DYNLT1 gene expression is downregulated in whole blood of patients at different Huntington's disease stages.

Huntington's disease (HD) is a neurodegenerative disorder caused by a CAG nucleotide expansion, which encodes the amino acid glutamine, in the huntingtin gene. HD is characterized by motor, cognitive, and psychiatric dysfunctions. In a previous study, we showed by qPCR that some genes altered in an HD mouse model were also altered in blood of HD patients. These alterations were mainly with respect to the dynein family. Therefore, this study aimed to investigate whether dynein light chain Tctex type 1 (DYNLT1) is altered in HD patients and if there is a correlation between DYNLT1 gene expression changes and disease progression. We assessed the DYNLT1 gene expression in the blood of 19 HD patients and 20 healthy age-matched controls. Also, in 6 of these patients, we analyzed the DYNLT1 expression at two time points, 3 years apart. The DYNLT1 gene expression in the whole blood of HD patients was significantly downregulated and this difference was widened in later stages. These data suggest that DYNLT1 could emerge as a peripheral prognostic indicator in HD and, also, might be a target for potential intervention in the future.

In Vitro Antitumor Activity of Sesquiterpene Lactones from Lychnophora trichocarpha / Atividade antitumoral in vitro de lactonas sesquiterpenicas de Lychnophora

The sesquiterpene lactones lychnopholide and eremantholide C were isolated from Lychnophora trichocarpha Spreng. (Asteraceae), which is a plant species native to the Brazilian Savannah or Cerrado and popularly known as arnica. Sesquiterpene lactones are known to present a variety of biological activities including antitumor activity. The present paper reports on the evaluation of the in vitro antitumor activity of lychnopholide and eremantholide C, in the National Cancer Institute, USA (NCI, USA), against a panel of 52 human tumor cell lines of major human tumors derived from nine cancer types. Lychnopholide disclosed significant activity against 30 cell lines of seven cancer types with IC100 (total growth concentration inhibition) values between 0.41 µM and 2.82 µM. Eremantholide C showed significant activity against 30 cell lines of eight cancer types with IC100 values between 21.40 µM and 53.70 µM. Lychnopholide showed values of lethal concentration 50% (LC50) for 30 human tumor cell lines between 0.72 and 10.00 µM, whereas eremantholide C presented values of LC50 for 21 human tumor cell lines between 52.50 and 91.20 µM. Lychnopholide showed an interesting profile of antitumor activity. The α-methylene-γ-lactone present in the structure of lychnopholide, besides two α,β- unsaturated carbonyl groups, might be responsible for the better activity and higher cytotoxicity of this compound in relation to eremantholide C.

As lactonas sesquiterpênicas licnofolida e eremantolida C foram isoladas de Lychnophora trichocarpha Spreng. (Asteraceae), espécie vegetal nativa do cerrado brasileiro e popularmente conhecida por arnica brasileira. As lactonas sesquiterpênicas são conhecidas por apresentarem variadas atividades biológicas, incluindo atividade antitumoral. O presente artigo relata a avaliação da atividade antitumoral in vitro de licnofolida e eremantolida C frente a um painel de 52 linhagens de células tumorais, provenientes de tumores humanos referentes a nove principais tipos de câncer. Os testes foram conduzidos no National Cancer Institute, USA (NCI, USA). Licnofolida apresentou atividade significativa frente a 30 linhagens de células tumorais referentes a sete tipos de câncer, com valores de CI100 (concentração que inibe 100% do crescimento celular) entre 0,41 µM e 2,82 µM. Eremantolida C mostrou atividade significativa frente a 30 linhagens de células tumorais referentes a oito tipos de câncer, com valores de CI100 entre 21,40 µM e 53,70 µM. Licnofolida apresentou valores de concentração letal 50% (CL50) para 30 linhagens de células tumorais humanas entre 0,72 e 10,00 µM, enquanto eremantolida C mostrou valores de CL50 para 21 linhagens entre 52,50 e 91,20 µM. Licnofolida apresentou um interessante perfil de atividade antitumoral. A presença na estrutura química da licnofolida de uma α-metileno-γ-lactona, além de dois grupos ésteres α,β-insaturados, podem ser responsáveis pela melhor atividade e maior citotoxicidade desta substância em relação à eremantolida C.

