Bhushan Kulkarni
@klepkbsrc.org
Scientist Grade II, Dr. Prabhakar Kore Basic Science Research Centre
KLE University
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Nehal Rami, Bhushan Kulkarni, Sandesh Chibber, Devendrasinh Jhala, Nilam Parmar, and Krupali Trivedi
International Journal of Experimental Research and Review
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.
Mithun Bhatt, Anshu Alok, and Bhushan B. Kulkarni
MDPI AG
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.
Geeta Hiremath, Madhu Singh, B. Kulkarni, Rajeev Potdar and B. Naik
Aim of the Study: This study aims to evaluate the antibiofilm activity of root end materials against Enterococcus faecalis. Materials and Methods: Mineral trioxide aggregate (MTA), MTA plus and Biodentine were conjugated with chitosan gel and tested against the 3-day biofilm of E. faecalis. The incubated plates were stained using crystal violet stain and the optical density of adherent stained biofilm was read at 590 nm using ELISA auto reader. Results: There was a mean clinical reduction in the biofilms of the conjugates as compared to their individual counter parts. There was a statistically significant difference seen between the groups (MTA Plus – Chitosan Conjugate) and (MTA – Chitosan Conjugate) with P = 0.0495. Conclusion: The conjugates did perform better in inhibiting the biofilm activity of E. faecalis. Although all conjugates formed were not statistically significant.
K. Hallikeri, K. Burde, Venktesh Anehosur, B. Kulkarni and S. V. Hiremath
Introduction: The tumor-suppressor p53 protein is inactivated by the human papillomavirus (HPV) E6 oncoprotein, causing polymorphism of the p53 at codon 72 of exon either proline (Pro) or arginine (Arg). Specific allele predisposition has been reported in the literature. The association between the p53 allele and HPV types has been reported. We analyzed the association between p53 polymorphism at codon 72 and HPV 16 and 18 genotypes in control, oral submucous fibrosis (OSF) and oral squamous cell carcinoma (OSCC). Materials and Methods: Of the total 90 cases, biopsy tissues of all groups (30 cases of OSF, OSCC and control each) were collected to extract DNA. Polymerase chain reaction was used to detect HPV 16 and 18 and alleles of codon 72 in p53 were evaluated in all the samples. Results: In control, OSF and OSCC samples showed the presence HPV 63.3%, 33.3% and 60%, respectively. In OSF, HPV 16 and 18 was detected in four and four cases, respectively, whereas in OSCC, HPV 16 and 18 was detected in ten and nine cases, respectively. In all three groups, predominantly, Arg/Arg protein was present followed by Pro/Pro and Arg/Pro. Among the control, Arg/Arg type protein was frequently seen followed by Arg/Pro, Pro/Pro in the presence of HPV. OSF and OSCC were associated homologous genes in the presence of HPV. Conclusion: The definite association between p53 codon 72, polymorphism and HPV 16 and 18 was seen in OSCC with low frequency in OSF. Frequency of homozygous genotype is at high risk in the presence of HPV 16 and 18 in developing OSCC.
Bhushan B. Kulkarni, Shivaprakash V. Hiremath, Suyamindra S. Kulkarni, Umesh R. Hallikeri, Basavaraj R. Patil, and Pramod B. Gai
Elsevier BV
B.B. Kulkarni, S.V. Hiremath, S.S. Kulkarni, U.R. Hallikeri, B.R. Patil, and P.B. Gai
Science Alert
Gururaj D. Kulkarni, Suyamindra S. Kulkarni, Gurushantappa S. Kadakol, Bhushan B. Kulkarni, Prakashgouda H. Kyamangoudar, Bhaskar V.K.S. Lakkakula, Kumarasamy Thangaraj, Tipperudra A. Shepur, Muralidhar L. Kulkarni, and Pramod B. Gai
Mary Ann Liebert Inc
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.