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25 results on '"Sethi, Dalip"'

11. Effects of Hypoxanthine Substitution in Peptide Nucleic Acids Targeting KRAS2Oncogenic mRNA Molecules: Theory and Experiment

Author

Sanders, Jeffrey M., Wampole, Matthew E., Chen, Chang-Po, Sethi, Dalip, Singh, Amrita, Dupradeau, François-Yves, Wang, Fan, Gray, Brian D., Thakur, Mathew L., and Wickstrom, Eric

Abstract

Genetic disorders can arise from single base substitutions in a single gene. A single base substitution for wild type guanine in the twelfth codon of KRAS2mRNA occurs frequently to initiate lung, pancreatic, and colon cancer. We have observed single base mismatch specificity in radioimaging of mutant KRAS2mRNA in tumors in mice by in vivohybridization with radiolabeled peptide nucleic acid (PNA) dodecamers. We hypothesized that multimutant specificity could be achieved with a PNA dodecamer incorporating hypoxanthine, which can form Watson–Crick base pairs with adenine, cytosine, thymine, and uracil. Using molecular dynamics simulations and free energy calculations, we show that hypoxanthine substitutions in PNAs are tolerated in KRAS2RNA:PNA duplexes where wild type guanine is replaced by mutant uracil or adenine in RNA. To validate our predictions, we synthesized PNA dodecamers with hypoxanthine, and then measured the thermal stability of RNA:PNA duplexes. Circular dichroism thermal melting results showed that hypoxanthine-containing PNAs are more stable in duplexes where hypoxanthine-adenine and hypoxanthine-uracil base pairs are formed than single mismatch duplexes or duplexes containing hypoxanthine-guanine opposition.

Published
2024
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12. Safety and Effectiveness of Bone Marrow Cell Concentrate in the Treatment of Chronic Critical Limb Ischemia Utilizing a Rapid Point-of-Care System

Author

Ponemone, Venkatesh, primary, Gupta, Saniya, additional, Sethi, Dalip, additional, Suthar, Manish, additional, Sharma, Monika, additional, Powell, Richard J., additional, Harris, Kenneth Lee, additional, Jungla, Nungshi, additional, Arambam, Priyadarshini, additional, Kaul, Upendra, additional, Seth, Ashok, additional, and Bukhari, Suhail, additional

Published
2017
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23. Strategies for preparation of oligonucleotide biochips and their applications

Author

Sethi, Dalip, Kumar, Pradeep, and Gupta, K.C.

Abstract

In the last few years, DNA-microarray technology has emerged as a powerful tool for gene discovery, detection of mutations, and mapping. Here, we have developed a simple and efficient single-step method for immobilization of phosphoryl-, mercaptoalkyl-, thiophosphoryl- and aminooxyalkylated oligonucleotides onto an epoxylated glass surface. It resulted in higher immobilization and hybridization efficiency in comparison to those obtained with the standard method. The probes immobilized were found to be stable over a range of temperature and pH, suggesting that the chemistry could be used in integrated PCR/microarray devices. Also the immobilized spots were found to be of superior quality in terms of spot morphology, homogeneity and signal reproducibility. The constructed microarrays were successfully used for the discrimination of nucleotide mismatches and detection of bacterial diseases.

Published
2009
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24. The monoculture of cord-blood-derived CD34 + cells by an automated, membrane-based dynamic perfusion system with a novel cytokine cocktail.

Author

Jones M, Cunningham A, Frank N, and Sethi D

Subjects
Humans, Hematopoietic Stem Cells metabolism, Cell Culture Techniques methods, Antigens, CD34 metabolism, Stem Cell Factor pharmacology, Stem Cell Factor metabolism, Perfusion, Cells, Cultured, Fetal Blood, Cytokines metabolism
Abstract

Human leukocyte antigen (HLA)-matched cord blood (CB) transplantation is a procedure for the treatment of certain hematological malignancies, hemoglobinopathies, and autoimmune disorders. However, one of the challenges is to provide a sufficient number of T cell-depleted hematopoietic stem and progenitor cells. Currently, only 4%-5% of the CB units stored in CB banks contain enough CD34 + cells for engrafting 70-kg patients. To support this clinical need, we have developed an automated expansion protocol for CB-derived CD34 + cells in the Quantum system's dynamic perfusion bioreactor using a novel cytokine cocktail comprised of stem cell factor (SCF), thrombopoietin (TPO), fms-like tyrosine kinase 3 ligand (Flt-3L), interleukin-3 (IL-3), IL-6, glial cell line-derived neurotrophic factor (GDNF), StemRegenin 1 (SR-1), and a fibronectin-stromal-cell-derived factor-1 (SDF-1)-coated membrane. In an 8-day expansion of a 2 × 10 6 positively selected CD34 + cell inoculum from 3 donor lineages, the mean cell harvest and cell viability were 1.02 × 10 8 cells and 95.5%, respectively, and the mean frequency of the CD45 + CD34 + immunophenotype was 54.3%. The mean differentiated cell frequencies were 0.5% for lymphocytes, 15.8% for neutrophils, and 15.4% for platelets. These results demonstrate that the automated monoculture protocol can support the expansion of CD34 + cells with minimal lymphocyte residual., Competing Interests: Conflict of interests M.J., A.C., N.F., and D.S. are all employees of Terumo Blood and Cell Technologies, Inc. (Lakewood, CO, USA) at the time of manuscript preparation., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Published
2022
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25. N-(Iodoacetyl)-N'-(anthraquinon-2-oyl)-ethylenediamine (IAED): a new heterobifunctional reagent for the preparation of biochips.

Author

Patnaik S, Swami A, Sethi D, Pathak A, Garg BS, Gupta KC, and Kumar P

Subjects
Molecular Structure, Oligonucleotides chemistry, Temperature, Microchip Analytical Procedures methods
Abstract

Design and synthesis of a new heterobifunctional reagent, N-(iodoacetyl)-N'-(anthraquinon-2-oyl)-ethylenediamine (IAED), have been described for the preparation of oligonucleotide-based biochips. The performance of the featured reagent is probed by the immobilization of thiolated and thiophosphorylated oligonucleotides on modified glass microslides via two routes (routes A and B). The immobilization procedure was accelerated by performing a chemical reaction between thiolated oligomers and the iodoacetyl moiety of the reagent under microwaves (MW), where it is completed in just 10 min. The quality of the constructed oligonucleotide microarrays was tested by performing a hybridization assay with a complementary target and subsequently used for the detection of base mismatches. The immobilized probes were found to be thermally stable.

Published
2007
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