Spatial distribution of marine crenarchaeota group I in the vicinity of deep-sea hydrothermal systems

Distribution profiles of marine crenarchaeota group I in the vicinity of deep-sea hydrothermal systems were mapped with culture-independent molecular techniques. Planktonic samples were obtained from the waters surrounding two geographically and geologically distinct hydrothermal systems, and the abundance of marine crenarchaeota group I was examined by 16S ribosomal DNA clone analysis, quantitative PCR, and whole-cell fluorescence in situ hybridization. A much higher proportion of marine crenarchaeota group I within the microbial community was detected in deep-sea hydrothermal environments than in normal deep and surface seawaters. The highest proportion was always obtained from the ambient seawater adjacent to hydrothermal emissions and chimneys but not from the hydrothermal plumes. These profiles were markedly different from the profiles of epsilon-Proteobacteria, which are abundant in the low temperatures of deep-sea hydrothermal environments.     

An RNA Binding Protein Negatively Controlling Differentiation in Fission Yeast

The fission yeast Schizosaccharomyces pombe starts sexual development when starved for nutrients and simultaneously activated by mating pheromones. We have identified a new gene regulating the onset of this process. This gene, called nrd1⁺, encodes a typical RNA binding protein that preferentially binds poly(U). Deletion of nrd1⁺ causes cells to initiate sexual development without nutrient starvation. We have found that the biological role of nrd1⁺ is to block the onset of sexual development by repressing the Ste11-regulated genes essential for conjugation and meiosis until cells reach a critical level of starvation.     

Distribution and Diversity of Symbiotic Thermophiles, Symbiobacterium thermophilum and Related Bacteria, in Natural Environments

Symbiobacterium thermophilum is a tryptophanase-positive thermophile which shows normal growth only in coculture with its supporting bacteria. Analysis of the 16S rRNA gene (rDNA) indicated that the bacterium belongs to a novel phylogenetic branch at the outermost position of the gram-positive bacterial group without clustering to any other known genus. Here we describe the distribution and diversity of S. thermophilum and related bacteria in the environment. Thermostable tryptophanase activity and amplification of the specific 16S rDNA fragment were effectively employed to detect the presence of Symbiobacterium. Enrichment with kanamycin raised detection sensitivity. Mixed cultures of thermophiles containing Symbiobacterium species were frequently obtained from compost, soil, animal feces, and contents in the intestinal tracts, as well as feeds. Phylogenetic analysis and denaturing gradient gel electrophoresis of the specific 16S rDNA amplicons revealed a diversity of this group of bacteria in the environment.     

Enzymatic and genetic characterization of carbon and energy metabolisms by deep-sea hydrothermal chemolithoautotrophic isolates of Epsilonproteobacteria

The carbon and energy metabolisms of a variety of cultured chemolithoautotrophic Epsilonproteobacteria from deep-sea hydrothermal environments were characterized by both enzymatic and genetic analyses. All the Epsilonproteobacteria tested had all three key reductive tricarboxylic acid (rTCA) cycle enzymatic activities--ATP-dependent citrate lyase, pyruvate:ferredoxin oxidoreductase, and 2-oxoglutarate:ferredoxin oxidoreductase--while they had no ribulose 1,5-bisphosphate carboxylase (RubisCO) activity, the key enzyme in the Calvin-Benson cycle. These results paralleled the successful amplification of the key rTCA cycle genes aclB, porAB, and oorAB and the lack of success at amplifying the form I and II RubisCO genes, cbbL and cbbM. The combination of enzymatic and genetic analyses demonstrates that the Epsilonproteobacteria tested use the rTCA cycle for carbon assimilation. The energy metabolisms of deep-sea Epsilonproteobacteria were also well specified by the enzymatic and genetic characterization: hydrogen-oxidizing strains had evident soluble acceptor:methyl viologen hydrogenase activity and hydrogen uptake hydrogenase genes (hyn operon), while sulfur-oxidizing strains lacked both the enzyme activity and the genes. Although the energy metabolism of reduced sulfur compounds was not genetically analyzed and was not fully clarified, sulfur-oxidizing Epsilonproteobacteria showed enzyme activity of a potential sulfite:acceptor oxidoreductase for a direct oxidation pathway to sulfate but no activity of AMP-dependent adenosine 5'-phosphate sulfate reductase for a indirect oxidation pathway. No activity of thiosulfate-oxidizing enzymes was detected. The enzymatic and genetic characteristics described here were consistent with cellular carbon and energy metabolisms and suggest that molecular tools may have great potential for in situ elucidation of the ecophysiological roles of deep-sea Epsilonproteobacteria.     

