Effects of quercetin-supplementation in NADH-diaphorase positive neurons subpopulations in the ileum of rats with experimental diabetes mellitus / Efectos de la suplementación con quercetina en subpoblaciones de neuronas NADH-diaforasa positivas en el Ileon de ratas con diabetes mellitus experimental

The effects of quercetin supplementation in NADH-diaphorase positive (NADH-d) neurons of streptozotocin-induced diabetic rats was carried in this study. Fifteen male rats were divided into three groups: normoglycemic (N), diabetic (D) and diabetic supplemented with quercetin (DQ). Whole mount preparations of the muscular layer of the ileum underwent NADH-d histochemistry for evidencing the NADH-d neuronal subpopulation. Quantitative analyzes were performed on 30 random fields, and morphometric analyzes in 100 neuronal bodies and nuclei per animal. The supplementation promoted a 44 % reduction in the neuronal density in D group when compared to N group (p <0.001); a 24.5 % reduction was observed in the DQ group when compared to N (p <0.01). Animals in D group presented an 18.7 % increase in the cell body areas of myenteric neurons when compared to N (p <0.001); DQ group showed a 14.2 % decrease in neuronal areas when compared to D (p <0.01); the nuclear area were similar among the three groups. We conclude that quercetin supplementation was positive for animals with diabetes mellitus.

Se estudiaron los efectos de la suplementación con quercetina en neuronas NADH-diaforasa positiva (NADH-d) de ratas diabéticas inducidas por estreptozotocina. Quince ratas machos se dividieron en tres grupos: normoglicémico (N), diabéticos (D) y diabéticos suplementados con quercetina (DQ). Las cortes montados de la capa muscular del íleon fueron sometidos a histoquímica de NADH-d para evidenciar la subpoblación neuronal NADH-d. Se realizaron análisis cuantitativos en 30 campos aleatorios y análisis morfométricos en 100 cuerpos y núcleos neuronales, por animal. La suplementación promovió una reducción del 44 % en la densidad neuronal en el grupo D cuando se comparó con el grupo N (p <0,001). Se observó una reducción del 24,5 % en el grupo DQ en comparación con N (p <0,01). Los animales del grupo D presentaron un aumento del 18,7 % en las áreas del cuerpo celular de las neuronas mientéricas cuando se compararon con N (p <0,001). El grupo DQ mostró una disminución de 14,2 % en las áreas neuronales en comparación con D (p <0,01). El área nuclear fue similar entre los tres grupos. Se concluye que la suplementación con quercetina fue positiva para animales con diabetes mellitus.

Effects of the Aqueous Extract from Tabebuia roseoalba and Phenolic Acids on Hyperuricemia and Inflammation.

Tabebuia species (Bignoniaceae) have long been used in folk medicine as anti-inflammatory, antirheumatic, antimicrobial, and antitumor. The aim of this study was to investigate if aqueous extract from the leaves (AEL) of Tabebuia roseoalba (Ridl.) Sandwith, Bignoniaceae, and its constituents could be useful to decrease serum uric acid levels and restrain the gout inflammatory process. HPLC analysis identified caffeic acid and chlorogenic acid in AEL. Antihyperuricemic effects and inhibition of liver XOD (xanthine oxidoreductase) by AEL and identified compounds were evaluated in hyperuricemic mice. Anti-inflammatory activity was evaluated on MSU (monosodium urate) crystal-induced paw edema. In addition, AEL antioxidant activity in vitro was evaluated. AEL, caffeic, and chlorogenic acids were able to reduce serum uric acid levels in hyperuricemic mice probably through inhibition of liver xanthine oxidase activity and significantly decreased the paw edema induced by MSU crystals. AEL showed significant antioxidant activity in all evaluated assays. The results show that the AEL of Tabebuia roseoalba can be a promising agent for treatment for gout and inflammatory diseases. We suggest that caffeic and chlorogenic acids may be responsible for the activities demonstrated by the species.

