Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy / Avaliação do dano celular e da modulação das citocinas TNF-α, IL-6 e IL-10 em macrófagos expostos à terapia fotodinâmica mediada pela PpIX

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10-30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

Resumo Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (MΦ), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos MΦ mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). MΦ peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-α, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF-α e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em MΦ.

Antiproliferative and photoprotective activities of the extracts and compounds from Calea fruticosa

In this paper, we complement our previous study on the antiproliferative activity of Calea fruticosa (Asteraceae) by isolating the compounds apigenin-4',7-dimethyl ether (1), budlein A (2), quercetin (3), and cichoriin (4) from the plant's aerial parts. The antiproliferative activity of these compounds was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method against human tumor cell lines. Compound 3 displayed moderate antiproliferative activity in three cell lines (HCT-116, PC-3, and SF-295, with cell growth inhibition values of 72.97, 74.55, and 68.94%) and high antiproliferative activity (90.86%) in the HL-60 cell line. The in vitro sun protection factor (SPF) of the extracts and compound 4, with and without sunscreen, was determined by a spectrophotometric method. The ethanol extract exhibited the highest SPF (9.67) at a concentration of 0.100 mg/mL, while compound 4, isolated from this extract, showed a SPF of 13.79 at the same concentration. A relative increased efficacy of SPF was observed for the extracts and compound 4 when sunscreen was also used. Compound 4 has not been reported previously from any species within the genus Calea. Compounds 1-4 were obtained from this species for the first time.

Physicochemical characterization, the Hirshfeld surface, and biological evaluation of two meloxicam compounding pharmacy samples / 药物分析学报

Meloxicam (MLX) is an anti-inflammatory drug susceptible to variations and crystalline transitions. In compounding pharmacies, the complete crystallographic evaluation of the raw material is not a routine procedure. We performed a complete crystallographic characterization of aleatory raw MLX samples from compounding pharmacies. X-ray diffraction indicated the presence of two crystalline forms in one sample. DSC experiments suggested that crystallization, or a crystal transition, occurred differently be-tween samples. The FTIR and 1H NMR spectra showed characteristic assignments. 13C solid-state NMR spectroscopy indicated the presence of more than one phase in a sample from pharmacy B. The Hirshfeld surface analysis, with electrostatic potential projection, allowed complete assignment of the UV spectra in ethanol solution. The polymorph I of meloxicam was more active than polymorph Ⅲ in an experi-mental model of acute inflammation in mice. Our results highlighted the need for complete crystal-lographic characterization and the separation of freely used raw materials in compounding pharmacies, as a routine procedure, to ensure the desired dose/effect.

Evaluation of cell damage and modulation of cytokines TNF-, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (M), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated M cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal M from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 1030%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF- and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in M.

Resumo Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (M), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos M mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). M peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF- e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em M.

"Candidatus Rickettsia andeanae" en Amblyomma tigrinum, San Luis, Argentina

El objetivo del estudio fue detectar especies del género Rickettsia en garrapatas de la especie Amblyomma tigrinum colectadas sobre carnívoros domésticos y en sangre de caninos domésticos de la provincia de San Luis (Argentina). Entre 2013 y 2015 se colectaron 56 garrapatas adultas de la especie A. tigrinum sobre caninos y felinos domésticos, y se obtuvieron 65 muestras sanguíneas de caninos. Tres garrapatas resultaron positivas mediante la amplificación de un fragmento del espacio intergénico 23S-5S ARNr del género Rickettsia, lográndose secuenciar uno de los productos positivos. La muestra positiva secuenciada también resultó positiva por PCRs de los fragmentos de los genes gltA y ompA. Las secuencias obtenidas resultaron tener una identidad del 100 % de identidad con "Candidatus Rickettsia andeanae". Todas las muestras sanguíneas resultaron negativas. "Ca. R. andeanae" no ha sido asociada con enfermedad en humanos o animales, sin embargo, es necesario realizar nuevas investigaciones para lograr un mayor conocimiento del riesgo potencial de transmisión de rickettsiosis en la región.

The aim of this study was to detect species of Rickettsia in Amblyomma tigrinum ticks collected from domestic carnivores and blood of domestic dogs of San Luis (Argentina). Between 2013 and 2015, 56 adults of A. tigrinum from dogs and cats and 65 blood from dogs were collected. Three ticks were positive by amplification of a 23S-5S rRNA fragment, and the sequence of one of the positive products was obtained. The positive sample sequenced was positive by PCRs of fragments of genes gltA and ompA. The sequences obtained were 100% identical with "Candidatus Rickettsia andeanae". All blood samples were negative. "Ca. R. andeanae" has not been associated with disease in humans or animals; however, further research is necessary to achieve greater awareness of the potential risk of transmission of rickettsial diseases in the region.

