Hepatic and renal damage by alcohol and cigarette smoking in rats.

Chronic use of alcohol and tobacco cigarettes is associated to millions of deaths per year, either by direct or indirect causes. However, few studies have explored the additional risks of the combined use of these drugs. Here we assessed the effect of the combined use of alcohol and cigarette smoke on liver or kidney morphology, and on biochemical parameters in chronically treated rats. Male Wistar rats were allocated to receive 2 g/kg alcohol orally, which was followed by the inhalation of smoke from six cigarettes during 2 h (ALTB group) for 28 days. Other groups received alcohol alone (AL) or were exposed to cigarette smoke (TB) alone and were compared to control (CT) rats, which received water followed by ambient air. On day 29, rats were euthanized and blood samples were collected for aminotransferase enzymes (AST and ALT), creatinine, and urea analysis. Liver and kidney were weighted, dissected, fixed, and stained with hematoxylin and eosin for morphological analysis. Our results showed that necrosis was elevated in the AL, TB, and mainly the ALTB group in both liver and kidney of rats. Serum levels of AST and ALT were reduced by cigarette smoke exposure, independently of alcohol use. Serum creatinine levels increased after tobacco smoke exposure. On the other hand, TB and AL groups decreased serum urea levels, and their association restored that decrease. Absolute liver and kidney weights were lower in the cigarette smoke exposure rats. Lastly, body weight gain was lower in TB group and combined use restored it. Thus, we may infer that the use of alcohol, exposure to tobacco cigarette smoke or, mainly, their association promotes liver and kidney injuries, and this damage is related with biochemical changes in rats.

Combined Exposure to Alcohol and Tobacco Smoke Changes Oxidative, Inflammatory, and Neurotrophic Parameters in Different Areas of the Brains of Rats.

Devastating effects of exposure to alcohol and tobacco smoke on health are extensively reported in the literature. However, few studies have attempted to elucidate the consequences of their combined use on the central nervous system. Here we studied the effect of this combined use on some oxidative, inflammatory, and neurotrophic parameters in the hippocampus, striatum, and frontal cortex of rats. Adult Wistar rats were allocated into control (CT), alcohol (AL), tobacco smoke (TB), or combined (ALTB) groups. Rats were exposed to environmental air (CT and AL groups) or to the smoke from six cigarettes (TB and ALTB groups) immediately after tap water (CT and TB) or 2 g of alcohol/kg (AL and ALTB) oral gavage administration, twice a day, for 4 weeks. On day 28, rats were euthanized and areas of the brain were dissected to evaluate some cellular redox parameters, pro-inflammatory cytokine levels, and brain-derived neurotrophic factor (BDNF) levels. A one-way analysis of variance showed that the ALTB combined treatment significantly increased oxidative stress levels in the hippocampus. ALTB also increased interleukin-1ß levels in the striatum and frontal cortex and tumoral necrosis factor-α levels in the frontal cortex compared with those of AL, TB, and CT rats. Combined treatment also decreased the BDNF levels in the frontal cortex of rats. Oxidative damage was found, more importantly, in the hippocampus, and inflammatory parameters were extended to all areas of the brain that were studied. Our results showed an interaction between alcohol and tobacco smoke according to the area of the brain, suggesting an additional risk of neural damage in alcoholics who smoke.

Nanoparticle formulation increases Syzygium cumini antioxidant activity in Candida albicans-infected diabetic rats.

CONTEXT: Syzygium cumini (L.) Skeels (Myrtaceae) is a medicinal plant widely used in folk medicine for the treatment of diabetes mellitus (DM). However, studies on the use of this plant and of nanoparticle formulations against DM-related fungal infections are scarce. OBJECTIVE: To evaluate the effect of the treatments with aqueous seed extract of S. cumini (ASc) and ASc-loaded polymeric nanoparticles (NPASc) on biochemical parameters in Candida albicans-infected diabetic rats. MATERIALS AND METHODS: Male Wistar rats were divided into eight groups: Control, DM, C. albicans, C. albicans + ASc, C. albicans + NPASc, DM + C. albicans, DM + C. albicans + ASc and DM + C. albicans + NPASc. Rats were daily treated with ASc or NPASc (100 mg/kg) for 21 days. Biochemical parameters in serum and urine, advanced oxidation protein product (AOPP) and TBARS levels in the serum, kidney, liver and pancreas and N-acetyl-ß-d-glucosaminidase (NAG) activities in kidney and urine were evaluated. RESULTS: Biochemical and oxidative stress parameters increased in rats with DM and/or candidiasis. NPASc was more effective than ASc in decreasing glucose (56%), cholesterol (33%) and creatinine (51%) levels; serum (16%) and pancreatic (46%) AOPP and renal (48%) TBARS levels when compared with DM + C. albicans group. In C. albicans group, both treatments decreased NAG activity but did not decrease creatinine levels. CONCLUSIONS: These data suggest that the use of nanotechnology is able to improve plant extract properties such as antioxidant activity that may be useful in diabetes-related complications.

