Effect of Energy Metabolism on NF-kB activity in Ovarian Cancer

NF-kB is a transcription factor involved in cancer cell growth and survival. The activation of NF-kB can be assessed by monitoring phosphorylation of RelA p65 at Ser-536, which is a surrogate of the NF-kB transcription factor activation. The objective of this study was to determine if the loss of ATP leads to NF-kB deficiency and thus, apoptotic cell death of “bad” cells in ovarian cancer cells. The independent variables were metformin (Met), an anti-diabetic medicine, another compound MinB functionally similar to Met and a glucose transporter inhibitor BAY-876. The dependent variables were the resulting effect of Met and MinB on phosphorylated AMPK at Thr-172 (marker of ATP loss) and RelA p65 at Ser-536 (marker of NF-kB activation). In each experiment, AMPK and RelA phosphorylation were tested by treatment of ovarian cancer cell lines with Met, MinB, BAY-876, Met+BAY-876, MinB+BAY-876. Western blotting was performed to determine the phosphorylation levels of AMPK and RelA p65. For two gels, the process was repeated. In each gel, Met or MinB treatment leads to thicker bands of AMPK-p, indicating decrease in cellular ATP levels following treatments. The effect of Met, MinB, or BAY-876 on RelA p65 was limited. However, co-treatment of Met or MinB with BAY-876 caused strong inhibition of NF-kB, as reflected by reduction in RelA p65-p. These results suggested that ATP deficiency together with inhibition of glucose transport cause inactivation of NK-kB. Future research will be conducted to study the effects of these compounds or their combinations on ovarian cancer cell growth and survival against from apoptosis.https://scholarscompass.vcu.edu/uresposters/1267/thumbnail.jp

The Role of Actin Depolymerizing Factor in Regulating Actin Dynamics in Toxoplasma gondii

Apicomplexan parasites utilize a unique process of rapid motility termed gliding, which is coupled to their invasion of host cells. Gliding and invasion are dependent on parasite actin filaments, yet parasite actin is mostly non-filamentous. Filaments have been detected only transiently during gliding, suggesting that parasite actin filaments are rapidly assembled and disassembled during gliding motility. Little is known about what regulates the turnover of parasite actin filaments. In higher eukaryotes the Actin Depolymerizing Factor: ADF)/Cofilin proteins are essential regulators of actin filament turnover. ADF is one of the few actin binding proteins conserved in apicomplexan parasites. To investigate the role of ADF in regulating actin dynamics in apicomplexan parasites, Toxoplasma gondii was used as a model apicomplexan, and the activities of T. gondii ADF: TgADF) were analyzed in vitro and in vivo. We found that TgADF engaged in dual activities. In contrast to most ADF/Cofilin proteins, TgADF was found to be a potent actin monomer sequestering protein that strongly inhibited actin polymerization, suggesting that it likely functions to maintain high G-actin concentrations in the cytoplasm of non-motile parasites. This role was reflected in its molecular structure, in which conserved G-actin binding sites were maintained, while key F-actin binding residues were absent. Despite this, TgADF demonstrated the ability to promote actin filament disassembly via a severing mechanism. Using a conditional knockout system we examined the function of TgADF in the parasite. TgADF was found to be essential for controlling productive gliding motility, and its absence lead to defects in host cell invasion, parasite egress, and parasite dispersal. Detailed analysis of motility revealed that parasites were unable to engage in sustained helical gliding, and moved at markedly reduced speeds. These defects are predicted to arise from the presence of more stable actin filaments in the parasite. Overall both the monomer sequestering and filament severing activities of TgADF are predicted to serve important functions in vivo for maintaining high G-actin concentrations for rapid filament assembly, and disassembling actin filaments for rapid filament turnover, respectively. These studies demonstrated that ADF is essential for regulating actin dynamics in T. gondii

Genetic diversity and gene expression analysis of Phytophthora pluvialis, a foliar pathogen of conifers : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy (PhD) in Genetics, Massey University, Manawatū, New Zealand

Phytophthora pluvialis is the causal agent of red needle cast on Pinus radiata in New Zealand. It was first isolated in 2008 but had previously been recovered from tanoak (Notholithocarpus densiflorus) and Douglas fir (Pseudotsuga menziesii) trees in Oregon, USA in 2002. Phytophthora pluvialis was subsequently described as a new species in 2013 and classified as a clade 3 Phytophthora species. The aims of this study were to (1) gain a better understanding of the genetic diversity and population structure of P. pluvialis and (2) examine gene expression profiles of P. pluvialis from naturally infected P. radiata seedlings. Studying the genetic diversity and population structure of P. pluvialis provided insight into the mode of reproduction of this pathogen and helped determine if P. pluvialis was introduced into New Zealand. This information is also important for the development of management strategies for P. pluvialis. Twenty-seven single nucleotide polymorphism (SNP) markers were designed to genotype a total of 360 isolates of P. pluvialis collected from New Zealand and the USA. The genotypic data showed that the population in New Zealand has lower diversity than the USA population. A minimum spanning network (MSN) showed two unique clusters in the New Zealand population, suggesting there may have been two separate introductions of P. pluvialis. For the second study, samples were collected from 45 P. radiata grafted plants that were part of a field trial, with the aim of identifying genes that are highly expressed and may be important for virulence. Interestingly, Phytophthora kernoviae was found in more of the samples than P. pluvialis. Needle samples were collected, RNA was extracted and sequenced, and the normalised reads that mapped to the genome of P. pluvialis were compared to those from P. pluvialis grown in culture. Differentially expressed genes (DEGs) of P. pluvialis that showed higher expression in the field trial included potential orthologs of sugar transporter, GH12 and effector genes with known pathogenicity functions in other species. This is the first study to examine the genetic diversity of P. pluvialis in New Zealand and the USA., and to examine the gene expression of a Phytophthora forest pathogen in the field. The results from these studies provide useful tools for forest disease management. The SNP markers can be used to monitor the population of P. pluvialis in New Zealand. The highly expressed genes can be used to help identify resistance genes in P. radiata that can be incorporated into future breeding programs

