MACUB (2021) Conference

Student Presentations

Biochemistry, Biophysics and Biotechnology (BBB-1)

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Dr. Kathleen, Nolan

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Dr. Martin, Hicks

Zoom Meeting

Time: 10/30/21, 11:05AM -

Meeting ID: 879 7350 3136 Meeting Password:


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1-1. Queensborough Community College, CUNY

A peek into the actions of "Molecular Scissors" of Biotechnology. (Piechowska, Sabina & Ghoshal, Sarbani).

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Abstract: The ability to fragment DNA became the cornerstone of molecular biology and biotechnology. In 1978, Arber, Nathans and Smith were awarded a Nobel Prize for their discovery of restriction enzymes (RE), which are also called molecular scissors. RE can cleave dsDNA at specific sequences called restriction sites. There are four types of RE, of which type II is most preferred during cloning procedures. In this presentation, we will discuss detailed procedures for restriction digestion of the vector, pUC19 with two RE, namely EcoRI and BglI. We are also going to discuss in detail how restriction maps are created from data obtained by analyzing DNA fragments from agarose gels. Restriction maps show the relative location of a selection of restriction sites along linear or circular DNA and this type of mapping has contributed immensely towards our knowledge of vectors and plasmids. Restriction mapping has been known to identify unknown genes without sequencing. Additionally, such mapping contributed heavily to our ability to genetically engineer organisms and recombinant DNA technology where an organism’s genes are manipulated, an example of this includes the generation of synthetic human insulin using transformed bacteria.

1-2. Queensborough Community College, CUNY


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Abstract: ELISA, or Enzyme-Linked Immunosorbent Assay is a diagnostic tool that detects and measures the antigens or, antibodies to determine whether a test subject is exposed to a disease or not. Additionally, if a test subject has a positive result for a disease, it is possible to determine the stage of the disease by using the ELISA method. In the ELISA technique, antigens, and antibodies or, in other words, immunocomplex is a very important factor. Antigens are referred to as "foreign substances" that causes the human immune system to respond and to be activated. In contrast, antibodies are a type of protein that contains four chains of amino acids and antigen-binding sites. One special characteristic of the antibody is that each antibody recognizes a very specific antigen that allows fighting against a very specific infection or, disease. ELISA can be performed in four different ways such as direct, indirect, sandwich, or, competitive. Qualitative ELISA allows deciding whether a patient has a positive or negative result in terms of a disease diagnosis whereas quantitative ELISA allows us to quantify or, measure the optical density of the sample using a spectrophotometer. To analyze the data of this honors project experiment, a standard plot was created by entering known antigen concentration on the Y-axis and corresponding absorbance value on the X-axis. ELISA was performed in triplicate for the unknown patient samples. Triplicate testing allows us to determine an intra-assay variation within one test. Additionally, testing each patient sample in triplicate allows avoiding false-positive and false-negative results. After plotting our patient samples we were able to determine if the patient tested positive or negative for HI. All data and log plots will be shown in detail for our conclusion. Even though this ELISA was being used to test the HIV patient samples, it is possible to test the sample of COVID- 19 patients by using the same methodology.

1-3. Queens College, CUNY.

Development inhibitors of Kinases via the synthesis of oxazepane compounds. (Cui, Chang & Choi, Junyong).

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Kinases is an enzyme that adds phosphate groups to other molecules, such as sugars or proteins. They cause other molecules in the cell to become either active or inactive. Kinases play essential roles. Many human diseases are caused by mis-regulated kinases, which can be activated or over-expressed in human diseases such as cancer. FDA has approved small molecule kinase inhibitors as anti-cancer drugs. The research focuses on the development of kinases inhibitors via the synthesis of oxazepane compounds such as 9-methoxy-7-(o-tolyl)-2,3,4,5-tetrahydrobenzo[f][1,4]oxazepane.

The start reagent tert-butyl(2-hydroxyethyl)carbamate was reacted with methanesulfonyl chloride and triethylamine in dichloromethane at room temperature. This reaction converted alcohol group to methanesulfonate. This mesylation reaction provided good yield (90%). Then through SN2 reaction and reductive amination reaction successively, the primary amine was converted to secondary amine compound, which is 7-bromo-9-methoxy-2,3,4,5-tetrahydrobenzo[f][1,4]oxazepane. Based on this compound, the intermediate compound was be obtained via Suzuki coupling reaction. Varieties of analogs have been synthesized from this intermediate compound. We will test these analogs to determine whether they inhibit kinases.

1-4. Columbia University

Uncovering the Mechanism of Heterotrimeric GTP-Binding Protein, Rap1a, in Hepatic Glucose Production. (Sarecha, Amesh; Wang, Yating & Ozcan, Lale).