Campomanesia velutina leaves extracts exert hypouricemic effects through inhibition of xanthine oxidase and ameliorate inflammatory response triggered by MSU crystals

ABSTRACT Gout is a destructive arthritis with a high prevalence worldwide. However, the available therapy is not able to increase life quality in many patients. Campomanesia velutina (Cambess) O. Berg, Myrtaceae, is used in Brazilian folk medicine to treat pain, inflammation and rheumatism. The aim of this study was to evaluate the potential of ethanolic and aqueous extracts from C. velutina leaves to treat hyperuricemia and inflammation in gout arthritis model. Ethanolic extract of leaves and aqueous extract of leaves were in vitro assayed on xanthine oxidase inhibitory effect and in vivo on an experimental model of oxonate-induced hyperuricemia in mice, liver xanthine oxidase inhibition and monosodium urate crystal-induced paw edema model. The extracts at both tested doses (100 and 300 mg/kg) reduced serum urate levels. They were also able to inhibit xanthine oxidase in vitro and in vivo, demonstrating that this might be the mechanism of action underlying the urate-lowering effects. In addition, the extracts showed significant anti-inflammatory activity on monosodium urate crystal-induced paw edema, especially aqueous extract (100 and 300 mg/kg) that reduced edema at all evaluated times. Rutin and myricitrin were identified in ethanolic and in aqueous extracts. In this study, myricitrin was able to reduce serum uric acid levels and inhibit liver xanthine oxidase at the dose of 15 mg/kg. The anti-hyperuricemic activity of rutin has been previously reported. Thus, rutin and myricitrin seem to contribute to the observed effects of ethanolic and aqueous extracts. The results demonstrated the ability of aqueous and ethanolic extracts to lower serum urate levels and to reduce edema induced by monosodium urate crystals. Therefore, they may contribute to the management of gout in the future.

Effects of anabolic steroid treatment associated with physical training in adipose tissue of male Wistar rats / Efeito do tratamento com anabolizante associado com atividade física no tecido adiposo de ratos machos Wistar

Anabolic androgenic-steroids (AAS) include a broad class of synthetic derivatives of testosterone, being nandrolone decanoate the most widely used in sports environment. The aim of this study was to evaluate the metabolic effects of nandrolone decanoate in sedentary and trained adult male rats. We established four experimental groups: sedentary control, sedentary treated, trained control and trained treated. The training had consisted of running on a treadmill for nine weeks. Treated animals received intramuscular injections of nandrolone decanoate (0.5 mg kg-1) during the last four weeks of physical training. The training time as the drug used were not sufficient to significantly reduce body weight gain, but caused a significative decrease on diameter of adipocytes and in the amount of adipose tissue stored, as well as decreased the plasma levels of glucose and total cholesterol.

Os esteróides anabólicos androgênios incluem uma ampla classe de derivados sintéticos da testosterona, sendo o decanoato de nandrolona um dos mais utilizados no meio esportivo. Este trabalho teve como objetivo avaliar os efeitos metabólicos desse anabolizante esteróide em ratos machos adultos sedentários e treinados. Foram estabelecidos quatro grupos experimentais: sedentário controle, sedentário tratado, treinado controle e treinado tratado. O treinamento consistiu de corrida em esteira ergométrica durante nove semanas. Os animais tratados receberam injeção intramuscular de decanoato de nandrolona (0.5 mg kg-1) durante as quatro últimas semanas de treinamento físico. Tanto o tempo de treinamento, quanto a dose de anabolizante utilizado não foi eficiente para reduzir significativamente o ganho de peso corporal, mas causou reduções significativas no diâmetro dos adipócitos e na quantidade de tecido adiposo armazenado, assim como diminuiu os valores plasmáticos de glicose e de colesterol total.

Assessment of acute toxicity of the ethanolic extract of Lychnophora pinaster (Brazilian arnica)