Extraction, characterization and biological activity of a (1,3)(1,6)-ß-d-glucan from the pathogenic oomycete Pythium insidiosum.

Pythiosis is a life-threatening infectious disease caused by the pathogenic oomycete Pythium insidiosum. This study is the first to evaluate the P. insidiosum glucan content and its biological activities. The enzymatic quantification of the glucans in P. insidiosum mycelia showed that the ß-glucan content was 18.99%±3.59. The cell wall polysaccharide extract consisted of ∼81.7% carbohydrates (exclusively glucose) and ∼18.3% residual amino acids and peptides. The results from monosaccharide composition, methylation and 1D/2D NMR spectroscopy analyses indicated the presence of a highly branched (1,3)(1,6)-ß-d-glucan, with (1,6)-ß-d-glucopyranosil side-branching unit on average every 1-2 repeat units. In vitro, the ß-d-glucan extract could significantly promote spleen lymphocyte proliferation in human, equine and mouse cell cultures. BALB/c mice that were subcutaneously pre-immunized with three doses of 0.5, 2.5 and 5.0mg of ß-glucan/mouse, showed a significant increase in IL-2, IL-6, IL-10, TNF-α and IL-17A production compared to non-immunized mice. These results suggested that ß-d-glucan extract induces significant and specific Th17 cellular immune response and provided the theoretical basis for further experiments.

Complex interaction of deferasirox and Pythium insidiosum: iron-dependent attenuation of growth in vitro and immunotherapy-like enhancement of immune responses in vivo.

Pythium insidiosum iron acquisition mechanisms are unknown. We previously showed that the iron chelator deferasirox had weak activity in vitro and in rabbits with experimental pythiosis. Here we show that deferasirox causes damage to P. insidiosum hyphae in vitro, but that activity is diminished in the presence of exogenous iron. The tissue activity of the proinflammatory enzyme adenosine deaminase and the histological pattern observed in pythiosis lesions of rabbits treated with deferasirox were similar to the ones in animals treated with immunotherapy.

Syzygium cumini is more effective in preventing the increase of erythrocytic ADA activity than phenolic compounds under hyperglycemic conditions in vitro.

Syzygium cumini (S. cumini) is a plant known for its antidiabetic properties. The aim of this study was to evaluate the effect of Sc aqueous leaf extract (ASc) on adenosine deaminase (ADA) activity in erythrocytes (RBCs) exposed to high glucose concentrations (30 mM) in vitro. We also investigated the effects of the main phenolic compounds found in ASc (gallic acid, rutin, and chlorogenic acid) and the effects of insulin, caffeine, and dipyridamole, which are substances involved in the adenosine metabolism, on ADA activity in vitro. Blood samples were obtained from healthy volunteers and a suspension of RBCs was used for the determination of ADA activity. The results showed that: (1) the effect of ASc on ADA activity was more significant than the combination of phenolic compounds; (2) insulin, caffeine, or dipyridamole prevented high glucose increase of ADA activity at doses as low as 50 µU/mL, 25 µM, and 1 µM, respectively; (3) the inhibitory effect caused by ASc on erythrocyte ADA activity remained practically the same after the combination of the extract with insulin or caffeine; (4) when RBCs were exposed to ASc plus dipyridamole, this chemical attenuated the effect of ASc on ADA activity, suggesting an antagonism or a competition with ASc by the same site of action. Therefore, ASc was more effective in preventing the increase in ADA activity than phenolic compounds, suggesting that ASc may collaborate to improve endothelial dysfunction, antioxidant, anti-inflammatory, and antithrombotic properties of adenosine by affecting its metabolism. The results of this study help to provide evidence of the empirically supported benefits of the use of S. cumini in diabetes.

