Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-β-D-(6"-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay / Análise do perfil toxicológico e farmacocinético de Kaempferol-3-O-β D-(6"-E-p-cumaril) glucopiranosídeo - Tilirosídeo: ensaio in silico, in vitro e ex vivo

Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as "pitó". This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6”-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como “pitó”. Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O-β-D- (6 ”-Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O--D-(6-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay

Abstract Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as pitó. This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O--D-(6-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

Resumo O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como pitó. Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O--D- (6 -Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-ß-D-(6"-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay / Análise do perfil toxicológico e farmacocinético de Kaempferol-3-O-ß-D- (6 "-E-p-cumaril) glucopiranosídeo - Tilirosídeo: ensaio in silico, in vitro e ex vivo

Abstract Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as "pitó". This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6"-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

Resumo O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como "pitó". Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O-β-D- (6 "-Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

In silico, in vitro, and ex vivo studies of the toxicological and pharmacological properties of the flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy

Phytochemical studies of the species Pavonia glazioviana were performed. Quercetin, kaempferol, acacetin, and trimethoxylated flavonoid compounds (which present biological activity) were isolated. We aimed to evaluate the in silico, in vitro, and ex vivo toxicity of flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy (Pg-1) obtained from P. glazioviana through chemical structure analyses, toxicity assessment, and predictive bioactive properties, using human samples in in vitro tests. In silico analysis suggested that Pg-1 presents a good absorption index for penetrating biological membranes (for oral bioavailability), while also suggesting potential antimutagenic, anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic, and apoptosis agonist bioactivities. Assessment of hemolytic and genotoxic effects revealed low hemolysis rates in red blood cells with no cellular toxicity in oral mucosa cells. The reduced cytotoxic activity suggested the safety of the concentrations used (500-1000 µg/mL), and demonstrated the varied interactions of Pg-1 with the analyzed cells. The data obtained in the present study suggested potential therapeutic application, and the non-toxic profile indicated viability for future studies.

Behavioral effects of endogenous or exogenous estradiol and progesterone on cocaine sensitization in female rats

Cocaine sensitization is a marker for some facets of addiction, is greater in female rats, and may be influenced by their sex hormones. We compared the modulatory effects of endogenous or exogenous estradiol and progesterone on cocaine-induced behavioral sensitization in 106 female rats. Ovariectomized female rats received progesterone (0.5 mg/mL), estradiol (0.05 mg/mL), progesterone plus estradiol, or the oil vehicle. Sham-operated control females received oil. Control and acute subgroups received injections of saline, while the repeated group received cocaine (15 mg/kg, ip) for 8 days. After 10 days, the acute and repeated groups received a challenge dose of cocaine, after which locomotion and stereotypy were monitored. The estrous cycle phase was evaluated and blood was collected to verify hormone levels. Repeated cocaine treatment induced overall behavioral sensitization in female rats, with increased locomotion and stereotypies. In detailed analysis, ovariectomized rats showed no locomotor sensitization; however, the sensitization of stereotypies was maintained. Only females with endogenous estradiol and progesterone demonstrated increased locomotor activity after cocaine challenge. Estradiol replacement enhanced stereotyped behaviors after repeated cocaine administration. Cocaine sensitization of stereotyped behaviors in female rats was reduced after progesterone replacement, either alone or concomitant with estradiol. The behavioral responses (locomotion and stereotypy) to cocaine were affected differently, depending on whether the female hormones were of an endogenous or exogenous origin. Therefore, hormonal cycling appears to be an important factor in the sensitization of females. Although estradiol increases the risk of cocaine sensitization, progesterone warrants further study as a pharmacological treatment in the prevention of psychostimulant abuse.

Characterization and anti-staphylococcal activity of the essential oil from Turnera subulata Sm