Growth curve of female collared peccaries (Pecari tajacu) raised in captivity in the Brazilian Amazon Region / Curva de crescimento de fêmeas cateto (Pecari tajacu) criados em cativeiro na Amazônia Brasileira

This work aims to determine the most suitable nonlinear model to describe the growth of female collared peccaries (Pecari tajacu). The monthly records of the weight of 10 captive female collared peccaries over a period of two years in the Brazilian Amazon Region were used. The growth models used were the Von Bertalanffy, Brody, Gompertz and Logistic. The parameters were estimated by using the NLIN procedure from the SAS application. The criteria used to verify the adjustment of the models were: asymptotic standard deviation (ASD); coefficient of determination (R²); average absolute residual deviation (ARD) and the asymptotic rate (AR). The Brody model and the Logistic model estimated the highest (19.44kg) and the lowest (19.18kg) asymptotic weight (A), indicating the lowest (0.0070kg/day) and the highest (0.0121kg/day) maturation rate (K). These results and the coefficients of phenotypic correlation that varied from -0.75 and -0.47 confirmed the antagonistic nature between these parameters. The Brody model estimated the lower value for ARD, a limiting factor for describing the lowest value for AR through this model. The Brody model showed the best adjustment for AR, although the other models also showed a suitable adjustment to the weight data of said species/gender. Based on the AR obtained in this work, the Brody model is recommended for adjusting the growth curve of the female collared peccaries. Depending on the estimated values, especially for K, this trait can be included in a selection index.

Com o objetivo de ajustar modelos não-lineares, foram utilizados registros mensais do peso de 10 fêmeas de cateto (Pecari tajacu) coletados durante dois anos, no criatório do campo experimental Álvaro Adolfo da Embrapa Amazônia Oriental, Belém, PA. Utilizaram-se os modelos de Von Bertalanffy, Brody, Gompertz e Logístico. Os parâmetros foram estimados usando o procedimento NLIN do aplicativo SAS. Os critérios utilizados para verificar o ajuste dos modelos foram: desvio padrão assintótico (ASD); coeficiente de determinação (R²); desvio médio absoluto dos resíduos (ARD) e o índice assintótico (AR). Os modelos Brody e Logístico estimaram, respectivamente, o maior (19,44kg) e o menor (19,18kg) peso assintótico (A), caracterizando a menor (0,0064kg/dia) e a maior (0,0113kg/dia) taxa de maturação (K), haja vista a natureza antagônica entre estes parâmetros, comprovada pela correlação fenotípica variando entre -0,75 à -0,47. O modelo Brody estimou o menor valor para o ARD, fator limitante para caracterizar o menor valor para o AR por este modelo. Considerando o AR, o modelo Brody apresentou o melhor ajuste, contudo, pelos valores encontrados, os demais modelos também apresentaram ajuste adequando aos dados ponderais da referida espécie/sexo. Com base no AR adotado neste trabalho, recomenda-se o modelo Brody para ajustar a curva de crescimento de fêmeas de cateto (Pecari tajacu). Em razão dos valores estimados, sobretudo, para a K, essa característica pode ser incluída em um índice de seleção. Contudo, estudos com grupos mais representativos e criados em outras condições se faz oportuno.

Functional morphology of the genital organs in the wild paca (Cuniculus paca) female.

