Elevated GABA levels in the medial prefrontal cortex and lower estrogen levels abolish cocaine sensitization behavior in ovariectomized female rats.

Animal models show that cocaine sensitization, a behavioral marker of addiction, is more significant in intact gonadal female than male rats and ovariectomy suppress this behavior in female rats. However, few studies explore changes in neurotransmission related to this phenomenon. Here we investigated the in vivo changes on GABA, glutamate, and taurine levels in the medial prefrontal cortex (mPFC) of gonadal intact or ovariectomized female rats after a cocaine challenge administration. Adult female rats were bilaterally ovariectomized (OVX), or sham-operated (SHAM) and randomly assigned to control (CTR), acute (ACT), or repeated (RPT) cocaine administration groups. In the challenge day, after eight days of daily cocaine (15 mg/kg) or saline administration and ten days of washout and stereotaxic surgery, RPT and ACT groups received cocaine, and the CTR group received saline. Horizontal locomotion was monitored concomitantly with microdialysate collection to determine extracellular GABA, glutamate, and taurine levels. Hormonal determination in blood samples confirmed the lower hormonal status of the OVX. Cocaine sensitization occurred in SHAM-RPT female rats after the challenge administration. Non-sensitized OVX-RPT rats showed a peak of GABA at 30 min after cocaine administration, with no change on glutamate and taurine levels. Therefore, elevated GABA levels in the mPFC and lower serum estrogen levels abolish cocaine sensitization behavior in ovariectomized female rats. We discuss some possible implications of these finding for future models of cocaine sensitization research lighting in the female hormonal influence.

Toxicological evaluation of a standardized hydroethanolic extract from leaves of Plantago australis and its major compound, verbascoside.

ETHNOPHARMACOLOGICAL RELEVANCE: Plantago australis is a perennial plant widely distributed in Latin America, and its seeds and leaves are used in folk medicine to treat many diseases and conditions. Among its various chemical compounds, verbascoside is one of the most present, and has several pharmacological activities described, but there is not much information about its toxicity. AIMS OF THE STUDY: The aims of this study were to optimize the extraction of verbascoside from P. australis leaves with ultrasound methods, to develop a validated HPLC method to quantify verbascoside, and to evaluate the toxicological safety of the extract and verbascoside using in vitro and in vivo assays. MATERIALS AND METHODS: Dried leaves of P. australis were submitted to different extraction methods (percolation and ultrasound). The optimization of the ultrasound extraction was carried out by complete factorial design (22) and response surface methodology (RSM), followed by HPLC analysis for marker compounds. HPLC analysis was performed to verify the presence of the marker compounds aucubin, baicalein, oleanolic acid, ursolic acid and verbascoside. Mutagenicity was assessed by Salmonella/microsome mutagenicity assay. Cytotoxicity and genotoxicity were evaluated in V79 cells by reduction of tetrazolium salt (MTT) and neutral red uptake (NRU) assays, and alkaline comet assay, respectively. Verbascoside phototoxicity was assessed in 3T3 cells by the NRU phototoxicity assay. Wistar rats were used to perform the acute and sub-chronic toxicity tests. RESULTS: Among the marker compounds, only verbascoside was found in the hydroethanolic extract of P. australis leaves (PAHE); its highest concentration was obtained with the ultrasound-assisted extraction (UAE) method, optimized in 40 min and 25 °C, and the method validation was successfully applied. Neither PAHE nor verbascoside showed mutagenic or genotoxic activities. Cytotoxicity assays demonstrated that both PAHE and verbascoside reduced cell viability only at the highest concentrations, and verbascoside had no phototoxic properties. The in vivo toxicity evaluation of PAHE suggested that the LD50 is higher than 5000 mg/Kg, indicating that this extract is safe for use. In addition, no signs of toxicity were found in subchronic exposure. CONCLUSION: The HPLC method to quantify verbascoside was validated, and the extraction of verbascoside from P. australis leaves through ultrasound method was optimized, yielding an extract with 6% verbascoside. Our results suggest the toxicological safety of PAHE and verbascoside, corroborating the use of P. australis in folk medicine, and also indicate verbascoside as a potential ingredient in topical formulations.

Wound healing and anti-inflammatory activities induced by a Plantago australis hydroethanolic extract standardized in verbascoside.

