Follicle Stimulating Hormone, or FSH, is a critical hormone involved in reproduction. In men, FSH signaling in Sertoli cells triggers spermatogenesis, and in women, FSH signaling in the granulosa cells is responsible for follicular development. Current hormonal contraceptives have been found to have incredibly high effectiveness but can also carry multiple side effects. In addition, their effectiveness can be altered due to an individual’s FSH, estrogen, progesterone , or GnRH levels. All FSH signaling happens through the FSH receptor, a member of the G-protein Coupled Receptor (GPCR) family. This receptor is known to interact with many other proteins along its complicated signaling pathway, and there are still many more associated proteins yet to be discovered. Using the APEX technique, the proteins proximal to the FSH receptor were labelled and purified. In the future, these proteins could be analyzed using HPLC-MS, and identified using various computer software. Small proximal proteins, like those isolated in this study, have been shown to allow for receptor activation and function modulation. Thus, this study opens the doors for much more work in the field of contraceptives for both men and women. Another potential benefit from identifying FSHR associated proteins would be in developing new approaches for In Vitro Fertilization (IVF). To perform IVF, FSH is given to women to cause rapid follicle stimulation and maturation. This produces multiple ovulation events and allows for a higher chance of pregnancy. The problem, however, is that FSH alone is expensive, and the chance of multiple pregnancies is vastly increased. Targeting the pathways downstream of the FSH receptor could allow for more careful modulation of the follicle growth and could be much less expensive, vastly lowering the cost of IVF and making it a more viable option for those seeking infertility treatment.
Around 10 million Americans suffer from osteoporosis, a disease that leads to over 1.5 million fractures every year and costs the national health care system between 17 to 20 billion dollars annually. Osteoporosis is a systemic disease that occurs when the rate of bone reabsorption (breakdown to retrieve calcium) surpasses the rate of bone regeneration. Our research stems from the observation that bone loss is more commonly seen in elderly women rather than in men: 80% of Americans with osteoporosis are women. Although previous studies suggest that the decrease in estrogen levels following menopause is the cause of bone loss, more recent research has proved that the highest rate of bone loss occurs during perimenopause when serum FSH levels spike which occurs before estrogen levels drop. In the past couple of years, research from Professor Cohen’s lab has concluded that osteoclast differentiation increases in the presence of FSH. If this is due to the presence of FSH receptors in osteoclasts, these would present as a critical target for pharmaceutical intervention. To explore this hypothesis a mouse osteoclast progenitor cell line was treated with RANK-Ligand to stimulate differentiation into osteoclasts. The osteoclasts were then treated with FSH in a time course. Some were pre-treated with negative allosteric modulators (NAMs) to prevent FSH activation of its receptor. The activation of the FSHRs in osteoclasts was measured using a western blot analysis. The data collected over the course of this experiment suggests that FSH receptors are present in osteoclasts. However, the presence of negative allosteric modulators did not seem to have particularly affected the hormone’s ability to activate signaling. Further investigation of the pathways through which FSH promotes the differentiation and proliferation of osteoblasts is needed to understand and identify new approaches for how they can be disrupted and prevent or minimize osteoporosis.
“ … 2004. Print. 7. Kehlbeck, Joanne D., Michael E. Hagerman, Brian D. Cohen, … ”
Abstract
The Department of Mechanical Engineering at Union College in conjunction with the Department of Chemistry and Biological Sciences are presently involved in the research and development of thin film solar nanocomposites. The thin film solar nanocomposite solar devices offer a cheaper alternative to solar technology, however the efficiencies of these thin film solar devices are very low compared to their silicon-based counterparts. The group’s ultimate goal is to produce a flexible solar cell with efficiencies that are comparable to traditional silicon-based solar cells. The collaboration of the Department of Mechanical Engineering at Union College has provided new insights that could help increase the development of solar nanocomposites. While there has been significant research on fabrication techniques, this project proposes to investigate the mechanical properties of the solar cell design, specifically the PEDOT:PSS/PET:ITO interface, to determine a correlation between interfacial interactions and overall efficiency.
