EPIGENETIC REGULATION OF HORMONE ACTION IN THE RED FLOUR BEETLE, \u3cem\u3eTRIBOLIUM CASTANEUM\u3c/em\u3e (HERBST)

Hormones are the chemical communication signaling molecules released into the body fluid to stimulate target cells of multicellular organisms. Two major hormones, ecdysteroids (20-hydroxyecdysone, 20E) and juvenile hormones (JH), regulate a wide variety of physiological and developmental processes in insects. Therefore, hormones have been extensively studied and are attractive targets for the development of target-specific insect control methods. Recent studies suggest that epigenetics adds another layer of regulation to explain multiple functions of the same circulating hormone in different tissues and at various time points. In my dissertation, I focused on a major post-translational modification, ‘acetylation,’ to elucidate the epigenetic mechanisms involved in the regulation of juvenile hormone action. Two highly conserved enzymes mediate the acetylation of histones, histone acetyltransferases (HATs) and histone deacetylases (HDACs), which mediate the addition or removal of the acetyl group to histones and other proteins. The first part of my dissertation identified the genes coding for HDACs in Tribolium castaneum and studied their function using RNA interference (RNAi). Knockdown of 12 HDAC genes showed variable phenotypes; the most severe phenotype was detected in insects injected with double-stranded RNA (dsRNA) targeting class I (HDAC1, HDAC3) and class IV (HDAC11) HDACs. The dsHDAC1 injected larvae showed arrested growth and development and eventually died during the larval stage. The larvae injected with dsHDAC11 also showed similar phenotype as dsHDAC1 injected larvae. The knockdown of the HDAC3 gene during the final instar larval stage resulted in a pupa that showed abnormally folded wings and mortality. Application of JH analog hydroprene to the T. castaneum larvae suppressed the expression of HDAC1, HDAC3, and HDAC11 genes. Sequencing of RNA isolated from control and dsHDACs injected larvae identified several differentially expressed genes, including those involved in JH action. The acetylation levels of core histones showed an increase in TcA cells exposed to JH III or dsHDACs. Knockdown of class III ‘sirtuins deacetylase’ did not cause significant developmental defects or mortality. The second part of my research focused on histone acetyltransferases (HATs), a superfamily of enzymes that acetylate histones and other proteins. Knockdown of four genes coding for GCN5-related N-acetyltransferases showed severe developmental defects and larval mortality. Interestingly, knockdown of an N-a-acetyltransferase, NAA40 (TC015921) induced severe developmental defects and mortality. 20E induced NAA40 expression in TcA Cells. Knockdown of NAA40 during the final instar larvae suppressed the expression of genes coding for proteins involved in ecdysone action. Overall, these data suggest that epigenetic modifications influence hormone action by modulating acetylation levels of histones and by affecting the recruitment of proteins involved in the regulation of hormone response genes

Histone Deacetylase 3 Is Required for Development and Metamorphosis in the Red Flour Beetle, \u3cem\u3eTribolium castaneum\u3c/em\u3e

Background Hormones are chemical communication signaling molecules released into the body fluids to stimulate target cells of multicellular organisms. We recently showed that histone deacetylase 1 (HDAC1) plays an important role in juvenile hormone (JH) suppression of metamorphosis in the red flour beetle, Tribolium castaneum. Here, we investigated the function of another class I HDAC member, HDAC3, and show that it is required for the normal development of T. castaneum. Results RNA interference-mediated knockdown of the HDAC3 gene affected development resulting in abnormally folded wings in pupae and adults. JH analog, hydroprene, suppressed the expression of HDAC3 in T. castaneum larvae. The knockdown of HDAC3 during the final instar larval stage resulted in an increase in the expression of genes coding for proteins involved in JH action. Sequencing of RNA isolated from larvae injected with dsRNA targeting malE (E. coli gene, control) or HDAC3 followed by differential gene expression analysis identified 148 and 741 differentially expressed genes based on the P-value \u3c 0.01 and four-fold difference, and the P-value \u3c 0.05 and two-fold difference, respectively. Several genes, including those coding for myosin-I heavy chain (Myosin 22), Shaven, and nuclear receptor corepressor 1 were identified as differentially expressed genes in HDAC3 knockdown larvae. An increase in histone H3 acetylation, specifically H3K9, H3K18, and H3K27, was detected in HDAC3 knockdown insects. Conclusion Overall, these data suggest that HDAC3 affects the acetylation levels of histones and influences the expression of genes coding for proteins involved in the regulation of growth, development, and metamorphosis

