Anethole supplementation during in vitro maturation increases in vitro goat embryo production in a concentration-dependent manner.

During in vitro maturation (IVM) cumulus-oocyte complexes (COCs) are exposed to conditions that can trigger oxidative stress, thus, reducing oocyte maturation and viability. Aiming to mitigate these detrimental conditions, the effects of IVM medium supplementation with anethole have been tested. Anethole, also known as trans-anethole (1-methoxy-4 [1-propenyl]-benzene), is a naturally occurring phenylpropanoid with various pharmacological properties, including antioxidant effects. However, no study has examined anethole effect on goat COCs during IVM. Thus, the aim of this study was to evaluate the effects of different anethole concentrations on oocyte maturation, oxidative stress, and in vitro development of caprine embryos after parthenogenetic activation. Goat COCs were selected and randomly distributed into the following treatments: TCM-199+ medium (control), or TCM-199+ medium supplemented with 30 µg/mL (AN30); 300 µg/mL (AN300) or 2000 µg/mL (AN2000) of anethole. After IVM, part of the COCs was chosen for oocyte viability and chromatin configuration, intracellular reactive oxygen species levels, and mitochondrial membrane potential assessment. Another part of COCs was parthenogenetically activated, and presumptive zygotes were cultured for 7 days. Results demonstrated that anethole at 30 µg/mL increased oocyte maturation and cleavage rates when compared to the other treatments (P < 0.05), as well as oocyte viability and in vitro embryo production when compared to the control treatment (P < 0.05). Additionally, treatment with anethole at 2000 µg/mL decreased oocyte nuclear maturation and cleavage rates when compared to other treatments (P < 0.05) and embryo production if compared to control and AN30 treatments (P < 0.05). Moreover, anethole at 2000 µg/mL increased mitochondrial membrane potential when compared to the other treatments (P < 0.05). In conclusion, anethole exerts a concentration-dependent effect during goat COCs IVM. For a more desirable outcome of oocyte viability and maturation, and in vitro embryo production, the use of anethole at 30 µg/mL is recommended.

Eugenol Improves Follicular Survival and Development During in vitro Culture of Goat Ovarian Tissue.

This study evaluated the effects of different concentrations (10, 20, or 40 µM) of eugenol (EUG 10, EUG 20, or EUG 40), ascorbic acid (50 µg/mL; AA) or anethole (300 µg/mL; ANE 300) on the in-vitro survival and development of goat preantral follicles and oxidative stress in the cultured ovarian tissue. Ovarian fragments from five goats were cultured for 1 or 7 days in Alpha Minimum Essential Medium (α-MEM+) supplemented or not with AA, ANE 300, EUG 10, EUG 20 or EUG 40. On day 7 of culture, when compared to MEM, the addition of EUG 40 had increased the rate of follicular development, as observed by a decrease in the proportion of primordial follicles alongside with an increase in the rate of normally developing follicles. Furthermore, EUG 40 significantly increased both follicular and oocyte diameters. Subsequently, ovarian fragments from three goats were cultured for 1 or 7 days in α-MEM+ supplemented or not with AA, ANE 300 or EUG 40. All tested antioxidants, except ANE 300, were able to significantly decrease the levels of reactive oxygen species in the ovarian tissue, but EUG 40 could most efficiently neutralize free radicals. All ovarian tissues cultured in the presence of antioxidants, especially EUG 40, presented a significant decrease in H3K4me3 labeling, indicating a silencing of genes that play a role in the inhibition of follicular activation and apoptosis induction. When compared to cultured control tissues, both EUG 40 and ANE 300 significantly increased the intensity of calreticulin labeling in growing follicles. The mRNA relative expression of ERP29 and KDM3A was significantly increased when the culture medium was supplemented with EUG 40, indicating a response to ER stress experienced during culture. In conclusion, EUG 40 improved in-vitro follicle survival, activation and development and decreased ROS production, ER stress and histone lysine methylation in goat ovarian tissue.

Crude glycerin in the supplementation of crossbred heifers in tropical pastures.

Crude glycerin (CG) is a biodiesel byproduct that has been tested as an alternative feed additive for use in beef production. After being absorbed, it is used in the liver to produce glucose, an important precursor of intramuscular fat in ruminants. This study aimed to evaluate the effect of CG (439 g/kg glycerol) on the performance and meat quality of crossbred heifers finished in Urochloa brizantha cv. Marandu. Thirty-six heifers with an initial BW of 301.5 ±â€¯23.02 kg were used. They were supplemented for 154 days with the following levels of CG: mineral mixture (without CG), 33.3, 66.6 and 99.9 g/kg CG in the DM of the supplement. Supplement or pasture DM intakes, slaughter BW and carcass traits were not influenced (P > 0.05) by increasing levels of CG. The total fat content of the meat, the vaccenic (18:1 n-7t) and conjugated linoleic acid (18:2 c9-t11) increased with the addition of CG in the diet (P < 0.05). Crude glycerin can be included up to 99.9 g/kg of the total diet without changing the performance of crossbred heifers finished in the tropical pasture.