“ … Michael Hagerman (Chemistry) and Brian Cohen (Biology). Students from the nanotechnology … Michael Hagerman (chemistry), Samuel Amanuel (physics) and Brian Cohen (biology). Professor Catravas, along with co … , … ”
Follicle stimulating hormone (FSH) and its receptor (FSHR) play important roles in reproduction and fertility in both men and women. Fertility problems arise when FSH, FSHR, or any downstream signaling components don't function properly. FSHR is a G-protein coupled receptor (GPCR) found on the cell surface of granulosa cells in women and Sertoli cells in men. When activated by FSH, FSHR initiates a signaling cascade that can result in downstream results such as ovarian follicular development and estrogen production in women and sperm production in men. To better understand how the receptor functions, the interactions between FSHR and effector proteins involved in signaling must be identified and understood. We plan to study the interactions between FSHR and its effector proteins using the APEX Assay. This assay utilizes an engineered ascorbate peroxidase (APEX) which is attached to the C-terminus of FSHR. APEX modifies proteins using biotin phenol and hydrogen peroxide, which creates a biotin-radical, thus labeling proteins within a small radius of FSHR because of the short lived nature of the radical. Using mass spectrometry, the biotinylated proteins can be quantitatively analyzed to give a better understanding of the proteins associated with FSHR and the changes in the proteins over time after hormone treatment. Using HEK293 cells, we created a stable cell line expressing an FSHR-APEX construct. We are currently using the FSHR-APEX expressing cells in the APEX Assay to analyze the proteins that are involved in FSHR signaling. This research will give us a better insight as to what effector proteins interact with FSHR and how these interactions change following hormone stimulation. This will provide us with a better understanding as to how the receptor signals and initiates downstream signaling and how this might affect fertility in humans.
There is currently a gap between what is known about genetic susceptibility to depression and how the condition presents itself. It is known that individuals with diseases that cause too much or too little of the stress hormone cortisol, are typically diagnosed with depression. This led our lab and others to the hypothesis that genetic factors related to cortisol action may put someone at an increased risk for developing depression. We used allele specific PCR to study the frequency of single nucleotide polymorphisms in genes associated with the function of different components of the hypothalamic-pituitary-adrenocortical axis (HPA-axis) to determine if dysregulation is associated with increased risk. Additionally, saliva samples were taken to quantify cortisol levels. Thus far, we have found significant relationships between the glucocorticoid receptor (GR) SNPs rs41423247 and rs56149945 and scores on the Beck Depression Inventory. The GR SNP rs10052957 and the mineralocorticoid receptor SNP rs5522 showed significant relationships with the State and Trait Anxiety Inventory. We have also found that SNPs rs12086634 and rs5522 may be related to increased risk. In addition to these SNPs we are currently investigating over ten others. Understanding how these changes in the HPA axis relate to altered function may be an important part of determining how to better diagnose and treat depression. Through the use of genomic data, this research could provide the means to improve current treatments of depression and be the next step to personalized clinical care.
Osteoporosis is an age-related chronic disease in which osteoclasts degrade minerals of the bone faster than osteoblasts can replace them. In addition to increased risk of fracture, osteoporosis patients experience oral, dental and prosthodontic complications due to degradation of the alveolar and jaw bone. Elderly women are disproportionately impacted by this disease. One potential culprit for the gender-based disparity is the increased levels of follicle stimulating hormone (FSH) during menopause. An isoform of the FSH receptor can be found on osteoclasts; when FSH binds to this receptor, osteoclast activity and differentiation are increased. Murine monocytes were differentiated into osteoclasts using RANK ligand, and FSH dependent signaling was measured through Western Blotting. Proteins specific to the FSH-receptor pathway were detected in osteoclasts. A better understanding of the osteoclast FSH receptor could allow for pharmaceutical intervention to interrupt the pathway. Negative allosteric modulators (NAMs) that inhibit FSH signaling on granulosa cells could be used for inhibiting FSH signaling on osteoclasts.
For thousands of women in the United States ovarian cysts, tumors, and hormonal imbalances contribute to reduced fertility and fecundity. In addition to these factors, many other unexplained physiological mechanisms may play a role in women's reproductive health. Specifically, human follicle stimulating hormone (hFSH) binds to a G protein coupled receptor (GPCR) called human follicle stimulating hormone receptor (hFSHR). This interaction helps to regulate the reproductive system in both males and females indicating that it may be a key player to target in order to promote fertility. Previous work in our lab has demonstrated that hFSHR, like other GPCR, has been shown to preferentially localize within microdomains in the cell membrane known as lipid rafts in order to facilitate proper signaling. This localization is thought to be mediated in part by a protein known as caveolin. In order to investigate the relationship hFSHR and caveolin, a cell line expressing hFSHR (HEK293-hFSHR) was transfected with a plasmid to transiently express caveolin. Immunoprecipitations were used to investigate the interaction between caveolin and full length hFSHR. Western blot analysis was subsequently performed to detect protein interactions. Based on the data generated from this experiment, the previously established relationship between FSHR and lipid raft residency was not confirmed because it was not shown that caveolin co-precipitated with FSHR. These findings may suggest that previous studies used methods that resulted in artifactual phenomenon. However, it is equally possible that the conditions, plasmids, or cells used in these experiments contributed to the inability to replicate the results found previously. Understanding the possible interaction between hFSHR and caveolin is crucial in the future development of therapeutics that will allow us to fine tune receptor function for infertility treatments.
