Insulin resistance does not influence gene expression in skeletal muscle

Insulin resistance is commonly observed in patients prior to the development of type 2 diabetes and may predict the onset of the disease. We tested the hypothesis that impairment in insulin stimulated glucose-disposal in insulin resistant patients would be reflected in the gene expression profile of skeletal muscle. We performed gene expression profiling on skeletal muscle of insulin resistant and insulin sensitive subjects using microarrays. Microarray analysis of 19,000 genes in skeletal muscle did not display a significant difference between insulin resistant and insulin sensitive muscle. This was confirmed with real-time PCR. Our results suggest that insulin resistance is not reflected by changes in the gene expression profile in skeletal muscle.     

Early Jurassic gastropods from England

Abstract: Twenty‐five gastropod taxa are reported from the Early Jurassic (Hettangian to Toarcian) of England. Of these, 14 are identified to species level, and the remaining 11 are treated in open nomenclature. One genus (Lensataphrus) and six species are introduced as new. The new species are Lensataphrus tatei, Lensataphrus tenuis, Tricarilda toddi, Cylindrobullina dorsetensis, Cylindrobullina ventricosa and Consobrinella greeni. The following new combinations are introduced: Cassianopsis hebertana (d’Orbigny, 1852) for Neritopsis hebertana; Cryptaulax abscisum (Terquem and Piette, 1868) for Cerithium abscisum; and Cylindrobullina avena (Terquem, 1855) for Striactaeonina avena. Most of the genera and some of the species are also known from Central Europe (Germany and France). Typical vetigastropod genera that are present in England and Central Europe are Colpomphalus, Costataphrus, Ooliticia, Eucycloscala and Eucycloidea. The caenogastropod genera Levipleura and Cryptaulax are present in both regions, as are the heterobranchs Tricarilda and Cylindrobullina. The heterobranch genus Consobrinella is reported from England for the first time. Gastropods seem to follow the diversity trends of other marine invertebrates during the Early Jurassic. They diversify until the Late Pliensbachian but decrease sharply in number around the Pliensbachian–Toarcian boundary. This may reflect both regional anoxia and a global mass extinction event in the Early Toarcian.     

Technique for cryopreservation of intestinal smooth muscle cells

Attempts were made to develop a simplified procedure for long-term cryopreservation of intestinal smooth muscle cells (ISMC). ISMC were collected from the ileum of Sprague-Dawley neonatal rats through cellular dissociation in trypsin. Cryopreservation method comprised of a rapid 1-step (protocol 1) and a slow 3-step (protocol 2) freezing of ISMC for 1 week. Preparations were thawed and single ISMC were assessed via the comet assay and damaged DNA was quantified through comet tail moment. The control unfrozen ISMC exhibited DNA damage of 2.34 ± 0.35 compared to ISMC cooled via protocol 2 (2.62 ± 0.36) and protocol 1 (10.15 ± 0.72). Thereafter, protocol 2 freezing method was adopted and ISMC were cryopreserved for 1-week, 1-month, and 4-months to analyse the temporal and long-term cryopreservation of ISMC. This revealed a DNA damage of 2.62 ± 0.36 (1-week), 3.81 ± 0.72 (1-month), and 5.1 ± 0.9 (4-months). Gradual cooling is suitable for continuing storage of ISMC and although fluctuation in cryoinjury is observed with time this is considered to reflect cell-to-cell variability.     

Phytophthora ramorum: a pathogen with a remarkably wide host range causing sudden oak death on oaks and ramorum blight on woody ornamentals

Phytophthora ramorum is an oomycete plant pathogen classified in the kingdom Stramenopila. P. ramorum is the causal agent of sudden oak death on coast live oak and tanoak as well as ramorum blight on woody ornamental and forest understorey plants. It causes stem cankers on trees, and leaf blight or stem dieback on ornamentals and understorey forest species. This pathogen is managed in the USA and Europe by eradication where feasible, by containment elsewhere and by quarantine in many parts of the world. Genomic resources provide information on genes of interest to disease management and have improved tremendously since sequencing the genome in 2004. This review provides a current overview of the pathogenicity, population genetics, evolution and genomics of P. ramorum. Taxonomy: Phytophthora ramorum (Werres, De Cock & Man in't Veld): kingdom Stramenopila; phylum Oomycota; class Peronosporomycetidae; order Pythiales; family Pythiaceae; genus Phytophthora. Host range: The host range is very large and the list of known hosts continues to expand at the time of writing. Coast live oak and tanoak are ecologically, economically and culturally important forest hosts in the USA. Rhododendron, Viburnum, Pieris, Syringa and Camellia are key ornamental hosts on which P. ramorum has been found repeatedly, some of which have been involved in moving the pathogen via nursery shipments. Disease symptoms: P. ramorum causes two different diseases with differing symptoms: sudden oak death (bleeding lesions, stem cankers) on oaks and ramorum blight (twig dieback and/or foliar lesions) on tree and woody ornamental hosts. Useful websites: http://nature.berkeley.edu/comtf/ , http://rapra.csl.gov.uk/ , http://www.aphis.usda.gov/plant_health/plant_pest_info/pram/index.shtml , http://genome.jgi‐psf.org/Phyra1_1/Phyra1_1.home.html , http://pamgo.vbi.vt.edu/ , http://pmgn.vbi.vt.edu/ , http://vmd.vbi.vt.edu./ , http://web.science.oregonstate.edu/bpp/labs/grunwald/resources.htm , http://www.defra.gov.uk/planth/pramorum.htm , http://www.invasive.org/browse/subject.cfm?sub=4603 , http://www.forestry.gov.uk/forestry/WCAS‐4Z5JLL     

