RESUMO
PURPOSE: This study aimed to investigate the effects of AGE on microvascular reactivity, systolic blood pressure (SBP), and diastolic blood pressure (DBP) in older individuals at high risk for cardiovascular disease (CVD). Urinary thiosulfate was also investigated as an indirect marker of endogenous hydrogen sulfide (H2S) synthesis. The study was conducted in a randomized, double-blind, crossover, and placebo-controlled way. METHODS: Twenty-eight participants (14 male), 67 ± 6 years old with CVD risk factors, ingested 2.4 g of AGE or placebo (PLA). Near-infrared spectroscopy evaluated tissue oxygen saturation (StO2) during a vascular occlusion test (30 s baseline, 5 min occlusion, and 2 min reperfusion). The upslope of StO2 signal after cuff release was calculated to measure microvascular reactivity. Urinary thiosulfate levels were measured using a high-performance liquid chromatography system. RESULTS: The upslope of StO2 was significantly faster after AGE (1.01 ± 0.37% s-1) intake compared to PLA (0.83 ± 0.35% s-1; P < 0.001; d = 0.50). Relative changes in Δ% SBP from pre- to post-AGE intake (- 5.17 ± 5.77%) was significantly different compared to Δ% PLA (0.32 ± 5.99%; P = 0.001; d = 0.93). No significant changes in urinary thiosulfate concentrations were observed between interventions. Moreover, no significant gender effect in any parameter assessed was found. CONCLUSION: This study demonstrated that a single dose of AGE improved microvascular reactivity in older adults at risk of CVD despite such an effect was not linked with urinary thiosulfate levels. This trial was registered at clinicaltrials.gov as NCT04008693 (May 19, 2020).
Assuntos
Doenças Cardiovasculares , Alho , Sulfeto de Hidrogênio , Idoso , Doenças Cardiovasculares/prevenção & controle , Humanos , Sulfeto de Hidrogênio/metabolismo , Masculino , Microcirculação/fisiologia , Pessoa de Meia-Idade , Músculo Esquelético/metabolismo , Consumo de Oxigênio/fisiologia , Extratos Vegetais/metabolismo , Poliésteres/metabolismo , Tiossulfatos/metabolismoRESUMO
Cyanide is a toxic compound that is converted to the non-toxic thiocyanate by a rhodanese enzyme. Rhodaneses belong to the family of transferases (sulfurtransferases), which are largely studied. The sulfur donor defines the subfamily of these enzymes as thiosulfate:cyanide sulfurtransferases or rhodaneses (TSTs) or 3-mercaptopyruvate sulfurtransfeases (MSTs). In Mycobacterium tuberculosis, the causative agent of tuberculosis, the gene Rv0815c encodes the protein CysA2, a putative uncharacterized thiosulfate:cyanide sulfurtransferase that belongs to the essential sulfur assimilation pathway in the bacillus and is secreted during infection. In this work, we characterized the functional and structural properties of CysA2 and its kinetic parameters. The recombinant CysA2 is a α/ß protein with two rhodanese-like domains that maintains the functional motifs and a catalytic cysteine. Sulfurtransferase activity was determined using thiosulfate and 3-mercaptopyruvate as sulfur donors. The assays showed Km values of 2.89 mM and 7.02 mM for thiosulfate and 3-mercaptopyruvate, respectively, indicating the protein has dual activity as TST and MST. Immunological assays revealed that CysA2 interacted with pulmonary cells, and it was capable to activate macrophages and dendritic cells, indicating the stimulation of the immune response, which is important for its use as an antigen for vaccine development and immunodiagnostic.
