Immunomodulatory activity of ß-glucan-containing exopolysaccharides from Auricularia auricular in phagocytes and mice infected with Cryptococcus neoformans.

Current antifungal drugs present poor effectiveness and there is no available vaccine for fungal infections. Thus, novel strategies to treat or prevent invasive mycosis, such as cryptococcosis, are highly desirable. One strategy is the use of immunomodulators of polysaccharide nature isolated from mushrooms. The purpose of the present work was to evaluate the immunostimulatory activity of ß-(1,3)-glucan-containing exopolysaccharides (EPS) from the edible mushrooms Auricularia auricula in phagocytes and mice infected with Cryptococcus neoformans. EPS triggered macrophages and dendritic cell activation upon binding to Dectin-1, a pattern recognition receptor of the C-type lectin receptor family. Engagement of Dectin-1 culminated in pro-inflammatory cytokine production and cell maturation via its canonical Syk-dependent pathway signaling. Furthermore, upon EPS treatment, M2-like phenotype macrophages, known to support intracellular survival and replication of C. neoformans, repolarize to M1 macrophage pattern associated with enhanced production of the microbicidal molecule nitric oxide that results in efficient killing of C. neoformans. Treatment with EPS also upregulated transcript levels of genes encoding products associated with host protection against C. neoformans and Dectin-1 mediated signaling in macrophages. Finally, orally administrated ß-glucan-containing EPS from A. auricular enhanced the survival of mice infected with C. neoformans. In conclusion, the results demonstrate that EPS from A. auricula exert immunostimulatory activity in phagocytes and induce host protection against C. neoformans, suggesting that polysaccharides from this mushroom may be promising as an adjuvant for vaccines or antifungal therapy.

Ureotelism is inducible in the neotropical freshwater Hoplias malabaricus (Teleostei, Erythrinidae).

Increased environmental pH decreases ammonia transport through the gills, impairing nitrogenous waste. The consequent toxicity is usually drastic to most fishes. A few species are able to synthesize urea as a way to detoxify plasma ammonia. We studied three teleosts of the family Erythrinidae living in distinct environments, and assumed the biochemical behaviors would be different in spite of their being closely related species. Adult fish collected in the wild were submitted to alkaline water and the urea excretion rate was determined. The specific activity of urea cycle enzymes was determined in liver samples of fish from neutral waters. The studied species Hoplias lacerdae, Hoplerithrynus unitaeniatus, and Hoplias malabaricus are ureogenic. Urea synthesis is not a metabolic way to detoxify ammonia in H. lacerdae and Hoplerithrynus unitaeniatus exposed to an alkaline environment. The plasma ammonia profile of both species showed two distinct biochemical responses. Urea excretion of H. malabaricus was high in alkaline water, and the transition to ureotelism is proposed. The nitrogen excretion rate of H. malabaricus was among the highest values reported and the high urea excretion leads us to include this species as ureotelic in alkaline water.

Ureotelism is inducible in the neotropical freshwater Hoplias malabaricus (Teleostei, Erythrinidae)

O aumento do pH ambiental diminui o transporte de amônia pelas brânquias, prejudicando a saída de nitrogênio. A conseqüente toxidade é normalmente drástica para muitos peixes. Poucas espécies são capazes de sintetizar uréia como forma de detoxificar a amônia plasmática. Estudamos três telósteos da família Erythrinidae que vivem em ambientes distintos e apresentam respostas bioquímicas diferentes apesar de serem espécies extremamente próximas. Peixes adultos foram coletados no ambiente, sendo submetidos a águas alcalinas, e o grau de excreção de uréia foi determinado. As atividades específicas das enzimas do ciclo da uréia foram determinadas em amostras de fígado de peixe de águas cujo pH é neutro. As espécies estudadas, Hoplias lacerdae, Hoplerithrynus unitaeniatus e Hoplias malabaricus, são ureogênicas. A síntese de uréia não é a rota metabólica utilizada para detoxificar a amônia em H. lacerdae e H. unitaeniatus expostas a ambiente alcalino. O perfil da amônia plasmática em ambas as espécies apresentaram duas respostas bioquímica distintas. A excreção de uréia em H. malabaricus foi superior em águas alcalinas, e transição para o ureotelismo é proposta. O grau de excreção nitrogenada em H. malabaricus foi superior aos valores citados, e o aumento da excreção de uréia sugere que essa espécie é ureotélica em águas alcalinas.

Biochemical responses of two Erythrinidae fish to environmental ammonia

A forma não ionizada da amônia é muito tóxica a vários organismos aquáticos, sendo particularmente importante em muitos aspectos da biologia dos peixes. Um amplo grupo de estratégias para enfrentar os estressores ambientais pode ser observado nos organismos vivos. Dentre estas, as respostas aos estressores químicos são bem estudadas. O presente trabalho compara respostas bioquímicas de duas espécies evolutivamente próximas Hoplias malabaricus e Hoplerythrinus unitaeniatus, expostas à amônia ambiental. Peixes adultos foram submetidos a 1.0 mg/L de cloreto de amônio por 24 horas e foram determinados os níveis plasmáticos de amônia e uréia. As atividades das enzimas do COU, OCT e ARG e a enzima acessória GS foram quantificadas em extrato de fígado e são expressas em mmol/min/mg de tecido úmido. Foi observado em H. malabaricus, exposto a 1,0 mg/L de cloreto de amônio, aumento (p < 0,05) nas enzimas: GS, de 1,14 para 2,43; OCT, de 0,81 para 1,72; ARG, de 3,15 para 4,23; na amônia plasmática, de 0,95 para 1,42 mmol/L, e na uréia plasmática, de 0,82 para 1,53. A GS de H. malabaricus aumentou de 1,43 para 1,84, todavia, OCT, ARG e uréia plasmática não variaram. Esses dados mostram que ambas as espécies, taxonomicamente próximas, respondem distintamente ao mesmo estressor ambiental.