ETHNOPHARMACOLOGICAL RELEVANCE: Plantago australis is a popular plant found to be widely spread in Latin America. In folk medicine, the seeds and leaves are used mainly for anti-inflammatory, wound healing, among others. The verbascoside, a phenolic glycoside, is an active chemical component described in this species of plant, which has antioxidant, anti-inflammatory and healing effects. PURPOSE: The aim of the present study was to evaluate whether P. australis hydroethanolic extract (PAHE) standardized in verbascoside could promote wound healing associated with anti-inflammatory action within both in vitro and in vivo models. METHODS: For the wound healing activity, we used a Scratch Test, an assay capable of evaluating the migratory ability of keratinocyte cells (HaCat) in vitro and thereby confirming the activity in rats. For the anti-inflammatory activity, the inflammation was induced with LPS in microglial murine cells (N9). Inflammatory mediators (IL-6, IL-10, IL-12p70, INFγ, MCP-1 and TNFα) were measured and the activity of superoxide dismutase (SOD), catalase (CAT), and mitochondrial membrane potential were evaluated. In addition, using paw edema induced by carrageenan in rats, the anti-inflammatory activity in vivo was analyzed. RESULTS: The PAHE and verbascoside, induced a significant increase in migration of keratinocytes, at all concentrations tested when compared to the negative control. The wound healing activity in vivo showed that the PAHE accelerated the process. The treatments with PAHE and verbascoside induce increases in the antioxidants enzymes, suggesting a possible activation of these enzymes. However, this did not result in an increase in the expression of inflammatory mediators in microglial cells. In LPS activated cells the verbascoside displayed a significant reduction of TNFα, IL-6, IL-12p70, MCP-1 and INFγ, while the PAHE only displayed statistically significant reduction in TNFα. Interestingly, both the compounds could reduce the oxidative parameters in N9 cells activated by LPS. Additionally, pretreatment with PAHE inhibited the paw edema in rats. CONCLUSION: The results suggest that PAHE has wound healing activity, improving cells migration and, as well as was able to reverse the oxidation effect in LPS-activated N9 cells. The wound-healing and anti-inflammatory activities of PAHE were confirmed in vivo. In addition, the presence of verbascoside can be related to PAHE effects, since this compound was capable of increase keratinocytes migration and inhibiting inflammation mediators.

Breast cancer i women: recurrence and survival at five years / Cácer de mama enn mujeres: recurrencia y supervivencia en cinco años / Câncer de mamaemm mulheres: recidiva e sobrevida em cinco anos

ABSTRACTWomen undergoing breast cancer treatment are still at risk of recurrence. This study investigated the association among survival and relapse with staging of breast cancer in women treated in an oncology reference hospital in Goiânia-GO, Brazil over a five-year period after diagnosis in 2008. This descriptive, retrospective, cross-sectional study was conducted with the medical records of 460 women. Data were analyzed using Fisher's Exact Test and the Kaplan Meier Curve. Overall, mortality, local/regional recurrence and metastasis were at 14.1%, 6.5% and 25.2%, respectively. There was an improvement in the survival of women with breast cancer at five years, but there is still a significant number of women diagnosed in advanced stages and experiencing recurrence, indicating there is a need to improve detection of the disease in its early stages and recurrences.

RESUMENLas mujeres que realizan tratamiento de cáncer de mama todavía presentan un riesgo de recurrencia de la enfermedad. Este estudio tiene como objetivo verificar a asociación entre sobrevivencia e recurencia del cáncer de mama en las mujeres atendidas en un hospital de referencia en Goiânia-GO cinco años después de del diagnóstico en 2008. Este es un estudio de cohorte, retrospectivo, descriptivo constituido por los expedientes de 460 mujeres. Los datos fueron analizados utilizando la Prueba Exacta de Fisher y la Curva de Kaplan Meier. Del total, la tasa de mortalidad, recurrencia regional/local y metástasis fue de 14.1%, 6.5% y 25.2%, respectivamente. Hubo una mejoría en la supervivencia de las mujeres con cáncer de mama, pero todavía existe un número considerable de mujeres que son diagnosticadas en estadios avanzados y que presentan recurrencia de la enfermedad, demostrando la necesidad de la detección precoz del cáncer en estadios iniciales y de casos de recurrencia.