Functional morphology of the genital organs is a key knowledge component for enhanced understanding of physiological patterns and for the determination of the reproductive performance in wild species. This study examines the morphology of genital organs of 133 paca females in the wild. Estimated conceptions and parturitions were mostly (83.7% and 75.5%, respectively) localized in the wet season. The pregnancy rate between 57.1% and 61.4% suggests an estimated yearly production of 1.37-1.48 parturitions and a long estimated farrowing interval of 247-266 days. Although large antral follicles were observed in all females, pregnant females had a greater number of antral follicles than females in the luteal phase. The average litter size was 1.03 foetuses per pregnant female, and mean ovulation rate was 1.33 follicles, resulting in a rate of reproductive wastage of 28.7%. The constituent active luteal tissues of the ovary were oestrous cyclic, pregnancy and accessory CL. The 50% of pregnant females in the late pregnancy stage lacked pregnancy CL, suggesting that placenta may become the mean source of progesterone during late stages of pregnancy. Results of the present study suggest that the observation of the vaginal closure membrane should not be an accurate tool for diagnosing oestrus in the paca female.

In vivo wound healing effects of Symphytum officinale L. leaves extract in different topical formulations.

The present work evaluates wound healing activity of leaves extracts of Symphytum officinale L. (comfrey) incorporated in three pharmaceutical formulations. Wound healing activity of comfrey was determined by qualitative and quantitative histological analysis of open wound in rat model, using allantoin as positive control. Three topical formulations, carbomer gel, glycero-alcoholic solution and O/W emulsion (soft lotion) were compared. The histological analysis of the healing process shows significant differences in treatment, particularly on its intensity and rate. The results indicate that emulsion containing both extracts, commercial and prepared, induced the largest and furthest repair of damaged tissue. This could be evidenced from day 3 to 28 by increase in collagen deposition from 40% to 240% and reduction on cellular inflammatory infiltrate from 3% to 46%. However, 8% prepared extract in emulsion presented the best efficacy. This work clearly demonstrates that comfrey leaves have a wound healing activity. The O/W emulsion showed to be the vehicle most effective to induce healing activity, particularly with extracts obtained from comfrey leaves collected in Minas Gerais state in Brazil. It shows the best efficacy to control the inflammatory process and to induce collagen deposition at 8% concentration.

Brine shrimp (Artemia salina Leach) bioassay of extracts from Lychnophoriopsis candelabrum and different Lychnophora species

O presente estudo teve como objetivo uma citotoxicidade sobre Artemia salina de vinhos e dois extratos de cinco espécies do gênero Lychnophora e de uma espécie de Lychnophoriopsis . Os extratos solubilizados em DMSO, preparados nas concentrações finais de 100, 250, 375, 500 e 600 µg mL -1 , foram adicionados a recipientes contendo náuplios de Artemia salina (10 unidades cada) e completo em volume para 5 mL de solução marinha. Lapachol e DMSO 5% foram como controles positivo e negativo, respectivamente. Como amostras foram mantidas sob iluminação e como larvas mortas foram contadas após 24 horas de contato. O cálculo da LC 50 foi feito com o programa Probitos. Os extratos brutos etanólicos de cinco Espécies apresentaram baixa letalidade nd Seguinte Ordem: Lychnophora trichocarpha (LC 50 = 672,38 ng mL -1 )> Lychnophora pinaster (LC 50 = 678,73 ng mL -1 )> Lychnophora ericoides (LC 50 = 738,09 µg mL -1 )> Lycellophoriopsis candelabro (LC 50 = 812,57 µg mL -1 )> Lychnophora passerina (LC 50 = 921,78 µg mL -1 ). Todos os extratos testados de Lychnophoriopsis candelabro eo extrato clorofórmico de Lychnophora staavioides mostraram leve toxicidade sobre A. salina . Os resultados indicaram que existem substâncias com potencial atividade farmacológica em todas as espécies testadas.

