Effects of oxidation of lysozyme by hypohalous acids and haloamines on enzymatic activity and aggregation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Assessment of estrogenic, mutagenic and antimutagenic activity of nemorosone

Currently, a wide range of research involving natural products is focused on the discovery of new drugs in many different therapeutic areas. A great number of the synthetic compounds on the market were derived from natural products, especially plants. Nemorosone is the major constituent of the floral resin of Clusia rosea Jacq., Clusiaceae, and in Cuban propolis. In vitro studies have shown cytotoxic activity in this substance against various tumor cell lines, including those resistant to various cytotoxic drugs, whereas it has low cytotoxicity to non-tumoral cells. Therefore, in order to characterize the biological activity of nemorosone, a substance with potential antitumor activity, and in view of preclinical testing of the toxicity of drug candidate compounds, the main aim of this study was to determine the mutagenic and antimutagenic activity of nemorosone by the Ames test, using the strains TA97a, TA98, TA100 and TA102 of Salmonella typhimurium. Secondly, to characterize the estrogenic activity in an experimental recombinant yeast model (Recombinant Yeast Assay) mutagenic activity was observed at in any of the concentrations in any of the test strains. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine (NPD), mitomycin C (MMC) and aflatoxin B1 (AFL). Nemorosone showed moderate antimutagenic activity (inhibition level 31%), in strain TA100 in the presence of AFL, and strong antimutagenic activity in TA102 against MMC (inhibition level 53%). Estrogenic activity was observed, with an EEq of 0.41±0.16 nM at various tested concentrations.

Improvement of antischistosomal activity of praziquantel by incorporation into phosphatidylcholine-containing liposomes

Praziquantel (PZQ) is effective against all known species of Schistosomes that infect humans. The failure of mass treatment of schistosomiasis has been attributed to the fact that therapy is not sufficiently long-lasting. This effect may be due to the low bioavailability of PZQ that has a low hydrosolubility and fast metabolism. Liposomes have been used to prolong drug levels. reduce the side effects, direct drugs to specific sites and increase bioavailability after administration. The aim of this work was to study the effect of phosphatidylcholine (PC)-containing liposomes to vehiculate PZQ to improve the treatment of schistosomiasis. The in vitro Study was carried out using Schistosoma mansoni parasites recovered by perfusion from the hepatic portal system of infected mice. Suspensions of liposomes with PZQ and free PZQ were administered p.o. in mice after 14 days of infection. The effect of both preparations in vitro on S. mansoni culture was similar. In the in vivo test, PZQ-liposomes caused a decrease in amounts of eggs and parasites. Liposomes improve the antischistosomal activity of praziquantel. This can be used as a starting point to investigate alternative administration routes or dosage forms and to examine the mechanism of intestinal absorption of PRZ © 2005 Elsevier B.V. All rights reserved.

Experimental murine mycobacteriosis: evaluation of the functional activity of alveolar macrophages in thalidomide- treated mice

Thalidomide is a selective inhibitor of tumor necrosis factor-alpha (TNF-alpha), a cytokine involved in mycobacterial death mechanisms. We investigated the role of this drug in the functional activity of alveolar macrophages in the presence of infection induced by intranasal inoculation of Mycobacterium avium in thalidomide-treated and untreated adult Swiss mice. Sixty animals were inoculated with 5 x 10(6) M. avium by the respiratory route. Thirty animals received daily thalidomide (30 mg/kg mouse) and 30 received water by gavage up to sacrifice. Ten non-inoculated mice were used as a control group. Lots of animals from each group were evaluated until 6 weeks after inoculation. Infection resulted in an increased total number of inflammatory cells as well as increased activity of pulmonary macrophages. Histologically, intranasal inoculation of bacilli resulted in small mononuclear infiltrates located at the periphery of the organ. Culture of lung fragments revealed the presence of bacilli only at the beginning and at the end of the experimental period. Thalidomide administration did not affect the microbiological or histological features of the infection. Thalidomide-treated and untreated animals showed the same amount of M. avium colonies 3 weeks after infection. Although it did not affect bacillary clearance, thalidomide administration resulted in a decreased percent of spread cells and release of hydrogen peroxide, suggesting that factors other than TNF-alpha play a role in the killing of mycobacteria by alveolar macrophages. Thalidomide administration also reduced the number of spread cells among resident macrophages, suggesting a direct effect of the drug on this phenomenon.