Around 10 million Americans suffer from osteoporosis, a disease that leads to over 1.5 million fractures every year and costs the national health care system between 17 to 20 billion dollars annually. Osteoporosis is a systemic disease that occurs when the rate of bone reabsorption (breakdown to retrieve calcium) surpasses the rate of bone regeneration. Our research stems from the observation that bone loss is more commonly seen in elderly women rather than in men: 80% of Americans with osteoporosis are women. Although previous studies suggest that the decrease in estrogen levels following menopause is the cause of bone loss, more recent research has proved that the highest rate of bone loss occurs during perimenopause when serum FSH levels spike which occurs before estrogen levels drop. In the past couple of years, research from Professor Cohen's lab has concluded that osteoclast differentiation increases in the presence of FSH. If this is due to the presence of FSH receptors in osteoclasts, these would present as a critical target for pharmaceutical intervention. To explore this hypothesis a mouse osteoclast progenitor cell line was treated with RANK-Ligand to stimulate differentiation into osteoclasts. The osteoclasts were then treated with FSH in a time course. Some were pre-treated with negative allosteric modulators (NAMs) to prevent FSH activation of its receptor. The activation of the FSHRs in osteoclasts was measured using a western blot analysis. The data collected over the course of this experiment suggests that FSH receptors are present in osteoclasts. However, the presence of negative allosteric modulators did not seem to have affected the hormone's ability to activate signaling. Further investigation of the pathways through which FSH promotes the differentiation and proliferation of osteoblasts is needed to understand and identify new approaches for how they can be disrupted and prevent or minimize osteoporosis.
Depression is one of the most prevalent diseases worldwide, afflicting approximately 17 million adults in the US in 2019. A prominent causal factor of depression is dysregulation of our body's response to stress when exposed to continuous stressors over long periods of time. Stress is regulated by the Hypothalamic-Pituitary-Adrenal (HPA) axis, with cortisol as its effector hormone. The effects of cortisol are exerted through the glucocorticoid and mineralocorticoid receptors (GR and MR, respectively) in the body and the brain, respectively. The HPA axis is regulated by a negative feedback loop where activation of the GR by circulating cortisol inhibits the production of additional cortisol. Our lab investigated the role of single nucleotide polymorphisms (SNPs), or mutations, in the genes of the MR, GR, and associated regulatory proteins that are hypothesized to be involved in the dysregulation of function leading to hypersensitivity or resistance to cortisol in the HPA axis. Previous literature has demonstrated that prominent GR and MR SNPs correlate with higher incidence of depression and/or depressive symptoms. Our study continued this investigation into the relationship between mutations and depression in a clinical sample of psychiatric patients from Albany Medical College. We were especially interested in the interplay between mutations of the GR and MR influencing cortisol levels and response in our bodies. DNA from patients was collected with buccal swabs and genotypes were analyzed through allele specific and quantitative polymerase chain reactions. We compared genotypes to the Beck Depression Inventory (BDI) score of patients, as well as other scales measuring anxiety and mood. No significant two-tailed correlation between individual SNPs or GR/MR SNP combinations and BDI scores was found. One SNP in the GR gene (rs33389) one one SNP on the 11?ü-hydroxysteroid dehydrogenase type 1 (rs12086634), an enzyme that regulates peripheral cortisol levels, demonstrated significant one-tailed correlations with the Mindful Attention Awareness Scale and State Trait Anxiety scale, respectively. Obtaining a greater sample size would yield more representation of mutant alleles and potentially allow for more significance correlating genetic mutations to the incidence of depression.
Bariatric surgery is an effective surgical option for treating severe obesity resistant to dieting and exercise. Bariatric surgeries have been shown to adjust the metabolic profile and structure of the patient, typically addressing the underlying issue contributing to the patient's obesity. However, some patients do not experience this efficacy, suggesting another, non-metabolic,etiology for their obesity. In order to provide adequate treatment for obesity, the non-metabolic mechanisms of feeding behavior, and their dysfunction, must be understood. The current study investigates single nucleotide polymorphisms (SNPs) in the dopamine and endocannabinoid brain pathways, responsible for reward and regulatory functions, respectively. These SNPs had been previously associated with drug addiction. It was predicted that the allelic frequency of the mutant genotypes for each SNP, rs324420 (endocannabinoid-related) and rs1076560 (dopamine-related), would be elevated in the bariatric population compared to the general population. SNP analysis was conducted using allele-specific polymerase chain reaction and quantitative real-time polymerase chain reaction. For both polymorphisms, the allelic frequency of the mutant allele was elevated in the bariatric population compared to the general population. The observed increase in mutant allelic frequency was much greater for rs324420 than for rs1076560. These results suggest that similar neuropathology may be at play in the manifestation of disordered eating as is in the etiology of substance use disorders. Future research should compare SNP analyses of populations with binge-eating disorder and populations with food addiction.
