Genetic differentiation among ten populations of the genus Neoplecostomus (Teleostei: Loricariidae) from the upper Parana River basin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of photodynamic therapy on clinical isolates of Staphylococcus spp

Staphylococcus spp. are opportunistic microorganisms known for their capacity to develop resistance against antimicrobial agents. The objective of this study was to evaluate the effect of photodynamic therapy (PDT) on 20 Staphylococcus strains isolated from the human oral cavity, including S. aureus, S. schleiferi, S. epidermidis, S. capitis, S. haemolyticus, and S. lentus. A suspension of each Staphylococcus strain (10(6) cells/mL) was submitted to PDT using methylene blue and a low power laser. The isolated effects of methylene blue, laser treatment and ciprofloxacin were also evaluated. After the experimental treatments, 0.1 mL aliquots of the suspensions were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were analyzed by analysis of variance and Tukey's test (p < 0.05). The mean reduction in bacterial counts of the strains submitted to PDT ranged from 4.89 to 6.83 CFU (log10)/mL, with the observation of a decreasing susceptibility to treatment of S. schleiferi, S. haemolyticus, S. epidermidis, S. capitis, S. aureus, and S. lentus. The results showed that PDT was effective in reducing the number of viable cells of all clinical Staphylococcus isolates studied.

Effects of Chamomilla recutita (L.) on oral wound healing in rats

Objectives. The purpose of this study was to evaluate the effect of Chamomilla recutita on the healing of ulcers in rats.Study Design. A 5-mm wound was inflicted on the tongue of 36 rats. Treatment group animals were treated topically with 0.04 mL/day of chamomile ointment, whereas control group animals were not treated. Animals were sacrificed after 3, 7 or 10 days. Semi-quantitative analysis of the degree of inflammation, fibroblast count and wound size was performed, as well as histometric analysis of re-epithelialization and percentage of collagen fibers of the lesion.Results. Animals treated with chamomile showed the best results regarding epithelialization and percentage of collagen fibers after 10 days. As expected, time had a statistically significant effect (p<0.05) on fibroblast count, epithelialization, inflammation and wound size; animals sacrificed at 3 days showed the worst results.Conclusions. Chamomile stimulated re-epithelialization and the formation of collagen fibers after 10 days of treatment; it did not, however, influence inflammation or fibroblast count.

Influence of artificial saliva in biofilm formation of Candida albicans in vitro

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

Vinegar as an antimicrobial agent for control of Candida spp. in complete denture wearers

The use of denture is known to increase the carriage of Candida in healthy patients, and the proliferation of Candida albicans strains can be associated with denture-induced stomatitis. The aim of this study was to evaluate the use of vinegar as an antimicrobial agent for control of Candida spp. in complete upper denture wearers. Fifty-five patients were submitted to a detailed clinical interview and oral clinical examination, and were instructed to keep their dentures immersed in a 10% vinegar solution ( pH less than 3) overnight for 45 days. Before and after the experimental period, saliva samples were collected for detection of Candida, counting of cfu/mL and identification of species by phenotypical tests ( germ tube formation, chlamidoconidia production, and carbohydrate fermentation and assimilation). The results were analyzed using Spearman's correlation and Student's t-test (p=0.05). Candida yeasts were present in 87.3% of saliva samples before the treatment. A significant reduction was verified in CFU/mL counts of Candida after treatment. A positive correlation between Candida and denture stomatitis was verified, since the decrease of cfu/mL counts was correlated with a reduction in cases of denture stomatitis. Although it was not able to eliminate C. albicans, the immersion of the complete denture in 10% vinegar solution, during the night, reduced the amounts (cfu/mL) of Candida spp. in the saliva and the presence of denture stomatitis in the studied patients.