RESUMOAs mulheres que realizam tratamento de câncer de mama ainda apresentam riscos de recorrência da doença. Objetivou-se verificar a associação entre sobrevida e recidiva com o estadiamento do câncer de mama em mulheres atendidas em um hospital de referência em Goiânia-GO cinco anos após o diagnóstico em 2008. Trata-se de um estudo de coorte, retrospectivo, descritivo constituído de prontuários de 460 mulheres. A análise dos dados foi realizada pelo Teste Exato de Fisher e Curva de Kaplan Meier. Do total, a taxa de óbito, recidiva local/regional e metástase foi de 14,1%, 6,5% e 25,2%, respectivamente. Verificou-se uma melhora da sobrevida de mulheres com câncer de mama, porém ainda há um número considerável de mulheres sendo diagnosticadas em estádios avançados e que apresentam recidiva da doença, demonstrando a necessidade de melhora na detecção precoce do câncer em estádios iniciais e de casos de recidiva.

[Emergency contraception: knowledge and attitudes of nursing students]. / Anticoncepção de emergência: conhecimento e atitude de acadêmicos de enfermagem.

This study aims to identify the knowledge and attitude towards emergency contraception among nursing students from a public university in Goiás--a state in Brazil. A descriptive and analytical research methodology with a quantitative approach was used, applying, from February to May 2011 a questionnaire on the sexual knowledge and attitude of students regarding emergency contraception. 178 students participated in the study. Knowledge was confirmed through the high frequency of correct answers to the questions, especially those concerning the correct time and the instructions for use (between 86%-96%). Although many students use this method, there are still some doubts about the mechanism of action, side effects and access availability (frequency of correct answers lower than 50%). We noticed the need to develop educational policies that encourage the promotion of sex education in schools and universities.

Emergency contraception: knowledge and attitudes of nursing students / Anticoncepción de emergencia: conocimientos y actitud de estudiantes de enfermería / Anticoncepção de emergência: conhecimento e atitude de acadêmicos de enfermagem

This study aims to identify the knowledge and attitude towards emergency contraception among nursing students from a public university in Goiás - a state in Brazil. A descriptive and analytical research methodology with a quantitative approach was used, applying, from February to May 2011 a questionnaire on the sexual knowledge and attitude of students regarding emergency contraception. 178 students participated in the study. Knowledge was confirmed through the high frequency of correct answers to the questions, especially those concerning the correct time and the instructions for use (between 86% - 96%). Although many students use this method, there are still some doubts about the mechanism of action, side effects and access availability (frequency of correct answers lower than 50%). We noticed the need to develop educational policies that encourage the promotion of sex education in schools and universities.

El objetivo fue identificar el conocimiento y la actitud frente a la anticoncepción de emergencia de estudiantes de enfermería de una universidad pública de Goiás utilizado la metodología descriptiva y analítica en la investigación con enfoque cuantitativo mediante un cuestionario aplicado de febrero a mayo de 2011 sobre el comportamiento sexual y el conocimiento de los estudiantes acerca de la anticoncepción de emergencia. Los participantes fueron 178 estudiantes. El conocimiento se lo confirma por la alta frecuencia de respuestas correctas a las preguntas, especialmente las relativas a la hora correcta, y a las instrucciones de uso (entre 86%-96%). A pesar de que muchos estudiantes utilizan este método, todavía hay algunas dudas sobre el mecanismo de acción, los efectos secundarios y la disponibilidad de acceso (frecuencia de aciertos inferiores al 50%). Tenga en cuenta la necesidad de desarrollar políticas educativas que fomenten la promoción de la educación sexual en las escuelas y universidades.

Objetivou-se identificar o conhecimento e a atitude em relação à anticoncepção de emergência de estudantes de enfermagem de uma universidade pública de Goiás. Utilizou-se a metodologia descritiva e analítica com abordagem quantitativa, com aplicação de questionário, de fevereiro a maio de 2011, sobre comportamento sexual e o conhecimento dos estudantes acerca da anticoncepção de emergência. Participaram do estudo 178 estudantes. O conhecimento se confirmou pela alta frequência de acertos às perguntas, principalmente as referentes ao tempo correto e as indicações de uso (entre 86% - 96%). Apesar de muitos estudantes utilizarem esse método, ainda persistem algumas dúvidas quanto ao mecanismo de ação, efeitos colaterais e disponibilidade de acesso (frequência de acertos inferior a 50%). Nota-se a necessidade de desenvolvimento de políticas educacionais que estimulem a promoção da educação sexual em escolas e universidades.