Species of the Lychnophora genus are plants native to Brazil, popularly known as "Brazilian arnica" and used in folk medicine as alcoholic and hydro-alcoholic preparations for the treatment of bruises, inflammation, pain, rheumatism and insect bites. The present study aimed to evaluate the safety of the use of Lychnophora pinaster Mart., Asteraceae. Acute toxicity of the crude ethanolic extract was evaluated by administration of the extract by oral route to male and female Swiss mice. A single extract dose of 125, 250 or 500 mg/kg was administered and the effects on spontaneous locomotor activity, exploratory behavior, muscle strength, body weight, food and water consumption, relative organ weight, histology, as well as hematological and biochemical parameters were evaluated. The three doses administered to the animals did not cause muscle tone alterations, but doses of 250 and 500 mg/kg induced a significant inhibition of the spontaneous locomotor activity and exploratory behavior of the animals in open-field test. There was no alteration to hematological parameters and consumption of water and food, body weight variation and organs relative weight. Changes were observed in AST and ALT during assessment of biochemical parameters. The histopathological evaluation showed that the extract provoked cellular alterations, such as vacuolar degeneration and inflammation in kidneys and liver at all doses. Liver morphometric analyses of male and female mice showed that the extract did not have dose-dependent effects. Although females showed a significant increase in inflammatory cells, the effect was not dose-dependent.

Testis morphophysiology of rats treated with nandrolone decanoate and submitted to physical training / Morfofisiologia testicular de ratos tratados com decanoato de nandrolona e submetidos ao treinamento físico

The mammalian testis is a complex organ with endocrine and exocrine functions. It consists of seminiferous tubules where the production of the male gametes called spermatogenesis occurs. This process is influenced by a number of factors including the use of physical performance-enhancing drugs. The objective of this study was to evaluate the effects of the anabolic steroid nandrolone decanoate on the morphofunctional structure of testes in sedentary rats and rats subjected to moderate aerobic exercise training. Twenty-four male rats were divided into four experimental groups: sedentary control, sedentary treated, trained control and trained treated.The training lasted eight weeks and consisted of running on a programmable ergometer treadmill, tailored to train eight rats simultaneously. Treated animals received intramuscular injections of nandrolone decanoate (0.5 mg kg-1 body weight) during the last four weeks of physical training, while the control groups received intramuscular injections of vehicle (vegetable oil).The male reproductive system morphology showed that treatment with nandrolone decanoate, in both sedentary and trained rats, promoted morphological and functional changes that result in reduced efficiency of spermatogenesis.

O testículo de mamíferos é um órgão complexo com funções endócrinas e exócrinas. É composto por túbulos seminíferos, local onde ocorre a espermatogênese, processo de produção dos gametas masculinos. Este processo é influenciado por uma série de drogas potencializadoras do desempenho físico, que são utilizadas para melhorar a performance. O objetivo deste estudo foi avaliar os efeitos do anabolizante esteróide decanoato de nandrolona sobre a estrutura morfofuncional de testículos de ratos sedentários e submetidos ao treinamento físico aeróbico moderado. Foram utilizados vinte e quatro ratos machos divididos em quatro grupos experimentais: sedentário controle, sedentário tratado, treinado controle e treinado tratado. O treinamento teve duração de oito semanas e consistiu de corrida em esteira ergométrica programável, adaptada para treinar oito ratos simultaneamente. Os grupos tratados receberam injeção intramuscular de decanoato de nandrolona (0,5 mg kg -1) durante as quatro últimas semanas de treinamento físico, enquanto os grupos controles receberam injeção intramuscular do veículo (óleo vegetal). Os resultados mostraram que o tratamento com o decanoato de nandrolona, tanto em ratos sedentários como submetidos ao exercício físico, promove alterações morfofuncionais que resultam na redução da eficiência da espermatogênese.

Anti-inflammatory sesquiterpene lactones from Lychnophora trichocarpha Spreng. (Brazilian Arnica).

The aerial parts of Lychnophora trichocarpha Spreng. (Asteraceae) are used macerated in water or ethanol to treat inflammation, pain, rheumatism, contusions, bruises and insect bites in Brazilian traditional medicine. In this study, anti-inflammatory activity of ethanol extract from aerial parts of L. trichocarpha and its ethyl acetate fraction was investigated. Sesquiterpene lactones, lychnopholide (Lyc) and eremantholide C (EreC), isolated of ethyl acetate fraction, were also assayed for in vitro and in vivo anti-inflammatory activity. Topical treatment with ointments containing ethanol extract, its ethyl acetate fraction and sesquiterpene lactones significantly reduced carrageenan-induced mice paw oedema. In vitro assays demonstrated that Lyc inhibited interferon -γ/lipopolysaccharide -stimulated nitric oxide (NO) production in J774A.1 macrophages and increased production of IL-10 anti-inflammatory cytokine. The reduction of tumor necrosis factor-α (TNF-α) production by EreC was accompanied by an increased production of IL-10 in a concentration-dependent manner in J774A.1 macrophages. The anti-inflammatory effect of Lyc seems to involve the inhibition of production of NO and increased production of IL-10. The mechanism of the effect of EreC on the reduction of carrageenan-induced paw oedema may be attributed to inhibition of production of TNF-α and stimulation of IL-10 production. The results corroborate the use of ethanol extract from Lychnophora trichocarpha in folk medicine for anti-inflammatory action and indicate that the topical route is suitable for use.