The present study is pioneer in analyzing the chemical composition of the essential oil from aerial parts of Turnera subulata Sm. and evaluates their antibacterial activity against a panel of drug-resistant strains of Staphylococcus aureus. The aerial parts were kiln-dried and then powdered in mechanical mill, and was subjected to hydrodistillation in a Clevenger-type apparatus. The components were analyzed using a mass spectrometrycoupled gas chromatography (GC-MS), and the identification of substances was performed by comparison of the mass spectra obtained with the mass spectra of the database of the GC-MS and retention indices . The essential oil was evaluated using the method of broth dilution at concentrations ranging from 3.125µg/mL to 3200µg/mL. It was possible to identify 45 substances (92.1%) of the essential oils, and the major components were trans-caryophyllene (6.7%), citronellol (5.6%), sphatulenol (5.3%), α-cadinol (4.3%), n-tricosano (4.3%), geraniol (4.1%) and trans-geranilacetone (3.7%), n-pentacosano (3.5%), globulol (3.4%), caryophyllene oxide (3.2%). The essential oil of T. subulata Sm. showed effective antibacterial activity for the various strains of S. aureus tested, with the MIC values between 25 µg/mL and 1600 µg/mL. The study of the essential oil of Turnera subulata Sm. showed that it consists of a complex mixture of several classes of compounds having the sesquiterpenes as major constituents followed by monoterpenes, and showed an antibacterial activity significant front the strains tested.

O presente estudo é pioneiro em analisar a composição química dos óleos essenciais das partes aéreas de Turnera subulata Sm. e sua atividade antibacteriana frente à Staphylococcus aureus resistentes a antibióticos. As partes aéreas da planta foram secas em estufa, pulverizadas em moinho mecânico e submetidas à hidrodestilaçao em aparato tipo Clevenger. A composição dos óleos essenciais foi analisada por cromatografia gasosa acoplada a espectrometria de massas (CG-EM), e a identificação realizada por comparação dos espectros de massas com a biblioteca do CG-EM e índices de retenção. A atividade antimicrobiana do óleo essencial foi avaliada usando o método de diluição em caldo, em concentrações que variaram de 3.125µg/mL to 3200µg/mL. Foi possível identificar 45 substâncias (92,1%) do óleo essencial, sendo os componentes majoritários: trans-cariofileno (6,7%), citronelol (5,6%), espatulenol (5,3%), α-cadinol (4,3%), n-tricosano (4,3%), geraniol (4,1%) e trans-geranilacetona (3,7%), n-pentacosano (3,5%), globulol (3,4%) e óxido de cariofileno (3,2%). O óleo essencial de T. subulata Sm. mostrou atividade antibacteriana eficaz para as várias cepas de S. aureus testadas, com valores de CIM entre 25 µg/mL e 1600 µg/mL. O estudo do óleo essencial de Turnera subulata Sm. evidenciou sua complexa mistura, contendo várias classes de substâncias, tendo os sesquiterpenos como constituintes majoritários seguido dos monoterpenos, e mostrou significativa atividade antibacteriana frente as cepas testadas.

Organic fertilization affecting pests and production of Lippia sidoides Cham. (Verbenacea) / Adubação orgânica afetando a ocorrência de pragas e a produção de Óleo essencial em Lippia sidoides Cham. (Verbenacea)

The objective of this paper was to study the effect of four levels of organic fertilization (0, 3, 6 and 9 kg m-2) to produce the essential oil of Lippia sidoides Cham. (Verbenacea) and its pests. A higher number of red mite Tetranychus sp. (Acari: Tetranychiidae) was observed on the abaxial face of the median and basal leaves of L. sidoides plants with eight weeks of age cultivated with 6 kg m-2 of organic fertilization. On the other hand, the smallest number of this mite was noted in control (0 kg m-2). L. sidoides cultivated with 6 kg m-2 of organic fertilization produced a greater quantity of essential oil. The highest number of mealy bug Phenacoccus sp. (Hemiptera: Pseudococcidae) was observed on the apical leaves (both leaf surfaces) of L. sidoides plants with nine and ten weeks of age cultivated with 3 kg m-2 of organic fertilization. No significant effect of the organic fertilization on Aphis gossypii Glover (Hemiptera: Aphididae) was detected. However, a greater number of this aphid and of its predator Scymnus sp. (Coleoptera: Coccinelidae) was observed on the abaxial face of the bottom leaves of L. sidoides plants with eight weeks of age. The dose of organic fertilization recommended for the production of essential oil of L. Sidoides is 6 kg m-2, however, one must pay attention to the fact this dosage favors the attack by Tetranychus sp.. And, when needed, pulverization must always be directed to the lower surface of the leaf.