Assuntos
Cisteína/análogos & derivados , Mycobacterium tuberculosis/enzimologia , Sulfurtransferases/química , Sulfurtransferases/metabolismo , Tiossulfatos/metabolismo , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Linhagem Celular , Cisteína/química , Cisteína/metabolismo , Células Dendríticas/citologia , Células Dendríticas/imunologia , Cinética , Macrófagos/citologia , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , Mycobacterium tuberculosis/genética , Ligação Proteica , Conformação Proteica , Especificidade por Substrato , Sulfurtransferases/genética , Sulfurtransferases/imunologiaRESUMO
Strain USBA-019T, an anaerobic and thermophilic strain, was identified as a new member of the genus Thermoanaerobacterium. USBA-019T cells are gram-positive, strictly anaerobic, thermophilic, chemoorganotrophic, moderately acidophilic, non-motile, endospore-forming, slightly curved, and rod-shaped. Cells measure 0.4×3.0-7.0µm. Optimal growth occurs at 50-55°C (35-65°C). Optimum pH is 5.0-5.5 (4.0-8.5). Thiosulfate, elemental sulfur and nitrate were utilized as electron acceptors. Fermentation of glucose, lactose, cellobiose, galactose, arabinose, xylose, starch and xylan primarily produced acetate and butyrate. Xylan, starch and cellobiose produced ethanol and starch, cellobiose, galactose, arabinose and mannose produced lactic acid. Phylogenetic analyses based on 16S rRNA gene sequence comparison and genomic relatedness indices show the close relation of USBA-019T to Thermoanaerobacterium thermostercoris and Thermoanaerobacterium aotearoense (similarity value: 99%). Hybridization of USBA-019T, Th. thermostercoris DSM22141T and Th. aotearoense DMS10170T found DNA-DNA relatedness of 33.2% and 18.2%, respectively. Based on phenotypic, chemotaxonomic and phylogenetic evidence, along with low identity at whole genome level, USBA-019T is a novel species of the genus Thermoanaerobacterium which we propose to name Thermoanaerobacterium butyriciformans sp. nov. The type strain is USBA-019T (=CMPUJ U-019T=DSM 101588T).
Assuntos
Fontes Termais/microbiologia , Thermoanaerobacterium/classificação , Thermoanaerobacterium/isolamento & purificação , Álcoois/metabolismo , Anaerobiose , Técnicas de Tipagem Bacteriana , Metabolismo dos Carboidratos , Ácidos Carboxílicos/metabolismo , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fermentação , Concentração de Íons de Hidrogênio , Nitratos/metabolismo , Hibridização de Ácido Nucleico , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Esporos Bacterianos/citologia , Enxofre/metabolismo , Temperatura , Thermoanaerobacterium/genética , Tiossulfatos/metabolismoRESUMO
The biogeochemical cycling of gold (Au), i.e. its solubilization, transport and re-precipitation, leading to the (trans)formation of Au grains and nuggets has been demonstrated under a range of environmental conditions. Biogenic (trans)formations of Au grains are driven by (geo)biochemical processes mediated by distinct biofilm consortia living on these grains. This review summarizes the current knowledge concerning the composition and functional capabilities of Au-grain communities, and identifies contributions of key-species involved in Au-cycling. To date, community data are available from grains collected at 10 sites in Australia, New Zealand and South America. The majority of detected operational taxonomic units detected belong to the α-, ß- and γ-Proteobacteria and the Actinobacteria. A range of organisms appears to contribute predominantly to biofilm establishment and nutrient cycling, some affect the mobilization of Au via excretion of Au-complexing ligands, e.g. organic acids, thiosulfate and cyanide, while a range of resident Proteobacteria, especially Cupriavidus metallidurans and Delftia acidovorans, have developed Au-specific biochemical responses to deal with Au-toxicity and reductively precipitate mobile Au-complexes. This leads to the biomineralization of secondary Au and drives the environmental cycle of Au.
Assuntos
Actinobacteria/metabolismo , Alphaproteobacteria/metabolismo , Biofilmes/crescimento & desenvolvimento , Cupriavidus/metabolismo , Delftia/metabolismo , Gammaproteobacteria/metabolismo , Ouro/metabolismo , Actinobacteria/crescimento & desenvolvimento , Alphaproteobacteria/crescimento & desenvolvimento , Austrália , Cupriavidus/crescimento & desenvolvimento , Delftia/crescimento & desenvolvimento , Gammaproteobacteria/crescimento & desenvolvimento , Nova Zelândia , América do Sul , Tiossulfatos/metabolismoRESUMO
One of the problems in waste water treatment plants (WWTPs) is the increase in emissions of hydrogen sulphide (H2S), which can cause damage to the health of human populations and ecosystems. To control emissions of this gas, sulphur-oxidizing bacteria can be used to convert H2S to sulphate. In this work, sulphate detection was performed by spectrophotometry, ion chromatography and atomic absorption spectrometry, using Paracoccus pantotrophus ATCC 35512 as a reference strain growing in an inorganic broth supplemented with sodium thiosulphate (Na2S2O3·5H2O), sodium sulphide (Na2S) or sodium sulphite (Na2SO3), separately. The strain was metabolically competent in sulphate production. However, it was only possible to observe significant differences in sulphate production compared to abiotic control when the inorganic medium was supplemented with sodium thiosulphate. The three methods for sulphate detection showed similar patterns, although the chromatographic method was the most sensitive for this study. This strain can be used as a reference for sulphate production in studies with sulphur-oxidizing bacteria originating from environmental samples of WWTPs.
