Assessment of underlying neurocircuitry in dysregulation of the Hypothalamic Pituitary Adrenal Stress Axis in HIV-1 Tat Male Mice

Overview: My name is Salahuddin Mohammed, a 2nd year PhD graduate student from Dr. Jason Paris’ Lab. My research dissertation focuses on investigating the neuroendocrine underpinnings of HIV-and opioid-mediated anxiety-/depression-like behavior and cognitive impairment via experiments utilizing humans and animal models. Through this work I strive to delineate the underlying mechanism for the neuroHIV symptomology and develop novel therapeutic agents which can ameliorate HIV-1 mediated Tat-neurotoxicity. Approximately 50% of the HIV infected patients suffer from HIV associated neurocognitive disorders (HAND) which can be exacerbated by drug use/abuse. Some neuroHIV symptoms include anxiety, major depressive disorder, neurocognitive impairment, and additional psychiatric comorbidities that impair the quality of life (QOL) like and increased abuse risk for opioids, alcohol, psychostimulants, etc. One protein, the HIV-1 regulatory protein, trans-activator of transcription (Tat), may contribute to neuropathology by promoting neuroinflammation and neurotoxicity which may be exacerbated by opioids like oxycodone. The underlying mechanisms for the neuroHIV symptomology are unknown. Moreover, the HIV+ population is commonly prescribed opioids like oxycodone (Merlin et al., 2016) which may further raise concerns about HIV-1 Tat and oxycodone interactions. Evidence shows that 20-80% of HIV+ patients may have a dysregulated hypothalamic-pituitary-adrenal (HPA) stress axis (usually characterized by high basal stress hormones with a lack of appropriate hormonal response to additional stressors). Preliminary findings from our lab use a mouse model of HIV-1 Tat exposure. These mice demonstrate higher circulating stress hormone (i.e. corticosterone) when exposed to the HIV-1 Tat protein and appear to recapitulate the human phenotype. These mice may be used to assess the underlying mechanisms within the brain that cause HPA dysregulation associated with HIV and, thus, may be used as a model with which to screen treatments for this disorder. Intellectual Merit: The present project will delineate the neurocircuitry involved in HIV-protein-mediated dysregulation of the stress axis. This work will further provide sites to be targeted to restore HPA stress axis homeostasis and reduce the overall burden of present and future neurological sequela in HIV+ patients. External Opportunity: The findings will be presented at the Society for Neuroscience (SfN) conference scheduled for this year November 13–21, 2021 at Chicago, IL, U.S.A. The conference is expected to be attended by approximately 30,000 scientists from over 80 countries. My attendance at this conference would greatly help me to garner structured feedback and insights from experts in this domain. Attending this meeting would also provide opportunities to learn relevant cellular and molecular advancements in the fields of neuroscience. Moreover, I would be enthusiastic to meet leaders in fields of sex differences and opioids/HIV research such as Drs. Pamela Knapp, Margaret McCarthy, Georgia Hodes, Oliver Hobert, Wendy Macklin, and others. I believe attendance at this conference will have a formative impact on my career. Additionally, this grant may also help me to apply for extramural Sigma Xi society funding which provides fellowship of $1000 to graduate students with application deadline of October 1 every year. This grant will advance my scientific fluency, professional development, and capacity for building my CV

Indigenous Toxin Affects Cell Viability, While an Artificial Proteolytic Queueing Causes the Upregulation of Specific Genes

Studying bacterial physiology is crucial to understand the fundamental mechanisms that govern bacterial growth, survival, and adaptation. This thesis combines two chapters investigating bacterial physiology by studying important cellular processes like bacterial toxin-antitoxin systems and proteolytic pathways. The first chapter involves studying bacterial toxin-antitoxin systems to understand the regulation and function of hypothetical toxin-antitoxin (TA) systems of bacteria and the effects of these TA systems on bacterial growth and survival. The emergence of these genetic modules in bacterial research and the unrevealing of some of their important roles in cell physiology in recent years has drawn much attention in scientific communities. This chapter looks for new TA systems using molecular techniques and conventional microbiology approaches. The results of this study show the effects of a toxin protein of the putative YfeD-YfeC TA system of Escherichia coli (E. coli). The second chapter utilizes a systems biology approach by using molecular biology, synthetic biology, and RNA-Seq techniques to understand the effect of artificial proteolytic queueing in E. coli by introducing synthetic degradation circuits into them. The goal of this project was to test if the formation of an artificial proteolytic queue alters gene regulation. Our results demonstrate that the artificial proteolytic queue causes a cellular burden. Furthermore, this burden does not slow growth because the cell mitigates the queueing effect by responding to it by upregulating specific genes; specifically, chaperones and proteases