Butyrylcholinesterase and γ-glutamyltransferase activities and oxidative stress markers are altered in metabolic syndrome, but are not affected by body mass index.

Metabolic syndrome (MetS) leads to changes in enzymatic activities, oxidative and inflammatory parameters. Adenosine deaminase (ADA), dipeptidyl peptidase IV (DPP-IV), butyrylcholinesterase (BuChE) and γ-glutamyltransferase (γ-GT) activities, C-reactive protein (hsCRP) and nitric oxide levels (NOx), as well as oxidative stress markers were analyzed in 39 subjects with MetS and 48 controls. Also, the influence of body mass index (BMI) and anthropometric measurements were evaluated. Disturbances in antioxidant defenses and higher γ-GT and BuChE activities, NOx and hsCRP levels were observed in subjects with MetS. These findings remained associated with MetS after adjustment for BMI, except for hsCRP. ADA was correlated with age, insulin levels and HOMA-IR index in MetS. DPP-IV and total cholesterol (TC), BuChE activity and TC, and VIT C and hsCRP levels also were correlated. The analyzed parameters may reflect the inflammatory state of the MetS, and could contribute to prevention and control of various aspects of this syndrome.

Exposição aguda ao Metilmercúrio em ratos em desenvolvimento: mini-revisão / Acute exposition of developing rats to Methylmercury: a mini-review

Mercúrio (Hg) está presente no ambiente em três diferentes formas químicas: elementar (Hg0), inorgânico e orgânico, sendo que a sua distribuição, toxicidade e metabolismo são dependentes de sua forma química. A exposição oral ao consumo de peixes e alimentos contaminados é a principal forma de exposição humana ao metilmercúrio (MeHg), um poluente ambiental que é absorvido por ingestão, inalação e através da pele. O MeHg é um potente neurotóxico, especialmente para o cérebro em desenvolvimento. Neste estudo, foram examinados os principais efeitos da exposição ao MeHg durante o desenvolvimento salientando os mecanismos bioquímicos envolvidos nestes processos. Também foram apresentados recentes resultados sobre o uso de extratos de plantas medicinais que atenuaram os efeitos adversos deste metal. Deste modo, estes dados reforçam a toxicidade do MeHg durante o desenvolvimento e sugerem possíveis caminhos para futuras intervenções terapêuticas.

Mercury (Hg) is present in the environment in three different chemical forms: elemental, inorganic and organic Hg. The distribution, toxicity and metabolism of Hg are linked to its chemical form. The oral exposition following fish and food contaminated is the main route of contamination of humans to the methilmercury (MeHg), an environmental pollutant which is absorbed by ingestion, inhalation and through the skin. MeHg is a strong neurotoxic molecule, especially for the developing brain. In this study, the main effects of the MeHg exposition and the biochemical parameters involved in this process were examined. The results of the use of plant extracts which attenuate the adverse effects of this metal are also presented here. Therefore, these data reinforce the MeHg toxicity during the development and suggest alternative ways for future therapeutic approaches.

Lymphocytic enzymes and lipid peroxidation in patients with metabolic syndrome.

OBJECTIVES: Metabolic syndrome (MetS) is considered a state of chronic inflammation. This study aimed to ascertain selected parameters of purinergic and cholinergic systems related to glucose metabolism and inflammation, as well as (γ)-glutamyltransferase (GGT) and N-acetyl-b-glucosaminidase (NAG) activities and lipoperoxidation in lymphocytes of patients with MetS. DESIGN AND METHODS: The adenosine deaminase (ADA), dipeptidyl peptidase IV (DPP-IV), acetylcholinesterase (AChE), GGT and NAG activities, as well as thiobarbituric acid reactive substances (TBARS) levels were investigated in lymphocytes of patients with MetS (n=38) and healthy volunteers (n=41). We also evaluated the insulin levels, anthropometric measurements and routine biochemical analyses. RESULTS: ADA (p<0.05), DPP-IV and AChE (p<0.0001) activities were higher in patients with MetS when compared to the control group. Furthermore, we observed correlations between ADA and DPP-IV activities (p=0.0002; r=0.5945), TBARS levels and ADA (p=0.0021; r=0.5172) and DPP-IV activities (p=0.0022; r=0.5010). CONCLUSIONS: Our findings showed that MetS might cause tissue distress that disturbed lymphocytic ADA, DPP-IV and AChE activities in response to inflammatory stimuli. These alterations evidence clinical abnormalities, since these enzymatic systems are able to regulate several aspects of adipose tissue function and inflammatory state of MetS and could be used successfully both for preventing and for halting the progression of MetS.

