Hydrogen sulfide augments neutrophil migration through enhancement of adhesion molecule expression and prevention of CXCR2 internalization: Role of ATP-sensitive potassium channels

In this study, we have addressed the role of H2S in modulating neutrophil migration in either innate (LPS-challenged naive mice) or adaptive (methylated BSA (mBSA)-challenged immunized mice) immune responses. Treatment of mice with H S synthesis inhibitors, DL-propargylglycine (PAG) or beta-cyanoalanine, reduced neutrophil migration induced by LPS or methylated BSA (mBSA) into the peritoneal cavity and by mBSA into the femur/tibial joint of immunized mice. This effect was associated with decreased leukocyte rolling, adhesion, and P-selectin and ICAM-1 expression on endothelium. Predictably, treatment of animals with the H2S donors, NaHS or Lawesson`s reagent, enhanced these parameters. Moreover, the NaHS enhancement of neutrophil migration was not observed in ICAM-1-deficient mice. Neither PAG nor NaHS treatment changed LPS-induced CD18 expression on neutrophils, nor did the LPS- and mBSA-induced release of neutrophil chemoattractant mediators TNF-alpha, keratinocyte-derived chemokine, and LTB4. Furthermore, in vitro MIP-2-induced neutrophil chemotaxis was inhibited by PAG and enhanced by NaHS treatments. Accordingly, MIP-2-induced CXCR2 internalization was enhanced by PAG and inhibited by NaHS treatments. Moreover, NaHS prevented MIP-2-induced CXCR2 desensitization. The PAG and NaHS effects correlated, respectively, with the enhancement and inhibition of MIP-2-induced G protein-coupled receptor kinase 2 expression. The effects of NaHS on neutrophil migration both in vivo and in vitro, together with CXCR2 internalization and G protein-coupled receptor kinase 2 expression were prevented by the ATP-sensitive potassium (K-ATP(+)) channel blocker, glybenclamide. Conversely, diazoxide, a K-ATP(+) channel opener, increased neutrophil migration in vivo. Together, our data suggest that during the inflammatory response, H`S augments neutrophil adhesion and locomotion, by a mechanism dependent on K-ATP(+) channels.

Dual role of hydrogen sulfide in mechanical inflammatory hypernociception

Hydrogen Sulfide (H2S) is an endogenous gas involved in several biological functions, including modulation of nociception. However, the mechanisms involved in such modulation are not fully elucidated. The present Study demonstrated that the pretreatment of mice with PAG, a H2S synthesis inhibitor, reduced LPS-induced mechanical paw hypernociception. This inhibition of hypernociception was associated with the prevention of neutrophil recruitment to the plantar tissue. Conversely, PAG had no effect on LPS-induced production of the hypernociceptive cytokines, TNF-alpha, IL-1 beta and CXCL1/KC and on hypernociception induced by PGE(2), a directly acting hypernociceptive mediator. In contrast with the pro-nociceptive role of endogenous H2S. systemic administration of NaHS, a H2S donor, reduced LPS-induced mechanical hypernociception in mice. Moreover, this treatment inhibited mechanical hypernociception induced by PGE(2), suggesting a direct effect of H2S on nociceptive neurons. The antinociceptive mechanism of exogenous H2S depends on K-(ATP)(+) channels since the inhibition of PGE(2) hypernociception by NaHS was prevented by glibenclamide (K-(ATP)(+) channel blocker). Finally, NaHS did not alter the thermal nociceptive threshold in the hot-plate test, confirming that its effect is mainly peripheral. Taken together, these results suggest that H2S has a dual role in inflammatory hypernociception: 1. an endogenous pro-nociceptive effect due to up-regulation of neutrophil migration. and 2. an antinociceptive effect by direct blockade of nociceptor sensitization modulating K-(ATP)(+) channels. (c) 2008 Elsevier B.V. All rights reserved.

