Vivid molecular divergence over volcanic remnants: the phylogeography of Megadromus guerinii on Banks Peninsula,New Zealand

Megadromus guerinii, an endemic carabid beetle (Carabidae), is the most common carabid throughout its restricted range on Banks Peninsula, a formation of extinct volcanoes in Canterbury, New Zealand. This study characterises the small-scale phylogeographic patterns of M. guerinii across the formerly volcanically active Banks Peninsula using mitochondrial and ribosomal genes. Between the eastern and western areas of the peninsula, the mitochondrial, but not nuclear, DNA has a well-defined geographic distribution. Specifically, mitochondrial cytochrome c oxidase subunit I (CO1) identifies two distinct groups (> 6% divergence between eastern and western beetles) while ribosomal genes show no discernible pattern. Whether such a pattern represents male-biased dispersal, Wolbachia infection, a recent range expansion of a divergent lineage, or a deeper historic separation is explored. There is potential that male-biased dispersal could have occurred. Wolbachia infection was not detected. We conclude that historical processes have likely separated taxa in the eastern and western peninsula.     

Climatic,ecological, and socioeconomic factors associated with West Nile virus incidence in Atlanta,Georgia, U.S.A.

The integrated effects of the many risk factors associated with West Nile virus (WNV) incidence are complex and not well understood. We studied an array of risk factors in and around Atlanta, GA, that have been shown to be linked with WNV in other locations. This array was comprehensive and included climate and meteorological metrics, vegetation characteristics, land use / land cover analyses, and socioeconomic factors. Data on mosquito abundance and WNV mosquito infection rates were obtained for 58 sites and covered 2009–2011, a period following the combined storm water – sewer overflow remediation in that city. Risk factors were compared to mosquito abundance and the WNV vector index (VI) using regression analyses individually and in combination. Lagged climate variables, including soil moisture and temperature, were significantly correlated (positively) with vector index as were forest patch size and percent pine composition of patches (both negatively). Socioeconomic factors that were most highly correlated (positively) with the VI included the proportion of low income households and homes built before 1960 and housing density. The model selected through stepwise regression that related risk factors to the VI included (in the order of decreasing influence) proportion of houses built before 1960, percent of pine in patches, and proportion of low income households.     

Association between chloroplast and mitochondrial lineages in oaks

Patterns of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) variation were studied in 378 populations of oak trees sampled throughout the southern half of France. Six cpDNA haplotypes detected in a previous European survey and three new cpDNA haplotypes were found in this region. Two mitochondrial polymorphisms detected earlier by restriction analysis of PCR-amplified fragments alone, or in combination with single-strand conformation polymorphism (SSCP), were compared with the cpDNA data. Sequencing revealed the nature of the two mitochondrial mutations: a single-base substitution and a 4-bp inversion associated with a 22-bp hairpin secondary structure. The single-base substitution was then analyzed by allele-specific amplification. Results for the two cytoplasmic genomes were combined, which allowed the identification of 12 cpDNA-mtDNA haplotypes. The 4-bp mtDNA inversion has appeared independently in different cpDNA lineages. Given the peculiar nature of this mtDNA mutation, we suggest that intramolecular recombination leading to repeated inversions of the 4-bp sequence (rather than paternal leakage of one of the two genomes) is responsible for this pattern. Furthermore, the geographic locations of the unusual cpDNA-mtDNA associations (due to the inversion) usually do not match the zones of contact between divergent haplotypes. In addition, in southern France, the groupings of populations based on the mtDNA substitution were strictly congruent with those based on cpDNA. Because many populations that are polymorphic for both cpDNA and mtDNA have remained in contact since postglacial recolonization in this area without producing any new combination of cytoplasms involving the mitochondrial substitution, we conclude that paternal leakage is not a significant factor at this timescale. Such results confirm and expand our earlier conclusions based on controlled crosses.      

Characterization ofl-leucine transport in the opportunistic yeastsMalassezia furfur andMalassezia pachydermatis

The lipophilic yeastsMalassezia furfur andM. pachydermatis show an initial rapid uptake ofl-leucine followed by slower steady-state rates. At least two independent transport systems forl-leucine were present in both species. The high-affinity system forM. furfur had a K_T of 0.047 µM with a J_max of 222 fM/min/10⁶ cells (65 pM/min/mg dry weight), whereas forM. pachydermatis the K_T was 0.067 µM with a J_max of 709 fM/min/10⁶ cells (89 pM/min/dry weight). The low-affinity system forM. furfur had a K_T of 646 µM with a J_max of 1.62 pM/min/10⁶ cells (0.5 nM/min/mg dry weight) and that ofM. pachydermatis had a K_T of 3.3 µM with a J_max of 9.97 pM/min/10⁶ cells (1.3 nM/min/mg dry weight). Both transport systems were energy-dependent. Cells incubated with Tween 80 showedl-leucine uptake via both transport systems. Cells incubated with a combination of glucose (1%) and Tween 80 (0.01%) showed decreased transport rates for the high-affinity system for both species as compared with cells incubated only with glucose. The low-affinity transport system of both species in the presence of glucose plus Tween 80 showed an initial rapid uptake followed by greater efflux than influx ofl-leucine.l-Leucine demonstrated binding to Tween 80, but the major effect of Tween 80 on membrane transport inMalassezia appears to be on the efflux of transported molecules.     

