Polysulfide as a possible substrate for sulfur-reducing bacteria

Because of its low solubility it is unlikely that elemental sulfur serves as the direct substrate for sulfur-reducing bacteria. To test the hypothesis that polysulfide may represent a soluble intermediate of sulfur reduction, the maximal polysulfide concentrations formed from elemental sulfur in aqueous sulfide solutions were measured at near neutral pH and at temperatures up to 90°C. The saturation concentrations decreased by two orders of magnitude when the pH was lowered from 7 to 6 at a given temperature, and increased about tenfold when the temperature was raised from 37°C to 90°C at a given pH. The dissolution of 0.1 mM zerovalent sulfur in 1 mM sulfide (H₂S+HS^–) required a pH of 7.5 at 20°C and of only 6.1 at 100°C. A comparison with the growth optima of sulfur-reducers suggests that polysulfide is present at sufficient concentration at the growth conditions of the Bacteria and the moderately acidophilic Archaea. Polysulfide is apparently not available at the growth conditions of the extremely acidophilic Archaea. Alternative mechanisms for the sulfur utilization under these conditions are discussed.Abbreviations MOPS Morpholinopropanesulfonate - PIPES 1,4 piperazine-N,N-bis(2-ethanesulfonate) - HEPES N-2-hydroxy-ethylpiperazine-N-ethanesulfonate     

Porcine spermatozoa inhibit post-breeding cytokine induction in uterine epithelial cells in vivo

Early endometrial cytokine responses after exposure to various inseminate components were investigated for a better understanding of the immunological reactions occurring in the porcine uterus after insemination. Baseline values were established for the mRNA concentrations of GM-CSF, IL-6, IL-10, CXCL8 (interleukin-8), Tumour Necrosis Factor α (TNF-α), TGF-β, cyclooxygenase-2 (COX-2) and arachidonate 5-lipooxygenase (ALOX-5) in periovulatory uterine endometrial tissue using quantitative RT-PCR. Synchronized gilts were inseminated with spermatozoa diluted either in the semen extender Androhep™ or seminal plasma. Uterine infusions of media without spermatozoa were used as controls. Three hours after insemination sows were slaughtered and the expression of the above mentioned cytokines was measured in uterine epithelial cells. Simultaneously, the influx of polymorphonuclear neutrophilic (PMN) granulocytes into the uterus was quantified. Compared to baseline values seminal plasma (SP) and Androhep™ (AH) respectively, if used alone, caused a significant increase in mRNA concentrations of IL-10 (SP: 1.5-fold), TGF-β (AH: 1.5-fold), CXCL8 (AH: 7.1-fold), TNF-α (AH: 1.9-fold) and COX-2 (AH: 7-fold). Surprisingly, in the presence of spermatozoa, none of the tested cytokines revealed mRNA concentrations higher than baseline values. The number of immigrated, intra-luminal PMN correlated only with mRNA concentrations of CXCL8 in presence of Androhep™ (r = 0.51). None of the other cytokines tested seemed to be involved in the regulation of neutrophil recruitment. However, the most interesting result was the sperm-induced down-regulation in the expression of TNF-α, TGF-β, IL-10, CXCL8 and COX-2 to mRNA concentration levels similar to or even below baseline values. In conclusion the results show that CXCL8 contributes significantly to uterine PMN recruitment and indicate a so far underestimated role of porcine spermatozoa in the general regulation of the uterine post-mating inflammatory response.     

Ferroglobus placidus gen. nov., sp. nov., a novel hyperthermophilic archaeum that oxidizes Fe2+ at neutral pH under anoxic conditions

A novel coccoid, anaerobic, Fe²⁺-oxidizing archaeum was isolated from a shallow submarine hydrothermal system at Vulcano, Italy. In addition to ferrous iron, H₂ and sulfide served as electron donors. NO₃ ^– was used as electron acceptor. In the presence of H₂, also S₂O₃ ^2– could serve as electron acceptor. The isolate was a neutrophilic hyperthermophile that grew between 65° C and 95° C. It represents a novel genus among the Archaeoglobales that we name Ferroglobus. The type species is Ferroglobus placidus (DSM 10642). Received: 7 March 1996 / Accepted: 4 September 1996     

Confocal Raman microspectroscopy of the activation of single neutrophilic granulocytes