Effective proliferation of low level Legionella pneumophila serogroup 1 cells using coculture procedure with Acanthamoeba castellanii

The multiplications of low level Legionella pneumophila serogroup 1 cells by the coculture procedure with Acanthamoeba castellanii were tested in five strains. The cells in all strains proliferated effectively for isolating. This procedure might be a useful means of improving the successful isolation from environmental and clinical specimens of low level Legionella cells, and pursuing the source of infection.     

Development of Geocoris varius and G. proteus (Hemiptera: Geocoridae) provided with Ephestia kuehniella (Lepidoptera: Pyralidae) eggs

We studied the development of Geocoris varius (Uhler) and Geocoris proteus Distant reared on Ephestia kuehniella Zeller eggs at 20, 24, 26, 30, 33, or 36?°C. The lower developmental thresholds (T ₀) and the thermal constants (K) of eggs and nymphs of G. varius were 13.3?°C, 151.1 degree-days and 13.4?°C, 433.0 degree-days, respectively; those of G. proteus were 16.1?°C, 98.3 degree-days and 16.9?°C, 226.9 degree-days, respectively. The hatch rate of G. varius eggs was significantly lower at 33?°C than at ??30?°C, and no eggs hatched at 36?°C. That of G. proteus was lowest at 20?°C and did not decline significantly at 36?°C. The survival rate throughout the nymphal period increased with temperature up to 30?°C in G. varius, and it was lowest at 20?°C in G. proteus. Thus, the optimal rearing temperatures for immature stages appear to be about 24?C30?°C for G. varius and 26?C33?°C for G. proteus. It might be possible to improve the efficiency of their mass production by controlling the rearing temperature in the above ranges. This would also make the developmental stages of nymphs more uniform and so prevent cannibalism in mass rearing.     

VAV2 and VAV3 as Candidate Disease Genes for Spontaneous Glaucoma in Mice and Humans

Background

Glaucoma is a leading cause of blindness worldwide. Nonetheless, the mechanism of its pathogenesis has not been well-elucidated, particularly at the molecular level, because of insufficient availability of experimental genetic animal models.

Methodology/Principal Findings

Here we demonstrate that deficiency of Vav2 and Vav3, guanine nucleotides exchange factors for Rho guanosine triphosphatases, leads to an ocular phenotype similar to human glaucoma. Vav2/Vav3-deficient mice, and to a lesser degree Vav2-deficient mice, show early onset of iridocorneal angle changes and elevated intraocular pressure, with subsequent selective loss of retinal ganglion cells and optic nerve head cupping, which are the hallmarks of glaucoma. The expression of Vav2 and Vav3 tissues was demonstrated in the iridocorneal angle and retina in both mouse and human eyes. In addition, a genome-wide association study screening glaucoma susceptibility loci using single nucleotide polymorphisms analysis identified VAV2 and VAV3 as candidates for associated genes in Japanese open-angle glaucoma patients.

Conclusions/Significance

Vav2/Vav3-deficient mice should serve not only as a useful murine model of spontaneous glaucoma, but may also provide a valuable tool in understanding of the pathogenesis of glaucoma in humans, particularly the determinants of altered aqueous outflow and subsequent elevated intraocular pressure.     

TGF-β induces surface LAP expression on murine CD4 T cells independent of Foxp3 induction

Background

It has been reported that human FOXP3⁺ CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP) after activation, based on the use of an anti-human LAP mAb. Murine CD4 Foxp3⁺ Tregs have also been reported to express surface LAP, but these studies have been hampered by the lack of suitable anti-mouse LAP mAbs.

Methodology/Principal Findings

We generated anti-mouse LAP mAbs by immunizing TGF-β^−/− animals with a mouse Tgfb1-transduced P3U1 cell line. Using these antibodies, we demonstrated that murine Foxp3⁺ CD4 Tregs express LAP on their surface. In addition, retroviral transduction of Foxp3 into mouse CD4⁺CD25⁻ T cells induced surface LAP expression. We then examined surface LAP expression after treating CD4⁺CD25⁻ T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3⁺ but also on T cells that remained Foxp3⁻ after TGF-β treatment. GARP expression correlated with the surface LAP expression, suggesting that surface LAP is GARP-anchored also in murine T cells.

Conclusions/Significance

Unlike human CD4 T cells, surface LAP expression on mouse CD4 T cells is controlled by Foxp3 and TGF-β. Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.