Harsh Gala

• Worked on a research project in PC2 Laboratory under the supervision of Dr. Ran Wang for a year.
• Project

1. Optimizing mouse naive CD4 T cell isolation process

2. Whether adhesion molecules directly promote mouse T helper cells in vitro

3. Examine T cell-intrinsic v/s microenvironment signalling driving colitis development

4. Whether adhesion molecule directly promotes human T helper cells in vitro.

• Research Contributions:

1. Established a robust in vitro model for studying T cell differentiation and cytokine secretion by isolating and culturing primary mouse CD4 T cells with or without endothelial adhesion molecules. Utilized T cell receptor stimulation and cytokine cocktails to induce differentiation, followed by measurement of cytokine levels in culture supernatants using the LegendPlex Mouse Inflammation Cytokine Kit.
2. Conducted flow analysis on T cells to assess their phenotype post-differentiation, providing insights into the functional characteristics of these cells under various conditions.
3. Contributed to advancing the understanding of immune cell interactions with endothelial cells, particularly in the context of inflammation, by manipulating endothelial adhesion molecules in the in vitro culture system.
4. Facilitated the exploration of novel therapeutic targets or interventions for immune-related disorders by elucidating the impact of endothelial adhesion molecules on T cell differentiation and cytokine production.

5. Employed bioinformatics and statistical methods such as FlowJo, PRISM Graphpad to analyze complex biological data, extracting meaningful insights and present implications from experimental findings.

• Skills Mastered:

1. Experimental Design: Developed a comprehensive experimental plan to investigate the impact of endothelial adhesion molecules on mucosal CD4 T cell function in the context of inflammatory bowel diseases (IBD).
2. Cell Culture Techniques: Proficiently isolated and cultured primary mouse CD4 T cells in vitro, maintaining cell viability and functionality for subsequent experiments.
3. Cytokine Analysis: Employed the LegendPlex Mouse Inflammation Cytokine Kit and Cytoflex machine to accurately measure cytokine levels in culture supernatants, providing crucial insights into the inflammatory response of CD4 T cells.
4. Flow Cytometry Analysis: Conducted flow analysis on T cells to assess their phenotype, enabling the characterization of T cell subsets and their activation status.
5. Data Interpretation: Analyzed experimental data to identify the impact of endothelial adhesion molecules on CD4 T cell activation and cytokine production, contributing to a deeper understanding of IBD pathogenesis.
6. Hypothesis Testing: Formulated and tested hypotheses regarding the role of endothelial adhesion molecules in modifying mucosal CD4 T cell function, leading to significant findings with potential therapeutic implications for IBD treatment.
7. Scientific Communication: Effectively communicated research findings, including the identification of novel therapeutic targets for IBD, through presentations and potentially forthcoming publications in peer-reviewed journals.

• Worked on research project in PC1 laboratory under the supervision of Prof. Hajra Gupta and Dr. Sejal Rathod for a year.
• Project: Isolation and Characterization of 6-APA Producing Bacteria for Cost-effective Antibiotic Production
• AIM
  1. Isolation of 6-APA producing bacteria and exploration of an alternative source of 6-aminopenicillanic acid.
  2. Reduction in the price and production time of antibiotics produced.
• OBJECTIVES
  1. Isolation of bacteria from leaf samples using Cetrimide, King’s B, and Nutrient media to identify 6-APA producing strains.
  2. Identification and characterization of the isolate through gram staining, biochemical tests, and selective media usage.
  3. Confirmation of the presence of 6-APA using thin layer chromatography.
• METHODS
  1. Isolation: Leaf samples from gardens and college environments will be cultured on Cetrimide, King’s B, and Nutrient agar media.
  2. Identification: Isolates showing potential for 6-APA production will undergo gram staining, biochemical tests, and characterization according to Bergey’s manual.
  3. Confirmation: Thin layer chromatography will be employed to confirm the presence of 6-APA in broth cultures of selected isolates.
• IMPACT
  The successful isolation and characterization of 6-APA producing bacteria could revolutionize antibiotic production, making it more cost-effective and efficient. By simplifying the production process and reducing reliance on mold cultures, this project has the potential to significantly lower the price of antibiotics while meeting the growing demand for these essential drugs.

o Testing of alcoholic beverages

· Gained hands-on experience with various instruments used in testing techniques, such as gas chromatography (GC), high-performance liquid chromatography (HPLC), and mass spectrometry.

o Analytical training for instrumentation

· Hands-on experience with various instruments used in analytical techniques, such as high-performance liquid chromatography (HPLC), gas chromatography (GC), electrophoresis, mass spectrometry (MS) and UV visible spectrophotometry.

o Training in techniques of virology and immunology

· Basic laboratory techniques, cell culture, virus isolation and identification, serological assays, and biology techniques.

· Gained hands-on experience with equipment such as centrifuges, microscopes, and PCR machines.

o Principles and Practices of Laboratory Animal Care

· The internship covered various topics, including animal welfare regulations, humane handling and restraint, experimental design, and health monitoring.

· I was also taught about the proper care and management of laboratory animals, including nutrition, housing, and sanitation.

o The covert existence of snakes

· Trained how to handle snakes.

o Recombinant DNA technology- Agrobacterium-mediated transformation

o Polymerase chain reaction, PCR amplification, Plant genomic DNA extraction.

o Plant tissue culture using Solanum Lycopersicon leaf for explantation, sterilisation, and subculturing of callus.

o Experience in synthetic seed preparation (Calcium Alginate method)

o Immuno-technology- Isolation of proteins by SDS-PAGE, Western Blotting. ELISA

o Experience in synthetic seed preparation (Calcium Alginate method)