“ … Fund ✵ Union College Student Research Grants ✵ Prof. Brian Cohen … ”
Abstract
Depression is one of the most common diseases worldwide with over 300 million people currently diagnosed. Currently, all depression is being treated the same even though depression can manifest itself in several different combinations. To be diagnosed with depression, a patient needs to have just 5 of the 9 symptoms defined by the American Psychiatric Association DSM-5. This means there are 256 possible combinations of depressive symptoms that could be presented by an individual. One observation that has been made is that individuals with diseases that cause too much or too little of the stress hormone cortisol often show signs of depression; this led us to our hypothesis that genetic factors related to cortisol levels, function, or activity may increase someone's risk for depression. In order to find better treatments for depression it is first necessary to have a better understanding of the different types of depression and the role of genetics in the hypothalamic-pituitary-adrenocortical axis (HPA-axis) that regulates cortisol levels and the different mechanisms by which cortisol action is regulated. Hypersensitivity is experienced when an individual's body reacts to a normal amount of cortisol as if it were an abundance of cortisol while resistance refers to someone who does not respond fully to normal amounts of cortisol. We can determine hypersensitivity and resistance by looking at single nucleotide polymorphisms (SNPs)- changes in the sequence of the DNA in some genes; our lab is interested in the contribution to depression of SNPs in several genes including the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). If a patient expresses GR resistance and MR sensitivity (or vice versa) the contrasting effects could result in dysregulation of the stress response. In partnership with the Union College Wicker Wellness Center DNA samples were collected from patients identified as having depression. Along with DNA samples, participants were asked to fill out both the Center for Epidemiologic Studies Depression Scale (CES-D) as well as the Mini Mood and Anxiety Symptom Questionnaire (Mini-MASQ). Both of these questionnaires were used in order to make measurements of depression for each of the participants to compare with the detected genotypes. By better understanding how dysregulation of the HPA-axis contributes to depression, we can begin to answer our research question: how do we better treat depression.
“ … by things that usually don’t bother me … Research Grant ● Brian D Cohen, PhD … ”
Abstract
Depression is one of the most common diseases worldwide with over 300 million people currently diagnosed. Currently, all depression is being treated the same even though depression can manifest itself in several different combinations. To be diagnosed with depression, a patient needs to have just 5 of the 9 symptoms defined by the American Psychiatric Association DSM-5. This means there are 256 possible combinations of depressive symptoms that could be presented by an individual. One observation that has been made is that individuals with diseases that cause too much or too little of the stress hormone cortisol often show signs of depression; this led us to our hypothesis that genetic factors related to cortisol levels, function, or activity may increase someone's risk for depression. In order to find better treatments for depression it is first necessary to have a better understanding of the different types of depression and the role of genetics in the hypothalamic-pituitary-adrenocortical axis (HPA-axis) that regulates cortisol levels and the different mechanisms by which cortisol action is regulated. Hypersensitivity is experienced when an individual's body reacts to a normal amount of cortisol as if it were an abundance of cortisol while resistance refers to someone who does not respond fully to normal amounts of cortisol. We can determine hypersensitivity and resistance by looking at single nucleotide polymorphisms (SNPs)- changes in the sequence of the DNA in some genes; our lab is interested in the contribution to depression of SNPs in several genes including the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). If a patient expresses GR resistance and MR sensitivity (or vice versa) the contrasting effects could result in dysregulation of the stress response. By better understanding how dysregulation of the HPA-axis contributes to depression, we can begin to answer our research question: how do we better treat depression.