Physiological responses to sulfide toxicity by the air-breathing catfish, Hoplosternum littorale (Siluriformes, Callichthyidae).

Hemolytic anemia accompanied by changes in the immunology system is one of the sulfide intoxication harmful effects on Hoplosternum littorale. Hematological parameters are considered as effective indicators of stress caused by this hydrogen sulfide. During sulfide exposure, H. littorale neither alters the methemoglobin concentration nor forms sulfhemoglobin in the presence of high levels of dissolved sulfide in the water. Cytochrome c oxidase shows little activity in the gills and blood of H. littorale when exposed to sulfide. Alternative metabolic routes are suggested through which the accumulation of pyruvate leads to the formation of an end product other than lactate.

Different biochemical strategies of two neotropical fish to cope with the impairment of nitrogen excretion during air exposure.

The exposure of fish to air is normally expected to interfere with the nitrogen excretion process. Hoplias malabaricus and Hoplerythrinus unitaeniatus, two teleost species, display distinct behaviors in response to decreases in natural reservoir water levels, although they may employ similar biochemical strategies. To investigate this point, plasma levels of ammonia, urea, uric acid, and the two urea cycle enzymes, ornithine carbamoyl transferase (OCT) and arginase (ARG), as well as glutamine synthetase (GS) were determined for both species after exposure to air. Plasma ammonia increased gradually during exposure to air, but only H. malabaricus showed increased concentrations of urea. Plasma uric acid remained very low in both fish. Enzymatic activities (mean +/- SD, micromol min(-1) g protein(-1)) of H. malabaricus showed significant increases (P<0.05, N = 6) in OCT from 0.84 +/- 0.05 to 1.42 +/- 0.03, in ARG from 8.07 +/- 0.47 to 9.97 +/- 0.53 and in GS from 1.15 +/- 0.03 to 2.39 +/- 0.04. The OCT and ARG enzymes remained constant in H. unitaeniatus (N = 6), but GS increased from 1.49 +/- 0.02 to 2.06 +/- 0.03. Although these species are very closely related and share the same environment, their biochemical strategies in response to exposure to air or to increased plasma ammonia are different.

Different biochemical strategies of two Neotropical fish to cope with the impairment of nitrogen excretion during air exposure

The exposure of fish to air is normally expected to interfere with the nitrogen excretion process. Hoplias malabaricus and Hoplerythrinus unitaeniatus, two teleost species, display distinct behaviors in response to decreases in natural reservoir water levels, although they may employ similar biochemical strategies. To investigate this point, plasma levels of ammonia, urea, uric acid, and the two urea cycle enzymes, ornithine carbamoyl transferase (OCT) and arginase (ARG), as well as glutamine synthetase (GS) were determined for both species after exposure to air. Plasma ammonia increased gradually during exposure to air, but only H. malabaricus showed increased concentrations of urea. Plasma uric acid remained very low in both fish. Enzymatic activities (mean ± SD, æmol min-1 g protein-1) of H. malabaricus showed significant increases (P<0.05, N = 6) in OCT from 0.84 ± 0.05 to 1.42 ± 0.03, in ARG from 8.07 ± 0.47 to 9.97 ± 0.53 and in GS from 1.15 ± 0.03 to 2.39 ± 0.04. The OCT and ARG enzymes remained constant in H. unitaeniatus (N = 6), but GS increased from 1.49 ± 0.02 to 2.06 ± 0.03. Although these species are very closely related and share the same environment, their biochemical strategies in response to exposure to air or to increased plasma ammonia are different

Blood parameters and metabolites in the teleost fish Colossoma macropomum exposed to sulfide or hypoxia.

Juvenile tambaqui, Colossoma macropomum, were exposed to sulfide and hypoxia for 12, 24, 48 and 96 h. Hemoglobin concentrations, red blood cell counts, and mean cell hemoglobin, were higher at 12 h in fish exposed to hypoxia. However, control fish and those exposed to sulfide and hypoxia had lower red blood cell count, hemoglobin concentration and hematocrit at 96 h. Methemoglobin was higher than in the controls, probably due to the hypoxemia induced by these stressors. Sulfhemoglobin was not detected in significant amounts in the blood of fish exposed to sulfide (in vivo), yet hemoglobin converted into sulfhemoglobin at 1-15 mM sulfide in vitro. Anaerobic metabolism seemed to be an important mechanism for adapting to sulfide exposure and blood pH returned to control values after 24 h of sulfide, preventing acidosis. The high sulfide tolerance in tambaqui is associated with its high tolerance to hypoxia.