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Diabetes Mellitus (DM) refers to a group of metabolic disorders characterized by chronic hyperglycemia and most commonly occurs from inadequate insulin production, inadequate sensitivity of cells to the action of insulin, or both. Diabetes falls under two main etiopathogenetic categories, I and II. Type I diabetes, which is called insulin-dependent diabetes mellitus (IDDM), is primarily due to a deficiency in insulin due to the autoimmune-mediated destruction of pancreatic β cells. Type II Diabetes Mellitus (T2DM), the most common form of diabetes, is a prevalent metabolic disorder and its pathogenesis is caused by a combination of peripheral insulin resistance and dysfunctional compensatory insulin secretory response from pancreatic β cells. Additionally, one of the mechanisms for why patients with T2DM have elevated fasting plasma glucose (FPG) levels is that there is aberrant activation of HGP in the post-absorptive state which fails to be properly suppressed by insulin.

Recent studies have provided insight on gluconeogenic signaling pathways in hepatocytes, which contribute to underlying abnormalities such as increased hepatic glucose production. However, little is known about the endogenous regulatory mechanisms that prevent aberrant activation of this pathway. With this project we focus on the protein and isoform Rap1a to show that Rap1a is a negative regulator of gluconeogenesis and prevents HGP's aberrant activation. Rap1a is a small G protein, which stands for Ras-related protein 1a and belongs to the Ras superfamily of GTPases and cycles between an inactive GDP-bound form and an active GTP-bound form. Upon binding to its receptors, glucagon activates adenylate cyclase and increases intracellular cAMP. GTPase-activating proteins, such as Rap1GAP, stimulate GTP hydrolysis and thereby inactivate Rap1. Through recent work, we have identified a heretofore unknown metabolic function of Rap1a in regulating plasma LDL-C metabolism and raised the possibility of targeted Rap1a activation as a new therapeutic strategy to treat hypercholesterolemia. Because Rap1a is downstream of glucagon and its role in glucose metabolism has never been studied, we became interested to see if Rap1a plays a role in glucose metabolism.

Mouse models relevant to diabetes and obesity-induced insulin resistance, such as Rap1afl/fl mice backcrossed onto C57BL/6J background, have been used to elucidate the role of, heterotrimeric GTP-binding protein, Rap1a in glucose metabolism. With the help of immunoblotting, real-time quantitative PCR, and novel assays, we have shown that Rap1a is a glucagon signaling effector, and activation of Rap1a suppresses hepatocyte gluconeogenic gene expression and glucose production. We identified Rap1a deficiency worsens glucose metabolism and insulin tolerance in lean and diet-induced obese mice and are currently elucidating on the mechanism of action behind Rap1a. Our mechanistic work revealed that Rap1a does not regulate gluconeogenesis through affecting intracellular cAMP/PKA levels and our further studies investigating the mechanism of action by which Rap1a functions has led to the identification of one mechanism by which Rap1a inhibition increases glucose production which may be through regulating the activity of insulin-sensitive gluconeogenic transcription factors, Foxo1,3,4. These studies could reveal new dimensions to the pathophysiology of diabetes and be leveraged to design new prevention treatments of cardiometabolic disease. 

1-5. New Jersey City University.

Effects of Stimulants and HIV Proteins on Pyroptosis and Apoptosis Pathways in Human Brain Microvascular Endothelial Cells (hBMEC). (Wei, Yufeng; Garzon, Luis & Joy Ikedife).

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Human Brain microvascular endothelial cells (hBMVEC) are a central element of the microvasculature that forms the blood-brain barrier (BBB) that shields the brain against toxins and immune cells via paracellular, transcellular, transporter, and extra cellular matrix proteins. Damaged hBMVEC cells may cause stroke, seizures and other ailments. The present experiment tested the effects of several stimulants (Cocaine, THC, and Amphetamine) and two HIV proteins (Tat-86 and tat-101) on hBMVEC cells to determine if Pyroptosis (cell inflammation) or Apoptosis pathways were induced. Western blot and Rt- qPCR techniques were used to determine if either pathway was induced. Through protein and cDNA gene Seeanalysis, it was determined that both pathways were induced, Apoptosis mostly. Four main protein markers were found: PEA-15, Casp1, Casp3, and Casp8.


Further research is needed to establish how other stimulants and organic compounds affect and influence the BBB penetrance to treat ailments such as Alzheimer’s and HIV.

1-6. Queensborough Community College, CUNY.