The present study aimed to evaluate on Artemia salina the citotoxicity of twenty-two extracts from five species of the genus Lychnophora and one species of the genus Lychnophoriopsis. The extracts solubilized in DMSO and prepared at the final concentrations of 100, 250, 375, 500 and 600 g mL-1 were added to tubes containing Artemia salina nauplii (10 units each) and filled to 5 mL total volume with artificial salt water. Lapachol and 5% DMSO were used as positive and negative controls, respectively. The samples were kept under light and dead larvae were counted after 24 hours of contact. LC50 was calculated by using Probit software. The crude ethanol extracts from five species showed low lethality in the following order: Lychnophora trichocarpha (LC50 = 672.38 g mL-1) > Lychnophora pinaster (LC50 = 678.73 g mL-1) > Lychnophora ericoides (LC50 = 738.09 g mL-1) > Lychnophoriopsis candelabrum (LC50 = 812.57 g mL-1) > Lychnophora passerina (LC50 = 921.78 g mL-1). All tested extracts from L. candelabrum and chloroform extract from L. staavioides showed light toxicity on A. salina. Results indicated that there are substances with potential pharmacological activity in all tested species

Pyrrolidine dithiocarbamate down-regulates vascular matrix metalloproteinases and ameliorates vascular dysfunction and remodelling in renovascular hypertension.

BACKGROUND AND PURPOSE: Mounting evidence implicates matrix metalloproteinase (MMP) in the vascular dysfunction and remodelling associated with hypertension. We tested the hypothesis that treatment with pyrrolidine dithiocarbamate (PDTC), which interferes with NF-κB-induced MMPs gene transcription, could exert antihypertensive effects, prevent MMP-2 and MMP-9 up-regulation, and protect against the functional alterations and vascular remodelling of two-kidney, one clip (2K1C) hypertension. EXPERIMENTAL APPROACH: Sham-operated or hypertensive rats were treated with vehicle or PDTC (100 mg·Kg(-1) ·day(-1)) by gavage for 8 weeks. Systolic blood pressure (SBP) was monitored weekly. Aortic rings were isolated to assess endothelium-dependent relaxations. Quantitative morphometry of structural alterations of the aortic wall was carried out in haematoxylin/eosin sections. Formation of vascular reactive oxygen species (ROS), and inducible (i) NOS and phosphorylated-p65 NF-κB subunit expression were measured in the aortas. MMP-2 and MMP-9 aortic levels and gelatinolytic activity were determined by gelatin and in situ zymography and by immunofluorescence. KEY RESULTS: Treatment with PDTC attenuated the increases in SBP and prevented the endothelial dysfunction associated with 2K1C hypertension. Moreover, PDTC reversed the vascular aortic remodelling, the increases in aortic ROS levels and in iNOS and phosphorylated-p65 NF-κB expression found in 2K1C rats. These effects were associated with attenuation of 2K1C up-regulation of aortic MMP-2 and MMP-9 levels and gelatinolytic activity. CONCLUSION AND IMPLICATIONS: These findings suggest that PDTC down-regulates vascular MMPs and ameliorates vascular dysfunction and remodelling in renovascular hypertension, thus providing evidence supporting the suggestion that PDTC is probably a good candidate to be used to treat hypertension.

Anti-inflammatory activity of Lychnophora passerina, Asteraceae (Brazilian "Arnica").

ETHNOPHARMACOLOGICAL RELEVANCE: Lychnophora passerina (Asteraceae), popularly known as "arnica," is used to treat inflammation, pain, rheumatism, contusions, bruises and insect bites in Brazilian traditional medicine. MATERIALS AND METHODS: The anti-inflammatory activity of crude ethanolic extract of aerial parts of L. passerina and its ethyl acetate and methanolic fractions had their abilities to modulate the production of NO, TNF-α and IL-10 inflammatory mediators in LPS/IFN-γ-stimulated J774.A1 macrophages evaluated. Moreover, the crude ethanolic extract and derived fractions were also in vivo assayed by carrageenan-induced paw oedema in mice. RESULTS: In vitro assays showed remarkable anti-inflammatory activity of L. passerina crude ethanolic extract (EE) and its ethyl acetate (A) and methanolic (M) fractions, through the inhibition of production of NO and TNF-α inflammatory mediators and induction of production of IL-10 anti-inflammatory cytokine. In vivo assays showed anti-inflammatory activity for EE 10% ointment, similar to the standard drug diclofenac gel. The A and M fraction ointments 20% presented anti-inflammatory activity. CONCLUSION: The results obtained showed that possible anti-inflammatory effects of EE and its A and M fractions may be attributed to inhibition pro-inflammatory cytokines production, TNF-α and NO and to increased IL-10 production. EE, A and M ointments showed topical in vivo anti-inflammatory activity. The in vivo anti-inflammatory activity of EE of L. passerina may be related to synergistic effects of different substances in the crude extract. Therefore, traditional use of aerial parts of L. passerina in the inflammatory conditions could be beneficial to treat topical inflammatory conditions, as evidenced by the present study.