O objetivo deste trabalho foi avaliar o efeito de quatro níveis de adubação orgânica (0, 3, 6, e 9 kg m-2) na produção de óleo essencial de Lippia sidoides Cham. (Verbenacea) e de suas pragas. Observou-se maior número de ácaro vermelho Tetranychus sp. (Acari: Tetranychiidae) na face inferior das folhas dos terços medianos e basais das plantas de L. sidoides com oito semanas de idade cultivadas com 6 kg m-2 de adubo orgânico. Por outro lado, a menor quantidade deste ácaro foi notada na testemunha (0 kg m-2). L. sidoides cultivada com 6 kg m-2 de adubo orgânico produziu maior quantidade de óleo essencial. Observou-se maior número de cochonilhas Phenacoccus sp. (Hemiptera: Pseudococcidae) nas folhas apicais, sem distinção de face foliar, das plantas de L. sidoides com nove e 10 semanas de idade cultivadas com 3 kg m-2 de adubo orgânico. Não se detectou efeito significativo de adubo orgânico sobre Aphis gossypii Glover (Hemiptera: Aphididae). Contudo, observou-se maior número desse pulgão bem como de seu predador Scymnus sp. (Coleoptera: Coccinelidae) nas faces abaxiais das folhas basais das plantas de L. sidoides com oito semanas de idade. A dose de adubo orgânico recomendada para produção de óleo essencial de L. sidoides é de 6 kg m-2. Contudo, essa dosagem favorece o ataque de Tetranychus sp. e, quando necessário, deve-se dirigir a pulverização sempre para a face inferior da folha, local preferencial de ataque dos artrópodes.

Avaliação da atividade antimicrobiana de Maytenus rigida Mart. (Celastraceae) / Evaluation of the antimicrobial activity of Maytenus rigida Mart. (Celastraceae)

A realização de estudos farmacológicos é fundamental para comprovar a eficácia do uso de plantas medicinais pela população para o tratamento de doenças e descobrir novos fitoterápicos. Esse estudo teve como objetivo avaliar o potencial antimicrobiano do extrato etanólico e fase acetato de etila do bom nome (Maytenus rigida Mart.) sobre Staphylococcus aureus ATCC 25923, 3 amostras de Staphylococcus aureus multirresistentes isoladas de pacientes com infecções nosocomiais, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 e Salmonella sp. (228-R-Tet, 118-R-Sut e 01-S) isoladas de ambiente aquático, utilizando o método de difusão em agar. Os testes revelaram que o extrato e fase de M. rigida apresentaram atividade antimicrobiana in vitro frente a todas as cepas de S. aureus testadas, apresentando concentração inibitória mínima (MIC) de 400 mg mL-1. Entretanto, estes produtos não apresentaram atividade frente às linhagens de bactérias Gram-negativas testadas, Escherichia coli, Pseudomonas aeruginosa e Salmonella sp.

Ppharmacological studies are essential to prove the effectiveness of using medicinal plants to treat diseases and discover new phytotherapics. This study aimed to evaluate the antimicrobial potential of ethanol and ethyl acetate extracts of "bom-nome" (Maytenus rigida Mart.) against Staphylococcus aureus ATCC 25923, three samples of multiresistant Staphylococcus aureus isolated from patients with nosocomial infections, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Salmonella sp. (228-R-Tet, 118-R-Sut and 01-S) isolated from water environment, using the agar diffusion test. Both extracts showed in vitro antimicrobial activity against all S. aureus strains, presenting 400 mg mL-1 minimum inhibitory concentration (MIC). However, these products did not show activity against strains of the Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa and Salmonella sp.

Produção de fitomassa e teor de óleo essencial de folhas de alecrim-pimenta (Lippia sidoides Cham. ) em diferentes espaçamentos de plantio / Phytomass production and essential oil yield of pepper-rosmarin (Lippia sidoides Cham. ) leaves in different planting spacings

