A New Column Perfusion Technique Using Isolated Rat Adrenal Cells to Study ACTH and Danazol

A thesis presented to the faculty of the School of Sciences and Mathematics at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science in Biology by Diane H. Johnson on August 15, 1982

ALMA Resolves 30 Doradus: Sub-parsec Molecular Cloud Structure Near the Closest Super-Star Cluster

We present ALMA observations of 30 Doradus -- the highest resolution view of molecular gas in an extragalactic star formation region to date (~0.4pc x 0.6pc). The 30Dor-10 cloud north of R136 was mapped in 12CO 2-1, 13CO 2-1, C18O 2-1, 1.3mm continuum, the H30alpha recombination line, and two H2CO 3-2 transitions. Most 12CO emission is associated with small filaments and clumps (<1pc, ~1000 Msun at the current resolution). Some clumps are associated with protostars, including "pillars of creation" photoablated by intense radiation from R136. Emission from molecular clouds is often analyzed by decomposition into approximately beam-sized clumps. Such clumps in 30 Doradus follow similar trends in size, linewidth, and surface density to Milky Way clumps. The 30 Doradus clumps have somewhat larger linewidths for a given size than predicted by Larson's scaling relation, consistent with pressure confinement. They extend to higher surface density at a given size and linewidth compared to clouds studied at 10pc resolution. These trends are also true of clumps in Galactic infrared-dark clouds; higher resolution observations of both environments are required. Consistency of clump masses calculated from dust continuum, CO, and the virial theorem reveals that the CO abundance in 30 Doradus clumps is not significantly different from the LMC mean, but the dust abundance may be reduced by ~2. There are no strong trends in clump properties with distance from R136; dense clumps are not strongly affected by the external radiation field, but there is a modest trend towards lower dense clump filling fraction deeper in the cloud.Comment: accepted to Ap

4-aminopyridyl-based lead compounds targeting CYP51 prevent spontaneous parasite relapse in a chronic model and improve cardiac pathology in an acute model of Trypanosoma cruzi infection.

BackgroundChagas disease, caused by the protozoan Trypanosoma cruzi, is the leading cause of heart failure in Latin America. The clinical treatment of Chagas disease is limited to two 60 year-old drugs, nifurtimox and benznidazole, that have variable efficacy against different strains of the parasite and may lead to severe side effects. CYP51 is an enzyme in the sterol biosynthesis pathway that has been exploited for the development of therapeutics for fungal and parasitic infections. In a target-based drug discovery program guided by x-ray crystallography, we identified the 4-aminopyridyl-based series of CYP51 inhibitors as being efficacious versus T.cruzi in vitro; two of the most potent leads, 9 and 12, have now been evaluated for toxicity and efficacy in mice.Methodology/principal findingsBoth acute and chronic animal models infected with wild type or transgenic T. cruzi strains were evaluated. There was no evidence of toxicity in the 28-day dosing study of uninfected animals, as judged by the monitoring of multiple serum and histological parameters. In two acute models of Chagas disease, 9 and 12 drastically reduced parasitemia, increased survival of mice, and prevented liver and heart injury. None of the compounds produced long term sterile cure. In the less severe acute model using the transgenic CL-Brenner strain of T.cruzi, parasitemia relapsed upon drug withdrawal. In the chronic model, parasitemia fell to a background level and, as evidenced by the bioluminescence detection of T. cruzi expressing the red-shifted luciferase marker, mice remained negative for 4 weeks after drug withdrawal. Two immunosuppression cycles with cyclophosphamide were required to re-activate the parasites. Although no sterile cure was achieved, the suppression of parasitemia in acutely infected mice resulted in drastically reduced inflammation in the heart.Conclusions/significanceThe positive outcomes achieved in the absence of sterile cure suggest that the target product profile in anti-Chagasic drug discovery should be revised in favor of safe re-administration of the medication during the lifespan of a Chagas disease patient. A medication that reduces parasite burden may halt or slow progression of cardiomyopathy and therefore improve both life expectancy and quality of life