Inositol Hexakisphosphate Kinase 1 (IP6K1) ameliorates diet induced obesity by enhancing energy expenditure pathway. (Wanderley, Mayra; Piechowska, Sabina & Ghoshal, Sarbani).

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The evolution of mankind resulted in physical inactivities and increased access to unhealthy and calorie dense food. This modern lifestyle is the principal cause of the silent and slow growth of the global pandemic called obesity. Obesity results when fat accumulation exceeds the storage-capacity of adipose tissue depots. Our research focuses on targeting IP6K1 to treat obesity and associated comorbidities. Previous research showed that insulin stimulates the inositol phosphate kinase IP6K1 to produce IP7 (5-diphosphoinositolpentakisphosphate), which in turn inhibits insulin sensitizing enzyme Akt, these result in obesity and type-2 diabetes. For this project, we reviewed extensively the effect of genetic deletion and pharmacologic inhibition of IP6K1 on obesity. Our review identified significant statistical differences in obese states among wild type mice, IP6K1 knockout mice, and mice which were treated with the inhibitor when fed high fat diet (HFD) or were allowed to age naturally. Our presentation will show IP6K1 deletion or inhibition leads to less body weight and less accumulation of body fat due to enhanced energy expenditure without altering food intake.

1-7. Queensborough Community College, CUNY

Ionic liquid-polymer gels for separations. (Nembhard, Shameir; Zmich, Nicole; Lall-Ramnarine, Sharon; Castner, Edward W. & Wishart, James F).

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Abstract: The need for more eco-friendly refrigerant gas alternatives to hazardous chlorofluorocarbons (CFCs) (due to their ozone-depleting properties) led to their replacement by mixtures of hydrofluorocarbons (HFCs). However, the required separation of these HFC blends before their recycling or disposal at the end of their life cycle is challenging. Ionic liquids (ILs), with their characteristic low vapor pressures and tunable properties, are potentially suitable for membrane-based separation of gases under vacuum conditions, but their viscosities are too low. Ion gels prepared from ionic liquid-polymer mixtures have shown promise as solid supports that facilitate the separation of gases while retaining IL properties. However, key attributes of ion gels are still poorly understood, and both the structure of the IL and the IL/polymer ratio need to be optimized to achieve good separation of gaseous mixtures. This project aims to develop improved and energy-efficient separation mechanisms that will reduce hazardous gas releases into our environment. We report on the preparation and physical characterization of selected ion gels. Ionic liquids based on tetraalkyl-phosphonium and ammonium cations and bis(trifluoromethylsulfonyl)amide anions were synthesized and purified in our labs. The alkyl groups on the IL cations were selected by design to form a significant non-polar region, and thus optimized for use as gas separation membranes. The polymeric material used in the ion gels is a common battery development diblock copolymer, PDVF-co-HFP. H-1 and C-13 Nuclear Magnetic Resonance (NMR) spectroscopy was used to confirm the structure of the ILs, and they were combined with five weight percent of the di-block co-polymer to produce ion gels. Preliminary results reveal soft, gel-like materials rather than thin membranes. The IL/polymer ratio will be varied to produce membranes optimized for gas separations and the ion gels will be characterized using differential scanning calorimetry, high-energy X-ray scattering and Pulse-Gradient Spin Echo NMR spectroscopy. These measurements will identify the best ion gel systems for gas separation tests.

1-8. Queensborough Community College, CUNY & Stony Brook University.

The Impact of Curcumin on H460 cells. (Daniels, Dontaye; Noel, Angela; Bustamante, Monic & Moloney, Daniel).

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Abstract: The objective of this research project is to explore the effects of curcumin on H460 lung cancer cells. In countries where the spice turmeric is a household staple and is regularly consumed in the diet, people have lower instances of colon, breast, and lung cancer. Previous studies have shown that curcumin, a component of turmeric, inhibits the proliferation and survival of many types of tumor cells. Curcumin may inhibit tumor growth by regulation of multiple signal pathways including cell proliferation (cyclin D1, c-myc), cell survival (Bcl-2, Bcl-xL, cFLIP, XIAP, c-IAP1), apoptosis activation (caspase-3, 8, 9), tumor suppressor (p53, p21) death receptor (DR4, DR5), and protein kinase pathways (JNK, Akt, and AMPK). Cell microscopy, cell proliferation assays, and a fluorescent apoptosis assay were used to measure Curcumin’s effectiveness on H460 cells. We found that curcumin inhibits cell proliferation and induces apoptosis with a 50% inhibitory concentration (IC50) value of 7.5 mM. Curcumin’s ability to selectively kill tumor cells and not normal cells makes it appealing as a preventive agent or anticancer therapy.