“ … References and Acknowledgements • Angela Stewart, Brian D. Cohen, … why do they go there?" Journal of Molecular Endocrinology. • Cohen Brian D., and Dias James A. (2019) Follitropin … … ”
Abstract
Follicle stimulating hormone is produced by the pituitary gland, regulating the function of both the ovaries and the testes. The inability of this hormone to bind to its receptor can cause infertility in both men and women. Human follicle stimulating hormone receptor (hFSHR) is a GPCR (g protein-coupled receptor) that plays a crucial role in the development of sperm and egg cells in the gonads. The successful interaction of FSH with its receptor results in activation of several downstream intracellular pathways including the production of cAMP. Currently, hFSHR signaling is thought to rely on the protein caveolin. Caveolin is a protein found in the caveolae of the cell membrane. Caveolae, a type of lipid raft, support a diverse group of receptors that are needed at the cell membrane. The ability of hFSHR to bind to the lipid raft is believed to be accomplished through an interaction with the transmembrane domain IV of the hFSHR with the caveolin protein. An interruption in the caveolin binding motif of the hFSHR could have a large effect on the development of sperm and egg cells and subsequently cause infertility. In this study, we introduced wild type and mutant peptides corresponding to the 4th transmembrane domain of the hFSHR into cells expressing the receptor. We expect the wild type peptide to interfere with the interaction between caveolin and its binding motif. Additionally, we expect the mutant peptide to not interfere with this interaction. Currently, we are using a quantitative fluorescent assay to measure cAMP production in order to determine if the peptide is interfering with the activation of the hFSHR. I hypothesize that if caveolin is prevented from binding to hFSHR, the receptor will become less active. I anticipate a decrease in levels of cAMP in the cells that are treated with the wild-type (interfering) peptide. There should be no difference in the level of cAMP in cells treated with the mutant peptide and the cells with no peptide. Preliminary trials have suggested that there is a difference in levels of cAMP produced between the two peptides. In the future, we plan on using scrambled peptides to validate our current observations.
“ … .ncbi.nlm.nih.gov/pmc/articles/PMC4480823/. • Brian D. Cohen, PhD … ”
Abstract
The Follicle Stimulating Hormone (FSH) and its receptor are critical in regulating fertility in humans. FSH is a gonadotropin that is produced in the anterior pituitary gland, and the FSHR is a G protein-coupled receptor (GPCR) found primarily on granulosa cells of the ovaries and Sertoli cells of the testes. When FSH binds to this receptor, it sets off intracellular signaling cascades that regulate gonadal functions and the maturation of gametes. Abnormalities in FSH levels and problems with the FSHR can lead to reproductive issues like delayed puberty, oligospermia, azoospermia, and other such problems. Because FSH and its receptor are so important in reproductive functions, scientists have looked to regulate this receptor in order to address issues of birth control and infertility. Controlling this receptor would essentially allow scientists to turn "on" and "off" reproductive functions in humans. According to prior research in our lab, the human FSH receptor has a high affinity to regions in the membrane called lipid rafts. Lipid rafts are specialized locations in the cell membrane that are critical in cell signaling. These regions are believed to be more rigid than the rest of the surrounding membrane, and this rigidity is critically important for signal transduction. Certain complexes can be kept together by a raft, which allows them to assemble and causes signals to be transmitted more quickly. My research focuses on confirming that the FSHR is located in the lipid rafts on the cell membrane using fluorescent imaging and Atomic Force Microscopy (AFM). I have been working to tag proteins in the lipid raft and the receptor itself with different fluorescent colors so that I can see if the locations of the fluorescences align. From there, I will be able to use the AFM to image the cell surface and determine if the region containing the receptor is stiffer than the surrounding membrane. If it is stiffer, this would confirm that it is a lipid raft. This information will be crucial in understanding how this receptor functions and subsequently being able to control it.
“ … Pathway Rachel Eisenberg ‘21, Dhara Shukla ‘22, Professor BrianCohen, Union College Biology Department, Schenectady, NY … ”
Abstract
The stress hormone cortisol is responsible for aspects of metabolism and visceral fat formation that are hypothesized to be linked to obesity. Cortisol directly and indirectly influences metabolism by stimulating lipolysis as well as gluconeogenesis, the breakdown of fat and creation of glucose respectively. By interacting with abdominal organs such as the liver and pancreas, cortisol also increases metabolism through downstream hormones, such as glucagon and epinephrine. Cortisol intersects a variety of pathways that influence the breakdown of sugar, making it an important target for metabolic studies. This study investigates the correlation of mutations in proteins responsible for cortisol action in target tissues, individually and collectively, to clinical measurements of patients with obesity who are seeking or have already sought out bariatric surgery. DNA samples and clinical information were collected from patients of the Ellis Hospital Bariatric Care who were recruited for the study under the supervision of the Ellis Hospital Institutional Review Board. Genotyping of patient samples was done by performing allele specific-polymerase chain reactions (AS_PCR) which were then analyzed through agarose gel electrophoresis to look for single nucleotide polymorphisms (SNPs). It was found that the mutations throughout the glucocorticoid pathway are present at comparable rates in both the bariatric population and general population. Therefore, it cannot be concluded that there is an increase of mutations in the study population compared to the values for the general public found in literature. There are strong correlations between a mutation in the glucocorticoid receptor (GR) and excess body weight. This mutation causes conversion of an aspartic acid to a serine (N363S) in the protein sequence which has previously been shown to cause increased sensitivity to cortisol. As more SNPs are analyzed in combination with each other, more conclusions may be drawn between SNPs and physiological parameters of the patients. High correlations may provide evidence that select bariatric surgeries may be more effective in patients with specific genotypes. If this is the case, bariatric surgery can adopt precision medicine that caters treatment to the patient.