Investigation of biological activities of dichloromethane and ethyl acetate fractions of Platonia insignis Mart. seed.

Platonia insignis Mart., a native species of the Brazilian Amazon more commonly known as bacuri, is a member of the Clusiaceae family. In this study, we evaluated the chemical composition and the antioxidant and toxicity activities of the dichloromethane and ethyl acetate fractions from P. insignis seed ethanolic extract using different experimental models. Our results demonstrate in vitro antioxidant effects, by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt and 1,1-diphenyl-2-picryl-hydrazyl assays, as well as in vivo effects in antioxidant-defective Saccharomyces cerevisiae strains to both fractions. Toxicity was evaluated against the micro-crustaceous Artemia salina Leach. and promastigote Leishmania amazonensis. The dichloromethane fraction was the most active fraction evaluated on A. salina and promastigote L. amazonensis (IC(50)  = 24.89 µg/mL and 2.84 µg/mL, respectively). In addition, a slight cytotoxicity was observed in mammalian V79 cells using ethyl acetate and dichloromethane fractions with MTT assays. Both fractions displayed genotoxicity up to 25 µg/mL (dichloromethane) and 10 µg/mL (ethyl acetate) in V79 cells, as evaluated by the alkaline comet assay. Thus, in this study, we demonstrate for the first time that ethyl acetate and dichloromethane fractions from P. insignis seeds display antioxidant effects, a toxic effect against A. salina and L. amazonensis and induce genotoxicity in V79 mammalian cells. The observed activities can be attributed to the phenolic compounds present in these fractions and to the presence of xanthones (alpha- and gamma-mangostin).

Genotoxic and mutagenic properties of Bauhinia platypetala extract, a traditional Brazilian medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: Bauhinia platypetala Burch. is a traditionally used Brazilian medicinal plant, although no evidence in the literature substantiates the safety of its use. AIM OF THE STUDY: The aim of this study was to investigate the safety of the ethanolic extract and the ethereal fraction of B. platypetala leaves. MATERIALS AND METHODS: The identification of chemical compounds from the B. platypetala ethanolic extract and its ethereal fraction was performed by GC/MS and ESI-MS/MS. The plant's toxicological, cytotoxic, mutagenic and genotoxic properties were determined in Saccharomyces cerevisiae strains and V79 cell culture by survival assays and comet assay. RESULTS: The major compound identified in the B. platypetala ethanolic extract is palmitic acid, kaempferitirin and quercitrin, while the B. platypetala ethereal fraction was found to be rich in phytol, gamma-sitosterol and vitamin E. Moreover, the results indicated that the B. platypetala ethanolic extract has an anti-oxidative effect against H(2)O(2) in yeast. In addition, the B. platypetala ethanolic extract did not induce mutagenic effects on the S. cerevisiae N123 strain, but the ethereal fraction of B. platypetala at higher concentrations (250-500 µg/mL) induced cytotoxicity and mutagenicity. A slight cytotoxic effect was observed in mammalian V79 cells; however, both the B. platypetala ethanolic extract and its ethereal fraction were able to induce DNA strand breaks in V79 cells, as detected by the alkaline comet assay. CONCLUSION: The B. platypetala ethanolic extract has antioxidant action and showed absence of mutagenic effects in yeast S. cerevisiae. On the other hand B. platypetala ethereal fraction is mutagenic and does not show antioxidant activity in yeast. In mammalian cells B. platypetala ethanolic extract and it's ethereal fraction induce cyotoxic and genotoxic action.

Comparison of soxhlet, ultrasound-assisted and pressurized liquid extraction of terpenes, fatty acids and Vitamin E from Piper gaudichaudianum Kunth.