Hydrogen Sulfide Improves Neutrophil Migration and Survival in Sepsis via K(ATP)(+) Channel Activation

Rationale Recovering the neutrophil migration to the infectious focus improves survival in severe sepsis. Recently, we demonstrated that the cystathionine gamma-lyase (CSE)/hydrogen sulfide (H(2)S) pathway increased neutrophil recruitment to inflammatory focus during sterile inflammation. Objectives: To evaluate if H(2)S administration increases neutrophil migration to infectious focus and survival of mice. Methods. Sepsis was induced by cecal ligation and puncture (CLP) Measurements and Main Results. The pretreatments of mice with H2S donors (NaHS or Lawesson`s reagent) improved leukocyte rolling/adhesion in the mesenteric microcirculation as well as neutrophil migration. Consequently, bacteremia levels were reduced, hypotension and lung lesions were prevented, and the survival rate increased from approximately 13% to approximately 80% Even when treatment was delayed (6 h after CLP), a highly significant reduction in mortality compared with untreated mice was observed Moreover, H(2)S pretreatment prevented the down-regulation of CXCR2 and L-selectin and the up-regulation of CD11b and G protein-coupled receptor kinase 2 in neutrophils during sepsis. H(2)S also prevented the reduction of intercellular adhesion molecule-1 expression in the endothelium of the mesenteric microcirculation in severe sepsis Confirming the critical role of H(2)S on sepsis outcome, pretreatment with dl-propargylglycine (a CSE inhibitor) inhibited neutrophil migration to the infectious focus, enhanced lung lesions, and induced high mortality in mice subjected to nonsevere sepsis (from 0 to similar to 80%). The beneficial effects of H(2)S were blocked by glibenclamide (a ATP-dependent K(+) channel blocker). Conclusions: These results showed that H(2)S restores neutrophil migration to the infectious focus and improves survival outcome in severe sepsis by an ATP-dependent K(+) channel-dependent mechanism.

Inhibition of hydrogen sulphide formation reduces cisplatin-induced renal damage

Background. Cisplatin (CP)-induced renal damage is associated with inflammation. Hydrogen sulphide (H(2)S) is involved in models of inflammation. This study evaluates the effect of DL-propargylglycine (PAG), an inhibitor of endogenous H(2)S formation, on the renal damage induced by CP. Methods. The rats were injected with CP (5 mg/kg, i.p.) or PAG(5 mg/kg twice a day, i.p.) for 4 days, starting 1 h before CP injection. Control rats were injected with 0.15 M NaCl or PAG only. Blood and urine samples were collected 5 days after saline or CP injections for renal function evaluation. The kidneys were removed for tumour necrosis factor (TNF)-alpha quantification, histological, immunohistochemical and Western blot analysis. The cystathionine gamma-lyase (CSE) activity and expression were assessed. The direct toxicity of H(2)S in renal tubular cells was evaluated by the incubation of these cells with NaHS, a donor of H(2)S. Results. CP-treated rats presented increases in plasma creatinine levels and in sodium and potassium fractional excretions associated with tubulointerstitial lesions in the outer medulla. Increased expression of TNF-alpha, macrophages, neutrophils and T lymphocytes, associated with increased H(2)S formation rate and CSE expression, were also observed in the outer medulla from CP-injected rats. All these alterations were reduced by treatment with PAG. A direct toxicity of NaHS for renal tubular epithelial cells was not observed. Conclusions. Treatment with PAG reduces the renal damage induced by CP. This effect seems to be related to the H2S formation and the restriction of the inflammation in the kidneys from PAG+CP-treated rats.

Minimização de odores em contentores de resíduos sólidos

Mestrado em Engenharia Química

Assessment of hydrogen sulfide removal from biogas using a field scale anoxic biotrickling filter.

Hydrogen sulphide is one of the most toxic and corrosive compound present in swine-derived biogas streams.In this study, afield scale biotrickling filter for the removal of hydrogen sulfide was investigated.A Biofilter packed with supporting biofilm materials was fed continuously with a proprietary nutrient solution and operatedfor over 73days. The system has been operating with a H2S inlet concentrations ranging from 1,000to 3,000 ppm.Significant removal efficiencies >95% was demonstrated. pH of the stock feeding solution decreased from 6.2 to as low as 3.5within couple days.The resulting drop in pH provided circumstantial evidence to support biological H2 Soxidation to sulphuric acid by sulfide-oxidizers. Sulfur precipitation was also observed to occur. The results suggested that H2S removal from biogas stream can be efficiently achieved using portable, low cost and maintenance free biotrickling filters.