“ … Zimmerman, BrianCohen, & Cay Anderson-Hanley ● There is a dementia … ”
Abstract
Objective: Researchers are focusing on behavioral interventions to counteract the cognitive decline associated with dementia. The interactive and Physical Cognitive Exercise System (iPACES v2.75) study at Union College is a single-bout multimodal intervention, consisting of pedaling an under-desk elliptical which controls progress while playing a video game, for patients with mild cognitive impairment (MCI). Our investigation focuses on the neurobiological mechanisms of cognitive decline by looking at the correlation between changes in biological markers and improvements in cognitive function. Participants and Methods: 27 older adults were recruited from multiple sites in NY and MA. Participants pedaled an under desk elliptical and used an iPad for iPACES, in which they traveled along a virtual bike path and completing errands. Two domains of cognitive function: executive function and verbal memory, were measured pre and post exercise with the paper and electronic Stroop (A:C ratio and CCII metric of BrainBaseline) and ADAS-COG. Salivary samples to analyze biomarker changes were taken before and after the exercise. The samples were assayed using the Enzyme-Linked Immunosorbent Assay (ELISA) for IGF-1, alpha-amylase, and cortisol. Paired t-tests were used to evaluate pre to post changes in IGF-1, Alpha-Amylase, cortisol, Stroop, and ADAS-Cog. Results: Significant improvements in the overall sample were noted on the ADAS-Cog (p = .05), Stroop CCII (p = .01) and alpha amylase (p = .01). There was a significantly greater improvement among MCI on the Stroop CCII (p = .04) and alpha amylase (p = .04). Conclusions: Our results support the idea that alpha-amylase is an effective biomarker for multi-modal interventions for patients with MCI. Since there was a significant improvement in the increase in alpha-amylase, it shows that there was an effective physiological response to the physical exercise that could be part of the broader neurobiological mechanisms leading to cognitive improvements. Further investigation is needed into single-bout interventions to understand how to maximize the effectiveness of the exercise to spur the neurobiological mechanism. Our study supports the notion that the combined physical and mental exercise benefits cognition, and should be the basis of more interventions and research for patients at risk for dementia. Authors: Kartik Nath, Professor Cay Anderson-Hanley
Powered wheelchairs are integral to the development of children with disabilities. However, commercial wheelchairs are very expensive, typically costing over $10,000, to be widely incorporated into physical therapy and rehabilitation programs. In 2017-18, two Union seniors designed a low-cost power mobility device (Version 1.0) by retrofitting a Fisher-Price children toy called the Wild Thing. This device was delivered to The Kevin G. Langan School in the spring of 2018. This senior capstone design project improves Version 1.0 to create Version 2.0 and addresses challenges in the field of assistive technologies.
The challenges in the field of assistive technologies lies in the need to customize hardware and/or software on each device for children with different motor skills. Current DIY assistive wheelchairs are not easily replicable and self-diagnosable upon system malfunctions. Poor hardware and software design documentation makes maintenance difficult.
To address these challenges, this senior capstone design project transforms a one-time project into an easily replicable open-source product. For Version 2.0 to be accessible to anyone who may wish to replicate the device, a public website (<a href="https://muse.union.edu/umobility">https://muse.union.edu/umobility</a>) was built as dedicated resource hub. On this site, users are able to access the construction manual of both the control system as well as the seating structure of Version 2.0.