Effects of cottonseed oil and ferrous sulfate on the performance and expression of antioxidant enzymes in broilers.

Aiming to reduce feed costs, cottonseed oil (CSO) has been used as an alternative component in diets for broilers. However, this oil contains gossypol, an antinutritional agent that impacts the use of mineral elements, inhibits glucose uptake, and has a direct inhibitory action on intestinal enzymes. Nevertheless, toxic effects of gossypol can be prevented by the addition of iron salts, such as ferrous sulfate (FS), to the diet. This work was conducted to evaluate performance and gene expression of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in the intestines of 21- and 42-day-old broilers fed 0, 2, 4, or 6% concentrations of CSO, with or without FS. All CSO diets led to weight gain (WG) at 21 D. At 42 D, an increase in WG and a decrease in feed conversion (FCR) in the diets containing FS were observed. In 21-day-old birds supplemented with 4% CSO and FS, an increase in GPx gene expression was observed when compared to the 6% level. Animals (42 day old) supplemented with 6% CSO and FS presented greater expression of SOD gene when compared to 2% CSO and FS. In addition, a higher GPx expression in broilers supplemented with 6% CSO and FS compared to 6% CSO without FS was achieved. In conclusion, including CSO in the diets of broiler favors WG in animals at 21 D of age, independent of the presence or absence of FS; and including 4% CSO and FS in the diet of these animals alters the expression of the GPx gene in the intestine, so it is not necessary to add FS at 21 D. On the other hand, in 42-day-old broilers, the addition of FS is indicated, due to increases WG, decreased FCR and at the 6% CSO level without FS increase in the expression of the SOD and GPx genes.

Lipid dynamics in LPS-induced neuroinflammation by DESI-MS imaging.

It is well-established that bacterial lipopolysaccharides (LPS) can promote neuroinflammation through receptor Toll-like 4 activation and induces sickness behavior in mice. This phenomenon triggers changes in membranes lipid dynamics to promote the intracellular cell signaling. Desorption electrospray ionization mass spectrometry (DESI-MS) is a powerful technique that can be used to image the distribution of lipids in the brain tissue directly. In this work, we characterize the LPS-induced neuroinflammation and the lipid dynamics in C57BL/6 mice at 3 and 24 h after LPS injection. We have observed that intraperitoneal administration of LPS (5 mg/kg body weight) induces sickness behavior and triggers a peripheral and cerebral increase of pro- and anti-inflammatory cytokine levels after 3 h, but only IL-10 was upregulated after 24 h. Morphological analysis of hypothalamus, cortex and hippocampus demonstrated that microglial activation was present after 24 h of LPS injection, but not at 3 h. DESI-MS revealed a total of 14 lipids significantly altered after 3 and 24 h and as well as their neuroanatomical distribution. Multivariate statistical analyzes have shown that ions associated with phosphatidylethanolamine [PE(38:4)] and docosatetraenoic acid [FA (22:4)] could be used as biomarkers to distinguish samples from the control or LPS treated groups. Finally, our data demonstrated that monitoring cerebral lipids dynamics and its neuroanatomical distribution can be helpful to understand sickness behavior and microglial activation after LPS administration.

Essential oil of Pterodon polygalaeflorus Benth attenuates nociception in mice.

Essential oils (EO) are volatile liquids responsible for the aroma of plants. Pterodon polygalaeflorus seeds have received widespread use in folk medicine for the treatment of inflammatory diseases. For this reason and because Pterodon polygalaeflorus seeds have great EO content, which is frequently pharmacologically active, the present study aimed to evaluate the antinociceptive effect of EO from Pterodon polygalaeflorus (EOPPgfl) and its acute toxic effects. The EEOPPgfl sample, which was extracted by steam distillation of the seeds, had a yield of 2.4% of the seeds weight and had, as major constituents, beta-elemene (48.19%), trans-caryophyllene (19.51%), and epi-bicyclosesquiphellandrene (12.24%). The EOPPgfl sample showed mild acute toxicity and its calculated median lethal dose (LD50) was 3.38 g/kg. EOPPgfl (20-60 mg/kg) showed antinociceptive activity as evidenced by several tests and inhibited writhing induced by acetic acid. The maximum effect was obtained with the 30 mg/kg dose and at 60 min after its administration. EOPPgfl also decreased formalin-induced nociception, as verified by the inhibition of the first and second phase of the formalin test. At 30 mg/kg, EOPPgfl also decreased thermally stimulated nociception. Nociception may be related to inflammatory and antiedematogenic activity and at doses ranging 10-100 mg/kg, EOPPgfl blocked dextran- and carrageenan-induced edema. The results demonstrated that EOPPgfl presented, at doses approximately 100 times smaller than LD50, an antinociceptive effect that probably was due to anti-inflammatory activities.

