Discriminatory performance of the pulpal inflammatory biomarkers; Interleukin-8 and TNF-α in patients with symptoms indicative of reversible and irreversible pulpitis: A diagnostic accuracy study.

AIM: The success of vital pulp treatment (VPT) procedures is dependent on an accurate diagnosis of the pulpal inflammatory condition. Compared with current subjective pulpal diagnostic tests, inflammatory molecular biomarkers involved in the pathogenesis of pulpitis represent potential objective indicators of the degree of pulpal inflammation. Therefore, the aim of this study was to quantify level of inflammatory biomarkers - Interleukin 8 (IL-8) and TNF-α in patients diagnosed with reversible pulpitis (RP), irreversible pulpitis (IR) and normal pulp (NP) and investigate their diagnostic accuracy in differentiating between healthy and inflamed conditions. METHODOLOGY: This prospective, cross-sectional study enrolled 72 patients aged 14-53 years with extremely deep carious lesions after establishing a clinical diagnosis of RP (n = 42), symptomatic IR (n = 22) and NP (n = 8). 50 µL of pulpal blood sample was collected from all the patients using a micropipette after pulpal exposure. The level of IL-8 and TNF-α was assessed in pg/mL using enzyme-linked immunosorbent assays. Mann-Whitney U test was applied to establish the association between IL-8/TNF-α level and degree of pulp inflammation. Receiver operating curve (ROC) analysis was carried out to calculate area under the curve (AUC) for RP versus IR. Cut-off values were established using Youden's index. RESULTS: IL-8 and TNF-α levels differed significantly between RP and IR groups (p ≤ .001). The median value of IL-8 in RP and IP groups was 259.8 pg/mL [187.5-310.0] and 1357.8 pg/mL [1036.7-2177.6] respectively. The AUC-ROC curve for RP versus IR was 0.997 with 95.5% sensitivity and 99.76% specificity. The median value of TNF-α in RP and IR groups was 75.4 pg/mL [62.7-95.8] and 157.6 pg/mL [94.1-347.3]. The AUC-ROC curve for TNF-α was 0.812 with a sensitivity and specificity of 59.1% and 92.1%, respectively. IL-8 and TNF-α levels were below detection levels for all NP samples. CONCLUSION: This study showed that pulpal blood could provide an excellent medium for establishing pulpal diagnosis under extremely deep carious lesions. The selected cytokines, IL-8 and TNF-α, demonstrated excellent discriminatory performance for reversible and irreversible pulpitis. Future studies should correlate the IL-8/TNF-α levels with VPT treatment outcomes.

Evidence of the presence of SARS-CoV-2 virus in atmospheric air and surfaces of a dedicated COVID hospital.

The present study was conducted from July 1, 2020 to September 25, 2020 in a dedicated coronavirus disease 2019 (COVID-19) hospital in Delhi, India to provide evidence for the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus in atmospheric air and surfaces of the hospital wards. Swabs from hospital surfaces (patient's bed, ward floor, and nursing stations area) and suspended particulate matter in ambient air were collected by a portable air sampler from the medicine ward, intensive care unit, and emergency ward admitting COVID-19 patients. By performing reverse-transcriptase polymerase chain reaction (RT-PCR) for E-gene and RdRp gene, SARS-CoV-2 virus was detected from hospital surfaces and particulate matters from the ambient air of various wards collected at 1 and 3-m distance from active COVID-19 patients. The presence of the virus in the air beyond a 1-m distance from the patients and surfaces of the hospital indicates that the SARS-CoV-2 virus has the potential to be transmitted by airborne and surface routes from COVID-19 patients to health-care workers working in COVID-19 dedicated hospital. This warrants that precautions against airborne and surface transmission of COVID-19 in the community should be taken when markets, industries, educational institutions, and so on, reopen for normal activities.

Is learning outcome after team based learning influenced by gender and academic standing?