“ … Receptor. Department of Biology, June 2019. ADVISOR: Brian D. Cohen Infertility affects millions of people in the … I would like to thank my advisor, Professor Brian D. Cohen, for his endless support and guidance throughout my research for this thesis. Prof. Cohen, thank you for your invaluable advice on everything … ”
Abstract
Infertility affects millions of people in the United States. In addition to being unable to conceive, patients can face a variety of challenges ranging from mental health issues to increased economic costs. One cause of infertility is a defect in follicle stimulating hormone (hFSH). hFSH is a peptide hormone that is crucial for fertility in both males and females. In males, hFSH is involved in spermatogenesis. In females, hFSH is involved in estrogen production and ovarian follicle development. To better understand the role hFSH plays in fertility, our lab is interested in investigating the function of the hFSH receptor (hFSHR). hFSHR is a G protein-coupled receptor (GPCR); signaling from the receptor is of particular interest to us, because if we know how hFSHR signals, we can identify potential molecules to investigate that may be targets to improve fertility. However, investigating the signaling of GPCRs is challenging. Ideal assays to investigate GPCR signaling should be sensitive, simple to use, high throughput, and non-radioactive. Although assays for investigating GPCR signaling have historically been used, they have significant drawbacks. Assays using radioactive ligands are sensitive and high throughput, but are not simple to use and generate radioactive waste. Other assays such as G protein-dependent functional assays are sensitive, but are only able to screen for the activity of one molecule. The APEX assay is a new assay for investigating GPCR signaling that has significant advantages over other assays. When cells expressing a GPCR-APEX fusion protein are incubated in culture media containing biotin-phenol, APEX tags signaling molecules 20 nm radius with biotin-phenoxy radicals. Furthermore, this reaction can be quenched. Thus, the iii APEX assay can be used to investigate GPCR signaling with high spatial and temporal resolution. To develop the APEX assay to investigate hFSHR signaling, it is necessary to clone a plasmid coding for the hFSHR-APEX fusion protein. Creation of this plasmid will allow us to use a high-throughput, time dependent assay to investigate new therapeutic targets for infertility.
“ … 25 ACKNOWLEDGEMENTS A special thank you to Professor Brian Cohen for all the help and guidance with the study. … a physical stressor that … Psychology, Union College BrianCohen, PhD, Department of Biology, Union College Lindsay C. … ”
Abstract
When the body perceives a stressor, the hypothalamic-pituitary-adrenal (HPA) axis is activated leading to synthesis and release of cortisol from the adrenal cortex. Exercise is a physical stressor that increases cortisol levels in the body. Studies have considered exercise as a self-regulatory strategy to adapt to stressful events. Performance athletes have shown to have higher baseline cortisol levels. The effects of exercise on cortisol levels in the general population is less clear. The aim of this study was to examine the relationship between exercise and physiological responses to stress in college students. Stress was induced in the undergraduates (with the approval of the Union College Human Subjects in Research Committee) using the Trier Social Stress Test (TSST). Saliva samples were taken four times during the study; pre TSST, immediately post TSST, and 5 min and 30 min after the TSST. Salivary cortisol was measured using an enzyme-linked immunosorbent assay. In addition to physiological measures, participants completed the brief mood introspection scale, the revised undergraduate hassles scale, and a self-reported exercise survey describing the frequency and intensity of their exercise habits. Students also completed a perceived functional ability questionnaire (PFA) rating their ability to jog, walk or run over a given distance which reflects participants� physical activity and fitness. Participants were divided into three groups based on the mean PFA score (8.7). The results showed that participants with a PFA score one standard deviation above the mean had lower baseline cortisol on average, compared to participants with a PFA score one standard deviation below the mean. The results from this study suggest that physically active participants have lower baseline cortisol levels when compared to their peers who are less physically active. Together the data indicates the need for specific stress-reduction programs for college students that include physical activity as one aspect of the intervention.
“ … population Department of Biochemistry, June 2019. ADVISOR: Brian Cohen The human body’s primary stress response is … glucocorticoid … . Acknowledgements: I would like to thank BrianCohen, my research Advisor, for this research experience and … ”
Abstract
The human body’s primary stress response is regulated through the secretion of the glucocorticoid hormone cortisol through the hypothalamic-pituitary-adrenal (HPA) axis. Utilizing glucocorticoid receptors (GR), cortisol regulates the expression of certain genes that play a role in metabolism regulation and obesity related conditions such as type 2 diabetes, insulin resistance, and elevated serum triglycerides. Based on this, our lab and others have identified the presence of mutations in the genes for the glucocorticoid receptor (GR), the closely related mineralocorticoid receptor (MR), and regulatory proteins associated with cortisol or GR function (heat shock protein 90, 11ß-hydroxysteroid dehydrogenase type 1, and FK506 binding protein (FKBP)) to be of higher frequency in obese populations. Our study continued to investigate the genotypic frequency of these single nucleotide polymorphisms in the clinical population of bariatric patients from Ellis Hospital in hopes to better understand the role of genotypic variation in cortisol regulation and obesity. Our study also investigated the correlation between one’s success in bariatric surgery (measured by percent excess body weight loss) and one’s genotype for each SNP. DNA samples were extracted from buccal swabs of Ellis Hospital participants and analyzed using allele specific polymerase chain reactions to determine the genotypic frequency of SNPs associated with hypersensitivity or resistance to cortisol. From the polymorphisms tested, a higher mutant allele frequency was seen in the bariatric population compared to the general population for SNPS: BclI, N363S, and rs120. A lower mutant allele frequency was seen in the bariatric population compared to the general population for SNPs: tthIII and FK506. Significant correlations could not be made between percent excess body weight loss and genotypes for each SNP due to small population size. In addition, our study designed a new multiplex assay in which wild type and mutant alleles can be tested in a single PCR reaction to greatly increase the efficiency of data collection. With a greater sample population, more significant correlations can be made between SNPs and obesity to aid in the steps towards personalized medicine based on one’s genetic information.
