Indian Journal of Clinical Biochemistry

Coomassie Brilliant Blue Can Visualize a Protein Band Without Destaining: A Quick Visualization Protocol on the Agarose Gel

Significance of Vitamin D Binding Protein in Assessing Vitamin D Status Among Under-Five Children Abstract Despite ample sunshine, 50–90% Indian children have Vitamin D deficiency (VDD). This enigma of widespread VDD needs exploration especially among under-fives as physiological variations in Vitamin D Binding Protein (VDBP) levels could be potential confounders in the interpretation of total 25-hydroxyvitamin D [25(OH)D]. However, there is scarce information about relevance of VDBP levels in under-five age group. We therefore, explored association of VDBP levels among 1–5 year old children with VDD. Serum levels of 25(OH)D, VDBP, calcium, parathyroid hormone (PTH) and alkaline phosphatase were estimated in 210 apparently healthy children in the age group of 1–5 years. VDD was defined as serum 25(OH)D levels < 20 ng/ml as per the IOM classification. VDBP levels were classified as low if levels were < 168 μg/ml as per the kit. The prevalence of VDD was 79.5% (n = 167) and VDBP levels were low in 48.6% (n = 102) of children. 25(OH)D levels correlated positively with VDBP (r = 0.298, p = 0.0001). A significant number of children (52.7%) with VDD had low VDBP (p = 0.015). and despite adequate sun exposure, 43% of children showed VDD and 56.6% had low VDPB levels. The low VDBP levels largely explain low 25OHD levels without necessarily implying VDD. It may add a new dimension for better understanding of widespread VDD among under-five children. It thus, points towards the need for redefining cut offs and complete evaluation of vitamin D status among under-fives including VDBP.

The Effects of Self-cleavage Intein-ELK16 Tag in the Transcript Steric Hindrance of IFN Abstract Intervening proteins (Inteins) are identified as protein domains in a precursor protein structure. Inteins can excise itself from precursor protein and join the remaining portions which result in forming an active protein. In this study, the transcript expression level of recombinant human Interferon beta (rhIFNβ) connected to the self-cleavage Intein-ELK16 (LELELKLKLELELKLK) tag was measured by real-time PCR in HEK293T cell line. First, the sequence of Mycobacterium tuberculosis RecA (Mtu recA) was obtained from the InBase database to do appropriate changes including adding the restriction sites, kozak sequence, signal peptide and ELK16 sequence by SnapGene software. The RNA secondary structure were also examined using the online RNA Fold 2.2 web server. Next, the construct was inserted into pUC19 plasmid. The sequence of rhIFNβ was also cloned into pBudCE4.1 vector. In the next step, the rhIFNβ was ligated into the construct (self-cleavage tag of ELK16) using T4 DNA ligase and the recombinant construct was transfected into HEK293T cell line. Finally, expression of the cassette was evaluated by real-time PCR. The analysis of secondary RNA structure indicates a minimum free energy of MEF − 261.10 kcal/mol. Our results indicate that IFNβ was upregulated (37.8-fold, p < 0.0001) in cells which transfected by rhIFNβ-ELK16 compared to the mock and un-transfected conditions. Altogether, our results show that the presence of mini self-cleavage Intein-ELK16 tag along with the rhIFNβ had no interference in transcription of rhIFNβ in the HEK293T cell line.

Increasing Use of Hair Dye and Associated Genotoxicity Needs to be Probed

Why Leadership Skills are Essential for the Future of Laboratory Medicine?

Copper and Zinc Status in Psoriasis: Correlation with Severity Abstract Psoriasis is a common and an inflammatory skin disease. Trace elements play an important role in the skin metabolism such as keratinisation and melanin formation as well as immunological and inflammatory reactions. Henceforth, the present study was aimed to evaluate the essential metals with special reference to Cu and Zn. In this study, 72 psoriatic patients and 50 controls were enrolled. On the basis of PASI score psoriatic group comprising of 48 mild and 24 severe psoriatic patients were grouped. The serum trace elements analysis in the subjects revealed that serum Cu levels and Cu/Zn ratio was significantly higher in the psoriatic patients as compared to controls. Interestingly, severity of the psoriasis was well correlated with the serum Cu levels. Taking together, all these findings suggest that Cu may be a major culprit in the pathogenesis as well as in the severity of the disease.

Adaptive Neuro-Fuzzy Inference System-Based Exploration of the Interrelationships of 25-Hydroxyvitamin D, Calcium, Phosphorus with Parathyroid Hormone Production Abstract The rationale of the current study was to assess the prevalence of 25-hydroxyvitamin D (25-OHD) deficiency and hyperparathyroidism in South Indian population and to explore interrelationships of 25-OHD, Ca, P towards parathyroid hormone (PTH) production using adaptive neuro-fuzzy inference system (ANFIS). A total of 407 subjects (228 men 179 women) with the mean age 56.8 ± 14.1 were tested for these parameters. In view of the skewed distribution of biochemical variables, data segregation was performed in tertiles and this data was trained to generate fuzzy interference system based on subclusters. The optimized model had 358 nodes and followed 44 fuzzy rules for prediction. This ANFIS model demonstrates that the deficiency of 25-OHD and Calcium triggers PTH production. PTH elevation is significant when Phosphorus is in the highest tertile. The associations observed by this model were consistent with the Kendall-Tau correlation matrix, which revealed inverse associations of Ca with P; and Ca with PTH and positive associations of P with PTH, and Ca with 25-OHD. Furthermore, the association statistics of the machine learning algorithm were also consistent, which suggested that depletion of Ca below 8.245 mg/dl was shown to elevate PTH levels greater than 167 pg/ml when P > 4.66. Subnormal depletion in 25-OHD (9.3–16.2 ng/ml) is associated with subnormal elevation in PTH (47–73.6 pg/ml). To conclude, ANFIS and machine learning algorithm are in agreement with each other in stating that 25-OHD deficiency triggers lower calcium levels, lower calcium and higher phosphorus trigger PTH production.

