Activity And Localization Of Maltodextrin Binding Site Mutants Of Glycogen Synthase In Saccharomyces Cerevisiae

Mentor: Wayne A. WilsonGlycogen is a glucose polymer formed by the enzyme glycogen synthase and is used in many organisms to store chemical energy. Saccharomyces cerevisiae (baker’s yeast) was used to study the activity and localization of glycogen synthase. Genes GSY1 and GSY2 encode glycogen synthase. GSY2 is responsible for the formation of Gsy2p, whose action accounts for ~90% of glycogen synthase activity; the remainder of total glycogen synthase activity stems from Gsy1p. Because glycogen synthase binds to glycogen, it can be used to determine glycogen localization. Glycogen synthase can appear in distinct patterns throughout the cell. Gsy2p has been shown to be regulated by phosphorylation. Phosphorylation of Gsy2p leads to inactivation of the enzyme, a decrease in glycogen storage, and a more localized pattern of glycogen synthase. Conversely, lowering the phosphorylation state of Gsy2p results in increased glycogen production and delocalization of glycogen synthase throughout the cell. Glucose-6-P (glucose-6-phosphate) activates glycogen synthase regardless of its phosphorylation state. We obtained a set of plasmids from a collaborator, encoding Gsy2p mutated at sites believed to be involved with maltodextrin binding. Maltodextrin is a chain of 20 or fewer dextrose molecules with α (1→4) glycosidic bonds. A protein sequence involved in maltodextrin binding likely would also bind to glycogen. Our task was to discover the localization pattern shown by the maltodextrin binding site mutants of glycogen synthase using a GFP tag on GSY2. The goal of this study was to determine the 16 effects of Gsy2p maltodextrin binding mutants on glycogen synthase activity, localization, and glycogen accumulation

A Mediterranean Diet and Walking Intervention to Reduce Cognitive Decline and Dementia Risk in Independently Living Older Australians:The MedWalk Randomized Controlled Trial Experimental Protocol, Including COVID-19 Related Modifications and Baseline Characteristics.

Background:Several clinical trials have examined diet and physical activity lifestyle changes as mitigation strategies for risk factors linked to cognitive decline and dementias such as Alzheimer’s disease. However, the ability to modify these behaviors longer term, to impact cognitive health has remained elusive.Objective:The MedWalk trial’s primary aim is to investigate whether longer-term adherence to a Mediterranean-style diet and regular walking, delivered through motivational interviewing and cognitive-behavioral therapy (MI-CBT), can reduce age-associated cognitive decline and other dementia risk factors in older, independently living individuals without cognitive impairment.Methods:MedWalk, a one-year cluster-randomized controlled trial across two Australian states, recruited 60–90-year-old people from independent living retirement villages and the wider community. Participants were assigned to either the MedWalk intervention or a control group (maintaining their usual diet and physical activity). The primary outcome is 12-month change in visual memory and learning assessed from errors on the Paired Associates Learning Task of the Cambridge Neuropsychological Test Automated Battery. Secondary outcomes include cognition, mood, cardiovascular function, biomarkers related to nutrient status and cognitive decline, MI-CBT effectiveness, Mediterranean diet adherence, physical activity, quality of life, cost-effectiveness, and health economic evaluation.Progress and Discussion:Although COVID-19 impacts over two years necessitated a reduced timeline and sample size, MedWalk retains sufficient power to address its aims and hypotheses. Baseline testing has been completed with 157 participants, who will be followed over 12 months. If successful, MedWalk will inform interventions that could substantially reduce dementia incidence and ameliorate cognitive decline in the community.<br/

Sorption and Photodegradation Processes Govern Distribution and Fate of Sulfamethazine in Freshwater−Sediment Microcosms