Peptide mimics targeting bacterial membranes

In recent years, society has been increasingly concerned with bacteria that are no longer susceptible to commercial antibiotics. Faced with a lack of tools, medical practitioners today are forced to prescribe medicines that, although effective, cause as much harm to the patient as the principal infection. The purpose of this research project is to develop novel antibacterials that remain potent against bacterial infections without being toxic to the patient

Computerized Working-Memory Training for Children Following Arterial Ischemic Stroke:A Pilot Study With Long-Term Follow-Up

Cognitive deficits in the domains of working memory (WM) and executive function are well documented following childhood arterial ischemic stroke (AIS). However, there are currently no evidence-based cognitive interventions for this population. Computerized, implicit WM training has been demonstrated to generate generalized cognitive gains for children with WM and attention deficits and for adults following brain injury. This study used a pilot design to investigate the efficacy and feasibility of such an intervention program (Cogmed WM Training) for a childhood AIS population. Outcomes were measured via psychometric assessment at preintervention and postintervention and again at 1-year follow-up. At longitudinal follow-up, participants were found to have significant and persistent cognitive difficulties, particularly with attention and response inhibition. Following the computerized, implicit WM intervention, a significant improvement in phonological-loop WM was seen; however, this improvement was not maintained after 12 months. No additional significant improvements on standardized psychometric outcome measures were seen either immediately or at 12-month follow-up. Findings of this pilot study therefore do not currently support Cogmed as an effective intervention for children with AIS but highlight the need for further research, including randomized, controlled trials, to investigate cognitive interventions for the childhood AIS population

Treating Fecal Incontinence: An Unmet Need in Primary Care Medicine

Fecal incontinence affects up to 36% of primary care patients. Although effective treatments are available, doctors rarely screen for this condition and patients seldom volunteer complaints of fecal incontinence. Conservative management yields 60% improvement in symptoms and continence in 20% of patients. Referrals are currently being accepted for studies seeking to improve case detection and to support conservative management or self-care

Biomarkers to distinguish functional constipation from irritable bowel syndrome with constipation

Treatments for functional constipation (FC) and irritable bowel syndrome with constipation (IBS-C) differ, but symptom criteria do not reliably distinguish between them; some regard FC and IBS-C as parts of a single constipation spectrum. Our goal was to review studies comparing FC and IBS-C to identify possible biomarkers that separate them. A systematic review identified 15 studies that compared physiologic tests in FC vs IBS-C. Pain thresholds were lower in IBS-C than FC for 3/5 studies and not different in 2/5. Colonic motility was decreased more in FC than IBS-C for 3/3 studies, and whole gut transit was delayed more in FC than IBS-C in 3/8 studies and not different in 5/8. Pelvic floor dyssynergia was unrelated to diagnosis. Sympathetic arousal, measured in only one study, was greater in IBS-C than FC. The most reliable separation of FC from IBS-C was shown by a novel new magnetic resonance imaging technique described in this issue of the journal. These authors showed that drinking one liter of polyethylene glycol laxative significantly increased water content in the small intestine, volume of contents in the ascending colon, and time to first evacuation in FC vs IBS-C; and resulted in less colon motility and delayed whole gut transit in FC compared to IBS-C. Although replication is needed, this well-tolerated, non-invasive test promises to become a new standard for differential diagnosis of FC vs IBS-C. These data suggest that FC and IBS-C are different disorders rather than points on a constipation spectrum

Update on Rome IV Criteria for Colorectal Disorders: Implications for Clinical Practice

Purpose of Review: The purpose of the review was to provide an update of the Rome IV criteria for colorectal disorders with implications for clinical practice. Recent Findings: The Rome diagnostic criteria are expert consensus criteria for diagnosing functional gastrointestinal disorders (FGIDs). The current version, Rome IV, was released in May of 2016 after Rome III had been in effect for a decade. It is the collective product of committees that included more than 100 leading functional GI experts. For functional bowel and anorectal disorders, the majority of changes relative to Rome III are relatively minor and will have little impact on clinical practice. However, notable changes with potential impact on clinical practice and research include the changes in the diagnostic criteria for IBS, the modified approach for subtyping of IBS, the view on functional bowel disorders as a spectrum of disorders, and the new definition of fecal incontinence. Summary: New features in the Rome IV diagnostic criteria for functional bowel and anorectal disorders will likely have modest influence on clinical practice, with a few exceptions

Balancing Clinical Objectives with Patient Centered Care

This poster is a reflection focuses on the challenges of balancing the medical needs of a patient and their preferences in an interdisciplinary health care setting. Students from UNE\u27s Physician Assistant, Osteopathic Medicine, Social Work, Dental Medicine, and Nursing programs collaborated with Allopathic Medicine and Podiatric Medicine students from Rosalind Franklin University Medical School as a virtual health care team to care for a patient with long-covid