New pharmacological insights of Galactia glaucescens.

Galactia glaucescens leaves are popularly used against snakebites in Brazil. The hydroethanolic extract from aerial parts of G. glaucescens (HEGg) was assayed against the neurotoxicity and myotoxicity induced by Bothrops jararacussu venom. A traditional myographic technique was applied for neurotoxicity and the resulting muscles were treated routinely by light microscopy analysis for myotoxicity. Additionally, the antimicrobial potential of HEGg was evaluated against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa strains, as well as Rutin was isolated for the first time in this specie using chromatographic and spectroscopic methods and its antiophidian property was assessed. HEGg totally prevents the neurotoxicity and myotoxicity effects caused by B. jararacussu, but did not show any antimicrobial effect. Concluding, HEGg and Rutin were able to counteract the toxic effects of the venom and confirmed the antiophidian potential, but not antimicrobial, of G. glaucescens as an alternative for neutralization of B. jararacussu venom.

An isoflavone from Dipteryx alata Vogel is active against the in vitro neuromuscular paralysis of Bothrops jararacussu snake venom and bothropstoxin I, and prevents venom-induced myonecrosis.

Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM), an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations) and myotoxicity (assessed by light microscopy) of Bothrops jararacussu snake venom in vitro. The toxicity of TM was assessed using the Salmonella microsome assay (Ames test). Incubation with TM alone (200 µg/mL) did not alter the muscle twitch tension whereas incubation with venom (40 µg/mL) caused irreversible paralysis. Preincubation of TM (200 µg/mL) with venom attenuated the venom-induced neuromuscular blockade by 84% ± 5% (mean ± SEM; n = 4). The neuromuscular blockade caused by bothropstoxin-I (BthTX-I), the major myotoxic PLA2 of this venom, was also attenuated by TM. Histological analysis of diaphragm muscle incubated with TM showed that most fibers were preserved (only 9.2% ± 1.7% were damaged; n = 4) compared to venom alone (50.3% ± 5.4% of fibers damaged; n = 3), and preincubation of TM with venom significantly attenuated the venom-induced damage (only 17% ± 3.4% of fibers damaged; n = 3; p < 0.05 compared to venom alone). TM showed no mutagenicity in the Ames test using Salmonella strains TA98 and TA97a with (+S9) and without (-S9) metabolic activation. These findings indicate that TM is a potentially useful compound for antagonizing the neuromuscular effects (neurotoxicity and myotoxicity) of B. jararacussu venom.

Implementation of the three Rs in the human hazard assessment of Brazilian medicinal plants: an evaluation of the cytotoxic and genotoxic potentials of Dipteryx alata Vogel.

In Brazil, medicinal plants are widely used by the indigenous people, which leads to a constant requirement for toxicity tests to be performed on the plant extracts. Although the current Brazilian Directive 90/2004 on the preclinical toxicity testing of phytotherapeutics recommends only in vivo tests, some Brazilian researchers would like to change this situation by implementing the Three Rs in the toxicological testing of medicinal plants. The present study evaluated the cytotoxic and genotoxic potentials of bark extracts from Dipteryx alata Vogel, a medicinal plant of the Brazilian cerrado, by using CHO-K1 (Chinese hamster ovary) cells. An IC50 value was obtained, which corresponded to 0.16mg/ml of plant extract, and from this the equivalent LD50 was determined as 705mg/kg. In order to determine the genotoxic potential of the sample, the frequency of micronucleus formation was assessed. CHO-K1 cells were exposed, during targeted mitosis, to different concentrations of plant extract and cytochalasin B, in the presence and absence of an appropriate metabolic activation system (an S9 mix). The results obtained indicated that it might be possible to implement the Three Rs in assessing the potential human hazard of medicinal plants. The publication of such data can increase awareness of the Three Rs by showing how to optimise the management of animal use, if in vivo toxicological experiments are required.

Chemical constituents of the bark of Dipteryx alata vogel, an active species against Bothrops jararacussu venom.

The effect of four sub-extracts prepared from the lyophilized hydroalcoholic bark of Dipteryx alata (Leguminosae-Papilionoideae) dissolved in a methanol-water (80:20) mixture through a liquid-liquid partition procedure has been investigated against the neuromuscular blockade of the venom of the snake Bothrops jararacussu. The active CH2Cl2 sub-extract has been extensively analyzed for its chemical constituents, resulting in the isolation of four lupane-type triterpenoids: lupeol, lupenone, 28-hydroxylup-20(29)-en-3-one, betulin, nine isoflavonoids: 8-O-methylretusin, 7-hydroxy-5,6,4'-trimethoxyisoflavone, afrormosin, 7-hydroxy-8,3',4'-trimethoxyisoflavone, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, odoratin, 7,8,3'-trihydroxy-4'-methoxyisoflavone, 7,8,3'-trihydroxy-6,4'-dimethoxyisoflavone, dipteryxin, one chalcone: isoliquiritigenin, one aurone: sulfuretin and three phenolic compounds: vanillic acid, vanillin, and protocatechuic acid. Their chemical structures were elucidated on the basis of spectroscopic analysis, including HRMS, 1D- and 2D-NMR techniques.

Calibration of low-level laser therapy equipment.