Screening antimycobacterial activity of Baccharis dracunculifolia, Centella asiatica, Lantana camara and Pterodon emarginatus / Atividade anti-micobacteriana de Baccharis dracunculifolia, Centella asiatica, Lantana camara e Pterodon emarginatus

ABSTRACT The permanent investigation of new antimycobacterial drugs is necessary for the eradication programs of tuberculosis and other mycobacterium-related diseases. The aim of the present study is to search for new sources of antimycobacterial drugs using plant materials. In this study, 11 plant materials (extracts, essential oils and some fractions) obtained from 4 species of medicinal plants traditionally used as general therapeutics for different illnesses and specifically as treatment of tuberculosis, were evaluated using the microplate resazurin assay against 2 species of the Mycobacterium tuberculosis Complex and 3 nontuberculous mycobacteria. The results showed the hexane extract and the essential oil from fruits of Pterodonemarginatus (Vogel) as potential sources of antimycobacterial drugs against 4 species of tested mycobacteria. The hexane fraction of methanol extract from leaves of Centella asiatica also presented significant mycobacterial growth inhibition, but against M. chelonae only. In conclusion, it was possible to contribute to the antimycobacterial investigations by presenting three new samples of plants with significant antimicrobial activity against four Mycobacteriumspp and suggest future studies about the antimycobacterial properties of fruits from P. emarginatus.

RESUMO A investigação permanente de novas drogas antimicobacterianas é necessária no programa de erradicação da tuberculose e de outras doenças relacionadas com micobactérias. O objetivo deste estudo foi buscar novas fontes de drogas antimicobacterianas usando material vegetal. Neste estudo, 11 materiais de base vegetal (extratos, óleos essenciais e algumas frações) foram avaliados contra 5 espécies de micobactérias. Estes materiais foram obtidos a partir de 4 espécies de plantas medicinais tradicionalmente utilizadas como terapêutica geral para diferentes doenças e, especificamente, no tratamento de tuberculose (Baccharis dracunculifolia, Centella asiatica, Lantana camara, Pterodon emarginatus). Os ensaios foram realizados em microplacas com resazurina contra duas espécies do Complexo Mycobacteriumtuberculosis e 3 espécies de micobactérias não tuberculosas. Os resultados mostraram o extrato hexânico e o óleo essencial de frutos de P.emarginatus como potenciais fontes para drogas antimicobacterianas contra quatro espécies de micobactérias testadas. A fração hexânica do extrato metanólico das folhas de C. asiatica também apresentou significativa inibição do crescimento de micobactérias apenas contra M.chelonae. Em conclusão, foi possível contribuir para as investigações de antimicobacterianos por apresentar três novas amostras de plantas com atividade antimicrobiana significativa contra quatro Mycobacterium spp e sugerir a realização de estudos futuros sobre as propriedades antimicobacterianas de frutos de P. emarginatus.

O uso do peritônio de paca conservado em solução supersaturada de açúcar a 300 por cento ou glicerina a 98 por cento implantados na parede abdominal de ratos / The use of paca peritoneum preserved in supersaturated sugar solution 300 percent or glycerine 98 percent implanted in the abdominal wall of rats

Na busca de material biológico alternativo para a realização de implantes, objetivou-se com o presente estudo avaliar comparativamente a implantação do peritônio de paca, uma nova opção de biomaterial, conservado em solução supersaturada de açúcar a 300 por cento e conservado em glicerina a 98 por cento na parede abdominal de ratos Wistar. Foram utilizados 60 ratos, machos, da linhagem Wistar, pesando entre 150 e 200 gramas, organizados nos seguintes grupos experimentais: grupo controle (GI), grupo peritônio conservado em solução supersaturada de açúcar a 300 por cento (GII) e grupo peritônio conservado em glicerina a 98 por cento (GIII), cada um com 20 animais. Os grupos GII e GIII receberam o enxerto de peritônio da paca conservado em solução de açúcar 300 por cento e glicerina 98 por cento, respectivamente, e o grupo GI não recebeu a membrana. Cinco ratos de cada grupo foram submetidos à eutanásia em quatro momentos distintos: sete, 15, 30 e 60 dias de pós-operatório para avaliações macroscópicas e microscópicas da interface implante-tecido nativo. Apesar de reações adversas observadas em 57,5 por cento dos animais do grupo GII e GIII, em 95 por cento dos animais desses grupos houve boa cicatrização da membrana. Na análise histológica, verificou-se a presença de grande infiltrado inflamatório nos períodos iniciais (sete e 15 dias) e grande presença de tecido conjuntivo nos momentos finais (30 e 60 dias). Concluiu-se que o peritônio da paca como membrana biológica conservado nos meios estudados pode ser utilizado com segurança na parede abdominal de ratos; ainda, que sua conservação em solução supersaturada de açúcar a 300 por cento permitiu melhor maleabilidade no ato cirúrgico...