Magnetic-field-free nonreciprocal transport in graphene multi-terminal Josephson junctions

Nonreciprocal superconducting devices have attracted growing interest in recent years as they potentially enable directional charge transport for applications in superconducting quantum circuits. Specifically, the superconducting diode effect has been explored in two-terminal devices that exhibit superconducting transport in one current direction while showing dissipative transport in the opposite direction. Here, we exploit multi-terminal Josephson junctions (MTJJs) to engineer magnetic-field-free nonreciprocity in multi-port networks. We show that when treated as a two-port electrical network, a three-terminal Josephson junction (JJ) with an asymmetric graphene region exhibits reconfigurable two-port nonreciprocity. We observe nonreciprocal (reciprocal) transport between superconducting terminals with broken (preserved) spatial mirror symmetry. We explain our observations by considering a circuit-network of JJs with different critical currents

Satellite chlorophyll concentration as an aid to understanding the dynamics of Indian oil sardine in the southeastern Arabian Sea

Coastal waters of Kerala, which form an integral part of the Malabar upwelling zone off the southwest coast of India, constitute an important fishing region for small pelagics. Satellite remote sensing data from 1998−2014 were used to test the hypothesis that fluctuations in the landings of Sardinella longiceps, the major pelagic fish landed in the area designated as the South Eastern Arabian Sea (SEAS), are influenced by seasonal variability in phytoplankton biomass (measured as chlorophyll a [chl a] concentration), under the changing strength of physical para - meters such as sea surface temperature (SST), alongshore wind stress, Ekman mass transport, sea level anomaly (SLA) and Kerala rainfall. Multiple linear regression analysis (MLRA) was used to assess the influence of physical forcing mechanisms on chl a concentration on monthly and seasonal scales. We found that SLA, alongshore wind stress, SST and rainfall were ranked 1 to 4, respectively, and the first 3 factors significantly influenced the chl a concentration of SEAS. Pearson’s correlation analysis between monthly chl a and sardine landing (with chl a leading) showed a maximum positive correlation (+0.26) at 2 and 3 mo lags, emphasizing that the influence of chl a on the fishery of S. longiceps is seasonal (r = 0.35 for seasonal lead−lag correlation) in the coastal waters of SEAS. Variation in phytoplankton biomass, as evidenced by chl a fluctuations, seems to have a decisive role in regulating the physiological condition of larvae spawned during the southwest monsoon season, their juveniles and finally the adults that are recruited into the fishery in the next season. Using the quantity of phytoplankton as a predictive tool will exploit the presumptive trophic link to aid understanding of sardine fishery dynamics in upwelling zones

The effect of cigarette smoke exposure on the development of inflammation in lungs, gut and joints of TNFΔARE mice

The inflammatory cytokine TNF-alpha is a central mediator in many immune-mediated diseases, such as Crohn's disease (CD), spondyloarthritis (SpA) and chronic obstructive pulmonary disease (COPD). Epidemiologic studies have shown that cigarette smoking (CS) is a prominent common risk factor in these TNF-dependent diseases. We exposed TNF Delta ARE mice; in which a systemic TNF-alpha overexpression leads to the development of inflammation; to 2 or 4 weeks of air or CS. We investigated the effect of deregulated TNF expression on CS-induced pulmonary inflammation and the effect of CS exposure on the initiation and progression of gut and joint inflammation. Upon 2 weeks of CS exposure, inflammation in lungs of TNF Delta ARE mice was significantly aggravated. However, upon 4 weeks of CS-exposure, this aggravation was no longer observed. TNF Delta ARE mice have no increases in CD4+ and CD8+ T cells and a diminished neutrophil response in the lungs after 4 weeks of CS exposure. In the gut and joints of TNF Delta ARE mice, 2 or 4 weeks of CS exposure did not modulate the development of inflammation. In conclusion, CS exposure does not modulate gut and joint inflammation in TNF Delta ARE mice. The lung responses towards CS in TNF Delta ARE mice however depend on the duration of CS exposure

Project #37: Management Guidelines for Patients with COVID-19: Rapid Cycle Improvement

Project’s purpose is to rapidly devise, continually improve, educate, and implement a live and changing COVID-19 management guideline based upon emerging best available evidence. This project also aims to optimize the care of patients with COVID-19 and improve patient outcomes.https://scholarlycommons.henryford.com/qualityexpo2022/1004/thumbnail.jp