Oral antioxidant therapy for prevention and treatment of preeclampsia: Meta-analysis of randomized controlled trials.

AIMS: To determine whether oral antioxidant therapies, of various types and doses, are able to prevent or treat women with preeclampsia. DATA SYNTHESIS: The following databases were searched: MEDLINE, CENTRAL, LILACS, and Web of Science. Inclusion criteria were: a) randomized clinical trials; b) oral antioxidant supplementation; c) study in pregnant women; d) control group, treated or not with placebo. Papers were excluded if they evaluated antioxidant nutrient supplementation associated with other non-antioxidant therapies. Data were extracted and the risk of bias of each study was assessed. Heterogeneity was analyzed using the Cochran Q test, and I2 statistics and pre-specified sensitivity analyses were performed. Meta-analyses were conducted on prevention and treatment studies, separately. The primary outcome was the incidence of preeclampsia in prevention trials, and of perinatal death in treatment trials. Twenty-nine studies were included in the analysis, 19 for prevention and 10 for treatment. The antioxidants used in these studies were vitamins C and E, selenium, l-arginine, allicin, lycopene and coenzyme Q10, none of which showed beneficial effects on the prevention of preeclampsia (RR: 0.89, CI 95%: [0.79-1.02], P = 0.09; I2 = 39%, P = 0.04) and other outcomes. The antioxidants used in the treatment studies were vitamins C and E, N-acetylcysteine, l-arginine, and resveratrol. A beneficial effect was found in intrauterine growth restriction. CONCLUSIONS: Antioxidant therapy had no effects in the prevention of preeclampsia but did show beneficial effects in intrauterine growth restriction, when used in the treatment of this condition.

Essential oil of Pterodon polygalaeflorus Benth attenuates nociception in mice

Essential oils (EO) are volatile liquids responsible for the aroma of plants. Pterodon polygalaeflorus seeds have received widespread use in folk medicine for the treatment of inflammatory diseases. For this reason and because Pterodon polygalaeflorus seeds have great EO content, which is frequently pharmacologically active, the present study aimed to evaluate the antinociceptive effect of EO from Pterodon polygalaeflorus (EOPPgfl) and its acute toxic effects. The EEOPPgfl sample, which was extracted by steam distillation of the seeds, had a yield of 2.4% of the seeds weight and had, as major constituents, beta-elemene (48.19%), trans-caryophyllene (19.51%), and epi-bicyclosesquiphellandrene (12.24%). The EOPPgfl sample showed mild acute toxicity and its calculated median lethal dose (LD50) was 3.38 g/kg. EOPPgfl (20-60 mg/kg) showed antinociceptive activity as evidenced by several tests and inhibited writhing induced by acetic acid. The maximum effect was obtained with the 30 mg/kg dose and at 60 min after its administration. EOPPgfl also decreased formalin-induced nociception, as verified by the inhibition of the first and second phase of the formalin test. At 30 mg/kg, EOPPgfl also decreased thermally stimulated nociception. Nociception may be related to inflammatory and antiedematogenic activity and at doses ranging 10-100 mg/kg, EOPPgfl blocked dextran- and carrageenan-induced edema. The results demonstrated that EOPPgfl presented, at doses approximately 100 times smaller than LD50, an antinociceptive effect that probably was due to anti-inflammatory activities.

Use of sugarcane molasses by Pycnoporus sanguineus for the production of laccase for dye decolorization.

Pycnoporus sanguineus is a white-rot basidiomycete that produces laccase as the only oxidoreductase; enzyme synthesis depends on cultivation variables, and fungal species and strain. Laccases have wide substrate specificity, oxidize a broad range of compounds, and show potential for use in dye decolorization. We evaluated laccase production in a recently isolated strain of P. sanguineus cultivated with sugarcane molasses as the only carbon source, and urea or yeast extract as the nitrogen source [at various nitrogen concentrations (0.4, 1.4, 2.4, 3.4, and 4.4 g/L)], supplemented with copper (0, 150, 200, 250, and 300 µM), with or without agitation. The enzymatic extract produced at laccase peak activity was tested for dye decolorization capability on Remazol Brilliant Blue R, Reactive Black 5, Reactive Red 195, and Reactive Yellow 145. The nitrogen source did not affect enzyme production and the higher nitrogen concentration (3.4 g/L nitrogen as urea) increased enzymatic activity. The addition of up to 300 µM of Cu did not affect laccase production, whereas cultivation with agitation increased the activity peak by 17%. The highest laccase activity was ~50,000 U/L on the ninth day of cultivation. After 24 h, decolorization was 80% for Remazol Brilliant Blue R, 9% for Reactive Yellow 145, 6% for Reactive Red 195, and 2% for Reactive Black 5. The enzymatic extract of P. sanguineus provides a potential alternative to wastewater treatment. A better understanding of the behavior of this fungus under various culture conditions would allow improvement of the enzyme production bioprocess.