In the search for alternative biological material to perform implants, this study aimed to compare the implantation of paca peritoneum, a new biomaterial option, preserved in supersaturated sugar solution 300 percent and preserved in glycerin 98 percent in the abdominal wall of Wistar rats. A total of 60 male rats from the Wistar strain weighing between 150 and 200 grams were housed into three different experimental groups: control group (GI), peritoneum preserved in supersaturated sugar solution 300 percent group (GII) and peritoneum preserved in glycerin 98 percent group (GIII), with 20 animals each. The GII and GIII received the paca peritoneum graft preserved in sugar solution 300 percent and glycerin 98 percent, respectively and group GI did not receive any membrane. Five rats from each group were euthanized at four different times: seven, 15, 30 and 60 days post-surgery for macroscopic and microscopic evaluations in graft-native tissue interface. Despite the adverse reactions observed in 57.5 percent of GII and GIII, there was good healing of the membrane in 95 percent of the animals in these groups. Upon histological examination there was a large presence of inflammatory infiltrates in the initial periods (seven and 15 days) and a large presence of connective tissue in the final stages (30 and 60). It was concluded that the paca peritoneum as biological membrane preserved as presented in this study can be used safely in the abdominal wall of rats, the preservation in supersaturated sugar solution 300 percent allowed better flexibility during surgery...

Levantamento sobre o uso de plantas medicinais com a terapêutica anticâncer por pacientes da Unidade Oncológica de Anápolis / Survey on medicinal plants used with anticancer therapy by patients from Anapolis Oncology Unit

Este estudo teve por objetivo conhecer o perfil dos pacientes em tratamento contra o câncer da Unidade Oncológica de Anápolis quanto ao uso de plantas medicinais. Foram entrevistados 59 pacientes (42,12% da população estudada) por meio de questionários avaliativos enfatizando características sócio-demográficas e dados referentes à utilização de plantas medicinais. Evidenciou-se o uso indiscriminado de plantas medicinais entre os pacientes. A maioria dos entrevistados compartilha a opinião errônea de que plantas medicinais não fazem mal. A orientação sobre a forma de utilização das plantas ocorre, principalmente, pela informação de familiares ou amigos, e os profissionais da saúde muitas vezes são ignorados neste processo. Constatou-se que as plantas medicinais são utilizadas para o tratamento de enfermidades de baixa a alta gravidade, como o câncer. Dentre as 14 espécies usadas pelos pacientes com finalidade antineoplásica as mais mencionadas foram as popularmente conhecidas como noni, babosa, graviola e romã. Apesar de alguns estudos relatarem atividade antineoplásica ou quimiopreventiva para algumas espécies vegetais, muitas delas podem ser tóxicas ou apresentar potencial risco quando usadas concomitantemente ao tratamento convencional. Desta forma, observa-se que é preciso mais profissionais especializados para orientação sobre o risco de reações adversas e interações medicamentosas no que se refere ao uso de espécies vegetais e a terapêutica do câncer.

This study aims to survey the profile ofcancer patients in the Oncology Unit of Anápolis in relation to the use of medicinal plants. To evaluate socio-demographic characteristics and to retrieve data about on the use of medicinal plants, a questionnaire was filled by 59 patients, or 42.12% of the population under study. A widespread use of plants was found. Most of patients share the wrong idea that medicinal plants are not harmful. Guidance on the use of medicinal plants is given primarily by relatives and friends, and health professional are often ignored in this process. Among the 14 species used by patients with antineoplastic purpose, the most mentioned were popularly species known as "noni", "aloe", "soursop" and "pomegranate". Although some studies have reported chemopreventive or antineoplastic activity for some plant species, many may be toxic or show potential risk when used concurrently with conventional treatment. Thus, we can observe that there is a need for more specialized professionals for the guidance on the risk of adverse reactions and drug interactions in relation to the use of plant species and cancer therapeutics.

Anomalous colour in Neotropical mammals: a review with new records for Didelphis sp. (Didelphidae, Didelphimorphia) and Arctocephalus australis (Otariidae, Carnivora) / Coloração anômala em mamíferos Neotropicais: uma revisão com novos registros para Didelphis sp. (Didelphidae, Didelphimorphia) e Arctocephalus australis (Otariidae, Carnivora)

Anomalous colourations occur in many tropical vertebrates. However, they are considered rare in wild populations, with very few records for the majority of animal taxa. We report two new cases of anomalous colouration in mammals. Additionally, we compiled all published cases about anomalous pigmentation registered in Neotropical mammals, throughout a comprehensive review of peer reviewed articles between 1950 and 2010. Every record was classified as albinism, leucism, piebaldism or eventually as undetermined pigmentation. As results, we report the new record of a leucistic specimen of opossum (Didelphis sp.) in southern Brazil, as well as a specimen of South American fur seal (Arctocephalus australis) with piebaldism in Uruguay. We also found 31 scientific articles resulting in 23 records of albinism, 12 of leucism, 71 of piebaldism and 92 records classified as undetermined pigmentation. Anomalous colouration is apparently rare in small terrestrial mammals, but it is much more common in cetaceans and michrochiropterans. Out of these 198 records, 149 occurred in cetaceans and 30 in bats. The results related to cetaceans suggest that males and females with anomolous pigmentation are reproductively successful and as a consequence their frequencies are becoming higher in natural populations. In bats, this result can be related to the fact these animals orient themselves primarily through echolocation, and their refuges provide protection against light and predation. It is possible that anomalous colouration occurs more frequently in other Neotropical mammal orders, which were not formally reported. Therefore, we encourage researchers to publish these events in order to better understand this phenomenon that has a significant influence on animal survival.

