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Abstract Microbial degradation of algal biomass following spring phytoplankton blooms has been characterised as a concerted effort among multiple clades of heterotrophic bacteria. Despite their significance to overall carbon turnover, many of these clades have resisted cultivation. One clade known from 16S rRNA gene sequencing surveys at Helgoland in the North Sea, was formerly identified as belonging to the genus Ulvibacter. This clade rapidly responds to algal blooms, transiently making up as much as 20% of the free-living bacterioplankton. Sequence similarity below 95% between the 16S rRNA genes of described Ulvibacter species and those from Helgoland suggest this is a novel genus. Analysis of 40 metagenome assembled genomes (MAGs) derived from samples collected during spring blooms at Helgoland support this conclusion. These MAGs represent three species, only one of which appears to bloom in response to phytoplankton. MAGs with estimated completeness greater than 90% could only be recovered for this abundant species. Additional, less complete, MAGs belonging to all three species were recovered from a mini-metagenome of cells sorted via flow cytometry using the genus specific ULV995 fluorescent rRNA probe. Metabolic reconstruction indicates this highly abundant species most likely degrades proteins and the polysaccharide laminarin. Fluorescence in situ hybridisation showed coccoid cells, with a mean diameter of 0.78 mm, with standard deviation of 0.12 μm. Based on the phylogenetic and genomic characteristics of this clade, we propose the novel candidate genus Candidatus Prosiliicoccus, and for the most abundant and well characterised of the three species the name Candidatus Prosiliicoccus vernus. [ABSTRACT FROM AUTHOR]

Abstract: The 23S rRNA gene was evaluated as target for the development of Sybr Green-based quantitative PCR (qPCR) for the analysis of nitrogen-fixing members of the genus Frankia or subgroups of these in soil. A qPCR with a primer combination targeting all nitrogen-fixing frankiae (clusters 1, 2 and 3) resulted in numbers similar to those obtained with a previously developed qPCR using nifH gene sequences, both with respect to introduced and indigenous Frankia populations. Primer combinations more specifically targeting three subgroups of the Alnus host infection group (cluster 1) or members of the Elaeagnus host infection group (cluster 3) were specific for introduced strains of the target group, with numbers corresponding to those obtained by quantification of nitrogen-fixing frankiae with both the 23S rRNA and nifH genes as target. Method verification on indigenous Frankia populations in soils, i.e. in depth profiles from four sites at an Alnus glutinosa stand, revealed declining numbers in the depth profiles, with similar abundance of all nitrogen-fixing frankiae independent of 23S rRNA or nifH gene targets, and corresponding numbers of one group of frankiae of the Alnus host infection only, with no detections of frankiae representing the Elaeagnus, Casuarina, or a second subgroup of the Alnus host infection groups. [Copyright &y& Elsevier]

Electron paramagnetic resonance (EPR) in a synthetic Si 0.975 Ge 0.025 O 2 single crystal of α–quartz structure has been studied. After fast electron irradiation previously studied and new Ge-related paramagnetic centers (PCs) have been recognized using computer simulation of the observed EPR spectra. Increasing radiation doses allow to distinguish four paramagnetic [GeO 4 /M+]0 centers A GeLi , C GeLi , D GeLi , and A GeNa as well as four oxygen-vacancy electron centers Ge E ˊ 1 , Ge E ˊ 2 , Ge(IV) and a new Ge(V) center. The selective UV bleaching and distinct thermal stability of different centers allow us to determine the spin Hamiltonian parameters for the Ge(V) center. Electron paramagnetic resonance (EPR) in a synthetic Si 0.975 Ge 0.025 O 2 single crystal of α–quartz structure has been studied. Detailed EPR measurements after different storage times, radiation doses, thermal annealing, and UV bleaching as well as computer simulations allow us to distinguish four [GeO 4 /M+]0 centers A GeLi , C GeLi , D GeLi , and A GeNa as well as four oxygen-vacancy electron centers Ge E ˊ 1 , Ge E ˊ 2 , Ge(IV) and Ge(V). The spin Hamiltonian parameters (g , A (73Ge) and P (73Ge) of the new Ge(V) center have been determined and are closely comparable to those of the Ge E ˊ 1 center. [Display omitted] • Single crystals of Ge-rich α-quartz were grown hydrothermally. • Ge-related paramagnetic defects are induced by fast electron irradiation. • Paramagnetic defects are identified using computer simulations of EPR spectra. • Spin Hamiltonian parameters for new Ge(V) oxygen vacancy defect are determined. [ABSTRACT FROM AUTHOR]

Abstract: Nitrogen (N2) fixation is a globally important process often mediated by diazotrophic cyanobacteria in the open ocean. In 2010, seawater was collected near Cape Verde to identify and measure N2 and carbon (C) fixation by unicellular diazotrophic cyanobacteria. The nifH gene abundance (104–106 nifH L−1) and nifH gene transcript abundance (102–104 cDNA nifH L−1) for two unicellular groups, UCYN-A and UCYN-B, were detected. UCYN-A was also identified and quantified (104–105 cellsL−1) by new probes (UCYN-A732 and UCYN-A159) using Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization (CARD-FISH) assays. The UCYN-A were observed as free cells or attached to a larger unidentified eukaryotic cell. A Halogen In Situ Hybridization-Secondary Ion Mass Spectrometry (HISH-SIMS) assay using the UCYN-A732 probe was applied on samples previously incubated with 13C-bicarbonate and 15N2. Free UCYN-A cells were enriched in both 13C and 15N and estimated C and N2 fixation rates for UCYN-A were lower compared to co-occurring unicellular cyanobacteria cells similar in size (3.1–5.6μm) and pigmentation to diazotroph Crocosphaera watsonii. Here, we identify and quantify two common co-occurring unicellular groups and measure their cellular activities by nanoSIMS. [Copyright &y& Elsevier]