Atividades biológicas de espécies amazônicas de Rubiaceae / Biological activities of Amazonian species of Rubiaceae

RESUMORubiaceae é uma família botânica com grande potencial químico e biológico Neste trabalho realizou-se a prospecção fitoquímica e avaliou-se as atividades citotóxica, antioxidante e antifúngica dos extratos diclorometânicos e metanólicos das folhas de Duroia saccifera, Ferdinandusa goudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera e P. guianensis. A avaliação da citotoxicidade foi realizada empregando-se o ensaio de toxicidade sobre Artemia salina. A atividade antifúngica foi determinada pelo método de difusão em ágar utilizando-se os fungos: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T. rubrum ATCC 28189, T. tonsurans 21/97, Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 e C. albicans U. 5/99. A atividade antioxidante foi determinada através dos ensaios de redução do radical livre 1,1-difenil-2-picrilhidrazila (DPPH) e de descoloração do cátion radical ácido-6-sulfônico-2,2-azinobis-3-etilbenzotiazolina (ABTS). Para o ensaio de toxicidade sobre A. salina observou-se maior toxicidade no extrato metanólico de F. goudotiana, que se mostrou tóxico até a concentração de 5 µg mL-1. A presença de princípios antifúngicos foi observada em F. hirsuta e F. paraensiscontra quatro dos fungos testados. Na avaliação da atividade antioxidante os extratos metanólicos mostraram maior atividade que os extratos diclorometânicos, sendo coincidente o resultado obtido através de ambos os métodos (redução do DPPH e descoloração do ABTS). Os resultados criam possibilidades para futuras investigações relacionadas à estrutura e atividade dos componentes de cada extrato ativo.

ABSTRACTRubiaceae is a plant family with great chemical and biological potential. In this work a phytochemical prospection was performed and it was assessed the cytotoxic, antioxidant and antifungal activities of dichloromethanic and methanolic extracts of the leaves of Duroia saccifera, Ferdinandusagoudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera and P. guianensis. The cytotoxicity evaluation was carried out using the test toxicity on Artemia salina. The antifungal activity was determined by agar diffusion method using fungi: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T.rubrum ATCC 28189, T. tonsurans 21/97 , Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 and C. albicans U. 5/99. The antioxidant activity was determined by testing the amount of free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the discoloration of the radical cation 6-sulfonic acid-2 ,2-azinobis-3-ethylbenzothiazoline (ABTS). For the toxicity test on A. salina greater toxicity in the methanol extract of F. goudotiana was observed, which proved to be toxic up to a concentration of 5 µg mL-1. The presence of antifungal principles was observed in the F. hirsuta and F. paraensis against four tested fungi. In the evaluation of the antioxidant activity, the methanol extracts showed greater activity than the dichloromethanic ones, being similar the obtained result through both methods (reduction of DDPH and ABTS decolorization) . The results create possibilities for future researches related to the structure and activity of the components of each active extract.

In vivo growth inhibition of sarcoma 180 by Kielmeyera rugosa Choisy (Calophyllaceae).

The plant Kielmeyera rugosa Choisy (family Calophyllaceae), popularly known as 'pau-santo', is traditionally used in Brazilian folk medicine. Recently, the dichloromethane extract-dichloromethane partition from stems of K. rugosa (KR) has shown positive results in our cytotoxic screening programme. Therefore, the aim of this study was to validate the antitumour activity of KR on sarcoma 180 tumour-bearing mice. KR showed antitumour activity with both administration routes: intraperitoneal (50 and 100 mg/kg/day) and oral (100 and 200 mg/kg/day). Tumour growth inhibition rates were 40.8-34.9% and 25.4-51.8% after intraperitoneal and oral administrations, respectively. Treatment with KR did not significantly affect body mass, macroscopy of the organs or blood leukocyte counts. In conclusion, KR exhibited an in vivo antitumour effect without substantial toxicity.