Potential of non-pathogenic Fusarium oxysporum isolates for control of Fusarium wilt of tomato

O trabalho avaliou a eficiência dos isolados (141/3, 233, 233/1, 245, 245/1, 251, 251/2, 251/5 e 257) de Fusarium oxysporum não patogênico ao tomateiro (Lycopersicon esculentum), no controle da murcha vascular causada por Fusarium oxysporum f. sp. lycopersici, raça 2 em plântulas de tomateiro cv. Viradoro. Para verificar o efeito dos isolados de F. oxysporum não patogênicos, o sistema radicular de plântulas de tomateiro, com 30 dias de idade, foi imerso na suspensão de conídios (10(6) ml-1) e as mudas transplantadas para substrato de cultivo. Após 35 dias do transplante foi verificado que esses isolados não foram patogênicos às plantas de tomateiro, nem afetaram o desenvolvimento das mudas. A eficiência dos isolados de Fusarium oxysporum não patogênicos no controle da murcha foi determinada imergindo-se as raízes de mudas de tomateiro em suspensão de conídios (10(6) conídios ml-1) e transplantando-as em substratos previamente infestados com os isolados de F. oxysporum f.sp. lycopersici, raça 2 (10(5) conídios ml-1 de substrato). Transcorridos 35 dias do transplante, foram realizadas as avaliações da severidade na escala de 1=planta sadia a 6=planta morta ou com vasos coloridos e folhas murchas até o ponteiro e altura das mudas. Os isolados de F. oxysporum não patogênicos foram eficientes em reduzir a severidade da doença e em manter normal o seu desenvolvimento. Esses resultados evidenciam a atividade antagônica dos isolados de F. oxysporum não patogênico no controle da murcha vascular do tomateiro, causada por Fusarium oxysporum f. sp. lycopersici raça 2.

Effects of acetylsalicylic acid and acetic acid solutions on VX2 liver carcinoma in rabbits: in vivo analysis

OBJETIVO: Analisar os efeitos das soluções de aspirina e de ácido acético, in vivo, em fígado de coelhos portadores de tumor hepático VX2, verificando o efeito histolítico e anatomo-patológico das soluções e eventuais alterações bioquímicas hepáticas. MÉTODOS: Utilizou-se 48 coelhos, divididos em 2 protocolos experimentais(3 e 4), subdivididos em 3 grupos cada. Após 4 dias da implantação do tumor no fígado, procedeu-se a laparotomia mediana, com injeção de 0,4 ml da solução de aspirina (5,0%), de ácido acético (5,0%) e solução salina; o sacrifício ocorreu apos 24 horas (protocolo 3) e 11 dias (protocolo 4); avaliou-se o peso, evolução clinica, dosagens bioquímicas, cavidade abdominal e torácica e microscopia do fígado. RESULTADOS: Não foram observadas alterações na evolução clinica, peso e nas dosagens bioquímicas, apenas elevação da fosfatase alcalina no grupo controle do protocolo 4. Observamos desaparecimento do tumor em ambos os protocolos. CONCLUSÃO: As soluções de ácido acético e ácido acetilsalicílico acarretam destruição do tumor hepático experimental.

Electrochemical noise analysis of chalcopyrite carbon paste electrodes by Acidithiobacillus ferrooxidans

The electrochemical response of chalcopyrite was studied using electrochemical noise analysis (ENA). The assay was carried out under constant aeration using 30 mL in two electrochemical cells containing iron-free mineral salts solution. These cells were initially monitored for 56 hours, After 72 hours, 7.25x 10(10) cells mL(-1) of A, ferrooxidans strain LR were added in both cells and monitored until 128 h. Subsequent to this period, 0.927 mmol L-1 of silver ions and 400 mmol L-1 of chloride ions were added each one separately. Both conditions were monitored until 168 hours. According to results obtained, it was observed that Cl- ions addition induced an accelerated corrosion process. However, there is a tendency of the system to reach the stationary state due to repassivation of the electrodic surface. In the other side, the Ag+ addition contributed for the maintenance of the oxidant atmosphere, in spite of controversial effect caused by considerable variations in the R-n values, resulting in a instability in the chalcopyrite reactivity.

Comparative studies of the anti-leishmanial activity of three Crotalus durissus ssp venoms