Toxicological evaluation of ethanolic extract of Lychnophora trichocarpha, Brazilian arnica

The species of the genus Lychnophora, Asteraceae, are popularly known as "arnica" and are native from Brazilian savana (Cerrado). They are widely used in Brazilian folk medicine as anti-inflammatory, to treat bruise, pain, rheumatism and for insect bites. For evaluation of acute toxicity, the ethanolic extract was given to albino female and male mice. In open-field test, the extract of Lychnophora trichocarpha (Spreng.) Spreng. (0.750 g/kg) induced a significant inhibition of the spontaneous locomotor activity and exploratory behavior of the animals were observed 1 and 4 h after administration. In traction test, the same dose reduced the muscular force 1 h after administration. The exploratory behavior reduced significantly in the group that received 0.50 g/kg, 1 and 4 h after administration of the extract. The animals that received the doses of 0.25, 0.50 and 0.75 g/kg did not show any change of blood biochemical parameters comparing to control group and showed some histopathological changes such as congestion and inflammation of kidney and liver. The dose of 1.5 g/kg caused the most serious signs of toxicity. Histopathological changes observed was hemorrhage in 62.5% and pulmonary congestion in 100% of the animals. Brain and liver congestion was found in 62.5% of the animals.

Maytenus salicifolia Reissek, Celastraceae: triterpenes isolated from stems and antioxidant property of extracts from aerial parts

Six pentacyclic triterpenes were isolated from hexane extract of stems of Maytenus salicifolia Reissek, Celastraceae: 30-hydroxyfriedelan-3-one (1), 3,16-dioxofriedelane (2), friedeline (3), lupeol (4), betuline (5) and lup-20(29)-en-3,30-diol (6). The structure each one was established on the basis of detailed ¹H and 13C NMR spectral investigation and by comparison with the respective literature values. For compound 1, the complete 2D NMR (HMBC, HMQC and NOESY) spectral data were herein reported for the first time. Compounds 1, 2, 5 and 6 were isolated for the first time from this plant. Antioxidant activity is described for some extracts from species of the Celastraceae family, then, the extracts from aerial parts of M. salicifolia were evaluated in relation to antioxidant potential using the DPPH method. Compared to quecertin, the AcEt extract (EAF) from leaves, AcEt (EAPF) and MeOH (EMPF) from pulp fruit and AcEt (EAT) and MeOH (EMT) from stems showed significant antioxidant property.

Effect of the nandrolone decanoate on the efficiency of spermatogenesis of sedentary rats and rats subjected to physical training / Efeito do decanoato de nandrolona sobre a eficiência da espermatogênese de ratos sedentários e ratos submetidos a treinamento físico

Spermatogenesis is a complex, highly organized and coordinated process that results in the production of male gametes. This process is influenced by a number of drugs that enhance physical performance, which have been used mainly by young people, athletes or not, seeking to gain athletic performance or only a prominent social position. The objective of this study was to evaluate the effects of the anabolic steroid nandrolone decanoate on the efficiency of spermatogenesis in sedentary rats and rats subjected to physical training. Twenty-four male rats were divided into four experimental groups: sedentary control, sedentary treated, trained control and trained treated. Treated animals received intramuscular injection of nandrolone decanoate (0.5 mg kg-1 body weight) during the last four weeks of physical training. The training program consisted of running on a programmable ergometer treadmill, tailored to train eight rats simultaneously, for nine weeks. We evaluated the morphological and morphoquantitative profiles of the male reproductive system. The results showed that the treatment causes a reduction in the efficiency of spermatogenesis; however, when associated with physical training, this compensates for the anabolic action, keeping the process of spermatogenesis within normality.