Use of swine waste-derived biogas to enhance microalgae productivity.

The effects of swine wastewater-derived biogas on microalgae productivity were determined. Experiments were conducted in a closed photobioreactor containing digestate effluent as culturing media and biogas in the headspaceas source of CO2. Experiments were carried out under mixothrophic and autothrophic conditions. Results showed that autotrophic growth rate (0.6 d-1)was twofoldfaster than mixotrophic. Frequent reinjections of biogas containing up to 2,000 ppm of hydrogen sulfide was not inhibitory to microalgae growth. The rapid removal of H2S in the system suggests photobioreactors can be an interesting alternative to biogas purification. A model to estimate microalgae productivity based on the amount of available CO2, inorganic and organic carbon was developedand showed good data fit correlation (r²= 0.99).

Contribuição para o estudo do tratamento de águas residuais provenientes da indústria de extração de petróleo

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente – Perfil Engenharia Sanitária

Tratamento biológico de odores em estações de tratamento de água residuais

O odor libertado em consequência da drenagem e tratamento de águas residuais gera consequências indesejáveis quer a nível do ecossistema quer a nível da saúde e bemestar das populações vizinhas. O tratamento de odores é por isso cada vez mais uma prioridade numa sociedade cada vez mais exigente. Para o correcto e eficiente tratamento dos gases odoríferos é necessário medir e caracterizar o odor, recorrendo muitas vezes à conjugação de medições analíticas e sensoriais, nomeadamente recorrendo ao electronic nose. Após a caracterização do odor e o conhecimento dos locais potências para a sua formação é necessário encaminhar e tratar o ar contaminado. O presente trabalho visa, através de uma perspectiva evolutiva, caracterizar os principais mecanismos de tratamento de odor em ETAR, com especial destaque para o tratamento biológico. Os mecanismos de tratamento podem ser agrupados genericamente em físicoquímico e térmico ou biológico. O tratamento biológico de odores é aplicado através de mecanismos como; biofiltros convencionais, biofiltros humidificados, biomassa em suspensão e um novo mecanismo apenas testado à escala laboratorial as biomembranas. Neste trabalho é feita uma exposição detalhada dos referidos mecanismos, apresentando as suas potencialidades e limitações.

A hidroquímica do Rio Solimões - Amazonas

Neste trabalho apresentamos os resultados dos estudos físicos-químicos do rio Solimões-Amazonas, em onze (11) locais e em trinta e um (31) afluentes e subafluentes, desde a fronteira do Brasil-Peru-Colômbia, em Tabatinga, até a amostragem, de frente a cidade de Santarém, no Estado do Pará, com uma distância de 2457 km. Os afluentes e subafluentes do rio Solimões-Amazonas ficam reduzidos a pequenos córregos nos seus altos cursos; apenas os rios Juruá, Purus, Negro e Madeira em alguns locais ficam com uma lâmina d'água de dois (2) metros, na região navegável, isto dependendo da estiagem. Todos os rios de água barrenta formam várzeas, que são ricas em nutrientes minerais, várzeas essas que possuem em toda a sua extensão lagos, paranás, igapós, forma-se grande abundância de capim flutuante que, em parte entre em decomposição, produzindo gases tóxicos, como H2S, CH4, CO2, etc. Os lagos e a várzea funcionam como se fossem uma esponja do rio, absolvendo sedimentos suspensos, nutrientes minerais e orgânicos, e liberando uma parte, no período de esvaziamento (seca). Os rios de água preta não formam várzeas e sim praias, e igapós no período da cheia, por possuírem pouco sedimentos; a cor escura é devida a substancias coloridas, como material húmico, que limitam a produção de fitoplancton. Os rios de água clara, também não formam várzeas e sim praias com poucos igapós; eles apresentam uma coloração verde azulada dada a grande formações de algas do tipo Cyanophyta. Os rios de água barrenta que apresentam maiores concentrações de sedimentos nos meses de novembro e abril tem como abastecedora a mobilização, com resuspensão, dos sedimentos, devidos ao aumento da vazão e, em parte, ao fenômeno das terras caldas. É possível que o rio Solimões-Amazonas receba suprimentos minerais e orgânicos dos rios de água barrenta, de pequenos rios que são represados na sua foz, de rios com pequenos afloramentos de calcáreo e de rios com elevados teores de substancias coloridas. As menores temperaturas no rio Solimões-Amazonas ocorrem no período de enchimento do Canal principal e as maiores no verão; quanto ao oxigênio as menores concentrações aparecem nos meses de abril a julho, são de vidas as águas oxidadas provenientes das várzeas e lagos.