“ … - 32 - Modified Western Blot Protocol- Created by Brian D. Cohen 1: Buffers and solutions • 1X Transfer Buffer … to express my gratitude to my thesis advisor Professor Brian Cohen for the invaluable … experiments will build off … … stimulating hormone. I like to thank the entire Team Cohen, for making our lab an organized and entertaining … ”
Abstract
Follicle stimulating hormone is involved in male and female fertility as a component of the HPG Axis. Its functions include spermatogenesis in males and ovarian follicle development in females. To better understand the physiology of this hormone and its effects, we focused on the receptor for hFSH (hFSHR) and its location on the cell membrane. Previous work in our lab has demonstrated that hFSHR localizes in lipid rafts. These are regions of the cellular membrane with high levels of cholesterol and sphingolipids. What is unclear is how the receptor gets into the lipid rafts. To study the interaction between hFSHR and caveolin of lipid rafts, I examined the properties of the lipid rafts and their sensitivity to various detergents. This was done to verify that regions of the membrane where hFSHR was located were detergent resistant. The results showed that lipid rafts are resistant to some mild detergents such as Triton but were disrupted by stronger ones such as SDS. I also used the methods of immunoprecipitation and western blot analysis to directly study the interaction between caveolin proteins and hFSHR. Experiments have suggested that the caveolin protein is closely linked with the receptor. Future experiments will build off these results and make use of caveolin mutants produced in the Cohen Lab to determine how changes to caveolin interfere with hFSHR function by measuring levels of downstream signaling molecules. This has potential implications in developing contraceptives and possible treatments for fertility treatments involving the action of FSH and its receptor.
“ … Hormone Receptor Action Using Small Molecules ADVISOR: Brian D. Cohen Follicle stimulating hormone (FSH) plays an … ”
Abstract
Follicle stimulating hormone (FSH) plays an integral role in the maturation of gametes in both males and females, making it a target for both contraceptive and fertility research. This heterodimeric protein hormone acts on a g protein-coupled receptor aptly named the follicle stimulating hormone receptor (FSHR), which is expressed in testicular Sertoli and ovarian granulosa cells. This receptor has been known to associate with cholesterol and sphingolipid-rich membrane domains called lipid rafts as well as the membrane protein caveolin, which act to regulate signaling. The binding of the hormone to its receptor activates an intracellular signaling cascade which as traditionally been considered to be mainly cAMP-dependent. However, recent research has indicated that the actual mechanism of action of FSHR's signaling may be far more complicated than previously considered. In this work, small molecule modulators were used to attempt to more clearly characterize the intricacies of receptor signaling. First, peptide mimetic of the receptor's caveolin interaction motif were used to examine the interaction between caveolin and the receptor complex. Second, downstream signaling after administration of negative allosteric modulators (NAMs) known to inhibit FSHR signaling was examined to determine where in the signaling cascade inhibition is evident. Western blotting was used to compare relative amounts of secondary messenger activation. Results suggest that caveolin has an inhibitory action on the receptor, and generally, small molecules have the ability to modulate receptor action. Better understanding of this signaling pathway may eventually enable the development of more targeted pharmaceuticals for both contraception and fertility.
“ … Cell line Annie Leamon Union College - Schenectady, NY Brian Cohen Union College - Schenectady, NY Follow this and … Recommended Citation Leamon, Annie and Cohen,Brian , "FSH Responsiveness by … ii ABSTRACT LEAMON, … ”
Abstract
Osteoporosis can be a devastating illness which causes large losses of both monetarily and in terms of quality of life. Interestingly, two of the major risk factors for developing osteoporosis are age and gender as osteoporosis commonly affects postmenopausal women. For decades, osteoporosis was thought be driven by estrogen loss following menopause, however recent research has shown that FSH, the upstream endocrine regulator of estrogen, drives the development of this disease. Using RAW 264.7 cells, we tested the changes that FSH induces in preosteoclastic cells. We noted significant changes in kinase activation and ROS production in an FSH-dependant manner. Similarly, FSH induced gross morphological changes in this cell line.