FORMULATION AND IN-VITRO EVALUATION OF DEFERASIROX ORO-DISPERSIBLE TABLETS

Objective: Deferasirox is a once daily, oral iron chelator approved for treatment of transfusional iron overload in adult and paediatric patients. In case of iron overdose or poisoning, immediate attention deferasirox is required in the systemic circulation to counter act the iron. Hence we feel that formulation of Oro-dispersible tablets of deferasirox is an ideal approach. Methods: FTIR showed that there is no interaction between drug and excipients. Mouth dissolving tablets of Deferasirox were prepared using crospovidone, croscarmellose and sodium starch glycolate as superdisintegrants by direct compression method. Dissolution of prepared Oro-dispersible tablets of Deferasirox was performed using USP type II apparatus in 6.8 pH phosphate buffer at 50 rpm with temperature being maintained at 37±0.5oC. Samples were withdrawn at 10, 15, 20, 30 and 45min time points and were analysed using UV- visible spectrophotometer at 245 nm. Results: The tablets prepared were evaluated for various parameters like weight variation, hardness, friability, in-vitro dispersion time, drug polymer interaction, drug content, water absorption ratio and wetting time and in-vitro release and were found to be within the limits. Conclusion: Formulation with crospovidone containing superdisintegrants has shown better in-vitro dissolution profile compared with other formulations. In conclusion, development of fast dissolving tablets using superdisintegrants gives rapid drug delivery and rapid onset of action

HIV-1 Tat Protein Promotes Neuroendocrine Dysfunction Concurrent with the Potentiation of Oxycodone’s Psychomotor Behavioral Effects in Female Transgenic Mice

Presenter: Salahuddin Mohammedhttps://egrove.olemiss.edu/pharm_annual_posters_2021/1005/thumbnail.jp

HIV-1 Tat Dysregulates the Hypothalamic-Pituitary-Adrenal Stress Axis and Potentiates Oxycodone-mediated Psychomotor and Anxiety-like Behavior of Male Mice

Graduate students: Salahuddin Mohammed, Department of BioMolecular Sciences, School of Pharmacy; Fakhri Mahdi, Department of BioMolecular Sciences, School of Pharmacy; Alaa N. Qrareya, Department of BioMolecular Sciences, School of PharmacyMajor/Minor: Major: Pharmaceutical Science, School of PharmacyFaculty advisor: Jason J. Paris, Department of BioMolecular Sciences,Research Institute of Pharmaceutical Science, School of Pharmacyhttps://egrove.olemiss.edu/neuro_showcase/1006/thumbnail.jp

Hiv-1 tat protein promotes neuroendocrine dysfunction concurrent with the potentiation of oxycodone’s psychomotor effects in female mice

Human immunodeficiency virus (HIV) is associated with neuroendocrine dysfunction which may contribute to co-morbid stress-sensitive disorders. The hypothalamic-pituitary-adrenal (HPA) or-gonadal (HPG) axes are perturbed in up to 50% of HIV patients. The mechanisms are not known, but we have found the HIV-1 trans-activator of transcription (Tat) protein to recapitulate the clinical phenotype in male mice. We hypothesized that HPA and/or HPG dysregulation contrib-utes to Tat-mediated interactions with oxycodone, an opioid often prescribed to HIV patients, in females. Female mice that conditionally-expressed the Tat1–86 protein [Tat(+) mice] or their counter-parts that did not [Tat(−) control mice] were exposed to forced swim stress (or not) and behaviorally-assessed for motor and anxiety-like behavior. Some mice had glucocorticoid receptors (GR) or cor-ticotropin-releasing factor receptors (CRF-R) pharmacologically inhibited. Some mice were ovari-ectomized (OVX). As seen previously in males, Tat elevated basal corticosterone levels and poten-tiated oxycodone’s psychomotor activity in females. Unlike males, females did not demonstrate adrenal insufficiency and oxycodone potentiation was not regulated by GRs or CRF-Rs. Rather OVX attenuated Tat/oxycodone interactions. Either Tat or oxycodone increased anxiety-like behavior and their combination increased hypothalamic allopregnanolone. OVX increased basal hypothalamic allopregnanolone and obviated Tat or oxycodone-mediated fluctuations. Together, these data provide further evidence for Tat-mediated dysregulation of the HPA axis and reveal the importance of HPG axis regulation in females. HPA/HPG disruption may contribute vulnerability to affective and substance use disorders