Deterioração bacteriológica do jaraqui Semaprochilodus spp. capturado no estado do Amazonas e conservado em gelo

Neste trabalho foram analisados 73 espécimes de jaraqui Semaprochilodus spp. conservados em caixas de poliestireno expandido entre camadas de gelo. Foram realizadas as seguintes análises: avaliação sensorial pela tabela de Torry modificada e pelo índice de qualidade por deméritos; determinação do pH e das bases voláteis totais (N-BVT); contagem total dos microrganismos aeróbios psicrófilos a 20 ºC por 4 dias, psicrotróficos a 7 ºC por 10 dias, dos mesófilos a 37 ºC por 2 dias; contagem, isolamento e identificação das bactérias Aeromonas sp. Bacillus sp. e Pseudomonas sp. a 20 ºC por 24 horas e de Plesiomonas sp. a 37 ºC por 2 dias. O jaraqui se manteve em condições de consumo, pela avaliação sensorial, por 18 e 21 dias. O pH e as bases voláteis totais não foram bons indicadores de qualidade; as contagens totais de psicrófilos, psicrotróficos e mesófilos não apresentaram diferença significativa e as bactérias não apresentaram comportamento deteriorador pela ausência da produção de H2S.

Paper del sulfur d'hidrogen com a neurotransmissor a nivell dels sistema nerviòs entèric

Al llarg dels últims anys l’òxid nítric (NO) i el monòxid de carboni (CO) han estat investigats com dos neurotransmissors gasosos del tracte gastrointestinal. Algunes dades demostren que el sulfur d’hidrogen (H2S) podria ser el tercer neurotransmissor gasos a nivell central i perifèric. Objectiu: valorar si el H2S compleix els requisits per ser considerat com el tercer neurotransmissor gasos a nivell del sistema nerviós entèric. Mètodes: la motilitat espontània de preparacions de múscul llis circular de mostres de còlon sigma humà i còlon mig de rata es van determinar in vitro mitjançant la tècnica de banys d’òrgans. El hidrogensulfur de sodi (NaHS) va ser utilitzat com donador de H2S. Resultats: 1- El NaHS va inhibir de forma concentració depenent la motilitat espontània en presència de bloqueig neural amb Tetrodotoxina (TTX 1 μM) (IC50 101±18 μM rata n=10; 329±86 μM humà n=18). 2- Aquesta relaxació es va veure reduïda per ODQ 10 μM, un inhibidor de la guanilat ciclasa, en còlon mig de rata (n=5). En humà a més de ser reduïda per ODQ 10μM (n=6), també ho va ser per SQ 22,536 100 μM (n=10), un inhibidor de l’adenilat ciclasa, i per L-NNA 1 mM (n=8), un inhibidor de la sintesi de NO. 3- En còlon de rata la incubació amb D,L-Propargilglicina 2 mM (PAG), un inhibidor de la síntesi de H2S, va incrementar la motilitat tant en situació control (10,05±2,06 vs 17,46±3,37 g•min-1 AUC, P&0,001, n=10) com en presència de TTX 1μM (14,45±7,28 vs 21,67±10,05 g•min-1 AUC, P&0,01, n=7). Conclusió: el H2S compleix part dels criteris per ser considerat un neurotransmissor gasos inhibitori en el tracte gastrointestinal: la inhibició de la seva síntesi incrementa la motilitat intestinal i la seva addició exògena produeix relaxació de la musculatura llisa intestinal.