Colorações anômalas ocorrem em muitos vertebrados tropicais. Entretanto, estas são consideradas raras em populações selvagens, havendo poucos registros para a maioria dos táxons. Reportam-se, neste estudo, dois novos casos de coloração anômala em mamíferos. Além disso, por meio de uma extensa revisão bibliográfica, foram compilados os casos publicados sobre coloração anômala em mamíferos neotropicais entre 1950 e 2010. Cada registro foi classificado como albinismo, leucismo, piebaldismo ou, eventualmente, como coloração indeterminada. Como resultados, reportou-se o registro de um espécime leucístico de gambá (Didelphis sp.) no sul do Brasil e de um espécime de lobo-marinho sul-americano (Arctocephalus australis) com piebaldismo no norte do Uruguai. Também foram analisados 31 artigos científicos, resultando em 23 registros de albinismo, 12 de leucismo, 71 de piebaldismo e 92 registros classificados como de pigmentação indeterminada. A coloração anômala aparentemente é rara em pequenos mamíferos terrestres, mas é muito mais comum em cetáceos e microquirópteros. Dos 198 registros encontrados, 149 ocorreram em cetáceos e 30 em morcegos. No caso dos cetáceos, este resultado sugere que machos e fêmeas com este padrão anômalo de pigmentação são reprodutivamente exitosos e, consequentemente, sua frequência está aumentando nas populações naturais. Com relação aos morcegos, este fenômeno pode estar relacionado ao fato de estes animais orientarem-se primariamente por meio de ecolocalização e seus refúgios oferecerem proteção contra luz e predação. É possível que a coloração anômala ocorra mais frequentemente em outras ordens de mamíferos neotropicais, as quais não foram formalmente reportadas. Desta forma, mostra-se importante encorajar os pesquisadores a publicar estes eventos em vida selvagem para um melhor entendimento deste fenômeno, que tem influência significativa na sobrevivência destes organismos.

Molecular detection of hemoplasma infection among cats from São Luís island, Maranhão, Brazil

Hemoplasmas are bacteria that infect erythrocytes, attaching to the red blood cell. There is a need for more reports of hemoplasma infection prevalence and molecular characterization among cats in Brazil since there are only few published reports. The present work aimed to detect and molecularly characterize the presence of hemotrophic mycoplasmas in domestic cats with outdoor access from São Luís, Maranhão, Brazil. Twenty cats (10%) were positive for Candidatus M. haemominutum, five (2.5%) for M. haemofelis, and four (2.%) for M. turicensis based on 16S rRNA gene PCRs. Five cats (2.5%) were co-positive for Candidatus M. haemominutum and M. haemofelis. PCR diagnosis was confirmed by sequencing; and phylogenetic analysis was based on 16S rRNA and rnpb genes.

Disponibilidade domiciliar de lipídeos para consumo e sua relação com os lipídeos séricos de adolescentes / Household availability of lipids for consumption and its relationship with serum lipids in adolescents / Disponibilidad domiciliar de lípidos para consumo y su relación con los lípidos séricos de adolescentes

Relacionar a disponibilidade domiciliar de lipídeos para consumo e o perfil lipídico sérico de adolescentes. MÉTODOS: Estudo transversal observacional. Avaliaram-se 95 adolescentes púberes de 10 a 13 anos de escolas públicas em Viçosa, Minas Gerais. Utilizou-se uma lista de compras mensal para avaliar a disponibilidade de alimentos per capita. Coletou-se amostra de sangue dos adolescentes, adotando-se o preconizado pela Sociedade Brasileira de Cardiologia como referência para adequação do perfil lipídico. Para as variáveis categóricas realizou-se o teste do qui-quadrado e calculou-se a Odds Ratio. Para comparar os grupos, aplicaram-se os testes Mann-Whitney ou t de Student, Kruskall-Wallis ou ANOVA. Verificou-se relação entre exames bioquímicos e disponibilidade de lipídeos pelas correlações de Spearmam ou de Pearson. RESULTADOS: Da energia total disponível, 27,2% foram provenientes de lipídeos. A disponibilidade de ácidos graxos saturados, monoinsaturados e poli-insaturados esteve de acordo com a recomendação de consumo. Em relação aos lipídeos séricos, 51,9% dos meninos e 55,8% das meninas apresentaram alterações de colesterol total (p=0,66); 18,6 e 25,6%, dos meninos e das meninas, respectivamente, apresentaram elevação de triglicerídeos, (meninas>meninos; p=0,02). Em relação ao LDL, 21,2 e 32,6% dos meninos e das meninas, respectivamente, apresentaram alterações (p=0,91). A prevalência de inadequação para HDL foi de 26,9 e 30,2% (p=0,93), para meninos e meninas, respectivamente. Não houve correlação entre os lipídeos disponíveis para consumo e os lipídeos séricos. Os adolescentes que possuíam mais que 35% de lipídeos disponíveis para consumo tiveram 9,1 vezes mais chances de apresentarem alterações de colesterol total (IC95% 1,81-61,74). CONCLUSÕES: É possível que adolescentes com maior disponibilidade de lipídeos para consumo apresentem colesterol total mais elevado.