We hypothesized that the phytosterols β-sitosterol (BSS), β-sitosterol glucoside (BSSG), and Moducare (MC; BSS:BSSG = 99:1) could modulate the growth of estrogen-dependent human breast cancer cells in vitro and in vivo. The present study evaluated the estrogenic and antiestrogenic effects of BSS, BSSG, and MC (0.001 to 150 μmol/L) on the proliferation of Michigan Cancer Foundation 7 (MCF-7) cells in vitro. Both BSS (>1 μmol/L) and MC (>50 μmol/L) increased MCF-7 cell proliferation. Treatment with 150 μmol/L of BSS and MC increased cell growth by 2.4 and 1.5 times, respectively, compared to the negative control (NC) group. However, BSSG had no effect at the concentrations tested. The effects of dietary BSS, BSSG, and MC on the growth of MCF-7 cells implanted in ovariectomized athymic mice were also evaluated. Estrogenic effects of the phytosterols were evaluated in the NC, BSS, BSSG, and MC treatment groups, and antiestrogenic effects were evaluated in the 17β-estradiol (E2), E2 + BSS, E2 + BSSG, and E2+ MC treatment groups. Mice were treated with dietary BSS (9.8 g/kg AIN93G diet), BSSG (0.2 g/kg diet), or MC (10.0 g/kg diet) for 11 wk. Dietary BSS, BSSG, and MC did not stimulate MCF-7 tumor growth. However, dietary BSS, BSSG, and MC reduced E2-induced MCF-7 tumor growth by 38.9% (P < 0.05), 31.6% (P = 0.08), and 42.13% (P < 0.05), respectively. The dietary phytosterols lowered serum E2 levels by 35.1, 30.2, and 36.5% in the E2 + BSS, E2 + BSSG, and E2+ MC groups, respectively (P < 0.05), compared to that of the E2 treatment group. Estrogen-responsive pS2 mRNA expression in tumors did not differ among groups, but expression of the antiapoptotic marker B-cell lymphoma/leukemia-2 (bcl-2) in tumors from the E2 + MC group was downregulated, compared to that of the E2 treatment group. In summary, BSS and MC stimulated MCF-7 cell growth in vitro. Although BSSG comprises only 1% of MC, BSSG made MC less estrogenic than BSS alone in vitro. However, dietary BSS and MC protected against E2-stimulated MCF-7 tumor growth and lowered circulating E2 levels. [ABSTRACT FROM AUTHOR]

Seawater contains free-living and particle-attached bacteria. Only a small fraction is cultivable on plates. As free-living and particle-associated bacteria differ in their physiological traits, their cultivability on plates may coincide with particle association. Using filtration and Imhoff sedimentation cones, particles were collected during a spring phytoplankton bloom off Helgoland (North Sea) in order to obtain particle-associated bacteria as inocula. Direct dilution plating resulted in 526 strains from 3 µm filtration retentates and 597 strains from settled particles. Motile Gammaproteobacteria from the genera Pseudoalteromonas , Shewanella , Psychrobacter , Vibrio and Colwellia , as well as particle-attached Flavobacteriia affiliating with the genera Tenacibaculum and Gramella , were frequently isolated. As a result, a diverse collection comprised of 266 strains was deposited. Two strains were most likely to represent novel genera and 78 strains were probably novel species. Recently, a high-throughput cultivation study from the same site using seawater as an inoculum had retrieved 271 operational phylogenetic units (OPUs) that represented 88% of the 4136 characterized strains at the species level. A comparison of 16S rRNA gene sequences revealed that the collection obtained matched 104 of the 271 seawater OPUs at the species level and an additional 113 at the genus level. This large overlap indicated a significant contribution of particle-associated bacteria to the cultivable microbiome from seawater. The presence of 49 genera not identified in the larger seawater study suggested that sample fractionation was an efficient strategy to cultivate rare members of the planktonic microbiome. The diverse collection of heterotrophic bacteria retrieved in this study will be a rich source for future studies on the biology of particle-associated bacteria. [ABSTRACT FROM AUTHOR]

The nitrogen mustard mechlorethamine (NH2) is a potent contact allergen, sensitizing up to 75% of those exposed. An attempt was made to prevent allergic contact sensitization, i.e. to induce specific immunologic tolerance, in 9 patients who were to be treated topically with NH2 for widespread and persistent psoriasis. Fourteen and 7 days before the first topical application of NH2, they were given intravenous injections of 3 trig NH2. Topical treatment with NH2 was repeated weekly for 8 to 10 weeks in 4 patients and daily for about 4 weeks in 5 patients. Two of the 9 patients became contact allergic to NH2. Despite the apparent failure to induce tolerance, further trials in a larger series of patients and trials with different "tolerogenic" schedules and different topical treatment schedules are warranted. [ABSTRACT FROM AUTHOR]