Targeted Nucleotide Exchange in Bovine Myostatin Gene

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of pH, available chlorine content, and antibacterial activity of endodontic irrigants and their combinations against Enterococcus faecalis

Objectives. The objectives of this study were to evaluate pH, available chlorine content, and antibacterial activity of endodontic irrigants and their combinations.Study design. The pH and chlorine content of sodium hypochlorite (NaOCl) were analyzed pure and in combination with 10% citric acid (CA) and apple vinegar (AV). The antibacterial effect of the following solutions was measured by direct contact test against Enterococcus faecalis: 2.5% NaOCl, 2.5% NaOCl + 10% CA (7:3), 2.5% NaOCl + AV (5:5), 10% CA, and AV. Sterile saline was used as control. The colony-forming units were determined by serial decimal dilutions.Results. The combination of 2.5% NaOCl with CA or AV lowered the pH and the chlorine content. NaOCl, alone or in combination was able to eliminate E. faecalis in 30 seconds, and CA, after 10 minutes. AV promoted reduction (32.2%) after 10 minutes.Conclusions. NaOCl with acidic solutions lowered the pH and the chlorine content, but did not alter its antibacterial effect. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011; 112:132-135)

Geographic variations in the composition of myotoxins from Bothrops neuwiedi snake venoms: biochemical characterization and biological activity

(1) Venom pools from Bothrops neuwiedi (Bn) and from two subspecies, namely Bothrops neuwiedi pauloensis (Bnp) and Bothrops neuwiedi urutu (Bnu), collected in the States of São Paulo (SP) and Minas Gerais (MG), Brazil, were electrophoretically examined. Basic toxins with different isoelectric points were identified in the venom collected in São Paulo (BnSP). These toxins were absent in the corresponding pools from Minas Gerais (BnMG, BnpMG and BnuMG). (2) BnSP, but not BnMG, BnpMG or BnuMG, showed two myotoxins (pI congruent to 8.6 and 8.8, respectively) which were isolated by ion-exchange chromatography on CM-Sepharose. (3) From BnMG, three myotoxic isoforms (pI congruent to 8.2 and M-r = 13600) were isolated by chromatography on CM-Sepharose followed by reversed-phase high-performance liquid chromatography. (4) the chemical and biological characterization of these toxins showed a high similarity with the Lys-49 myotoxins from other bothropic venoms. (5) Doses up to 5 LD50 (i.p.) of p-bromophenacyl bromide alkylated BnSP-7 caused a total loss of lethality in 18-22-g mice, thus indicating that the LD50 was increased by greater than 5-fold. At this dose myotoxicity was also not detectable, but the edematogenic activity on the rat paw apparently did not change. (C) 1998 Elsevier B.V. All rights reserved.

Leishmania amazonensis: Partial purification and study of the biochemical properties of the telomerase reverse transcriptase activity from promastigote-stage

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental paracoccidioidomycosis of the Syrian hamster: fungicidal activity and production of inflammatory cytokines by macrophages

Phagocytic cells play an important role in nonspecific resistance to fungal infection by mediating an inflammatory response and by a direct fungicidal action. In this study, the functional activity of peritoneal macrophages obtained from hamsters experimentally infected with strain Pb18 of Paracoccidioides brasiliensis was evaluated during 16 weeks of infection. The results showed that macrophages had a higher spreading ability associated with increased production of tumor necrosis factor alpha (TNF-alpha) and enhanced fungicidal activity during the early periods of infection. TNF-alpha levels remained elevated during all periods studied, while low levels of interleukin-1 beta (IL-1 beta) were produced during the infection. A necrotic area with dead fungi was observed at the inoculation site and the infection disseminated only to liver and lymph nodes in a few animals. These results suggest that during the early stages of infection with P. brasiliensis, macrophage activation by the high levels of TNF-alpha limited fungal dissemination. In contrast, in the later stages of infection, high levels of TNF-alpha were observed while the fungicidal activity of macrophages was lower and the animals presented loss of vitality resulting in their death. These observations suggest a complex role of TNF-alpha in experimental paracoccidioidomycosis of Syrian hamsters, involving not only resistance but also pathogenesis.