O alecrim-pimenta, espécie nativa do nordeste brasileiro, é uma planta medicinal de porte arbustivo, com folhas aromáticas, que possuem óleo essencial rico em timol e carvacrol. Essas substâncias conferem grande importância à planta, uma vez que apresentam atividades farmacológicas comprovadas cientificamente. Desta forma, o estudo das condições de cultivo da espécie é essencial para obtenção de maiores produtividades. O espaçamento entre plantas destaca-se como um dos fatores mais importantes relativos ao cultivo, de forma que a produtividade e o rendimento da cultura são afetados pela densidade de plantas. Dada a necessidade de otimizar técnicas de cultivo do alecrim-pimenta (Lippia sidoides Cham.), objetivou-se avaliar a produção de fitomassa e óleo essencial em quatro espaçamentos de plantio. O trabalho foi realizado em campo, no Instituto de Ciências Agrárias da Universidade Federal de Minas Gerais (ICA/UFMG), no município de Montes Claros/MG. Utilizou-se o delineamento em blocos casualizados, com quatro tratamentos e cinco repetições. Os tratamentos foram definidos pelos espaçamentos de plantio: 1,0 x 0,5 m; 1,0 x 1,0 m; 1,5 x 1,0 m; e fileiras duplas (1,0 x 0,8 x 0,5m), sendo 1,0 m entre fileiras duplas, 0,8 m entre fileiras simples e 0,5 m entre plantas na fileira. Após 150 dias do plantio no campo, foram avaliadas as variáveis produção de fitomassa fresca e seca das folhas, a produção e o teor de óleo essencial. Observou-se que o espaçamento de 1,0 x 0,5 m possibilitou uma maior produção de fitomassa e de óleo essencial, sendo, portanto, o mais indicado para o cultivo do alecrim-pimenta (Lippia sidoides Cham.).

Native to Northeastern Brazil, pepper-rosmarin (Lippia sidoides Cham.) is a shrubby medicinal plant with aromatic leaves and essential oil rich in thymol and carvacrol. These components have great importance to the species since they present scientifically proven pharmacological activities. Thus, the study of cultivation conditions for this species is essential to obtain higher productivity. Spacing between plants is one of the most important factors related to cultivation, since crop productivity and yield are affected by plant density. Based on the need of optimizing pepper-rosmarin cultivation techniques, this work aimed to evaluate the phytomass production and the essential oil yield in four planting spacings. The experiment was carried out under field conditions in the Agricultural Sciences Institute, Federal University of Minas Gerais (ICA/UFMG), Montes Claros Municipality, Minas Gerais State, Brazil. Experimental design was in randomized blocks, with four treatments and five replicates. Treatments consisted of the planting spacings 1.0 x 0.5 m; 1.0 x 1.0 m; 1.5 x 1.0 m; and double rows (1.0 x 0.8 x 0.5 m), with 1.0m between double rows, 0.8m between single rows and 0.5m between plants in the row. After 150 days of planting in the field, leaf fresh and dry phytomass production and essential oil yield were evaluated. The spacing 1.0 x 0.5 m led to higher phytomass production and essential oil yield, being therefore the most recommended for pepper-rosmarin cultivation.

Calagem e adubação orgânica na produção de biomassa e óleo essencial em Lippia citriodora (Kunth) / Liming and organic fertilization on Lippia citriodora (Kunth) phytomass and essential oil production

O presente trabalho objetivou avaliar a calagem e adubação orgânica na produção de biomassa e óleo essencial em Lippia citriodora Kunth. O delineamento experimental utilizado constou de fatorial 7 x 2, sendo sete tratamentos (testemunha; adição de sulfato de Ca e Mg; calcário dolomítico; silicato de Ca e Mg; sulfato de Ca e Mg + esterco de curral; calcário dolomítico + esterco de curral; silicato de Ca e Mg + esterco de curral) e duas épocas de colheita, com quatro repetições, inteiramente casualizado (DIC). Verificou-se que a correção do solo mostrou-se prática necessária para o desenvolvimento da Lippia citriodora. Independentemente da época de colheita, a produção de massa fresca e seca foi maior com a aplicação do esterco de curral (32 t ha-1), no entanto, isso não refletiu em maior rendimento de óleo essencial.

The present work aimed to evaluate the effect of liming and bovine fertilization on Lippia citriodora Kunth phytomass and essential oil production. The experimental design was completely randomized with seven treatments (control; Ca and Mg sulfate; limestone; Ca and Mg silicate; Ca and Mg sulfate + manure; limestone + manure; Ca and Mg silicate + manure) and two harvest seasons, with four replicates. Soil adjustment showed to be a necessary procedure for Lippia citriodora development. Independently of the harvest season, fresh and dry matter yields were higher under treatments with bovine manure (32 t ha-1); however, no effect of treatments was observed on essential oil concentration.

