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Monoclonal gammopathy (MG) is rare in dogs and multiple myeloma (MM) is its main cause. This study reports the case of a female dog which presented MG associated with a MM and consequent blood hyperviscosity syndrome (HVS). Patient presented systemic hypertension, renal disease, chorioretinitis and secondary glaucoma due to HVS. Serological tests for leishmaniasis and for ehrlichiosis demonstrated negative and score 4 from 5, respectively. Non-regenerative anemia, thrombocytopenia, hyperproteinemia, hypoalbuminemia, hyperglobulinemia, creatinine and blood urea nitrogen (BUN) increases, hypercalcemia, hyperphosphatemia, proteinuria, decreased urinary density, and monoclonal peak of gamma globulins were observed. First myelogram identified 81% of medullary plasmacytosis which suggested MM. Plasmapheresis and chemotherapy with melphalan and prednisolone were performed with positive results. The treatment was effective with complete remission of HVS signs, medullary plasmocytosis reduction (21,8%) and malignant criteria decrease, as well as neoplastic control.

Gamopatia monoclonal (GM) é rara nos cães, e o mieloma múltiplo (MM) é a sua principal causa. Este estudo descreve o caso de uma cadela que apresentou GM associada ao MM e consequente síndrome da hiperviscosidade sanguínea (SHS). A paciente apresentou hipertensão arterial sistêmica, doença renal, coriorretinite e glaucoma secundário decorrentes da SHS. Testes sorológicos para leishmaniose e para erliquiose apresentaram baixos títulos e escore 4 e 5, respectivamente. Anemia arregenerativa, trombocitopenia, hiperproteinemia com hipoalbuminemia e hiperglobulinemia, aumentos de ureia e creatinina, hipercalcemia, hiperfosfatemia, proteinúria, diminuição da densidade urinária e pico monoclonal de gamaglobulinas foram identificados. Foi consignado um primeiro mielograma, com identificação de 81% de plasmocitose medular, sugestivo de MM. Dessa forma, foi instituída plasmaférese e quimioterapia com melfalano e prednisolona, com resultados positivos. O tratamento instituído foi efetivo, com auxílio na completa redução dos sinais da SHS, na diminuição da plasmocitose medular (21,8%) e dos critérios de malignidade, bem como no controle da neoplasia.

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Anticancer therapy often leads to premature ovarian insufficiency (POI) and infertility due to the extreme sensitivity of the ovarian follicle reserve to the effects of chemotherapy. Withanolides are known for their cytotoxic effect on cancer cells and low cytotoxicity on non-malignant or healthy cells. Therefore, this study aimed to investigate the in vivo effects of three withanolides derivatives: 27-dehydroxy-24,25-epoxywithaferin A (WT1), 27-dehydroxywithaferin A (WT2), and withaferin A (WTA) on fertility, and the ovarian preantral follicles of young female mice. To achieve this, mice received 7 intraperitoneal doses of WT1, WT2, or WTA at a concentration of 2 mg/kg (Experiment I) and 5 or 10 mg/kg (Experiment II) over 15 alternate days. In experiment I, two days after administration of the last dose, half of the mice were mated to evaluate the effects of withanolides on fertility. The other half of the mice, as well as all mice from experiment II, were sacrificed for histological, inflammation, senescence, and immunohistochemical analyses of the follicles present in the ovary. Regardless of the administered withanolide, the concentration of 2 mg/kg did not show toxicity on the follicular morphology, ovarian function, or fertility of the mice. However, at concentrations of 5 and 10 mg/kg, the three derivatives (WT1, WT2, and WTA) increased follicular activation, cell proliferation, and ovarian senescence without affecting inflammatory cells. Furthermore, at a concentration of 10 mg/kg, the three withanolides showed intensified toxic effects, leading to DNA damage as evidenced by the labeling of γH2AX, activated Caspase 3, and TUNEL. We conclude that the cytotoxic effect of the tested withanolide derivatives (WT1, WT2, and WTA) in the concentration of 2 mg/kg did not show toxicity on the ovary. However, in higher concentrations, such as 10 mg/kg, toxic effects are potentiated, causing DNA damage.(AU)