Activities of adenine nucleotide and nucleoside degradation enzymes in platelets of rats infected by Trypanosoma evansi.

Nucleotide and nucleoside-degrading enzymes, such as nucleoside triphosphate diphosphohydrose (NTPDase), 5'-nucleotidase and adenosine deaminase (ADA) are present in the surface membranes of platelets, involved in clotting disturbances of Trypanosoma evansi-infected animals. Thus, this study was aimed at evaluating the activities of these enzymes in platelets of rats experimentally infected with T. evansi. Animals were divided into four groups, according to the level of parasitemia. Blood samples were collected on days 3 (group A: at the beginning of parasitemia), 5 (group B: high parasitemia) and 15 (group C: chronic infection), post-infection. Group D (control group) was composed of non-infected animals for platelet count, separation and enzymatic assays. Animals from groups A and B showed marked thrombocytopenia, but platelet count was not affected in chronically infected rats. NTPDase, 5'-nucleotidase and ADA activities decreased (p<0.05) in platelets from rats of groups A and B, when compared to the control group. In group C, only NTPDase and 5'-nucleoside activities decreased (p<0.001). The correlations between platelet count and nucleotide/nucleoside hydrolysis were positive and statistically significant (p<0.05) in groups A and B. Platelet aggregation was decreased in all infected groups, in comparison to the control group (p<0.05). It is concluded that the alterations observed in the activities of NTPDase, 5'-nucleotidase and ADA in platelets of T. evansi-infected animals might be related to thrombocytopenia, that by reducing the number of platelets, there was less release of ATP and ADP. Another possibility being suggested is that changes have occurred in the membrane of these cells, decreasing the expression of these enzymes in the cell membrane.

Activity of the enzyme adenosine deaminase in serum, erythrocytes and lymphocytes of rats infected with Trypanosoma evansi.

In Trypanosoma evansi infections changes in the haemogram are commonly observed, and the enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. Thus, the aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with T. evansi compared to non-infected rats. Thirty adult rats were used, divided into 3 uniform groups. The animals in groups A and B were infected intraperitoneally with 2 x 106 trypomastigotes/rat. Rodents from group C (control group), were not-infected. Blood collection was performed on days 4 and 20 post-infection (p.i.) in order to obtain acute and chronic infection stages of disease. The blood was used to assess the activity of ADA. In the blood, reduced haematocrit and increased lymphocytes were correlated with ADA activity in erythrocytes and lymphocytes. We observed reduction of ADA activity in serum and erythrocytes in rats infected with T. evansi compared to non-infected rats (P < 0.05). ADA activity in lymphocytes was decreased after 4 days, when the parasitaemia was high and increased after 20 days, when the number of circulating parasites was low. In conclusion, our results showed that the ADA activity was altered in serum, lymphocytes and erythrocytes of rats, concomitantly with haematological parameters, in experimental infection by T. evansi.

Trypanosoma evansi: adenosine deaminase activity in the brain of infected rats.

The study was undertaken to evaluate changes in the activity of adenosine deaminase (ADA) in brains of rats infected by Trypanosoma evansi. Each rat was intraperitoneally infected with 10(6) trypomastigotes either suspended in fresh (group A; n = 13) and cryopreserved blood (group B; n = 13). Thirteen animals were used as control (group C). ADA activity was estimated in the cerebellum, cerebral cortex, striatum and hippocampus. No differences (P > 0.05) in ADA activity were observed in the cerebellum between infected and non-infected animals. Significant (P < 0.05) reductions in ADA activity occurred in cerebral cortex in acutely (day 4 post-infection; PI) and chronically (day 20 PI) infected rats. ADA activity was significantly (P < 0.05) decreased in the hippocampus in acutely infected rats, but significantly (P < 0.05) increased in the chronically infected rats. Significant (P < 0.05) reductions in ADA activity occurred in the striatum of chronically infected rats. Parasites could be found in peripheral blood and brain tissue through microscopic examination and PCR assay, respectively, in acutely and chronically infected rats. The reduction of ADA activity in the brain was associated with high levels of parasitemia and anemia in acute infections. Alterations in ADA activity of the brain in T. evansi-infected rats may have implications for pathogenesis of the disease.