Team based learning (TBL) is a time tested teaching-learning (T-L) tool involving collaborative learning but has hardly been tested for teaching clinical biochemistry to undergraduate medical students. The present study was designed to (a) compare problem solving skills of first year MBBS students after attending a TBL session on 'organ function test' with that of the students taught the same topic by didactic lecture, (b) assess their perception towards TBL and (c) evaluate if difference in academic standing and gender influence the learning outcome of TBL.One Hundred first professional MBBS students were divided by stratified randomization into two groups. Group I was exposed to TBL to teach 'organ function tests', while group II was taught the same topic by traditional lecture. The outcome of the T-L sessions was assessed by a test for problem solving skills. Student perception towards TBL was assessed from students' response to a questionnaire. No significant difference between the two groups in problem solving skills could be discerned. High achievers performed better after TBL session, while the low achievers were more benefited by traditional lecture method. The female students showed better academic performance after TBL in comparison to male students. The students gave positive feedback for TBL as an instructional technique. We conclude that TBL gives satisfaction, is not inferior to lecture in effectiveness and hence should be used as a T-L method for undergraduate medical students. Moreover, being more effective for female students and high achievers, it is judicious to utilize TBL more frequently for them in an attempt to provide the best individualized teaching. © 2018 International Union of Biochemistry and Molecular Biology, 47(1):58-66, 2018.

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.

Oxidative stress in perinatal asphyxia in relation to outcome.

OBJECTIVE: To study the serum levels of oxidative stress markers - malondialdehyde (MDA) and protein carbonyl in babies with perinatal asphyxia and to correlate their levels with the outcome in terms of mortality and neurodevelopmental sequelae. METHODS: A group of 40 term AGA (appropriate for gestational age) infants with perinatal asphyxia were selected as cases and same number of healthy babies as controls. Serum levels of oxidative stress markers - malondialdehyde and protein carbonyl were determined in cord blood and at 48 hours of life. Their levels were correlated with the outcome of perinatal asphyxia in terms of mortality and the long term neurological outcome. RESULTS: MDA and protein carbonyl, in cord blood were significantly higher among cases (5.88+/-1.40 micromol/L and 1.50+/-0.48 nmol/mg of protein respectively) than controls (3.11+/-0.82 micromol/L and 0.83+/-0.19 nmol/mg of protein respectively). Among the cases, MDA and protein carbonyl values at 48 hours of life (7.52 +/- 1.06 micrommol/L and 2.91 +/- 0.62 nmol/mg of protein respectively) were significantly higher than those at birth. MDA at birth and 48 hours was significantly higher among babies who had seizures than those who remained seizure free. These values were also significantly higher in babies who expired as compared to those who survived. Protein carbonyl values though higher in those who had seizures and in those who expired, were not statistically significant from controls. MDA and protein carbonyl at birth and 48 hours were higher in babies with developmental delay but the association was not statistically significant. CONCLUSIONS: In hypoxic ischemic encephalopathy (HIE), oxidative stress markers MDA and protein carbonyl are high at birth and rise further at 48 hours and the values correlate with the morbidity and mortality. Therefore, determining the serum levels of oxidative stress markers MDA and protein carbonyl will be of benefit in predicting the outcome in perinatal asphyxia.

Lymphoplasmacytic lymphoma with IgA paraproteinemia mimicking Waldenstrom's Macroglobulinemia.

Central to the diagnosis of Waldenstrom's Macroglobulinemia is the demonstration of bone marrow infiltration by lymphoplasmacytic lymphoma with Ig M monoclonal gammopathy. We describe a patient who presented with a clinical and haematological picture, highly suggestive of Waldenstrom's Macroglobulinemia, but whose serum monoclonal immunoglobulin belonged to Ig A class. Ig A secreting lymphoplasmacytic lymphoma undoubtedly exist but are exceedingly uncommon and their relationship to Waldenstrom's Macroglobulinemia needs to be clarified.