Investigation of Postmortem Gases: A Forensic Imaging and Analytical Chemistry Approach

Background: Distinguishing postmortem gas accumulations in the body due to natural decomposition and other phenomena such as gas embolism can prove a difficult task using purely Multi-Detector Computed Tomography (MDCT). The Radiological Alteration Index (RAI) was created with the intention to be able to identify bodies undergoing the putrefaction process based on the quantity of gas detected within the body. The flaw in this approach is the inability to absolutely determine putrefaction as the origin of gas volumes in cases of moderate alteration. The aim of the current study is to identify percentage compositions of O2, N2, CO2 and the presence of gases such as H2 and H2S within these sampling sites in order to resolve this complication. Materials and methods: All cases investigated in our University Center of Legal Medicine are undergoing a Post-Mortem Computed Tomography (PMCT)-scan before external examination or autopsy as a routine investigation. In the obtained images, areas of gas were characterized as 0, I, II or III based on the amount of gas present according to the RAI (1). The criteria for these characterizations were dependent of the site of gas, for example thoracic and abdominal cavities were graded as I (1 - 3cm gas), II (3 - 5cm gas) and III (>5cm gas). Cases showing gaseous sites with grade II or III were selected for this study. The sampling was performed under CT-guidance to target the regions to be punctured. Luer-lock PTFE syringes equipped with a three-way valve and needles were used to sample the gas directly (2). Gaseous samples were then analysed using gas chromatography coupled to a thermal conductivity detector (GC-TCD). The components present in the samples were expressed as a percentage of the overall gas present. Results: Up to now, we have investigated more than 40 cases using our standardized procedure for sampling and analysis of gas. O2, N2 and CO2 were present in most samples. The following distributions were found to correlate to gas origins of gas embolism/scuba diving accidents, trauma and putrefaction: ? Putrefaction → O2 = 1 - 5%; CO2 > 15%; N2 = 10 - 70%; H2 / H2S / CH4 variable presence ? Gas embolism/Scuba diving accidents → O2 and N2= varying percentages; CO2 > 20% ? Trauma → O2 = small percentage; CO2 < 15%; N2 > 65% H2 and H2S indicated levels of putrefaction along with methane which can also gauge environmental conditions or conditions of body storage/burial. Many cases showing large RAI values (advanced alteration) did reveal a radiological diagnosis which was in concordance with the interpretation of the gas composition. However, in certain cases (gas embolism, scuba divers) radiological interpretation was not possible and only chemical gas analysis was found to lead to the correct diagnosis, meaning that it provided complementary information to the radiological diagnosis. Conclusion: Investigation of postmortem gases is a useful tool to determine origin of gas generation which can aid the diagnosis of the cause of death. Levels of gas can provide information on stage of putrefaction and help to perform essential medico-legal diagnosis such as vital gas embolism.