Metabolic consequences of a species difference in Gibbs free energy of Na+/Ca2+ exchange: rat versus guinea pig

The Gibbs free energy of the sarcolemmal Na+/Ca2+ exchanger (DeltaG(Na/Ca)) determines its net Ca2+ flux. We tested the hypothesis that a difference of diastolic DeltaG(Na/Ca) exists between rat and guinea pig myocardium. We measured the suprabasal rate of oxygen consumption (VO2) of arrested Langendorff-perfused hearts of both species, manipulating DeltaG(Na/Ca) by reduction of extracellular Na+ concentration, [Na+](o). Hill equations fitted to the resulting VO2-[Na+](o) relationships yielded Michaelis constant (K(m)) values of 67 and 25 mM for rat and guinea pig, respectively. We developed and tested a simple thermodynamic model that attributes this difference of K(m) values to a 7.84 kJ/mol difference of DeltaG(Na/Ca). The model predicts that reversal of Na+/Ca2+ exchange, leading to diastolic Ca2+ influx, should occur at a value of [Na+](o) about three times higher in rat myocardium. We verified this quantitative prediction using fura 2 fluorescence to index intracellular Ca2+ concentration in isolated ventricular trabeculae at 37 degrees C. The postulated difference in free energy of Na+/Ca2+ exchange explains a number of reported disparities of Ca2+ handling at rest between rat and guinea pig myocardia.     

A rapid method for assessing the suitability of quenching agents for individual biocides as well as combinations

AIMS: To develop a novel, rapid method for testing the ability of quenching agents to neutralize disinfectants. METHODS AND RESULTS: Tests were performed to determine the suitability of different neutralizers for a range of disinfectants, using a new method based on the Bioscreen optical density analyser. Results showed that during disinfection tests, efficacy could be over-estimated due to poor, or no, neutralization of the disinfectant after a specified time of exposure to the bacteria. The failure to distinguish adequately between bacteriostatic and bactericidal effects can lead to false results during disinfectant testing. Experiments also showed that dilution of the disinfectant, following exposure to the bacteria, was not always sufficient to stop the activity of the disinfectant for chemicals with low dilution coefficients. CONCLUSIONS: The quench test proved to be very quick and easy to perform, with results being available within 18 h. Using the Bioscreen, the test is automated and determines whether dilution into a particular neutralizer is able to inactivate a disinfectant within 30 s. SIGNIFICANCE AND IMPACT OF THE STUDY: This new approach allows the efficacy of quenching agents to be determined, prior to undertaking each disinfection study, and can help in the development of more suitable quenching solutions. The test has also been used to find suitable neutralizers for mixtures of disinfectants which might be used during studies on synergistic biocide combinations.     

Small mammalian herbivore determines vegetation response to patchy nutrient inputs

Nutrient inputs to plant communities are often spatially heterogeneous, for example those deriving from the dung and urine of large grazing animals. The effect of such localised elevation of nutrients on plant growth and composition has been shown to be modified by the grazing of large herbivores. However, there has been little work on interactions between small mammalian herbivores and such patchy nutrient inputs, even though these interactions are potentially of major significance for plant performance and community structure.
We examined the effect of simulated cattle urine deposition on the vegetation structure, above-ground biomass and species composition of chalk grassland within enriched patches. Short-term exclosures were used to determine whether a small herbivore (rabbit) would preferentially graze the vegetation in enriched patches and what impact this interaction would have on the performance of plants in such patches. Rabbit grazing pressure determined whether nutrient inputs had a negative or positive effect on plant biomass. Nutrients increased plant biomass in the absence of grazing, but when exposed to grazing, plants in nutrient-rich patches had more biomass consumed by herbivores than neighbouring plants. Further, nutrients increased the relative palatability of a less preferred forage species ( Brachypodium pinnatum ), contributing to changes in plant community composition. We conclude that a small herbivore can drive plant responses to patchily distributed nutrients.