The antibiotic sulfamethazine can be transported from manured fields to surface water bodies. We investigated the degradation and fate of sulfamethazine in pond water using 14C-phenyl-sulfamethazine in small pond water microcosms containing intact sediment and pond water. We found a 2.7-day half-life in pond water and 4.2-day half-life when sulfamethazine was added to the water (5 mg L–1 initial concentration) with swine manure diluted to simulate runoff. Sulfamethazine dissipated exponentially from the water column, with the majority of loss occurring via movement into the sediment phase. Extractable sulfamethazine in sediment accounted for 1.9–6.1% of the applied antibiotic within 14 days and then declined thereafter. Sulfamethazine was transformed mainly into nonextractable sediment-bound residue (40–60% of applied radioactivity) and smaller amounts of photoproducts. Biodegradation, as indicated by metabolite formation and 14CO2 evolution, was less significant than photodegradation. Two photoproducts accounted for 15–30% of radioactivity in the water column at the end of the 63-day study; the photoproducts were the major degradates in the aqueous and sediment phases. Other unidentified metabolites individually accounted for \u3c7% of radioactivity in the water or sediment. Less than 3% of applied radioactivity was mineralized to 14CO2. Manure input significantly increased sorption and binding of sulfamethazine residues to the sediment. These results show concurrent processes of photodegradation and sorption to sediment control aqueous concentrations and establish that sediment is a sink for sulfamethazine and sulfamethazine-related residues. Accumulation of the photoproducts and sulfamethazine in sediment may have important implications for benthic organisms

Mapping adipose and muscle tissue expression quantitative trait loci in African Americans to identify genes for type 2 diabetes and obesity

Relative to European Americans, type 2 diabetes (T2D) is more prevalent in African Americans (AAs). Genetic variation may modulate transcript abundance in insulin-responsive tissues and contribute to risk; yet published studies identifying expression quantitative trait loci (eQTLs) in African ancestry populations are restricted to blood cells. This study aims to develop a map of genetically regulated transcripts expressed in tissues important for glucose homeostasis in AAs, critical for identifying the genetic etiology of T2D and related traits. Quantitative measures of adipose and muscle gene expression, and genotypic data were integrated in 260 non-diabetic AAs to identify expression regulatory variants. Their roles in genetic susceptibility to T2D, and related metabolic phenotypes were evaluated by mining GWAS datasets. eQTL analysis identified 1,971 and 2,078 cis-eGenes in adipose and muscle, respectively. Cis-eQTLs for 885 transcripts including top cis-eGenes CHURC1, USMG5, and ERAP2, were identified in both tissues. 62.1% of top cis-eSNPs were within ±50kb of transcription start sites and cis-eGenes were enriched for mitochondrial transcripts. Mining GWAS databases revealed association of cis-eSNPs for more than 50 genes with T2D (e.g. PIK3C2A, RBMS1, UFSP1), gluco-metabolic phenotypes, (e.g. INPP5E, SNX17, ERAP2, FN3KRP), and obesity (e.g. POMC, CPEB4). Integration of GWAS meta-analysis data from AA cohorts revealed the most significant association for cis-eSNPs of ATP5SL and MCCC1 genes, with T2D and BMI, respectively. This study developed the first comprehensive map of adipose and muscle tissue eQTLs in AAs (publically accessible at https://mdsetaa.phs.wakehealth.edu) and identified genetically-regulated transcripts for delineating genetic causes of T2D, and related metabolic phenotypes

Dysregulation of the Norepinephrine Transporter Sustains Cortical Hypodopaminergia and Schizophrenia-Like Behaviors in Neuronal Rictor Null Mice

A novel animal model highlights the link between Akt dysfunction, reduced cortical dopamine function, norepinephrine transporters, and schizophrenia-like behaviors

Increased expression of receptor phosphotyrosine phosphatase-β/ζ is associated with molecular, cellular, behavioral and cognitive schizophrenia phenotypes