Assuntos
Paracoccus pantotrophus/metabolismo , Sulfatos/metabolismo , Sulfetos/metabolismo , Sulfitos/metabolismo , Tiossulfatos/metabolismoRESUMO
Sulfite oxidase (SO) deficiency is biochemically characterized by tissue accumulation and high urinary excretion of sulfite, thiosulfate and S-sulfocysteine. Affected patients present severe neurological symptoms and cortical atrophy, whose pathophysiology is still poorly established. Therefore, in the present work we investigated the in vitro effects of sulfite and thiosulfate on important parameters of energy metabolism in the brain of young rats. We verified that sulfite moderately inhibited the activity of complex IV, whereas thiosulfate did not alter any of the activities of the respiratory chain complexes. It was also found that sulfite and thiosulfate markedly reduced the activity of total creatine kinase (CK) and its mitochondrial and cytosolic isoforms, suggesting that these metabolites impair brain cellular energy buffering and transfer. In contrast, the activity of synaptic Na(+),K(+)-ATPase was not altered by sulfite or thiosulfate. We also observed that the inhibitory effect of sulfite and thiosulfate on CK activity was prevented by melatonin, reduced glutathione and the combination of both antioxidants, as well as by the nitric oxide synthase N(ω)-nitro-l-arginine methyl ester, indicating the involvement of reactive oxygen and nitrogen species in these effects. Sulfite and thiosulfate also increased 2',7'-dichlorofluorescin oxidation and hydrogen peroxide production and decreased the activity of the redox sensor aconitase enzyme, reinforcing a role for oxidative damage in the effects elicited by these metabolites. It may be presumed that the disturbance of cellular energy and redox homeostasis provoked by sulfite and thiosulfate contributes to the neurological symptoms and abnormalities found in patients affected by SO deficiency.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/complicações , Encefalopatias Metabólicas/etiologia , Encéfalo/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Sulfito Oxidase/deficiência , Sulfitos/farmacologia , Tiossulfatos/farmacologia , Erros Inatos do Metabolismo dos Aminoácidos/genética , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/fisiopatologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/fisiologia , Encefalopatias Metabólicas/genética , Encefalopatias Metabólicas/metabolismo , Transporte de Elétrons/efeitos dos fármacos , Transporte de Elétrons/genética , Transporte de Elétrons/fisiologia , Metabolismo Energético/fisiologia , Masculino , Oxirredução/efeitos dos fármacos , Ratos , Ratos Wistar , Sulfito Oxidase/genética , Sulfito Oxidase/metabolismo , Sulfitos/metabolismo , Tiossulfatos/metabolismoRESUMO
Sulphur Oxidizing Bacteria (SOB) is a group of microorganisms widely used for the biofiltration of Total Reduced Sulphur compounds (TRS). TRS are bad smelling compounds with neurotoxic activity which are produced by different industries (cellulose, petrochemical). Thiobacillus thioparus has the capability to oxidize organic TRS, and strains of this bacterium are commonly used for TRS biofiltration technology. In this study, two thiosulphate oxidizing strains were isolated from a petrochemical plant (ENAP BioBio, Chile). They were subjected to molecular analysis by real time PCR using specific primers for T. thioparus. rDNA16S were sequenced using universal primers and their corresponding thiosulphate activities were compared with the reference strain T. thioparus ATCC 10801 in batch standard conditions. Real time PCR and 16S rDNA sequencing showed that one of the isolated strains belonged to the Thiobacillus branch. This strain degrades thiosulphate with a similar activity profile to that shown by the ATCC 10801 strain, but with less growth, making it useful in biofiltration.