In this study, we compared the anti-leishmanial activity of three crotalic venoms (Crotalus durissus terrificus-Cdt, Crotalus durissus cascavella-Cdca, and Crotalus durissus collilineatus-Cdcol). Different concentrations of each venom incubated with Leishmania (Leishmania) amazonensis promastigotes were used. Cdt venom exhibited a higher anti-leishmanial activity (Inhibitory concentration-IC50-value of 4.70 +/- 1.72 mu g/ml) in comparison with that of Cdca venom (IC50 value of 9.41 +/- 1.21 mu g/ml), while Cdcol venom increased parasite numbers in 50% at a concentration of 44.30 +/- 2.18 mu g/ml. In addition, this venom showed a low anti-leishmanial activity in higher concentrations (IC50 value of 281.00 +/- 9.50 mu g/ml). The main fractions of Cdca venom were isolated and assayed under similar conditions used for assessing crude venom. The most active fractions were gyroxin and crotamine that had IC50 values of 3.80 +/- 0.52 mu g/ml and 19.95 +/- 4.21 mu g/ml, respectively. Convulxin also inhibited parasite growth rate, although this effect was not dose-dependent. Crotoxin was the least effective fraction with an IC50 value of 99.80 +/- 2.21 mu g/ml. None of the protein fractions presented cytotoxic effects against J774 cells in culture. In vivo assays using BALB/c mice revealed that crotoxin and crotamine were the main toxic fractions. In conclusion, C. durissus cascavella venom has three main fractions with anti-leishmanial activity. These results open new possibilities to find proteins that might be used as possible agents against cutaneous leishmaniasis.

ANTINOCICEPTIVE EFFECT OF AMITRAZ IN MICE AND RATS

Amitraz, a formamidine insecticide and acaricide used in veterinary practice, presents side effects related to its pharmacological activity on az-adrenergic receptors. The present study was undertaken to investigate the antinociceptive effect of amitraz in rats and mice. The tail-flick test was used to determine the duration of the antinociceptive effect of the intraperitoneal tip) administration of amitraz (1 and 2 mg/kg, 10 animals per group) in male Wistar rats weighing 180-220 g. The writhing test (using 10 ml/kg of a 0.6% acetic acid solution as a painful stimulus). was performed in 140 male Swiss mice weighing 20-30 g, divided into 14 groups that received ip injections of saline, amitraz (0.5, 1.0, 1.5, 2.0 and 4.0 mg/kg), xylazine or detomidine (1.0, 1.5, 2.0 and 4.0 mg/ kg), in order to compare the effect of amitraz to that caused by xylazine and detomidine, and to investigate the participation of alpha(2)-adrenergic receptors which were blocked by idazoxan (1 mg/kg). Amitraz induced a significant antinociceptive effect in both rats and mice. This effect is blocked in mice by idazoxan.

Partial purification and characterization of pectin methylesterase from acerola (Malpighia glabra L.)

The enzyme pectin methylesterase (PME) is present in acerola fruit and was partially purified by gel filtration on Sephadex G-100. The results of gel filtration showed different PME isoforms. The total PME (precipitated by 70% salt saturation) and one of these isoforms (fraction from Sephadex G-100 elution) that showed a molecular mass of 15.5 +/- 1.0 kDa were studied. The optimum pH values of both forms were 9.0. The total and the partially purified PME showed that PME specific activity increases with temperature, the total acerola PME retained 13.5% of its specific activity after 90 min of incubation at 98 degreesC. The partially purified acerola (PME isoform) showed 125.5% of its specific activity after 90 min of incubation at 98 degreesC. The K-m values of the total PME and the partially purified PME isoform were 0.081 and 0.12 mg/mL, respectively. The V-max values of the total PME and the partially purified PME were 2.92 and 6.21 mumol/min/mL/mg of protein, respectively.

Electrochemical study of hydroxychloroquine and its determination in plaquenil by differential pulse voltammetry

Hydroxychloroquine (HCQ) is a halogenated aminoquinoline that presents wide biological activity, often being used as an antimalarial drug. The electrochemical reduction of HCQ was investigated by cyclic voltammetry and chronoamperometry using glassy carbon electrodes. By cyclic voltammetry, in acid medium, only the cathodic peak was observed. The electrochemical behavior of this peak is dependent on pH and the electrodic process occurs through an ErCi mechanism. The electron number (le) consumed in the reduction of HCQ was obtained by chronoamperometry. A method for the electrochemical determination of HCQ in pharmaceutical tablets was developed using differential pulse voltammetry. The detection limit reached was 11.2 mug ml(-1) of HCQ with a relative standard deviation of 0.46%. A spectrophotometric study of HCQ has been also carried out utilizing a band at 343 nm. The obtained detection limit and the relative standard deviation were 0.1 mug ml(-1) and 0.36%, respectively. The electrochemical methods are sufficiently accurate and precise to be applied for HCQ determination, in laboratorial routine, which can be used to determine the drug at low level. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Release of intermediate reactive hydrogen peroxide by macrophage cells activated by natural products