To analyze the correlation between household availability of lipids to be consumed and the serum lipid profile in adolescents. METHODS: This cross sectional study enrolled 95 adolescents aged 10-13 years, selected by random raffle in public schools of Viçosa, Southeast Brazil. A monthly shopping list was used to evaluate the availability of food per capita. A blood sample was collected from each adolescent and Brazilian Cardiology Society guideline was adopted as a reference for lipid profile adequacy. For categorical variables, the chi-square was applied and the Odds Ratio calculated. For comparison between groups, Mann-Whitney or Student's t test, Kruskall-Wallis test or ANOVA were applied. Association between biochemical parameters and lipid availability was evaluated by Spearman or Pearson correlation tests. RESULTS: 27.2% of the total available energy came from lipids. The availability of saturated fatty acids, monounsaturated and polyunsaturated agreed with the recommendation for their consumption. Regarding serum lipids, 51.9% of boys and 55.8% of girls showed alteration in total cholesterol (p=0.66); 18.6 and 25.6% of boys and girls, respectively, showed increased triglycerides level (girls>boys; p=0.02). In relation to LDL, 21.2 and 32.6% of boys and girls, respectively, showed alteration (p=0.91). The prevalence of inadequacy for HDL was of 26.9 and 30.2% of boys and girls, respectively (p=0.93). There was no correlation between the available lipids for consumption and the serum lipids. The teenagers with more than 35% of lipids to be consumed had 9.1-fold higher levels of total cholesterol (95%CI 1.81-61.74). CONCLUSIONS: It is possible that adolescents with higher lipid availability at home show higher total cholesterol.

Relacionar la disponibilidad domiciliar de lípidos para consumo y el perfil lipídico sérico de adolescentes. MÉTODOS: Estudio transversal, observacional. Se evaluó a 95 adolescentes púberes de 10 a 13 años, de escuelas públicas en Viçosa, Minas Gerais (Brasil). Se utilizó una lista de compras mensual para evaluación de la disponibilidad de alimentos per capita. Se recogió muestra de sangre de los adolescentes, adoptándose lo preconizado por la Sociedad Brasileña de Cardiología como referencia para adecuación del perfil lipídico. Para las variables categóricas, se utilizó la prueba de chi cuadrado y se calculó la Odds Ratio. Para comparar los grupos, se aplicaron las pruebas de Mann-Whitney o t de Student, Kruskall-Wallis o ANOVA. Se verificó la relación entre exámenes bioquímicos y disponibilidad de lípidos por las correlaciones de Spearman o de Pearson. RESULTADOS: De la energía total disponible, el 27,2% fue proveniente de lípidos. La disponibilidad de ácidos grasos saturados, monoinsaturados o poli-insaturados, estuvo en conformidad con la recomendación de consumo. Respecto a los lípidos séricos, 51,9% de los muchachos y 55,8% de las muchachas presentaron alteraciones de colesterol total (p=0,66); 18,6 y 25,6% de los muchachos y muchachas, respectivamente, presentaron aumento de los triglicéridos (muchachas>muchachos; p=0,02). Respecto al LDL, 21,2 y 32,6% de los muchachos y muchachas, respectivamente, presentaron alteraciones (p=0,91). La prevalencia de inadecuación para HDL fue de 26,9 y 30,2% (p=0,93) respectivamente para muchachos y muchachas. No hubo correlación entre lípidos disponibles para consumo y lípidos séricos. Los adolescentes que poseían más que 35% de lípidos disponibles para consumo tuvieron 9,1 veces más posibilidad de presentar alteraciones de colesterol total (IC95% 1,81-61,74). CONCLUSIONES: Es posible que adolescentes con mayor disponibilidad de lípidos para consumo presenten colesterol total más elevado.