Avaliação toxicológica pré-clínica do chá das folhas de Morus nigra L. (Moraceae) / Pre-clinical toxicological evaluation of tea from the leaves of Morus nigra L. (Moraceae)

O objetivo desse estudo foi realizar um ensaio toxicológico pré-clínico para analisar a toxicidade do chá das folhas de Morus nigra L. (Moraceae). A toxicidade subcrônica do chá (CF-Mn) foi avaliada durante 30 dias por via oral em ratos. Ao grupo controle foi administrado água, para comparação. Durante o período experimental foi avaliada a presença de sinais de toxicidade, variação do peso corporal, e o consumo de líquido e alimento. Ao final do experimento o sangue dos animais foi retirado para análise de parâmetros hematológicos e bioquímicos. Não foram observados mortalidade e sinais de toxicidade indicando baixa toxicidade da planta. Não houve alterações nos parâmetros hematológicos e bioquímicos. Nas condições do estudo, o CF-Mn pode ser considerado de baixa toxicidade, pois não produziu efeitos tóxicos nos animais tratados.

The aim of this study was to carry out a pre-clinical toxicological assay to analyze the toxicity of tea from the leaves of Morus nigra L. (Moraceae). The subchronic toxicity of this tea (CF-Mn) was orally evaluated during 30 days in rats. The control group was given water for comparison. During the experimental period, signs of toxicity, body weight variation, and water and food consumption were assessed. At the end of the experiment, the blood of animals was removed for analysis of hematological and biochemical parameters. No mortality and no toxicity signs were observed, indicating low toxicity of the plant. There was no alteration in the hematological and biochemical parameters. Under the study conditions, CF-Mn can be considered of low toxicity since it did not produce toxic effects in treated animals.

New magnetic fluid developed with natural organic compounds biocompatible.

This work was developed with an aqueous suspension of maghemite nanoparticles and colloidal emulsions with nanoparticles of magnetite. The nanoparticles were synthesized by co-precipitation method. The first was the magnetic emulsion nanoparticles of maghemite dispersed in the aqueous extract obtained from the leaf embauba (Cecropia Obtusifolia), whose tree is native to Central and South America. Thereby achieving the magnetic fluid extract embauba stabilized with ionic buffer solution pH 7.4. A second emulsion was prepared with colloidal magnetite nanoparticles with surfaces previously coated with oleic acid as a means of dispersing and using the oil extracted from in nature seed Andiroba (Carapa Guianensis), tree of the Brazilian Amazon. These new magnetic fluids the nanoparticles were characterized by Photoacoustic spectroscopy (PAS) to determine the coating layer of molecules on the surfaces of nanoparticles. In aqueous ionic magnetic fluid Cecropia Obtusifolia (MFCO) chlorogenic acid contributes to the electron density in the presence of four groups alcohols, a ketone group and a carboxylic group. In magnetic fluid-based oil andiroba MFAD PAS spectra show that oleic acid molecules are tightly linked on the surface of the nanoparticles.

Treatment of advanced hepatocellular carcinoma with very low levels of amplitude-modulated electromagnetic fields.

BACKGROUND: Therapeutic options for patients with advanced hepatocellular carcinoma (HCC) are limited. There is emerging evidence that the growth of cancer cells may be altered by very low levels of electromagnetic fields modulated at specific frequencies. METHODS: A single-group, open-label, phase I/II study was performed to assess the safety and effectiveness of the intrabuccal administration of very low levels of electromagnetic fields amplitude modulated at HCC-specific frequencies in 41 patients with advanced HCC and limited therapeutic options. Three-daily 60-min outpatient treatments were administered until disease progression or death. Imaging studies were performed every 8 weeks. The primary efficacy end point was progression-free survival 6 months. Secondary efficacy end points were progression-free survival and overall survival. RESULTS: Treatment was well tolerated and there were no NCI grade 2, 3 or 4 toxicities. In all, 14 patients (34.1%) had stable disease for more than 6 months. Median progression-free survival was 4.4 months (95% CI 2.1-5.3) and median overall survival was 6.7 months (95% CI 3.0-10.2). There were three partial and one near complete responses. CONCLUSION: Treatment with intrabuccally administered amplitude-modulated electromagnetic fields is safe, well tolerated, and shows evidence of antitumour effects in patients with advanced HCC.

Evaluation of the genotoxicity of Orthosiphon stamineus aqueous extract.