Confocal Raman micro-spectroscopy has been applied to investigate the activation process of single, living neutrophilic granulocytes. Both resting cells as well as activated cells were measured. The activation of cells was performed with phorbol-12-myristate-13-acetate activator and Escherichia Coli bacteria. Raman microspectroscopy combines a high spatial resolution inside a single, living cell with detailed material information. Using this approach it can be concluded that activation of the cells with phorbol-12-myristate-13-acetate causes a change in the redox state of cytochrome b₅₅₈. This protein is a part of the NADPH-oxidase complex that neutrophilic granulocytes employ to generate O₂ ^–, superoxide anion. Additionally a change in the redox state of myeloperoxidase can be observed. Myeloperoxidase is known to react with O₂ ^–. Activation of the cells with bacteria gives rise to corresponding changes in the Raman spectra. From this single cell study it can be concluded that the enzymes cytochrome b₅₅₈ and myeloperoxidase are present inside the cytoplasm of the living cell, while participating in the redox processes. Activation causes an intra-cellular release of oxygen metabolites. Activation with bacteria of neutrophilic granulocytes from a patient with chronic granulomatous disease, that contain no cytochrome b₅₅₈, led to typical changes in the redox state of myeloperoxidase. This indicates that in the bacterium/neutrophilic granulocyte system oxygen metabolites are generated that are capable of reacting with MPO. Received: 1 September 1998 / Revised version: 20 February 1998 / Accepted: 22 February 1998     

The human neutrophilic myelocyte

Summary The developmental changes in the neutrophilic myelocyte from normal human bone marrow have been analyzed by means of phase contrast and electron microscopy. This developmental stage is characterized principally by the elaboration of secondary (specific) granules. In addition, there is a modest decrease in cell size, a decrease in the number and mean size of primary (azurophil) granules, a decrease in the number of polysomes, free ribosomes and mitochondria, a depletion of rough endoplasmic reticulum, an increase in cytoplasmic glycogen, an increase in chromatin aggregations and a loss of nucleoli, and the formation of a markedly indented nucleus. The myelocyte stage has been subdivided into three arbitrary phases based upon morphological and functional characteristics which relate to the onset, active production and cessation of secondary granulogenesis.Supported by Grant No. AM-HE-12084-13 from the National Institutes of Health, Bethesda, Maryland.Appreciation is expressed to Dr. Arthur Sagone who performed the bone marrow aspirations and to Anita Topson, Barbara Jordan and Marjorie Griffith for their technical assistance.     

The human neutrophilic promyelocyte

Summary The developmental changes in the neutrophilic promyelocytes from normal human bone marrow have been analyzed by means of phase contrast and electron microscopy. This developmental phase is characterized by the elaboration of primary (azurophillysosomal) granules and the entire intracellular machinery is directed principally toward this goal. The promyelocyte stage has been subdivided into three arbitrary stages based upon morphological, histochemical and functional characteristics which relate to the onset, active production and cessation of primary granulogenesis.Supported by Grant No. AM-HE-12084-12 from the National Institutes of Health, Bethesda, Maryland.Appreciation is expressed to Dr. Arthur Sagone who performed the bone marrow aspirations and to Marjorie Griffith, Anita Topson and Barbara Jordan for their technical assistance.     

Role of the p70 S6 kinase cascade in neutrophilic differentiation and proliferation of HL-60 cells-a study of transferrin receptor-positive and -negative cells obtained from dimethyl sulfoxide- or retinoic acid-treated HL-60 cells

Previously, we suggested that p70 S6 kinase (p70 S6K) plays an important role in the regulation of neutrophilic differentiation of HL-60 cells; this conclusion was based on our analysis of transferrin receptor (Trf-R) positive (Trf-R(+)) and negative (Trf-R(-)) cells that appeared after treatment with dimethyl sulfoxide (Me(2)SO). In this study, we analyzed the upstream of p70 S6K in relation to the differentiation and proliferation of both cell types. The granulocyte colony-stimulating factor (G-CSF)-induced enhancement of phosphatidylinositol 3-kinase (PI3K) activity in Trf-R(+) cells was markedly higher than that in Trf-R(-) cells. Wortmannin, a specific inhibitor of PI3K, partially inhibited G-CSF-induced p70 S6K activity and G-CSF-dependent proliferation, whereas rapamycin, an inhibitor of p70 S6K, completely inhibited these activities. The wortmannin-dependent enhancement of neutrophilic differentiation was similar to that induced by rapamycin. From these results, we conclude that the PI3K/p70 S6K cascade may play an important role in negative regulation of neutrophilic differentiation in HL-60 cells. For the G-CSF-dependent proliferation, however, p70 S6K appears to be a highly important pathway through not only a PI3K-dependent but also possibly an independent cascade.