Cardiac and renal effects induced by different exercise workloads in renovascular hypertensive rats

We examined the effect of exercise training (Ex) without (Ex 0 percent) or with a 3 percent workload (Ex 3 percent) on different cardiac and renal parameters in renovascular hypertensive (2K1C) male Fisher rats weighing 150-200 g. Ex was performed for 5 weeks, 1 h/day, 5 days/week. Ex 0 percent or Ex 3 percent induced similar attenuation of baseline mean arterial pressure (MAP, 119 ± 5 mmHg in 2K1C Ex 0 percent, N = 6, and 118 ± 5 mmHg in 2K1C Ex 3 percent, N = 11, vs 99 ± 4 mmHg in sham sedentary (Sham Sed) controls, N = 10) and heart rate (HR, bpm) (383 ± 13 in 2K1C Ex 0 percent, N = 6, and 390 ± 14 in 2K1C Ex 3 percent, N = 11 vs 371 ± 11 in Sham Sed, N = 10,). Ex 0 percent, but not Ex 3 percent, improved baroreflex bradycardia (0.26 ± 0.06 ms/mmHg, N = 6, vs 0.09 ± 0.03 ms/mmHg in 2K1C Sed, N = 11). Morphometric evaluation suggested concentric left ventricle hypertrophy in sedentary 2K1C rats. Ex 0 percent prevented concentric cardiac hypertrophy, increased cardiomyocyte diameter and decreased cardiac vasculature thickness in 2K1C rats. In contrast, in 2K1C, Ex 3 percent reduced the concentric remodeling and prevented the increase in cardiac vasculature wall thickness, decreased the cardiomyocyte diameter and increased collagen deposition. Renal morphometric analysis showed that Ex 3 percent induced an increase in vasculature wall thickness and collagen deposition in the left kidney of 2K1C rats. These data suggest that Ex 0 percent has more beneficial effects than Ex 3 percent in renovascular hypertensive rats.

Excepcional y compleja lesión duodenal por ingesta de objetos metálicos / Duodenal perforation caused by metallic objects: case report

We report a 20 years old schizophrenic male, with a history of ingestion of four metallic tubes through which mate tea is drunk, that was admitted to the hospital with abdominal pain. A plain abdominal film showed the metallic objects in the abdominal cavity. The patient was operated. Two of the tubes were perforating the second portion of duodenum and the other two were in the large bowel. The tubes were extracted and the bowel wall was repaired. Due to a bad postoperative evolution, on a second operation, an antrectomy and duodenal excision was carried out. A duodenal fistula appeared that closed after 30 days, when the patient was discharged.

Las lesiones duodenales graves por ingestión de cuerpos extraños son excepcionales y cursan con alta morbimortalidad a pesar de los avances en el diagnóstico, su manejo quirúrgico y endoscópico. Actualmente, para su resolución definitiva se requiere a veces de un tratamiento por etapas y mediante la combinación de diversos procedimientos. Se presenta un caso clínico de lesión multivisceral a partir de una perforación duodenal (tipo III) provocada por la ingesta de varios objetos metálicos (bombillas de mate), que asocia compromiso hepático y colónico y complicada con una fístula duodeno cutánea. Su manejo requirió de varios procedimientos de reparación duodenal, que se combinaron en etapas sucesivas. El excepcional tipo de agente lesional, la compleja situación que provocó y el análisis de la literatura, otorgan particular interés al tema.

Proteinogramas séricos de ratos Wistar experimentalmente infectados com Trypanosoma evansi / Serum protein concentrations in Wistar rats experimentally infected with Trypanosoma evansi

Estabeleceu-se o perfil eletroforético de proteínas séricas de ratos Wistar experimentalmente infectados com Tripanosoma evansi, utilizando-se 40 ratos, distribuídos em oito grupos de cinco animais cada. Um grupo foi mantido como testemunho (G1), e os demais (G2 a G8) foram inoculados, via intraperitoneal, com cerca de 10³tripomastigota de T. evansi. Amostras de sangue para obtenção de soro foram coletadas no quinto (G2), 10º (G3), 15º (G4), 30º (G5), 45º (G6), 60º (G7) e 75º (G1 e G8) dia após as inoculações. O fracionamento das proteínas foi realizado pela técnica SDS-PAGE. Foram identificadas 31 proteínas, sendo sete de fase aguda: ceruloplasmina (101KD), hemopexina (83KD), transferrina (75KD), albumina (66KD), antitripsina (60KD), haptoglobina (44KD) e glicoproteína ácida (38KD). As proteínas com pesos moleculares 12KD; 22KD; 25KD; 28KD; 32,5KD; 35KD; 53,5KD; 63KD e 72KD apareceram apenas nos ratos inoculados com T. evansi.

This study established the electrophoretic profile of serum proteins of Wistar rats experimentally infected with Tripanosoma evansi. For such, 40 rats were allocated into eight groups of five animals. A group was kept as control (G1) and the others (G2 to G8) were intraperitoneally inoculated with 1.0 x 10³ tripomastigote of T. evansi. Blood samples were collected at 5th (G2), 10th (G3), 15th (G4), 30th (G5), 45th (G6), 60th (G7), and 75th (G1 and G8) days after inoculation (DAI). The serum protein concentrations were determined by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-one distinct proteins were identified, seven of these were identified as acute phase proteins: ceruloplasmin (110KD), hemopexin (83KD), transferrin (75KD), albumin (66KD), antitrypsin (60KD), haptoglobin (44KD), and acid glycoprotein (38KD). The proteins with molecular weights 12KD; 22KD; 25KD; 28KD; 32,5KD; 35KD; 53,5KD; 63KD, and 72KD were found only in infected rats.