Smoking Enhances Risk for New External Genital Warts in Men

Repeat episodes of HPV-related external genital warts reflect recurring or new infections. No study before has been sufficiently powered to delineate how tobacco use, prior history of EGWs and HIV infection affect the risk for new EGWs. Behavioral, laboratory and examination data for 2,835 Multicenter AIDS Cohort Study participants examined at 21,519 semi-annual visits were evaluated. Fourteen percent (391/2835) of men reported or were diagnosed with EGWs at 3% (675/21,519) of study visits. Multivariate analyses showed smoking, prior episodes of EGWs, HIV infection and CD4+ T-lymphocyte count among the infected, each differentially influenced the risk for new EGWs

Geochemistry and oxygen isotope composition of main-group pallasites and olivine-rich clasts in mesosiderites: Implications for the "Great Dunite Shortage" and HED-mesosiderite connection

Evidence from iron meteorites indicates that a large number of differentiated planetesimals formed early in Solar System history. These bodies should have had well-developed olivine-rich mantles and consequentially such materials ought to be abundant both as asteroids and meteorites, which they are not. To investigate this “Great Dunite Shortage” we have undertaken a geochemical and oxygen isotope study of main-group pallasites and dunitic rocks from mesosiderites. Oxygen isotope analysis of 24 main-group pallasites (103 replicates) yielded a mean Δ17O value of −0.187 ± 0.016‰ (2σ), which is fully resolved from the HED Δ17O value of −0.246 ± 0.014 (2σ) obtained in our earlier study and demonstrates that both groups represent distinct populations and were derived from separate parent bodies. Our results show no evidence for Δ17O bimodality within the main-group pallasites, as suggested by a number of previous studies. Olivine-rich materials from the Vaca Muerta, Mount Padbury and Lamont mesosiderites, and from two related dunites (NWA 2968 and NWA 3329), have Δ17O values within error of the mesosiderite average. This indicates that these olivine-rich materials are co-genetic with other mesosiderite clasts and are not fragments from an isotopically distinct pallasite-like impactor. Despite its extreme lithologic diversity the mesosiderite parent body was essentially homogeneous with respect to Δ17O, a feature best explained by an early phase of large-scale melting (magma ocean), followed by prolonged igneous differentiation. Based on the results of magma ocean modeling studies, we infer that Mg-rich olivines in mesosiderites formed as cumulates in high-level chambers and do not represent samples of the underlying mantle. By analogy, recently documented Mg-rich olivines in howardites may have a similar origin. Although the Dawn mission did not detect mesosiderite-like material on Vesta, evidence linking the mesosiderites and HEDs includes: (i) their nearly identical oxygen isotope compositions; (ii) the presence in both of coarse-grained Mg-rich olivines; (iii) both have synchronous Lu-Hf and Mn-Cr ages; (iv) there are compositional similarities between the metal in both; and (v) mesosiderite-like material has been identified in a howardite breccia. The source of the mesosiderites remains an outstanding question in meteorite science. The underrepresentation of olivine-rich materials amongst both asteroids and meteorites results from a range of factors. However, evidence from pallasites and mesosiderites indicates that the most important reason for this olivine shortage lies in the early, catastrophic destruction of planetesimals in the terrestrial planet-forming region and the subsequent preferential loss of their olivine-rich mantles

A human MAP kinase interactome.

Mitogen-activated protein kinase (MAPK) pathways form the backbone of signal transduction in the mammalian cell. Here we applied a systematic experimental and computational approach to map 2,269 interactions between human MAPK-related proteins and other cellular machinery and to assemble these data into functional modules. Multiple lines of evidence including conservation with yeast supported a core network of 641 interactions. Using small interfering RNA knockdowns, we observed that approximately one-third of MAPK-interacting proteins modulated MAPK-mediated signaling. We uncovered the Na-H exchanger NHE1 as a potential MAPK scaffold, found links between HSP90 chaperones and MAPK pathways and identified MUC12 as the human analog to the yeast signaling mucin Msb2. This study makes available a large resource of MAPK interactions and clone libraries, and it illustrates a methodology for probing signaling networks based on functional refinement of experimentally derived protein-interaction maps

Variation of organic matter quantity and quality in streams at Critical Zone Observatory watersheds