AIM OF THE STUDY: Orthosiphon stamineus, Benth, also known as Misai Kucing in Malaysia and Java tea in Indonesia, is traditionally used in Southeastern Asia to treat kidney dysfunctions, diabetes, gout and several other illnesses. Recent studies of Orthosiphon stamineus pharmacological profile have revealed antioxidant properties and other potentially useful biological activities thereby lending some scientific support to its use in folk medicine. So far the genotoxicity of Orthosiphon stamineus extracts has not been evaluated. In this study the genotoxic potential of Orthosiphon stamineus aqueous extract was investigated by the Salmonella/microsome mutation assay and the mouse bone marrow micronucleus test. MATERIALS AND METHODS: Chemical composition of Orthosiphon stamineus aqueous extract was analyzed by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD). The Salmonella/microsome assay (TA97a, TA98, TA100 and TA1535; plate incorporation method) was performed in the presence or in the absence of extrinsic metabolic activation (S9 mixture). In the mouse micronucleus assay, Orthosiphon stamineus extract was administered by gavage (0, 500, 2000 and 4000 mg/kg body weight/day for 3 days) to male and female Swiss Webster mice (N=6 per dose per sex) and bone marrow cells were harvested 24 h after the last dose. Ethoxy-resorufin-O-dealkylase (EROD) and benzyloxy-resorufin-O-dealkylase (BROD) activities were determined in mouse liver microsomes. RESULTS: The chemical analysis revealed that the Orthosiphon stamineus extract contained flavonoids (sinensetin, eupatorin), caffeic acid, and rosmarinic acid (44.00±1.879 µg/mg), the latter seemed to be one of its major constituent. Tested at doses up to 5000 µg/plate, the Orthosiphon stamineus extract was not toxic to Salmonella tester strains and did not increase the number of revertant colonies over the background incidence. In the mouse bone marrow assay, the extract did not alter the polychromatic:normochromatic erythrocytes (PCE:NCE) ratio, nor did it increase the incidence of micronucleated polychromatic erythrocytes (MNPEs). No overt toxicity and no change of CYP1A (EROD) and 2B9/10 (BROD) activities were noted. CONCLUSIONS: Based on the aforementioned findings, it is concluded that the use of Orthosiphon stamineus in the traditional medicine poses no genotoxic risk.

Micropropagação e aclimatização de gerânio (Pelargonium graveolens L. ) / Micropropagation and acclimatization of geranium (Pelargonium graveolens L. )

O gerânio (Pelargonium graveolens L.) é uma planta medicinal e aromática nativa do sul da África, cujo óleo essencial é amplamente empregado nas indústrias de perfumaria e cosmético, além de ser usado como terapêutico. Avaliou-se o efeito da luminosidade e dos reguladores de crescimento BAP e ANA na multiplicação in vitro e o uso de diferentes substratos na aclimatização de mudas micropropagadas. Foram avaliadas as concentrações 0; 0,5; 1,0 e 2,0 mg L-1 de BAP, 0; 0,1 e 0,5 mg L-1 de ANA e duas condições de luminosidade (ausência e presença de luz), em esquema fatorial 4x3x2; para a aclimatização foram utilizados quatro substratos, pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) (PCBC); pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) + vermiculita (1:1) (PCBCV 1:1); pó de coco + Biosafra® (3-12-6) (12 g L­-1) + calcário (1 g L-1) + vermiculita (PCBCV 2:1) e vermiculita e adição semanal de sais MS (VS). Para a micropropagação de P. graveolens a utilização de 1,3 mg L-1 de BAP e 0,5 mg L-1 de ANA é eficiente na regeneração direta de plantas de gerânio, sendo a condição de escuro a mais indicada por proporcionar o maior número de brotos por explante. Para a aclimatização das mudas de gerânio, os melhores resultados foram obtidos em mudas aclimatizadas no substrato vermiculita com adição semanal de sais de MS seguido do substrato PCBCV 1:1.

Geranium (Pelargonium graveolens L.) is a medicinal and aromatic plant native to the south of Africa and whose essential oil is widely used by perfume and cosmetic industries, as well as in therapeutics. The effect of luminosity and growth regulators BAP and NAA on in vitro multiplication was evaluated, together with the use of different substrates on the acclimatization of micropropagated seedlings. The evaluated concentrations were 0, 0.5, 1.0 and 2.0 mg L-1 BAP; 0, 0.1 and 0.5 mg L-1 NAA; and two luminosity conditions (absence and presence of light), in a 4 x 3 x 2 factorial arrangement. For acclimatization, four substrates were tested: coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) (CDBL); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (1:1) (CDBLV 1:1); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (CDBLV 2:1); and vermiculite with weekly addition of MS salts (VS). For P. graveolens micropropagation, the use of 1.3 mg L-1 BAP and 0.5 mg L-1 NAA is efficient for the direct regeneration of geranium plants, and the dark condition is the most suitable since it provides a larger number of shoots per explant. For the acclimatization of geranium seedlings, the best results were obtained with seedlings acclimatized in the substrate vermiculite with weekly addition of MS salts, followed by the substrate CDBLV 1:1.