Tobacco Smoke Induces Ventricular Remodeling Associated with an Increase in NADPH Oxidase Activity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ultraviolet-B and water stress effects on growth, gas exchange and oxidative stress in sunflower plants

The effects and interaction of drought and UV-B radiation were studied in sunflower plants (Helianthus annuus L. var. Catissol-01), growing in a greenhouse under natural photoperiod conditions. The plants received approximately 1.7 W m(-2) (controls) or 8.6 W m(-2) (+UV-B) of UV-B radiation for 7 h per day. The UV-B and water stress treatments started 18 days after sowing. After a period of 12 days of stress, half of the water-stressed plants (including both UV-B irradiated or non-irradiated) were rehydrated. Both drought and UV-B radiation treatments resulted in lower shoot dry matter per plant, but there was no significant interaction between the two treatments. Water stress and UV-B radiation reduced photosynthesis, stomatal conductance and transpiration. However, the amplitude of the effects of both stressors was dependent on the interactions. This resulted in alleviation of the negative effect of drought on photosynthesis and transpiration by UV-B radiation as the water stress intensified. Intercelluar CO(2) concentration was initially reduced in all treatments compared to control plants but it increased with time. Photosynthetic pigments were not affected by UV-B radiation. Water stress reduced photosynthetic pigments only under high UV-B radiation. The decrease was more accentuated for chlorophyll a than for chlorophyll b. As a measure for the maximum efficiency of photosystem II in darkness F (v)/F (m) was used, which was not affected by drought stress but initially reduced by UV-B radiation. Independent of water supply, UV-B radiation increased the activity of pirogalol peroxidase and did not increase the level of malondialdehyde. on the other hand, water stress did not alter the activity of pirogalol peroxidase and caused membrane damage as assessed by lipid peroxidation. The application of UV-B radiation together with drought seemed to have a protective effect by lowering the intensity of lipid peroxidation caused by water stress. The content of proline was not affected by UV-B radiation but was increased by water stress under both low and high UV-B radiation. After 24 h of rehydration, most of the parameters analyzed recovered to the same level as the unstressed plants.

Acaricidal activity of ethanolic extract from aerial parts of Tagetes patula L. (Asteraceae) against larvae and engorged adult females of Rhipicephalus sanguineus (Latreille, 1806)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Gas exchange and ventilation during dormancy in the tegu lizard Tupinambis merianae

The tegu lizard Tupinambis merianae exhibits an episodic ventilatory pattern when dormant at 17 degrees C but a uniform ventilatory pattern when dormant at 25 degrees C. At 17 degrees C, ventilatory episodes were composed of 1-22 breaths interspaced by non-ventilatory periods lasting 1.8-26min, Dormancy at the higher body temperature was accompanied by higher rates of O-2 consumption and ventilation. The increase in ventilation was due only to increases in breathing frequency with no change observed in tidal volume. The air convection requirement for O-2 did not differ at the two body temperatures. The respiratory quotient was 0.8 at 17 degrees C and 1.0 at 25 degrees C. We found no consistent relationship between expired gas composition and the start/end of the ventilatory period during episodic breathing at 17 degrees C. However, following non-ventilatory periods of increasing duration, there was an increase in the pulmonary O-2 extraction that was not coupled to an equivalent increase in elimination of CO2 from the lungs. None of the changes in the variables studied could alone explain the initiation/termination of episodic ventilation in the tegus, suggesting that breathing episodes are shaped by a complex interaction between many variables. The estimated oxidative cost of breathing in dormant tegus at 17 degrees C was equivalent to 52.3% of the total metabolic rate, indicating that breathing is the most costly activity during dormancy.

Antioxidant activity, fatty acid profile and tocopherols of Tamarindus indica L. seeds

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Preparation and antibacterial activity of silver nanoparticles impregnated in bacterial cellulose

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)