BACKGROUND: Despite the increase in the use of low-level laser therapy (LLLT), there is still a lack of consensus in the literature regarding how often the equipment must be calibrated. OBJECTIVE: To evaluate the real average power of LLLT devices in the Greater São Paulo area. METHODS: For the evaluation, a LaserCheck power meter designed to calibrate continuous equipment was used. The power meter was programmed with data related to the laser's wavelength to gauge the real average power being emitted. The LLLT devices were evaluated in two ways: first with the device cooled down and then with the device warmed up for 10 minutes. For each condition, three tests were performed. The laser probe was aligned with the power meter, which provided the real average power being emitted by the LLLT device. All of the data and information related to the laser application were collected with the use of a questionnaire filled in by the supervising therapists. RESULTS: The 60 devices evaluated showed deficit in real average power in the cooled-down and warmed-up condition. The statistical analysis (ANOVA) showed a significant decrease (p<0.05) in the real average power measured in relation to the manufacturer's average power. On average, the most common dose in the clinics was 4 J/cm², and the most desired effects were healing and anti-inflammatory effects. According to the World Association for Laser Therapy (WALT), 1 to 4 J of final energy are necessary to achieve these effects, however only one device was able to reach the recommended therapeutic window. CONCLUSION: The LLLT devices showed a deficit in real average power that emphasized a lack of order in the application of this tool. The present study also showed the need for periodical calibration of LLLT equipment and a better technical knowledge of the therapists involved.

Aferição dos equipamentos de laser de baixa intensidade / Calibration of low-level laser therapy equipment

CONTEXTUALIZAÇÃO: A laserterapia de baixa intensidade (LBI) vem sendo cada vez mais utilizada, porém ainda não há consenso na literatura quanto ao tempo em que os equipamentos devem ser submetidos à aferição ou calibragem. OBJETIVO: Analisar a potência média real (PmR) dos equipamentos de LBI na região da Grande São Paulo. MÉTODOS: Para análise dos equipamentos, utilizou-se um potenciômetro (Lasercheck), próprio para aferição de equipamentos contínuos, o qual foi programado com dados referentes ao comprimento de onda do laser a ser avaliado, obtendo-se assim a PmR emitida. Os equipamentos foram analisados de duas formas: uma, com o LBI desaquecido, e outra, após 10 minutos de uso (aquecido), sendo que três análises foram feitas para cada condição. A caneta emissora foi acoplada ao potenciômetro, o qual fornecia a PmR emitida pelo LBI. Todos os dados e informações referentes à aplicação do laser foram coletados por um questionário respondido pelos responsáveis. RESULTADOS: Os 60 equipamentos avaliados mostraram déficit na PmR com os equipamentos desaquecidos e aquecidos. A análise estatística (ANOVA) mostrou diminuição significativa (P<0,05) da PmR aferida em relação à potência média do fabricante (PmF). Em média, a dose mais empregada nas clínicas foi de 4 J/cm², tendo os efeitos de cicatrização e anti-inflamatório como os mais desejados. Segundo a World Association for Laser Therapy (WALT), para atingir esse efeito, necessita-se de 1 a 4 J de energia final, sendo que apenas um dos 60 aparelhos conseguiria atingir a janela terapêutica preconizada. CONCLUSÃO: Os equipamentos de LBI apresentam um déficit acentuado na PmR, o que mostra uma desordem na utilização desse recurso. Neste estudo, observou-se a necessidade de aferição periódica dos aparelhos de LBI bem como melhor conhecimento técnico dos profissionais envolvidos.

BACKGROUND: Despite the increase in the use of low-level laser therapy (LLLT), there is still a lack of consensus in the literature regarding how often the equipment must be calibrated. OBJECTIVE: To evaluate the real average power of LLLT devices in the Greater São Paulo area. METHODS: For the evaluation, a LaserCheck power meter designed to calibrate continuous equipment was used. The power meter was programmed with data related to the laser's wavelength to gauge the real average power being emitted. The LLLT devices were evaluated in two ways: first with the device cooled down and then with the device warmed up for 10 minutes. For each condition, three tests were performed. The laser probe was aligned with the power meter, which provided the real average power being emitted by the LLLT device. All of the data and information related to the laser application were collected with the use of a questionnaire filled in by the supervising therapists. RESULTS: The 60 devices evaluated showed deficit in real average power in the cooled-down and warmed-up condition. The statistical analysis (ANOVA) showed a significant decrease (p<0.05) in the real average power measured in relation to the manufacturer's average power. On average, the most common dose in the clinics was 4 J/cm², and the most desired effects were healing and anti-inflammatory effects. According to the World Association for Laser Therapy (WALT), 1 to 4 J of final energy are necessary to achieve these effects, however only one device was able to reach the recommended therapeutic window. CONCLUSION: The LLLT devices showed a deficit in real average power that emphasized a lack of order in the application of this tool. The present study also showed the need for periodical calibration of LLLT equipment and a better technical knowledge of the therapists involved.

Cuticular hydrocarbons of Heterotermes tenuis (Isoptera: Rhinotermitidae): analyses and electrophysiological studies.

Termites have become an important pest of Eucalyptus and Pinus reforestations, sugarcane and other cultures. An alternative for the control of this pest would be the use of attractive traps that take in account the social behavior of these insects. Diverse factors are important for the insects in the localization of the habitat and the choice of the food and specific odors can facilitate this. Studies referring to Heterotermes tenuis (Isoptera: Rhinotermitidae) are scarce. The objective of this work was to analyze the tergal cuticular extract of H. tenuis and determine the selectivity and sensitivity of its antennae to the components of this extract by electroantennography (EAG). The composition of the cuticular extract was determined by GC-MS analysis. The hydrocarbons found were restricted to linear alkanes, being most abundant C24 to C27 that comprises ca. 65% of the total. Olefins were not detected. EAG and behavioral test responses to the cuticular hydrocarbons were greater and significantly different from the control and the high selectivity of the antennae to the extract indicates its potential as chemical messenger. Cuticular hydrocarbons mixture is species-specific and can be used to identify a given taxon without the diagnostic castes, soldiers or imagoes Difference in the composition appears to relate with the type of habitat of specie.