Emprego do óleo de Melaleuca alternifolia Cheel (Myrtaceae) na odontologia: perspectivas quanto à utilização como antimicrobiano alternativo às doenças infecciosas de origem bucal / Use of Melaleuca alternifolia Cheel (Myrtaceae) oil in dentistry: perspectives on its use as alternative antimicrobial to infectious diseases of oral origin

O interesse por medicamentos alternativos, principalmente daqueles provenientes de extratos naturais, tem aumentado nas últimas décadas. A Melaleuca alternifolia é um arbusto pertencente ao gênero Melaleuca, popularmente conhecida como "árvore de chá", cujo principal produto é o óleo essencial (TTO - tea tree oil), de grande importância medicinal por possuir comprovada ação bactericida e antifúngica contra diversos patógenos humanos. Em virtude da atividade terapêutica em diversas especialidades médicas, o TTO passou a ser empregado na área odontológica. Esta revisão de literatura foi realizada com o objetivo de discutir os ensaios já realizados com o TTO contra microrganismos relacionados à doença cárie, doença periodontal e problemas pulpares. O óleo de Melaleuca tem demonstrado boa ação antibacteriana in vitro contra microrganismos bucais, porém, pesquisas envolvendo o estudo do mecanismo de ação sobre as células microbianas ou estudos in vivo ainda são escassos e precisam ser realizados, já que esse produto pode ser útil na odontologia, seja na manutenção química da higiene ou prevenção de doenças bucais.

The interest in alternative medicines, especially those from natural extracts, has increased in recent decades. Melaleuca alternifolia is a shrub belonging to the genus Melaleuca, popularly known as "tea tree", the main product of which is its essential oil (TTO - tea tree oil), of great medicinal importance for its proven bactericidal and antifungal activity against several human pathogens. By virtue of its therapeutic activity in various medical specialties, TTO is now used in dentistry. This literature review was conducted in order to discuss the tests already carried out with TTO against microorganisms related to dental caries, periodontal disease and pulpal problems. Melaleuca oil has shown good in vitro antibacterial activity against oral microorganisms; however, research involving the study of its mechanism of action on the microbial cells or in vivo studies are still scarce and need to be done since this product may be useful in dentistry, either in the chemical maintenance of hygiene or in the prevention of oral diseases.

Produção de mudas de dois genótipos de alecrim-de-tabuleiro (Lippia gracilis Schauer) em função de fertilizante mineral, calcário, substratos e recipientes / Seedling production of two "alecrim-de-tabuleiro" (Lippia gracilis Schauer) genotypes under mineral fertilizer, limestone, substrates and containers

Lippia gracilis Schauer popularmente conhecida como alecrim-de-tabuleiro possui moderada atividade antibacteriana, antimicrobiana e antiséptica. Objetivando avaliar o efeito de doses de fertilizante mineral, calcário, substratos e recipientes na produção de mudas de dois genótipos de alecrim-de-tabuleiro realizou-se experimentos com estacas de dois genótipos de L. gracilis. No experimento 1, foram utilizadas estacas apicais distribuídas em bandejas de poliestireno expandido de 128 células com o substrato pó-de-coco + areia (1:1), três repetições e oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4x2x2, sendo quatro doses do fertilizante (6-24-12+micronutrientes) (1, 2, 3 e 4 g L-1), duas doses de calcário (0 e 1 g L-1) e dois genótipos (LGRA106 e LGRA201). Aos 35 após plantio, foram avaliadas a sobrevivência ( por cento), enraizamento ( por cento), comprimento de raiz (cm) e massa seca de raiz (mg). No experimento 2, foram utilizadas três repetições com oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema de parcelas subdivididas onde foram testados nas parcelas dois recipientes (bandeja de poliestireno expandido de 128 alvéolos e tubetes de 110 cm³, na sub parcela foram testadas as combinações de quatro substratos (PC - pó de coco; PCA (1:1) - pó de coco + areia (1:1); PCA (2:1) - pó de coco + areia (2:1) e PCA (3:1) - pó de coco + areia (3:1) e dois genótipos (LGRA106 e LGRA201). Aos 35 dias após o plantio foram analisadas as mesmas variáveis do experimento 1, além de altura de parte aérea (cm) e massa seca de parte aérea (mg). A utilização de 1 g L-1 do fertilizante na ausência de calcário foi efetivo para sobrevivência de plantas e enraizamento de estacas de alecrim-de-tabuleiro. Com base nesses experimentos, concluiu-se que o genótipo LGRA106 é superior ao LGRA201 nas variáveis analisadas e recomenda-se a bandeja de poliestireno expandido para produção de mudas de L. gracilis via estaquia.