A espermatogênese é um processo complexo, altamente organizado e coordenado, que resulta na produção dos gametas masculinos. Este processo é influenciado por uma série de drogas potencializadoras do desempenho físico, as quais têm sido utilizadas principalmente, por jovens, atletas ou não, que buscam adquirir maior desempenho esportivo ou mesmo apenas uma posição de destaque na sociedade. O objetivo deste estudo foi avaliar os efeitos do anabolizante esteroide decanoato de nandrolona na eficiência da espermatogênese de ratos sedentários e submetidos ao treinamento físico. Foram utilizados 24 ratos machos divididos em quatro grupos experimentais: sedentário-controle, sedentário tratado, treinado-controle e treinado tratado. Os animais tratados receberam injeção intramuscular de decanoato de nandrolona (0,5 mg kg-1) durante as quatro últimas semanas de treinamento físico. O programa de treinamento consistiu de corrida em esteira ergométrica programável, adaptada para treinar oito ratos simultaneamente, durante nove semanas. Foi avaliado o perfil morfológico e morfoquantitativo do sistema reprodutor masculino. Os resultados demonstraram que o tratamento causa redução da eficiência da espermatogênese, no entanto, o treinamento físico quando associado compensa a ação do anabolizante, mantendo o processo de espermatogênese normal.

Eunotiaceae (Bacillariophyceae) em igarapés da Amazônia Central, Manaus e Presidente Figueiredo, Brasil / Eunotiaceae (Bacillariophyceae) from central Amazon rivers, Manaus and Presidente Figueiredo districts, Brasil

Estudos florísticos e taxonômicos envolvendo diatomáceas são escassos para a região amazônica. As publicações existentes incluem registros de diatomáceas da Amazônia brasileira, do Equador, da Colômbia e do Peru e comumente mostram que Eunotia e Actinella (Eunotiaceae) são gêneros bem representados nessa região. A maioria dos igarapés amazônicos costuma apresentar potencial hidrogeniônico (pH) ácido, característica aquática que promove o desenvolvimento de uma comunidade típica de diatomáceas, dominada por espécimes de Eunotiaceae. O objetivo deste trabalho foi providenciar um levantamento florístico das espécies de Eunotiaceae presentes em igarapés da Amazônia Central brasileira e registrar os morfotipos de algumas espécies. Amostras fitoplanctônicas e perifíticas foram coletadas em cinco igarapés na rodovia BR-174, em Manaus e Presidente Figueiredo, em setembro e outubro de 1996 e fevereiro e março de 1997. Lâminas permanentes foram preparadas de acordo com a técnica de oxidação lenta para o estudo qualitativo. Vinte e três espécies pertencentes ao gênero Eunotia e seis ao gênero Actinella foram determinadas. Chaves dicotômicas de identificação, descrição detalhada, comentários relevantes e ilustrações foram providenciadas para cada táxon determinado. Morfotipos foram documentados para Eunotia zygodon. Espécies raramente citadas na literatura foram registradas, tais como, Eunotia falcifera e Eunotia rostellata.

Floristic and taxonomical studies about diatoms to Amazonian region are commonly well represented by Eunotia and Actinella (Eunotiaceae). The available works include diatom assemblage from Brazilian, Ecuadorian, Colombian and Peruvian Amazonia. The local streams often present acid pH which provides environmental conditions to develop a very particular diatom community dominated by specimens of Eunotiaceae. Thus, the aim of this paper is to give a floristic survey of Eunotiaceae from central Amazonian rivers and to register the typical morphologic frustule variations of some species. Planktonic and periphytic samples were collected from five rivers located at the BR-174 highway, near Manaus and Presidente Figueiredo districts, during 1996 and 1997. Slides were prepared in accordance with the technique of slow oxidation for the qualitative determination. Twenty-three species of Eunotia and six of Actinella were identified. Dicotomic keys, taxonomic comments and photographic illustrations were added. Morphotypes of Eunotia zygodon were registered. Species scarcelly mentioned on diatom literature were registered, such as, Eunotia falcifera and Eunotia rostellata.

