased with elevated serum creatinine by 15.6% and uric acid by 21.7% ( p < 0.001). The HR for hypoalbuminemia was 0.254 (95% CI: 0.196-0.33; p < 0.001) and CAR was 1.319 (95% CI: 1.246-1.397; p < 0.001). Saturation of oxygen ( p < 0.001), lactate dehydrogenase ( p = 0.006), ferritin ( p = 0.004), hyperuricemia ( p = 0.027), hyperkalemia ( p < 0.001), hypoalbuminemia ( p = 0.002), and high CAR values (0.031) served as potential predictors for mortality. Conclusion Adjusting for all the predictor variables, serum uric acid, potassium, albumin, and CAR values at the time of admission were affirmed as the potential biomarkers for mortality.

Uncommon antinuclear antibody patterns as diagnostic indicators.

INTRODUCTION: The clinical significance of common antinuclear antibody (ANA) patterns, such as nuclear homogenous and nuclear speckled patterns with their corresponding specific antibodies, has already been established. However, the clinical relevance of these uncommon ANA patterns have not been well elucidated and these patterns are therefore not reported by most clinical laboratories. We herein report some retrospective data analysis linking patients' clinical status to several uncommon ANA patterns. METHODS: We retrieved and assessed the patient records for ANA reports generated in our hospital over a period of two years. All testing had been performed using the gold standard Indirect Immunofluorescence Assay. RESULTS: Records of 1235 consecutive patients tested for ANA were reviewed. ANA was positive in 330 of these patients with 6.39% found to have uncommon nuclear, cytoplasmic or mitotic sub-patterns. The mitotic spindle (0.89%), cytoplasmic anti-mitochondrial antibodies (0.80%), followed by discrete nuclear dots-multiple (0.72%) were the dominating patterns, with a higher prevalence in females than in males. Systemic lupus erythematosus and rheumatoid arthritis were the two most common autoimmune disorders associated with mitotic spindle fibers and nuclear centromere and nuclear large/coarse speckled ANA patterns. CONCLUSION: The prevalence of these relatively uncommon ANA patterns was higher than expected. Further evaluation of these patterns along with their corresponding antibodies and their clinical utility must be encouraged. We trust this endeavour will provide diagnostic information in autoimmune and other disease conditions.