By determining the hydrogen peroxide (H2O2) released in cultures of peritoneal macrophage cells from Swiss mice, we evaluated the action of 27 vegetable compounds (pristimerin, tingenone, jatrophone, palustric acid, lupeol, cladrastin, ocoteine, boldine, tomatine, yohimbine, reserpine, escopoletin, esculine, plumericin, diosgenin, deoxyschizandrin, p-arbutin, mangiferin, and others) using a 2 mg/ml solution of each compound (100 mug/well). Macrophages are cells responsible for the development of the immunological response reaction, liberating more than one hundred compounds into the extracellular environment. Among these are the various cytokines and the intermediate compounds of nitrogen (NO) and oxygen (H2O2). This coordinated sequence of biochemical reactions is known as the oxidative burst. When we compared the results with those obtained with zymosan (an important stimulator of H2O2) we observed that the compounds showing the highest activity were substances 2 (tingenone), 16 (reserpine) and 20. Other substances such as compounds 1, 4, 5, 6, 8, 12, 13, 14, 15, 17, 19, 23, 24, 26, and 27 also showed a certain activity, but with less intensity than the aforementioned ones. Compounds 3, 7, 9, 10, 11, 18, 21, 22 and 25 presented no activity. These results suggest that natural products (mainly tingenone and reserpine and others) with different chemical structures are strong immunological modulators. However, further tests are needed to determine the 'oxidative burst' in future studies.

Effect of porcine reproductive and respiratory syndrome virus on subsequent Pasteurella multocida challenge in pigs

This trial was conducted to evaluate the effect of Porcine reproductive and respiratory syndrome virus (PRRSv) on a subsequent challenge with Pasteurella multocida in pigs. Sixteen, 3-4 week-old piglets, from a PRRSv and Aujeszky disease virus (ADV) free herd were used. Animals were equally and randomly allocated in four groups which were treated according the following schedule: Group I: negative controls; Group II: inoculation with only PRRSV; Group III: inoculation with PRRSV and P. multocida; Group IV: inoculation with ADV and multocida (positive controls), PRRSV and ADV were inoculated intranasally, at the doses of 10(4.6) and 10(4.5) TCID50/ml, respectively. Five days later, pigs from groups III and IV were inoculated intranasally, with two ml of a 10(9) CFU/mL suspension of equal parts of P. multocida, strains A52 and A24. No lesions were observed in piglets of group I. Microscopically, interstitial pneumonia was identified in all piglets of groups II and III and 3/4 piglets from group IV. Bronchopneumonia was detected in 3/4 of the piglets from group III and in all animals of group TV which, additionally, showed meningo-encephalitis and purulent rhinitis. Macroscopically, only piglets of groups III and IV had lung consolidation. However, much lower pneumonic scores (2.3%) were observed in group III, where 3 of 4 piglets were affected. on the other hand, all piglets of group IV showed some degree of pulmonary consolidation, with a mean score of 13.7%. Based on these results, it appears that the role of PRRSV as a initiator of secondary diseases is still undefined, but is probably mild, There was no clear interaction between PRRSV and Pasteurella multocida under the conditions and strains tested here. (C) 1997 Elsevier B.V. B.V.

Determination of the vinylsulphone azo dye, remazol brilliant orange 3R, by cathodic stripping voltammetry

Remazol brilliant orange 3R shows only a voltammetric peak for the reduction of the azo group. No peak was observed for the reduction of the sulfatoethylsulfone or vinylsulfone reactive groups. The reduction of a pre-protonated ate group involving a two-electron process, gives a hydrate derivative in acidic solution. In alkaline solution the reduction process occurs at more negative potential with the formation of an unstable hydrate compound which decomposes via HN-NH bond cleavage and loss of a sulfate group. Optimum conditions are given for the cathodic stripping voltammetric determination of dir: dye in aqueous solution. The optimum accumulation potential and time were 0 V and up to 60 s, respectively. Linear calibration graphs were obtained from 30 to 300 ng ml(-1) in pH 4 and 6.2 to 62 ng ml(-1) in pH 10. The limit of determination obtained was 1.5 ng ml(-1) (pH 10). The coefficient of variation was 2.6% (n = 7) at 62 ng ml(-1) of the reactive dye. (C) 1999 Elsevier B.V. B.V. All rights reserved.