The complete nucleotide sequence and genomic organization of Citrus Leprosis associated Virus, Cytoplasmatic type (CiLV-C).

The Citrus leprosis disease (CiL) is associated to a virus (CiLV) transmitted by Brevipalpus spp. mites (Acari: Tenuipalpidae). CiL is endemic in Brazil and its recently spreading to Central America represents a threat to citrus industry in the USA. Electron microscopy images show two forms of CiLV: a rare nuclear form, characterized by rod-shaped naked particle (CiLV-N) and a common cytoplasmic form (CiLV-C) associated with bacilliform-enveloped particle and cytoplasmic viroplasm. Due to this morphological feature, CiLV-C has been treated as Rhabdovirus-like. In this paper we present the complete nucleotide sequence and genomic organization of CiLV-C. It is a bipartite virus with sequence similarity to ssRNA positive plant virus. RNA1 encodes a putative replicase polyprotein and an ORF with no known function. RNA2 encodes 4 ORFs. pl5, p24 and p61 have no significant similarity to any known proteins and p32 encodes a protein with similarity to a viral movement protein. The CiLV-C sequences are associated with typical symptoms of CiL by RT-PCR. Phylogenetic analysis suggests that CiLV-C is probably a member of a new family of plant virus evolutionarily related to Tobamovirus.

Assessment of four different detergents used to extract membrane proteins from Xylella fastidiosa by two-dimensional electrophoresis

O objetivo deste trabalho foi comparar a eficiência da solubilização de quatro detergentes comercialmente disponíveis, ASB 14, SB 3-10, CHAPS e Triton X100, na extração de proteínas de membrana da bactéria Xylella fastidiosa para estudos proteômicos. Estas proteínas foram solubilizadas em duas etapas em tampões diferenciados pelos detergentes e submetidas à eletroforese bidimensional (2-DE) em uma faixa de pH não linear de 3-10. Os detergentes ASB 14 e SB 3-10 foram os mais efecientes, revelando 221 e 157 proteínas, respectivamente, enquanto que o CHAPS e o Triton X100 resultaram somente 72 e 43 proteínas, respectivamente. A identificação das proteínas foi feita por 'peptide mass fingerprinting' em espectrometria de massa MALDI-TOF, através de peptídeos obtidos por digestão com tripsina in gel. Os 18 spots de proteínas do gel com tratamento com ASB 14 mostrou que 83% eram proteínas de membrana. Este estudo concluiu que o detergente ASB-14 foi o mais eficiente na solubilização de proteínas de membrana de Xylella fastidiosa.

Proteome analysis of the plant pathogen Xylella fastidiosa reveals major cellular and extracellular proteins and a peculiar codon bias distribution.

The bacteria Xylella fastidiosa is the causative agent of a number of economically important crop diseases, including citrus variegated chlorosis. Although its complete genome is already sequenced, X. fastidiosa is very poorly characterized by biochemical approaches at the protein level. In an initial effort to characterize protein expression in X. fastidiosa we used one- and two-dimensional gel electrophoresis and mass spectrometry to identify the products of 142 genes present in a whole cell extract and in an extracellular fraction of the citrus isolated strain 9a5c. Of particular interest for the study of pathogenesis are adhesion and secreted proteins. Homologs to proteins from three different adhesion systems (type IV fimbriae, mrk pili and hsf surface fibrils) were found to be coexpressed, the last two being detected only as multimeric complexes in the high molecular weight region of one-dimensional electrophoresis gels. Using a procedure to extract secreted proteins as well as proteins weakly attached to the cell surface we identified 30 different proteins including toxins, adhesion related proteins, antioxidant enzymes, different types of proteases and 16 hypothetical proteins. These data suggest that the intercellular space of X. fastidiosa colonies is a multifunctional microenvironment containing proteins related to in vivo bacterial survival and pathogenesis. A codon usage analysis of the most expressed proteins from the whole cell extract revealed a low biased distribution, which we propose is related to the slow growing nature of X. fastidiosa. A database of the X. fastidiosa proteome was developed and can be accessed via the internet (URL: www.proteome.ibi.unicamp.br).