Sulphate-reducing bacteria associated with biocorrosion: a review

Biocorrosion means any process of corrosion in wich microorganisms are somehow involved. As far as the petroleum industry is concerned, the anaerobic type is the more important, with Sulphate-Reducing Bacteria (SRB) accouting for half of the described processes. SRB are obligate anaerobs that use sulphur, sulphate or other oxidized sulphur compounds as oxidizing agents when decomposing organic material. A typical product of SRB metabolism, hydrogen sulphide -H2S-, is extremely toxic. In the present work we review the literature on mechanisms underlying biocorrosive process in wich SRB are involved and summarize some of the ultrastructural and eletrochemical work developed using SRB obtained from water injection flow in wells located on PETROBRAS offshore marine plataforms, sampled directly in the field over metallic probes, or cultured under laboratory conditions. Biofilms develop when SRB adhere to inert surfaces. A high diversity of morphological types is found inside these biofilms. Their extracellular matrix is highly hydrated and mainly anionic, as shown by its avid reaction with cationic compounds like ruthenium red. We have noted that variations in iron contet lead to interesting changes in the ultrastructure of the bacterial cell coat and also in the rate of corrosion induced in metallic test cupons. Since routine methods to prevent and treat SRB contamination and biodeterioration involve the use of biocides that are toxic and always have some environmental impact, an accurate diagnosis of biocorrosion is always required prior to a treatment decision. We developed a method that detects and semi-quantifies the presence of living or dead SRB by using free silver potentials as an indicator of corrosive action by SRB-associated sulphides. We found a correlation between sulphide levels (determined either by spectrophotometry, or using a silver electrode -E(Ag)- that measured changes in free potentials induced by the presence of exogeneously added sulphide) and SRB concentration (enumerated by a culturing method). E (Ag) was characterized under a variety of conditions andwas found to be relatively immune to possible interference resulting from aeration of media or from the psence of iron corrosion products. The method offers a simple, rapid, and effective means of diagnosing biocorrosive processes prior to their control.

Hydrocarbon biomarkers in the Topla-Mezica zinc-lead deposits, northern Karavanke/Drau Range, Slovenia: Paleoenvironment at the site of ore formation

The Mississippi Valley-type zinc and lead deposits at Topla (250,150 metric tons (t) of ore grading 1.0 wt % Zn and 3.3 wt % Pb) and Mezica (19 million metric tons (Mt) of ore grading 5.3 wt % Pb and 2.7 wt % Zn) occur within the Middle to Upper Triassic platform carbonate rocks of the northern Karavanke/Drau Range geotectonic units of the Eastern Alps, Slovenia. The ore and host rocks of these deposits have been investigated by a combination of inorganic and organic geochemical methods to determine major, trace, and rare earth element (REE) concentrations, hydrocarbon distribution, and stable isotope ratios of carbonates, kerogen, extractable organic matter, and individual hydrocarbons. These data combined with sedimentological evidence provide insight into the paleoenvironmental conditions at the site of ore formation. The carbonate isotope composition, the REE patterns, and the distribution of hydrocarbon biomarkers (normal alkanes and steranes) suggest a marine depositional environment. At Topla, a relatively high concentration of redox sensitive trace elements (V, Mo, U) in the host dolostones and REE patterns parallel to that of the North American shale composite suggest that sediments were deposited in a reducing environment. Anoxic conditions enhanced the preservation of organic matter and resulted in relatively higher total organic carbon contents (up to 0.4 wt %). The isotopic composition of the kerogen (delta C-13(kerogon) = -29.4 to -25.0 parts per thousand, delta N-15(kerogen) = -.13.6 to 6.8 parts per thousand) suggests that marine algae and/or bacteria were the main source of organic carbon with a very minor contribution from detrital continental plants and a varying degree of alteration. Extractable organic matter from Topla ore is generally depleted in C-13 compared to the associated kerogen, which is consistent with an indigenous source of the bitumens. The mineralization correlates with delta N-15(kerogen) values around 0 per mil, C-13 depleted kerogen, C-13 enriched n-heptadecane, and relatively high concentrations of bacteria] hydrocarbon biomarkers, indicating a high cyanobacterial biomass at the site of ore formation. Abundant dissimilatory sulfate-reducing bacteria, feeding on the cyanobacterial remains, led to accumulation of biogenic H2S in the pore water of the sediments. This biogenic H2S was mainly incorporated into sedimentary organic matter and diagenetic pyrite. Higher bacterial activity at the ore site also is indicated by specific concentration ratios of hydrocarbons, which are roughly correlated with total Pb plus Zn contents. This correlation is consistent with mixing of hydrothermal metal-rich, fluids and local bacteriogenic sulfide sulfur. The new geochemical data provide supporting evidence that Topla is a low-temperature Mississippi Valley-type deposit formed in an anoxic supratidal saline to hypersaline environment. A laminated cyanobacterial mat, with abundant sulfate-reducing bacteria was the main site of sulfate reduction.