In vitro culture, PCR, and nested PCR for the detection of Theileria equi in horses submitted to exercise / Cultivo in vitro, PCR e nested PCR na detecção de Theileria equi em eqüinos submetidos a exercícios

This study compared the usefulness of in vitro culture, PCR, and nested PCR for the diagnosis of Theileria equi in horses submitted to stress during exercise. Blood samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken prior to and after exercise. The animals were divided into two experimental groups: 30-day training schedule (G1) and 90-day training schedule (G2). Statistical analysis was performed using a chi-square test and kappa statistic was used in order to assess agreement. No significant difference was observed between samples collected at resting or after exercise. In G1, merozoites of T. equi were detected in the blood smears of four horses before in vitro culture, whereas 14 samples were positive, confirmed by culture. In G2, five and 11 horses were positive before and after culture, respectively. No PCR amplified product was observed in any of the tested animals although the PCR system based on the 16S rRNA gene of T. equi detected DNA in blood with an equivalent 8x10-5 percent parasitaemia. The nested PCR based on the T. equi merozoite antigen gene (EMA-1) allowed the visualization of amplified products in all the horses. Therefore, nested PCR should be considered as a means of detection of sub-clinical T. equi infections and in vitro culture could be used as a complement to other methods of diagnosis.

Comparou-se a utilização do cultivo in vitro, PCR e nested PCR no diagnóstico de Theileria equi em eqüinos submetidos ao estresse induzido por exercícios. Amostras de sangue foram obtidas de 15 eqüinos submetidos a treinamento em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Os animais foram divididos em dois grupos experimentais: 30 dias de treinamento (G1) e 90 dias de treinamento (G2). O teste do qui-quadrado foi empregado para as análises estatísticas e o índice kappa utilizado para avaliar a concordância. Não houve diferença significativa entre as amostras obtidas em repouso e após exercícios. Merozoítas de T. equi foram detectados em apenas quatro eqüinos do G1 pela microscopia direta realizada antes do cultivo, enquanto 14 animais apresentaram-se positivos pelo cultivo in vitro. No G2, cinco e 11 eqüinos foram positivos antes e após o cultivo, respectivamente. Nenhum produto de amplificação foi observado pela técnica de PCR, apesar do PCR baseado no gene 16S rRNA de T. equi ter detectado DNA em sangue com parasitemia equivalente a 8x10-5 por cento. O nested PCR, baseado na seqüência do gene do antígeno de merozoíta de T. equi (EMA-1,) permitiu a visualização de produtos de amplificação em todos os eqüinos testados. O nested PCR deve ser considerado para a detecção de infecções subclínicas de T. equi e o cultivo in vitro pode ser utilizado como uma ferramenta complementar a outros métodos de diagnóstico.

Frequency of the HFE C282Y and H63D polymorphisms in Brazilian malaria patients and blood donors from the Amazon region

Malaria is an endemic parasitosis and its causitive agent, Plasmodium, has a metabolism linked to iron supply. HFE is a gene with the polymorphisms C282Y and H63D, which are associated with a progressive iron accumulation in the organism leading to a disease called hereditary hemochromatosis. The aim of the present study was to determine the allelic and genotypic frequencies of the HFE gene polymorphisms in malaria patients and blood donors from the Brazilian Amazon region. We screened 400 blood donors and 400 malaria patients for the HFE C282Y and H63D polymorphisms from four states of the Brazilian Amazon region by polymerase chain reaction and restriction fragment length polymorphism analysis. We did not find any C282Y homozygous individuals, and the only five heterozygous individuals detected were from Pará State. The most frequent genotype in the North region of Brazil was the H63D heterozygote, in both study groups. Our results contribute to the concept that the Brazilian Amazon region should not be regarded as a single entity in South America. These polymorphisms did not influence the symptoms of malaria in the population studied, as neither severe signs nor high parasitemia were observed. Therefore, different hereditary hemochromatosis diagnostic and control measures must be developed and applied within its diverse locations. Investigations are currently being carried out in our laboratory in order to determine the importance of the coexistence of hereditary hemochromatosis in patients affected by parasitic diseases, such as malaria.

Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7

Toxoplasma gondii is an intracellular obligate protozoan, which infects humans and warm-blooded animals. The aim of the present study was to clone the rop2, gra5 and gra7 genes from T. gondii RH strain and to produce recombinant proteins. The rop2, gra5 and gra7 gene fragments produced by polymerase chain reaction were cloned into the pET102/D-TOPO® vector which contains thioredoxin and polyhistidine tags at the C- and N-ends, respectively, and is expressed in Escherichia coli BL21(DE-3). The expression fusion proteins were found almost entirely in the insoluble form in the cell lysate. These recombinant proteins were purified with an Ni-NTA column. Concentrations of the recombinant antigens produced in the E. coli BL21-star ranged from 300 to 500 μg/ml growth media, which was used to immunize rabbits. We observed an identity ranging from 96 to 97% when nucleotide sequences were compared to GenBank database sequences. Immunocharacterization of proteins was made by indirect immunofluorescence assay. These proteins will be used for serodiagnosis and vaccination.