The quantity and chemical composition of dissolved organic matter (DOM) in surface waters influence ecosystem processes and anthropogenic use of freshwater. However, despite the importance of understanding spatial and temporal patterns in DOM, measures of DOM quality are not routinely included as part of large-scale ecosystem monitoring programs and variations in analytical procedures can introduce artifacts. In this study, we used consistent sampling and analytical methods to meet the objective of defining variability in DOM quantity and quality and other measures of water quality in streamflow issuing from small forested watersheds located within five Critical Zone Observatory sites representing contrasting environmental conditions. Results show distinct separations among sites as a function of water quality constituents. Relationships among rates of atmospheric deposition, water quality conditions, and stream DOM quantity and quality are consistent with the notion that areas with relatively high rates of atmospheric nitrogen and sulfur deposition and high concentrations of divalent cations result in selective transport of DOM derived from microbial sources, including in-stream microbial phototrophs. We suggest that the critical zone as a whole strongly influences the origin, composition, and fate of DOM in streams. This study highlights the value of consistent DOM characterization methods included as part of long-term monitoring programs for improving our understanding of interactions among ecosystem processes as controls on DOM biogeochemistry

Tracing chemical evolution over the extent of the Milky Way's Disk with APOGEE Red Clump Stars

We employ the first two years of data from the near-infrared, high-resolution SDSS-III/APOGEE spectroscopic survey to investigate the distribution of metallicity and alpha-element abundances of stars over a large part of the Milky Way disk. Using a sample of ~10,000 kinematically-unbiased red-clump stars with ~5% distance accuracy as tracers, the [alpha/Fe] vs. [Fe/H] distribution of this sample exhibits a bimodality in [alpha/Fe] at intermediate metallicities, -0.9<[Fe/H]<-0.2, but at higher metallicities ([Fe/H]=+0.2) the two sequences smoothly merge. We investigate the effects of the APOGEE selection function and volume filling fraction and find that these have little qualitative impact on the alpha-element abundance patterns. The described abundance pattern is found throughout the range 5<R<11 kpc and 0<|Z|<2 kpc across the Galaxy. The [alpha/Fe] trend of the high-alpha sequence is surprisingly constant throughout the Galaxy, with little variation from region to region (~10%). Using simple galactic chemical evolution models we derive an average star formation efficiency (SFE) in the high-alpha sequence of ~4.5E-10 1/yr, which is quite close to the nearly-constant value found in molecular-gas-dominated regions of nearby spirals. This result suggests that the early evolution of the Milky Way disk was characterized by stars that shared a similar star formation history and were formed in a well-mixed, turbulent, and molecular-dominated ISM with a gas consumption timescale (1/SFE) of ~2 Gyr. Finally, while the two alpha-element sequences in the inner Galaxy can be explained by a single chemical evolutionary track this cannot hold in the outer Galaxy, requiring instead a mix of two or more populations with distinct enrichment histories.Comment: 18 pages, 17 figures. Accepted for publication in Ap

The Impact of Nocturia on Mortality: A Systematic Review and Meta-Analysis

Purpose: Nocturia (waking from sleep at night to void) is a common cause of sleep disruption associated with increased comorbidity and impaired quality of life. However, its impact on mortality remains unclear. We performed a systematic review and meta-analysis to evaluate the association of nocturia with mortality as a prognostic factor and a causal risk factor. Materials and Methods: We searched PubMed (R), Scopus (R), CINAHL (R) (Cumulative Index of Nursing and Allied Health Literature) and major conference abstracts up to December 31, 2018. Random effects meta-analyses were done to address the adjusted RR of mortality in people with nocturia. Metaregression was performed to explore potential determinants of heterogeneity, including the risk of bias. We applied the GRADE (Grades of Recommendation, Assessment, Development and Evaluation) framework to rate the quality of evidence for nocturia as a prognostic risk factor for mortality and separately as a cause of mortality. Results: Of the 5,230 identified reports 11 observational studies proved eligible for inclusion. To assess nocturia 10 studies used symptom questionnaires and 1 used frequency-volume charts. Nocturia was defined as 2 or more episodes per night in 6 studies (55%) and as 3 or more episodes per night in 5 (45%). Pooled estimates demonstrated a RR of 1.27 (95% CI 1.16-1.40, I-2=48%) with an absolute 1.6% and 4.0% 5-year mortality difference in individuals 60 and 75 years old, respectively. The pooled estimates of relative risk did not differ significantly across varying age, gender, followup, nocturia case definition, risk of bias or study region. We rated the quality of evidence for nocturia as a prognostic factor as moderate and as a cause of mortality as very low. Conclusions: Nocturia is probably associated with an approximately 1.3-fold increased risk of death.Peer reviewe