Schizophrenia is a serious and chronic mental disorder, in which both genetic and environmental factors have a role in the development of the disease. Neuregulin-1 (NRG1) is one of the most established genetic risk factors for schizophrenia, and disruption of NRG1 signaling has been reported in this disorder. We reported previously that NRG1/ErbB4 signaling is inhibited by receptor phosphotyrosine phosphatase-β/ζ (RPTP β/ζ) and that the gene encoding RPTPβ/ζ (PTPRZ1) is genetically associated with schizophrenia. In this study, we examined the expression of RPTPβ/ζ in the brains of patients with schizophrenia and observed increased expression of this gene. We developed mice overexpressing RPTPβ/ζ (PTPRZ1-transgenic mice), which showed reduced NRG1 signaling, and molecular and cellular changes implicated in the pathogenesis of schizophrenia, including altered glutamatergic, GABAergic and dopaminergic activity, as well as delayed oligodendrocyte development. Behavioral analyses also demonstrated schizophrenia-like changes in the PTPRZ1-transgenic mice, including reduced sensory motor gating, hyperactivity and working memory deficits. Our results indicate that enhanced RPTPβ/ζ signaling can contribute to schizophrenia phenotypes, and support both construct and face validity for PTPRZ1-transgenic mice as a model for multiple schizophrenia phenotypes. Furthermore, our results implicate RPTPβ/ζ as a therapeutic target in schizophrenia

NuSTAR and Chandra observations of new X-ray transients in the central parsec of the Galaxy

We report NuSTAR and Chandra observations of two X-ray transients, SWIFT J174540.7$-$290015 (T15) and SWIFT J174540.2$-$290037 (T37), which were discovered by the Neil Gehrels Swift Observatory in 2016 within $r\sim1$ pc of Sgr A*. NuSTAR detected bright X-ray outbursts from T15 and T37, likely in the soft and hard states, with 3-79~keV luminosities of $8\times10^{36}$ and $3\times10^{37}$ erg/s, respectively. No X-ray outbursts have previously been detected from the two transients and our Chandra ACIS analysis puts an upper limit of $L_X \lesssim 2 \times10^{31}$ erg/s on their quiescent 2-8 keV luminosities. No pulsations, significant QPOs, or type I X-ray bursts were detected in the NuSTAR data. While T15 exhibited no significant red noise, the T37 power density spectra are well characterized by three Lorentzian components. The declining variability of T37 above $\nu \sim 10$ Hz is typical of black hole (BH) transients in the hard state. NuSTAR spectra of both transients exhibit a thermal disk blackbody, X-ray reflection with broadened Fe atomic features, and a continuum component well described by Comptonization models. Their X-ray reflection spectra are most consistent with high BH spin ($a_{*} \gtrsim 0.9$) and large disk density ($n_e\sim10^{21}$ cm$^{-3}$). Based on the best-fit ionization parameters and disk densities, we found that X-ray reflection occurred near the inner disk radius, which was derived from the relativistic broadening and thermal disk component. These X-ray characteristics suggest the outbursting BH-LMXB scenario for both transients and yield the first BH spin measurements from X-ray transients in the central 100 pc region.Comment: 15 pages, 7 figures, accepted for publication in Ap

Two systems of resting state connectivity between the insula and cingulate cortex.

Abstract: The insula and cingulate cortices are implicated in emotional, homeostatic/allostatic, sensorimotor, and cognitive functions. Non-human primates have specific anatomical connections between sub-divisions of the insula and cingulate. Specifically, the anterior insula projects to the pregenual anterior cingulate cortex (pACC) and the anterior and posterior mid-cingulate cortex (aMCC and pMCC); the mid-posterior insula only projects to the posterior MCC (pMCC). In humans, functional neuroimaging studies implicate the anterior insula and pre/subgenual ACC in emotional processes, the mid-posterior insula with awareness and interoception, and the MCC with environmental monitoring, response selection, and skeletomotor body orientation. Here, we tested the hypothesis that distinct resting state functional connectivity could be identified between (1) the anterior insula and pACC/aMCC; and (2) the entire insula (anterior, middle, and posterior insula) and the pMCC. Functional connectivity was assessed from resting state fMRI scans in 19 healthy volunteers using seed regions of interest in the anterior, middle, and posterior insula. Highly correlated, low-frequency oscillations (&lt; 0.05 Hz) were identified between specific insula and cingulate subdivisions. The anterior insula was shown to be functionally connected with the pACC/aMCC and the pMCC, while the mid/posterior insula was only connected with the pMCC. These data provide evidence for a resting state anterior insula-pACC/ aMCC cingulate system that may integrate interoceptive information with emotional salience to form a subjective representation of the body; and another system that includes the entire insula and MCC, likely involved in environmental monitoring, response selection, and skeletomotor body orientation