Behavioral deficits in sepsis-surviving rats induced by cecal ligation and perforation

Sepsis and its complications are the leading causes of mortality in intensive care units, accounting for 10-50 percent of deaths. Intensive care unit survivors present long-term cognitive impairment, including alterations in memory, attention, concentration, and/or global loss of cognitive function. In the present study, we investigated behavioral alterations in sepsis-surviving rats. One hundred and ten male Wistar rats (3-4 months, 250-300 g) were submitted to cecal ligation and puncture (CLP), and 44 were submitted to sham operation. Forty-four rats (40 percent) survived after CLP, and all sham-operated animals survived and were used as control. Twenty animals of each group were used in the object recognition task (10 in short-term memory and 10 in long-term memory), 12 in the plus-maze test and 12 in the forced swimming test. Ten days after surgery, the animals were submitted individually to an object recognition task, plus-maze and forced swimming tests. A significant impairment of short- and long-term recognition memory was observed in the sepsis group (recognition index 0.75 vs 0.55 and 0.74 vs 0.51 for short- and long-term memory, respectively (P < 0.05). In the elevated plus-maze test no difference was observed between groups in any of the parameters assessed. In addition, sepsis survivors presented an increase in immobility time in the forced swimming test (180 vs 233 s, P < 0.05), suggesting the presence of depressive-like symptoms in these animals after recovery from sepsis. The present results demonstrated that rats surviving exposure to CLP, a classical sepsis model, presented recognition memory impairment and depressive-like symptoms but not anxiety-like behavior.

Síntese e modelagem molecular do novo derivado indolinônico como candidato a antiinflamatório COX-2 seletivo / Synthesis and molecular modeling of a novel indolinone derivative as a candidate anti-inflammatory selectively inhibiting COX-2

Os fármacos antiinflamatórios são sabidamente os mais comercializados em todo o mundo, e apesar disto apresentam sérios efeitos colaterais, sobretudo no que se refere ao trato gastrintestinal. A descoberta de novos protótipos com atividade e segurança terapêutica melhoradas continua sendo uma busca constante. Com o adventoda química computacional torna-se mais fácil o estudo teórico do comportamento fisiológico de uma nova substância bem como a compreensão do possível mecanismo de ação destas novas moléculas. Assim, através de modelos matemáticos de moléculas e receptores estudou-se neste trabalho o composto I (1-(2,6- diclorofenil)indolin-2-ona) quanto à sua possibilidade deinibir seletivamente a isoforma COX-2 da enzima prostaglandina endoperóxido sintase (PGHS), e também as melhores posições para a introdução de grupamentos químicos e modificações moleculares.

Experimental Trypanosoma evansi infection in donkeys: hematological, biochemical and histopathological changes

Five adult donkeys were experimentally infected with Brazilian strain of Trypanosoma evansi originally isolated from a naturally infected dog to study the hematological biochemical and histopathological alterations during the evolution of the disease. The course of the experimental infection was followed up to 145 days. Hematological analyses of the infected donkeys revealed a marked decline in hemoglobin, packed-cell volume, and erythrocyte count. Anemia was observed after successive peaks of parasitemia. Biochemical analyses showed increased levels of icterus index, serum globulins and decreased serum albumin and glucose values. All infected donkeys revealed enlargement of spleen and its white pulp, enlargement of mediastinal lymph nodes and lungs congestion. The main histopathological features consisted of meningoencephalitis. Demyelination in some areas of the cerebellum pediculus and neuropil vacuolization were observed. This study showed that donkeys infected with a Brazilian strain of T. evansi developed a chronic disease.

Cinco jumentos, adultos foram infectados experimentalmente com cepa brasileira de Trypanosoma evansi, isolada de um cão naturalmente infectado, com o intuito de observar as alterações hematológicas, bioquímicas e histopatológicas durante a evolução da enfermidade. O curso da infecção experimental foi de 145 dias. Análise hematológica dos jumentos infectados revelou declínio nos valores de hemoglobina, hematócrito e contagem total de eritrócitos. Notou-se anemia após sucessivos picos de parasitemia. Análise bioquímica indicou aumento dos níveis de índice ictérico, globulinas séricas e diminuição dos valores séricos de albumina e glicose. Todos os jumentos infectados apresentaram aumento do baço e de sua polpa branca, aumento de linfonodos mediastínicos e congestão pulmonar. Meningoencefalite foi o principal achado histopatológico. Em algumas áreas do pedículo cerebelar foram observadas desmielinização, além de vacuolização do neurópilo. O estudo mostrou que jumentos infectados com a cepa brasileira do T. evansi desenvolveram doença crônica.