In β-thalassemia, point mutations in the β-globin gene are largely responsible for either decreased or no β-globin synthesis. The β-globin gene has three exons and two introns. The molecular characterization of β-thalassemia is absolutely necessary for carrier screening, for genetic counseling, and to offer prenatal diagnosis. The objective of the present study was to identify the rare mutations in β-globin gene of β-thalassemia patients. We have sequenced the entire β-globin gene in 36 clinically identified thalassemia patients from the Karnataka region using polymerase chain reaction and sequencing. Our analysis revealed 11 β-thalassemia variants. The most common being IVSII-16 G>C, IVSI-5G>C, IVSII-74 T>G, codon 3 (T>C), and Poly A site (T>C). In addition, we have also documented a novel deletion at codon 6 (-CT) (HBB:c.16delCT). These data are useful in future molecular screening of the population for implementing a thalassemia prevention and control program. Further it is found that family studies and comprehensive hematological analyses would provide useful insights for accurate molecular diagnosis of thalassemia phenotype and offers an interesting subject for further investigations in the Indian populations.

Various ailments have long been treated with plant-based remedies. This research aims to determine which compounds are present in Annona squamosa L. leaves and seeds extracts, as well as their antioxidant and anticancer potential. Annona squamosa L. hexane, methanol, and aqueous extracts were analysed for their phytochemical content, and results showed the existence of many useful compounds. The DPPH assay was used to measure the antioxidant activity. In particular, the low IC50 values revealed that the plant extracts under study possessed remarkable antioxidant activities. Antioxidant phytochemicals such alkaloid, flavonoid, phenol, oil, phytosterol, coumarin, and saponins may be responsible for these benefits. This research shows that extracts of Annona squamosa L. have a high antioxidant activity and promise as a dietary supplement and possible medicinal agent. MTT assay was used to test for anticancer characteristics. Cell viability was assessed for each extract using a negative control and a positive control (Cisplatin). Both the Annona squamosa L. (seeds) methanolic extract and the Annona squamosa L. (leaves) water extract showed substantial efficacy in this investigation against the MCF-7 breast cancer cell line. It follows that extracts of Annona squamosa L. (leaves) and (seeds) could be used to create a natural medicine for breast cancer treatment.

Post-translational modifications (“PTMs”) in monoclonal antibodies (mAbs) contribute to charge variant distribution, which will affect biological efficacy and safety. For the characterization of mAbs, charge variants are used as a critical quality attributes for product quality, stability consistency and effectiveness. Charge variants in mAbs are characterized by a time-consuming and a multistep process starting from cation/anion exchange chromatography, acidic/basic fractions collection and subsequent reverse phase (RP) liquid chromatography, coupled with mass spectrometry (MS) analysis. Hence, an alternative characterization approach that would be highly selective for ion exchange chromatography-based charge variant analysis, which is compatible with on-line MS detection, is needed in the biopharma industry. Against this backdrop, multiple studies are being conducted to develop a simple straight on-line charge variant analysis method. In this regard, we apply the current study, which aims to develop a charge variant analytical method, based on volatile buffers with low ionic strength that can be used for on-line MS detection of charge variants of mAbs. This would enable the detection on “PTMs” using low ionic strength mobile phase compatible with MS. Hence, fruitful data can be obtained with a single chromatography run without any test sample preparation, eliminating the need for multiple steps of analysis, time-consuming process and multiple sample preparation steps. Thus, Charge Variant Analysis-MS technique will allow the characterization of charge-related PTMs on the intact protein stage. In this regard, this study is about development of a method having combination of chromatography and volatile mobile phase for mass spectrometry detection of mAbs being analyzed in native form. The method is qualified considering pharmacopeia guidelines because the ultimate aim is to transfer this method for Quality Control (QC) release testing of a monoclonal antibody, which is critical for batch release and the regulatory point of view. Acidic and basic variants have been separated with high resolution peak profile. Furthermore, there was no matrix interference and good separation selectivity in terms of specificity was obtained using this method. The experimental data suggested for the linearity of the method are 2.4 mg/mL to 3.6 mg/mL with % RSD below 2.0%. Additionally, Limit of Quantitation is found to be 0.15 mg/mL, which is 5% of loading amount. Consistently, the data show that the method is precise under the same operating conditions with a short time interval. Overall a simple, accurate, robust and precise pH gradient cation exchange chromatography method was developed and qualified for the characterization of a therapeutic native mAb. Additionally, this method can be used to claim a biosimilar product profile of an in-house product compare to an innovator.

To evaluate salivary telomere length as a potential non-invasive exploratory biomarker for early molecular alterations associated with oral leukoplakia and oral squamous cell carcinoma. Forty-five participants were initially recruited: 15 healthy controls, 15 leukoplakia cases, and 15 oral squamous cell carcinoma (OSCC) cases. Two milliliters of unstimulated saliva was collected, and genomic DNA was isolated using the QIAamp DNA Mini Kit (Qiagen, Hilden, Germany). Relative telomere length (RTL) was assessed using quantitative real-time PCR via the comparative 2 - Δ Δ C t method. Given the non-normal distribution of RTL values (Shapiro-Wilk p < 0.001 in all groups), group variations were evaluated using the non-parametric Kruskal-Wallis test, with Bonferroni-corrected Mann-Whitney U tests for pairwise comparisons. The mean relative telomere length (RTL) ratios 2 - Δ Δ C t were 0.41 ± 1.00 for healthy controls, 0.48 ± 0.80 for leukoplakia, and 0.61 ± 0.87 for OSCC. The Kruskal-Wallis test confirmed no significant difference in RTL ranks across the cohorts (H(2) = 2.68, p = 0.261), and Bonferroni-corrected post hoc Mann-Whitney U comparisons showed no significant pairwise differences between any two groups (all p > 0.35), revealing a highly variable, non-significant upward trend in salivary relative telomere length from health to malignancy. Salivary telomere length exhibits substantial inter-individual variability and a non-significant increasing trend from healthy controls to oral leukoplakia and OSCC. These preliminary findings suggest that relative telomere length may not be a reliable standalone biomarker in its current form and warrants further validation in larger, longitudinal studies.