This paper describes a comparative study of extraction methods of terpenes (terpenic alcohols and phytosterols), fatty acids and Vitamin E from leaves of Piper gaudichaudianum Kunth. The analysis of extracts was done by gas chromatography with mass spectrometric detection. The identification and quantification was made by co-injections of the extract with certified standards. The use of pressurized liquid extraction (PLE; Dionex trade name: ASE, for accelerated solvent extraction) decrease significantly the total time of extraction, the amount of solvent and the manipulation of sample and solvents in comparison with soxhlet (SE) and ultrasound-assisted (USE). In addition, PLE was more effective for the extractions of terpenes (terpenic alcohols and phytosterols), fatty acids and Vitamin E.

Optimization of pressurized liquid extraction of Piper gaudichaudianum Kunth leaves.

Piperaceae family is original from tropical regions and it shows more than 700 species around the world. Piper gaudichaudianum Kunth is the specie more abundant in Brazil, occurring from Northeast to South Brazil. In this paper, it was investigated the influence of some experimental parameters on the pressurized liquid extraction (PLE) of P. gaudichaudianum Kunth leaves, using petroleum ether as extractor solvent. The optimization of the main variables involved in the PLE process (extraction temperature and time) has been done by response surface methodology (RSM) using, as responses, the extraction yield and the chromatographic profile (GC/MS) of the extracts. The optimized procedure employed 3 g of ground leaves, 10 min of extraction and one cycle of extraction at 85 degrees C. The major compounds present in the petroleum ether extracts were: palmitic acid, stearic acid and nerolidol. The results presented in this work show the possibility of using a fast and easy process to recover compounds from P. gaudichaudianum Kunth.

Heart transplantation across the antibodies against HLA and ABO.

We have intentionally performed heart transplantation in a 5-year-old child, despite the most unfavourable risk factors for patient survival; the presence of high level of antibodies against donor's human leucocyte antigen (HLA) class I/II and blood group antigens. Pretransplant treatment by mycophenolate mofetil, prednisolone, tacrolimus, intravenous immunoglobulin, rituximab, protein-A immunoadsorption (IA) and plasma exchange reduced antibody titres against the donor's lymphocytes from 128 to 16 and against the donor's blood group antigen from 256 to 0. The patient was urgently transplanted with a heart from an ABO incompatible donor (A(1) to O). A standard triple-drug immunosuppressive protocol was used. No hyperacute rejection was seen. Antibodies against the donor's HLA antigens remained at a low level despite three acute rejections. Rising anti-A(1) blood group antibodies preceded the second rejection and were reduced by two blood group-specific IAs and remained at a low level. The patient is doing well despite the persistence of donor-reactive antibodies.

Optimization of the sonication extraction method of Hibiscus tiliaceus L. flowers.

The influence of several experimental parameters on the ultrasonic extraction of Hibiscus tiliaceus L. flowers were investigated: extraction time, solvent polarity, sample amount, solvent volume and sample particle size. It was concluded that the most influential variables were extraction time and solvent polarity. The optimized procedure employed 5 g of ground flowers, 150 mL of methanol and 140 min of extraction. The extracts were fractionated using preparative silica columns and the resulting fractions were analyzed by GC/MS. Some saturated hydrocarbons, fatty acids, fatty acid methyl esters, phytosterols, and vitamin E were identified in the plant extracts.

Chemical composition of mate tea leaves (Ilex paraguariensis): a study of extraction methods.

The objective of this work was to investigate the extraction of Ilex paraguariensis leaves by means of three extraction techniques: pressurized liquid extraction (PLE, also called accelerated solvent extraction--ASE), maceration, and sonication. Samples of mate tea leaves were collected from an experiment conducted under agronomic control at Indfistria e Comércio de Erva-Mate Barão LTDA, Brazil. Six solvents with increasing polarities (n-hexane, toluene, dichloromethane, ethyl acetate, acetone, and methanol) were used in this investigation. Chemical analysis of the extracts was performed by GC coupled with a mass spectrometer detector. The identification and quantification were accomplished by coinjections of certified standards. The results showed that no significant differences in the qualities of the extracts were noticed regarding the extraction methods. On the other hand, the PLE technique was found to be more effective for the extractions of caffeine, phytol, palmitic, and stearic acid. The use of PLE led to a significant decrease in the total extraction time, amount of solvent consumption, and manipulation of samples compared to maceration and ultrasound-assisted extraction methods.