EVALUATION OF RATIONAL USE OF ANTIBIOTIC DRUGS IN SURGERY DEPARTMENT AT A TERTIARY CARE HOSPITAL

Objective: Antibiotics are the only drug where use in one patient can impact the effectiveness in another, so antibiotic misuse adversely impacts the patients and society. Improving antibiotic use improves patient outcomes and saves money. Antibiotic resistance has been identified as a major threat by the WHO due to the lack of development of new antibiotics and the increasing infections caused by multidrug resistance pathogens became untreatable. Methods: A prospective observational study was conducted for a period of 6 months. Data were collected from prescriptions and inpatient record files at the surgery department of the tertiary care hospital. Patients above age of 18 years of either gender whose prescription containing the antibiotics and patients who are willing to participate in the study were included in the study. Microsoft Excel was used for recording and analyzing the data of recruited subjects. Results: During our study period, we have collected 100 cases as per inclusion criteria, in total collected 100 cases, 52% are male and 48% are female. The mean age and standard deviation of the study population were found to be 46.61±16.12. The most commonly prescribed classification before and after the surgery is cephalosporin’s that is 57%. Results show that in pre-surgery, almost 93% of prescriptions have chosen the drugs as per ASHP guidelines, whereas in post-surgery, 95% of drugs have selected the drug as per ASHP guidelines. Conclusion: Our study has observed that some of the prescriptions are irrationally prescribed so the pharmacist has to take the responsibility to improve the awareness regarding rational prescribing of antibiotics. The national wide monitoring of antibiotics use, national schemes to obtain rational use of antibiotics, reassessing the prescriptions, education to practitioners, and surveys on antibiotics should be implemented

Psychometric analysis of the perceived stress scale among healthy university students

© 2020 Anwer et al. Background: There is a gradual increase in the prevalence of stress during professional courses. Previous studies reported a high incidence of stress among university students. The psychometric properties of the perceived stress scale-10 (PSS-10) have been established in different populations. The current study aimed to assess psychometric properties of the PSS10 in Saudi university students. Methods: Healthy university students (n= 192) participated in this cross-sectional study. All the participants were explained about the aim and procedures of the study. Participants were requested to complete the English version of the PSS, the generalized anxiety disorder-7 (GAD-7), the sleep hygiene index (SHI), and demographic details. Results: The range of the PSS-10 total score was 0–35; 1% reported minimum score of 0, but none reported maximum score of 40. Therefore, there was no issue of ceiling or floor effect in the PSS-10 total score. Positive and significant correlations of the PSS total and the PSS Factor-1 (distress perception) with the GAD-7 total score, SHI item-8 and SHI item-13 score support its convergent validity. Negative or no correlation of the PSS Factor-2 score (coping perception) with the GAD-7 total score, SHI item-8 and SHI item-13 scores demonstrate its divergent validity. The internal homogeneity test indicated moderate to strong positive correlations (r=0.60–0.82) between the PSS Factors and the items loading on them. The internal consistency test showed a good agreement for the PSS Factor-1 and the PSS Factor-2 scores (Cronbach’s alpha 0.78 and 0.71, respectively), suggesting an acceptable level of consistency. Factor analysis favored a 2-Factor model of the PSS in the Saudi students. Conclusion: The current study supported the use of the PSS-10 to assess the perceived stress among Saudi university students

Substation system simulation models for transformer risk assessment analysis

This paper comprises a study which is carried out to investigate and evaluate the effect of lightning stresses on the 132 kV substation in the way to improve its reliability in the event of active lightning activities. The paper also detailed the modelling parameters of substation for this transient analysis in order to evaluate the performance and to recommend such configuration to optimize its design to be not only to withstand the stresses but to be more cost effective. The modelling and simulation are carried out using one of the most powerful power system simulations tools that is PSCAD-EMTDC and the substation layout design is adapted from 132/11 kV Simpang Renggam -- Ayer Hitam substation, courtesy of TNB. The model is based on single phase line model as it was suggested by the IEEE to be adequate to represent the substation in transient analysis simulation. The outcome of this paper would be the results of lightning stresses in term of voltage level measured at particular points in substation. The results are then compared with the suggested BIL for assessment of transformer failure