Assuntos
Animais , Halothiobacillus , Halothiobacillus/genética , Thiobacillus , Thiobacillus/enzimologia , Thiobacillus/genética , Tiossulfatos/metabolismo , Tiossulfatos/química , Indústria de Petróleo e Gás/métodosRESUMO
This work describes the design and performance of a thiosulfate-oxidizing bioreactor that allowed high elemental sulfur production and recovery efficiency. The reactor system, referred to as a Supernatant-Recycling Settler Bioreactor (SRSB), consisted of a cylindrical upflow reactor and a separate aeration vessel. The reactor was equipped with an internal settler and packing material (structured corrugated PVC sheets) to facilitate both cell retention and the settling of the formed elemental sulfur. The supernatant from the reactor was continuously recirculated through the aerator. An inlet thiosulfate concentration of 100 mmol l(-1) was used. The reactor system was fed with 89 mmol l(-1) d(-1) thiosulfate reaching 98 to 100% thiosulfate conversion with an elemental sulfur yield of 77%. Ninety-three percent of the produced sulfur was harvested from the bottom of the reactor as sulfur sludge. The dry sulfur sludge contained 87% elemental sulfur. The inclusion of an internal settler and packing material in the reactor system resulted in an effective retention of sulfur and biomass inside the bioreactor, preventing the oxidation of thiosulfate and elemental sulfur to sulfate in the aerator and, therefore, improving the efficiency of elemental sulfur formation and recovery.
Assuntos
Reatores Biológicos , Enxofre/metabolismo , Tiossulfatos/metabolismo , Biomassa , Oxirredução , Oxigênio/metabolismo , Esgotos , Eliminação de Resíduos Líquidos/métodosRESUMO
In this paper, we report the chromium(VI) reduction by filtrates of Acidithiobacillus and Thiobacillus cultures. Chromium(VI) reduction by filtrates of A. ferrooxidans cultures under acidic conditions was higher than that observed for A. thiooxidans. However, at pH close to 7, chromium(VI) reduction by filtrates of T. thioparus cultures was as high as that by filtrates of A. thiooxidans cultures and much higher than that observed for A. ferrooxidans cultures at the same pH. The capability of these cultures to reduce chromium(VI) was associated specifically with the fraction of cultures (cells, sulphur and associated sulphur compounds) retained by filtration through a 0.45mum filter. In the fraction that comes from A. thiooxidans culture, polythionates (S(x)O(6)(2-)) with 3-7 sulphur atoms were detected and identified (by HPLC with MS as detector). The model of vesicles containing polythionates, sulphur and water agrees with our results.
Assuntos
Acidithiobacillus/metabolismo , Técnicas de Cultura de Células/métodos , Cromo/metabolismo , Thiobacillus/metabolismo , Tiossulfatos/metabolismo , Acidithiobacillus/crescimento & desenvolvimento , Biodegradação Ambiental , Concentração de Íons de Hidrogênio , Oxirredução , Especificidade da Espécie , Thiobacillus/crescimento & desenvolvimentoRESUMO
Eight nucleotide sequences containing a single rhodanese domain were found in the Acidithiobacillus ferrooxidans ATCC 23270 genome: p11, p14, p14.3, p15, p16, p16.2, p21, and p28. Amino acids sequence comparisons allowed us to identify the potentially catalytic Cys residues and other highly conserved rhodanese family features in all eight proteins. The genomic contexts of some of the rhodanese-like genes and the determination of their expression at the mRNA level by using macroarrays suggested their implication in sulfur oxidation and metabolism, formation of Fe-S clusters or detoxification mechanisms. Several of the putative rhodanese genes were successfully isolated, cloned and overexpressed in E. coli and their thiosulfate:cyanide sulfurtransferase (TST) and 3-mercaptopyruvate/cyanide sulfurtransferase (MST) activities were determined. Based on their sulfurtransferase activities and on structural comparisons of catalytic sites and electrostatic potentials between homology- modeled A. ferrooxidans rhodaneses and the reported crystal structures of E. coli GlpE (TST) and SseA (MST) proteins, two of the rhodanese-like proteins (P15 and P16.2) could clearly be defined as TSTs, and P14 and P16 could possibly correspond to MSTs. Nevertheless, several of the eight A. ferrooxidans rhodanese-like proteins may have some different functional activities yet to be discovered.