Glutathione S-Transferase P1 313 (A > G) Ile105Val Polymorphism Contributes to Cancer Susceptibility in Indian Population: A Meta-analysis of 39 Case–Control Studies Abstract GSTP1 involved in the metabolism of carcinogens and toxins, reduces damage of DNA and act as a suppressor of carcinogenesis. Many studies have reported that 313 A > G polymorphism is associated with different cancer in Indian population, but the results remain conflicting rather than conclusive. Therefore, we have performed meta-analysis to clarify the more precise association of GSPT1 313 A > G polymorphism with cancer risk in Indian population. We retrieved all relevant published literature from PubMed (Medline) and Google scholar web database and included those study only based on the established inclusion criteria. Pooled ORs and 95% CIs were used to appraise the strength of association. Publication bias and sensitivity analysis was also evaluated. A total of 6581 confirmed cancer cases and 8218 controls were included from eligible thirty nine case–controls studies. Pooled analysis suggested that the variant genotypes significantly increased the risk of cancer in allele (G vs. A: OR 1.266, 95% CI 1.129–1.418, p = 0.001), heterozygous (AG vs. AA: OR 1.191, 95% CI 1.047–1.355, p = 0.008), homozygous (GG vs. AA: OR 1.811, 95% CI 1.428–2.297, p = 0.001), dominant (GG + AG vs. AA: OR 1.276, 95% CI 1.110–1.466, p = 0.001) and recessive (GG vs. AG + AA: OR 1.638, 95% CI 1.340–2.002, p = 0.001) genetic models. The stability of these observations was confirmed by a sensitivity analysis. Begger’s funnel plot and Egger’s test did not reveal any publication bias. This meta-analysis suggests that the GSTP1 313 A > G polymorphism may contribute to genetic susceptibility to cancer in Indian population. However, larger studies and randomized clinical trial will be required to elucidate the biological and molecular mechanism of GSTP1 gene in cancer.

Differential Iron Status and Trafficking in Blood and Placenta of Anemic and Non-anemic Primigravida Supplemented with Daily and Weekly Iron Folic Acid Tablets Abstract The molecular mechanism of iron transfer across placenta in response to maternal anemic status/ iron supplementation is not clear. We hypothesized that maternal iron/ anemia status during early trimesters can be utilized as a biomarker tool to get estimates of placental iron status. Early interventions can be envisaged to maintain optimum placental/ foetal iron levels for healthy pregnancy outcomes. One hundred twenty primigravida were recruited and divided into non-anemic and anemic group on the basis of hemoglobin levels. The groups were randomly allocated to receive daily and weekly iron folic acid (IFA) tablets till six weeks postpartum. Hematological and iron status markers in blood and placenta were studied along with the delivery notes. Weekly IFA supplementation in anemic primigravidas resulted in significantly reduced levels of hematological markers (p < 0.01); whereas non-anemic primigravidas showed lower ferritin and iron levels, and higher soluble transferrin receptor levels (p < 0.05). At baseline, C-reactive protein and cortisol hormone levels were also significantly lower in non-anemic primigravidas (p < 0.05). A significantly decreased placental ferritin expression (p < 0.05); and an increased placental transferrin expression was seen in anemic primigravidas supplemented with weekly IFA tablets. A significant positive correlation was observed between serum and placental ferritin expression in anemic pregnant women (r = 0.80; p < 0.007). Infant weight, gestational length and placental weight were comparable in both the supplementation groups. To conclude, mother’s serum iron / anemia status switches the modulation in placental iron transporter expression for delivering the optimum iron to the foetus for healthy pregnancy outcomes. Trial Registration Clinical Trial Registry-India: CTRI/2014/10/005135.

Evaluation of COMT Gene rs4680 Polymorphism as a Risk Factor for Endometrial Cancer Abstract Catechol-O-methyletransferase (COMT) enzyme is involved in the inactivation of catecholamine and catechol estrogens. Catechol estrogens have carcinogenic potential and DNA damaging ability. Several studies investigated COMT Val158Met polymorphism as risk factor for endometrial cancer but the results were inconclusive. Hence the objective of present study was to find out exact association between COMT gene Val158Met polymorphism and endometrial cancer by a meta-analysis. Pubmed, Google Scholar, Springer Link and Science Direct databases were searched for case–control articles which investigated COMT Val158Met polymorphism in endometrial cancer cases. All statistical analysis was performed using MetaAnalyst and Mix programs. The results of meta-analysis suggested that there were no association between COMT Val158Met polymorphism and endometrial cancer risk (allele contrast model—ORA vs. G = 0.97, 95% CI = 0.86–1.10, p = 0.67; co-dominant model—ORAG vs. GG = 0.91, 95% CI = 0.77–1.06, p = 0.23; homozygote model—ORAA vs. GG = 1.01, 95% CI = 0.84–1.19, p = 0.29; dominant model—ORAA+AG vs. GG = 0.93, 95% CI = 0.77–1.11, p = 0.43; recessive model—ORAA vs. AG+GG = 1.02, 95% CI = 0.89–1.20, p = 0.62). Publication bias was absent. Subgroup analysis based on source of controls was also performed. In conclusion, results of present meta-analysis showed no association between COMT Val158Met polymorphism and susceptibility to endometrial cancer.