Influence of progesterone on GAD65 and GAD67 mRNA expression in the dorsolateral striatum and prefrontal cortex of female rats repeatedly treated with cocaine

Female rats are intensely affected by cocaine, with estrogen probably playing an important role in this effect. Progesterone modulates the GABA system and attenuates the effects of cocaine; however, there is no information about its relevance in changing GABA synthesis pathways after cocaine administration to female rats. Our objective was to investigate the influence of progesterone on the effects of repeated cocaine administration on the isoenzymes of glutamic acid decarboxylase (GAD65 and GAD67) mRNA in brain areas involved in the addiction circuitry. Ovariectomized, intact and progesterone replacement-treated female rats received saline or cocaine (30 mg/kg, ip) acutely or repeatedly. GAD isoenzyme mRNA levels were determined in the dorsolateral striatum (dSTR) and prefrontal cortex (PFC) by RT-PCR, showing that repeated, but not acute, cocaine decreased GADs/β-actin mRNA ratio in the dSTR irrespective of the hormonal condition (GAD65: P < 0.001; and GAD67: P = 0.004). In the PFC, repeated cocaine decreased GAD65 and increased GAD67 mRNA ratio (P < 0.05). Progesterone replacement decreased both GAD isoenzymes mRNA ratio after acute cocaine in the PFC (P < 0.001) and repeated cocaine treatment reversed this decrease (P < 0.001). These results suggest that cocaine does not immediately affect GAD mRNA expression, while repeated cocaine decreases both GAD65 and GAD67 mRNA in the dSTR of female rats, independently of their hormonal conditions. In the PFC, repeated cocaine increases the expression of GAD isoenzymes, which were decreased due to progesterone replacement.

Flavonóides glicosilados de Herissantia tiubae (K. Schum) Brizicky (Malvaceae) e testes farmacológicos preliminares do canferol 3,7-di-O-a-L-ramnopiranosídeo / Glycosyl flavonoids from Herissantia tiubae (K. Schum) Brizicky (Malvaceae) and preliminary tests of kaempferol 3,7-di-O-a-L-rhamnopyranoside

Das partes aéreas de Herissantia tiubae (K. Schum.) Brizicky foram isolados, através de métodos cromatográficos, dois flavonóides glicosilados, canferol 3,7-di-O-a-L-ramnopiranosídeo e canferol 3-b-O-D-(6’’-E-p-cumaroil) glicosídeo. As estruturas foram identificadas com o uso de técnicas espectroscópicas de IV, RMN ¹H e 13C incluindo métodos bidimensionais, além de comparações com dados da literatura. O canferol 3,7-di-O-a-L-ramnopiranosídeo foi submetido a testes farmacológicos preliminares com a finalidade de avaliar o seu efeito sobre o sistema cardiovascular.

From the aerial parts of Herissantia tiubae (K. Schum.) Brizicky two flavonol glycosides were isolated by means of chromatographic methods, kaempferol 3,7-di-O-a-L-rhamnopyranoside and kaempferol 3-O-b-D-(6’’-E-p-coumaroyl) glucoside. Their structural identification was made by IV, ¹H and 13C NMR spectroscopy, including two dimensional techniques, together with comparison with literatura data. Preliminary tests were carried out with kaempferol 3,7-di-O-a-L-rhamnopyranoside in order to study its possible cardiovascular effect.

The impact of multimedia on teaching tropical medicine

This study evaluated the impact of multimedia in the information transfer of subjects specific to Tropical Diseases - Tetanus and Snake Envenoming. we evaluated the autonomous learning process of 76 fourth-year medical students at Botucatu School of Medicine of UNESP, using printed matter, video, and CD-ROM. The students were submitted to a specific test, which was repeated approximately one week later. They were divided into groups and received a kit containing a textbook, a video, and a CD-ROM. These materials were used for out-of-class study. Before the second test, the students gave a seminar, where they discussed and resolved their doubts with their professor. The results of the first test showed averages between 4.27±1.41 and 6,41±1.61. The second test, given after the seminar, presented averages that increased to 8.41±0.76 and 9.52±0.42, significance alfa = 5 per cent. At the end of the course, the students answered a questionnarie, which evaluated the material quality and acquired knowledge. The students concluded that multimedia was a more efficient and quick means for knowledge building than traditional teaching materials. They said that the active participation and interactivity with the CD-ROM were the major differences. The authors continue to study the associated use of printed matter, video, and CD-ROM as a faster alternative to the traditional method of information transfer, which may be of help in the knowledge building process in medical education.