Hydrodynamics and mass transfer in two-phase aqueous extraction using spray columns

Spray columns can be used to isolate and purify proteins using the two-phase aqueous extraction technique based on polyethylene glycol (PEG) and dextran. The fractional dispersed phase (PEG) holdup and overall mass transfer coefficients were measured in a 9.7 mm i.d. spray column. We found that the dispersed phase holdup increased with increasing PEG phase velocity. The overall mass transfer coefficients for bovine serum albumin, normalized for the PEG holdup, were found to be independent of the PEG phase velocity. This result was expected, since true mass transfer coefficients do not vary with phase velocity.     

Role of GhHDA5 in H3K9 deacetylation and fiber initiation in Gossypium hirsutum

Cotton fibers are single‐celled trichomes that initiate from the epidermal cells of the ovules at or before anthesis. Here, we identified that the histone deacetylase (HDAC ) activity is essential for proper cotton fiber initiation. We further identified 15 HDAC s homoeologs in each of the A‐ and D‐subgenomes of Gossypium hirsutum . Few of these HDAC homoeologs expressed preferentially during the early stages of fiber development [?1, 0 and 6 days post‐anthesis (DPA )]. Among them, GhHDA 5 expressed significantly at the time of fiber initiation (?1 and 0 DPA). The in vitro assay for HDAC activity indicated that GhHDA 5 primarily deacetylates H3K9 acetylation marks. Moreover, the reduced expression of GhHDA 5 also suppresses fiber initiation and lint yield in the RNA interference (RNA i) lines. The 0 DPA ovules of GhHDA 5 RNA i lines also showed alterations in reactive oxygen species homeostasis and elevated autophagic cell death in the developing fibers. The differentially expressed genes (DEG s) identified through RNA ‐seq of RNA i line (DEP 12) and their pathway analysis showed that GhHDA 5 modulates expression of many stress and development‐related genes involved in fiber development. The reduced expression of GhHDA 5 in the RNA i lines also resulted in H3K9 hyper‐acetylation on the promoter region of few DEG s assessed by chromatin immunoprecipitation assay. The positively co‐expressed genes with GhHDA 5 showed cumulative higher expression during fiber initiation, and gene ontology annotation suggests their involvement in fiber development. Furthermore, the predicted protein interaction network in the positively co‐expressed genes indicates HDA 5 modulates fiber initiation‐specific gene expression through a complex involving reported repressors.     

Immunohistochemical localization of irisin in mole rats (Spalax leucodon)

Irisin was first identified in skeletal muscle cells. It is an exercise protein that is secreted into the circulation; it causes conversion of white adipose tissue to brown adipose tissue. We investigated irisin immunoreactivity in mole rat (Spalax leucodon) tissues. We examined cerebellum, pituitary, heart, liver, pancreas, spleen, uterus, kidney and striated muscle in female adult mole rats. Tissues were processed, embedded in paraffin, sectioned at 5 μm and stained immunohistochemically for irisin. Irisin immunostaining was detected in the cytoplasm of stained cells; the cytoplasm of Purkinje cells was unstained. We found that irisin may be synthesized in many tissues. The function of locally synthesized irisin currently is unknown.     

gamma-aminobutyric acid increases the water accessibility of M3 membrane-spanning segment residues in gamma-aminobutyric acid type A receptors

gamma-Aminobutyric acid type A (GABA(A)) receptors are members of the ligand-gated ion channel gene superfamily. Using the substituted cysteine accessibility method, we investigated whether residues in the alpha(1)M3 membrane-spanning segment are water-accessible. Cysteine was substituted, one at a time, for each M3 residue from alpha(1)Ala(291) to alpha(1)Val(307). The ability of these mutants to react with the water-soluble, sulfhydryl-specific reagent pCMBS(-) was assayed electrophysiologically. Cysteines substituted for alpha(1)Ala(291) and alpha(1)Tyr(294) reacted with pCMBS(-) applied both in the presence and in the absence of GABA. Cysteines substituted for alpha(1)Phe(298), alpha(1)Ala(300), alpha(1)Leu(301), and alpha(1)Glu(303) only reacted with pCMBS(-) applied in the presence of GABA. We infer that the pCMBS(-) reactive residues are on the water-accessible surface of the protein and that GABA induces a conformational change that increases the water accessibility of the four M3 residues, possibly by inducing the formation of water-filled crevices that extend into the interior of the protein. Others have shown that mutations of alpha(1)Ala(291), a water-accessible residue, alter volatile anesthetic and ethanol potentiation of GABA-induced currents. Water-filled crevices penetrating into the interior of the membrane-spanning domain may allow anesthetics and alcohol to reach their binding sites and thus may have implications for the mechanisms of action of these agents.