Commonly known as "alecrim-de-tabuleiro", Lippia gracilis Schauer (Verbenaceae) has moderate antibacterial, antimicrobial and antiseptic activity. This study aimed to evaluate the effect of doses of mineral fertilizer, limestone, substrates and containers on seedling production of two "alecrim-de-tabuleiro" genotypes, using cuttings. In experiment 1, apical cuttings were distributed on expanded polystyrene trays of 128 cells containing coir + sand (1:1), using three replicates of eight cuttings each. Experimental design was in randomized blocks, in a 4x2x2 factorial arrangement, i.e. four doses of fertilizer (6-24-12 + micronutrients) (1, 2, 3 and 4 g L-1), two doses of limestone (0 and 1 g L-1) and two genotypes (LGRA106 and LGRA201). At 35 days after planting, we evaluated survival ( percent), rooting ( percent), root length (cm) and root dry matter (mg). In experiment 2, three replicates of eight cuttings each were used. Experimental design was in randomized blocks, in a split-plot scheme. In the plots, we tested two containers (expanded polystyrene tray of 128 cells and 110 cm³ tubets). In the subplots, we tested four substrate combinations [coir, coir + sand (1:1), coir + sand (2:1) and coir + sand (3:1)] and two genotypes (LGRA106 and LGRA201). At 35 days after planting, the same variables of experiment 1 were evaluated, besides shoot length (cm) and shoot dry matter (mg). The use of 1 g L-1 fertilizer without limestone was effective for plant survival and cutting rooting. Based on these experiments, the genotype LGRA106 is superior to LGRA201 as to the evaluated variables and the expanded polystyrene tray is recommended for the production of L. gracilis seedlings through cuttings.

Antinociceptive effects of the essential oil of Mentha x villosa leaf and its major constituent piperitenone oxide in mice.

Mentha x villosa Huds (Labiatae) is an aromatic herb widely used in folk medicine. Since the essential oil of the herb has many pharmacological activities, including antispasmodic effects, we determined whether the oil and its major constituent, piperitenone oxide (PO), have antinociceptive activity. The essential oil of M. x villosa (EOMV) and PO administered orally at 200 mg/kg (vehicle: 0.1% Tween 80 in water) significantly reduced the writhings induced by acetic acid from control values of 59.5 +/- 3.1 s (N = 10) to 31.9 +/- 2.8 s (N = 10) and 23.8 +/- 3.4 s (N = 10), respectively. When administered at 100 and 200 mg/kg, EOMV reduced the paw licking time for the second phase of the formalin test from the control value of 20.6 +/- 2.1 s (N = 13) to 5.3 +/- 2.2 s (N = 12) and 2.7 +/- 1.2 s (N = 18), respectively. At 100 and 200 mg/kg, PO reduced this second phase to 8.3 +/- 2.7 s (N = 12) and 3.0 +/- 1.2 s (N = 10), respectively. This effect of EOMV and PO was not reversed by naloxone. EOMV and PO had no significant effect on the first phase of the formalin test. As evaluated by the hot-plate and tail immersion test, EOMV and PO, at doses up to 200 mg/kg, showed no analgesic activity. These results show that EOMV and PO have antinociceptive activity and suggest that this effect is probably an indirect anti-inflammatory effect, which does not involve the central nervous system.

Antinociceptive effects of the essential oil of Mentha x villosa leaf and its major constituent piperitenone oxide in mice

Mentha x villosa Huds (Labiatae) is an aromatic herb widely used in folk medicine. Since the essential oil of the herb has many pharmacological activities, including antispasmodic effects, we determined whether the oil and its major constituent, piperitenone oxide (PO), have antinociceptive activity. The essential oil of M. x villosa (EOMV) and PO administered orally at 200 mg/kg (vehicle: 0.1% Tween 80 in water) significantly reduced the writhings induced by acetic acid from control values of 59.5 ± 3.1 s (N = 10) to 31.9 ± 2.8 s (N = 10) and 23.8 ± 3.4 s (N = 10), respectively. When administered at 100 and 200 mg/kg, EOMV reduced the paw licking time for the second phase of the formalin test from the control value of 20.6 ± 2.1 s (N = 13) to 5.3 ± 2.2 s (N = 12) and 2.7 ± 1.2 s (N = 18), respectively. At 100 and 200 mg/kg, PO reduced this second phase to 8.3 ± 2.7 s (N = 12) and 3.0 ± 1.2 s (N = 10), respectively. This effect of EOMV and PO was not reversed by naloxone. EOMV and PO had no significant effect on the first phase of the formalin test. As evaluated by the hot-plate and tail immersion test, EOMV and PO, at doses up to 200 mg/kg, showed no analgesic activity. These results show that EOMV and PO have antinociceptive activity and suggest that this effect is probably an indirect anti-inflammatory effect, which does not involve the central nervous system.