Weight of preterm newborns during the first twelve weeks of life

A longitudinal and prospective study was carried out at two state-operated maternity hospitals in Belo Horizonte during 1996 in order to assess the weight of preterm appropriate-for-gestational-age newborns during the first twelve weeks of life. Two hundred and sixty appropriate-for-gestational-age preterm infants with birth weight <2500 g were evaluated weekly. The infants were divided into groups based on birth weight at 250-g intervals. Using weight means, somatic growth curves were constructed and adjusted to Count's model. Absolute (g/day) and relative (g kg-1 day-1) velocity curves were obtained from a derivative of this model. The growth curve was characterized by weight loss during the 1st week (4-6 days) ranging from 5.9 to 13.3 percent (the greater the percentage, the lower the birth weight), recovery of birth weight within 17 and 21 days, and increasingly higher rates of weight gain after the 3rd week. These rates were proportional to birth weight when expressed as g/day (the lowest and the highest birth weight neonates gained 15.9 and 30.1 g/day, respectively). However, if expressed as g kg-1 day-1, the rates were inversely proportional to birth weight (during the 3rd week, the lowest and the highest weight newborns gained 18.0 and 11.5 g kg-1 day-1, respectively). During the 12th week the rates were similar for all groups (7.5 to 10.2 g kg-1 day-1). The relative velocity accurately reflects weight gain of preterm infants who are appropriate for gestational age and, in the present study, it was inversely proportional to birth weight, with a peak during the 3rd week of life, and a homogeneous behavior during the 12th week for all weight groups

Evaluation of the impact of printed matter, video, and multimedia on the learning/teaching process in tropical diseases

This study evaluated the impact of printed matter, video, and multimedia on the learning/teaching process in Tropical Diseases. Eighty-four of 90 fourth-year medical students at Botucatu School of Medicine of UNESP were evaluated. The students received a kit containing a textbook, a video, and a CD-ROM on the Clinical Study of Tetanus to prepare a seminar on the subject. They were then asked to complete a questionnaire, which led to the following conclusions: 67.86 per cent read the textbook, 91.66 per cent watched the video, and 77.38 per cent explored the CD-ROM. These results were obtained observing the total number of students using each different media. When asked which of these media contributed most, the CD-ROM came out on top. The authors stress that this learning teaching process motivated the students by opening possibilities for new teaching alternatives in medicine.

Medium optimization fot the production of Actinomycin-D by Streptomyces parvulus

The present work aims to optimize the complex medium for the production of actinomycin-D by S.parvulus. The quantitative influence of the medium components was investigated in order to maximize the antibiotic synthesis. In the original medium, concentrations of actinomycin-D not higher than 245 ug/ml were achieved. In contrast, with the optimized medium the antibiotic concentration reached a value of approximately 530 ug/ml

Anti-anaphylactic and anti-inflammatory effects of ternatin, a flavonoid isolated from Egletes viscosa Less

Ternatin, a tetramethoxy flavone isolated from Egletes viscosa Less (Compositae), was tested for its efficacy in modulating mouse passive cutaneous anaphylaxis (PCA) and rat carrageenan-induced pleurisy. Ternatin (12.5, 25 and 50 mg/kg, ip) caused a dose-dependent inhibition of IgG antibody-mediated 1.5-h homologous PCA as well as IgE antibody-mediated 48-h homologous PCA in 2-month old mice (N = 5 per group). The inhibitory activity of ternatin was more potent on IgE-mediated PCA (47-79%) than on IgG (45-59%). In the rat carrageenan pleurisy test, ternatin (25 and 50 mg/kg, ip) reduced the response to carrageenan at 5 h both by decreasing exudate volume (33-40%) and leukocyte number (60%) in 5-6-month old rats (N = 6 per group). In contrast, indomethacin (2 mg/kg, po), a known cyclooxygenase inhibitor, showed greater potency in the inhibition of exudate volume (57%) and leukocyte number (77%). These results show that ternatin has both anti-inflammatory and anti-anaphylactic properties and suggest that it may be a useful alternative to anti-allergic drugs of the disodium cromoglycate (DSCG) type for use in bronchial asthma