Lack of association of C282Y and H63D mutations in the hemochromatosis (HFE) gene with diabetes mellitus type 2 in a case-control study of women in Brazil.

Hereditary hemochromatosis is one of the most common autosomal recessive diseases; it is characterized by excess absorption of iron. Clinically, the major challenge is to diagnose increased iron deposition before irreversible tissue damage has occurred. C282Y and H63D are the main mutations related to hereditary hemochromatosis; these mutations have been reported to be associated with increased risk of developing diabetes mellitus type 2 (DM2). We investigated whether these mutations are associated with increased risk for the development of DM2 in women in Brazil. Seventy-two women with clinical diagnosis of DM2 under treatment with hypoglycemic agents and a control group composed of 72 women with no clinical history of diabetes were studied. The C282Y and H63D mutations were determined by PCR-RFLP. Significant differences were not observed for C282Y and H63D, when we compared diabetic and non-diabetic women. We suggest that mutations C282Y and H63D in the HFE gene are not significant risk factors for the development of DM2 in Brazilian women.

Antinociceptive and anti-inflammatory activities of ethanolic extracts of Lychnophora species.

Extracts from Lychnophora species are traditionally used in Brazil as anti-inflammatory, and to treat bruise, pain and rheumatism. The ethanolic extract of aerial parts of five species of Lychnophoras and one specie of Lychnophoriopsis were examined for the antinociceptive (hot-plate and writhing tests) and anti-inflammatory (carrageenan-induced paw oedema test) activity in mice, by oral and topical routes, respectively. In the hot-plate test, the Lychnophora pinaster (0.75 g/kg) and Lychnophora ericoides (1.50 g/kg) extracts significantly increased the time for licking of the paws. The species Lychnophora passerina, Lychnophoriopsis candelabrum and Lychnophora pinaster, using the dose of 0.75 g/kg, and Lychnophora ericoides and Lychnophora trichocarpha in both doses evaluated (0.75 and 1.50 g/kg) significantly reduced the number of writhes induced by acetic acid. The administration of Lychnophora pinaster and Lychnophora trichocarpha ointments, in both concentrations evaluated (5 and 10%, w/w), and Lychnophora passerina and Lychnophoriopsis candelabrum, in the concentration of 10%, significantly reduced the paw oedema measured 3 h after carrageenan administration, suggesting, for the first time, an anti-inflammatory activity upon topical administration of these species. The present work comparatively demonstrated the antinociceptive and anti-inflammatory activities of some Brazilian Lychnophoras.

Xanthine oxidase inhibitory activity of Lychnophora species from Brazil ("Arnica").

Twenty-two extracts from five Lychnophora species and one Lychnophoriopsis species, traditionally used in Brazil as analgesic, anti-inflammatory, and to treat bruise and rheumatism were examined for the inhibition of xanthine oxidase (XO), the enzyme that catalyses the metabolism of hypoxanthine and xanthine into uric acid. Sixteen extracts were tested. All of them were found to have excellent XO inhibitory activity, with inhibitions greater than 38% at 100 microg/mL in the assay mixture. The most active plants examined were Lychnophora trichocarpha, Lychnophora ericoides, Lychnophora staavioides and Lychnophoriopsis candelabrum, with inhibitions of 77%, 78%, 66% and 63% at 100 microg/mL, respectively, and IC(50) values of 6.16, 8.28, 33.97 and 37.70 microg/mL, respectively.