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Purpose: The limitations in cancer treatment and the inadequacy of classical methods have made it necessary to discover therapeutics in cancer treatment. The cytotoxicity of thymoquinone, which has quite different properties in terms of biological activities, in ovarian cancer cells, and the changes in the expression levels of apoptotic genes (p53/caspase-3 (casp-3)) were investigated. Methods: In the study, thymoquinone (5, 50, 100, 250 and 500 µM and 24, 48, 72 hours) were applied to ovarian adenocarcinoma cancer cell line (OVCAR3), at different concentrations. Cytotoxic effect of thymoquinone on OVCAR-3 cells were analyzed by MTT method, and apoptotic and pro-apoptotic gene expression levels (p53, Casp-3) of thymoquinone in cancer cells were analyzed by quantitative real-time polymerase chain reaction. Results: Thymoquinone, whose effect has been revealed in many types of cancer, was shown to significantly reduce the viability of OVCAR3 cancer cells depending on the dose and time (p < 0.05). It was also determined that Casp-3 and p53 gene expressions increased in OVCAR3 cells. Conclusions: Considering the in-vitro cytotoxic activity and apoptotic gene expressions of thymoquinone, an important treatment agent, since it is a promising agent for the future of cancer treatment, more comprehensive studies may pave the way for its clinical use.

Resumo

The present work was showed to assess the effect of administration of rosemary extract on etoposide-induced toxicity, injury and proliferation in male rats were investigated. Forty male albino rats were arranged into four equal groups. 1st group, control; 2nd group, etoposide; 3rd group, co-treated rosemary & etoposide; 4th group, rosemary alone. In comparison to the control group, etoposide administration resulted in a significant increase in serum ALT, AST, ALP, total bilirubin, total protein, and gamma GT. In contrast; a significant decrease in albumin level in etoposide group as compared to G1. G3 revealed a significant decrease in AST, ALT, ALP, total protein and total bilirubin levels and a significant rise in albumin level when compared with G2. Serum levels of urea, creatinine, potassium ions, and chloride ions significantly increased; while sodium ions were significantly decreased in G2 when compared with G1. Also, there was an increase of MDA level for etoposide treated group with corresponding control rats. However, there was a remarkable significant decrease in SOD, GPX and CAT levels in G2 as compared to G1. There was a significant increase in serum hydrogen peroxide (H2O2) and Nitric oxide (NO) levels in group treated with etoposide when compared to control group. It was noticeable that administrated by rosemary alone either with etoposide had not any effect on the levels of H2O2 and Nitric oxide. Serum level of T3 and T4 was significantly increased in etoposide-administered rats in comparison with G1. The administration of rosemary, either alone or with etoposide, increased the serum levels of T3 and T4 significantly when compared to control rats. The gene expression analysis showed significant downregulation of hepatic SOD and GPx in (G2) when compared with (G1). The treatment with rosemary extract produced significant upregulation of the antioxidant enzymes mRNA SOD and GPx. MDA gene was increased in (G2) when contrasted with (G1). Treatment of the etoposide- induced rats with rosemary extract delivered significant decrease in MDA gene expression when compared with etoposide group. Rats treated with etoposide showed significant decline in hepatic Nrf2 protein expression, when compared with G1. While, supplementation of Etoposide- administered rats with the rosemary produced a significant elevation in hepatic Nrf2 protein levels. Additionally, the liver histological structure displayed noticeable degeneration and cellular infiltration in liver cells. It is possible to infer that rosemary has a potential role and that it should be researched as a natural component for etoposide-induced toxicity protection.

O presente trabalho foi apresentado para avaliar o efeito da administração de extrato de alecrim na toxicidade, lesão e proliferação induzidas por etoposídeos em ratos machos. Quarenta ratos albinos machos foram organizados em quatro grupos iguais: 1º grupo, controle; 2º grupo, etoposídeo; 3º grupo, alecrim e etoposídeo cotratados; 4º grupo, alecrim sozinho. Em comparação com o grupo controle, a administração de etoposídeo resultou em aumento significativo da ALT, AST, ALP, bilirrubina total, proteína total e gama GT séricas. Em contraste, houve diminuição significativa do nível de albumina no grupo etoposídeo em relação ao G1. O G3 revelou diminuição significativa dos níveis de AST, ALT, ALP, proteína total e bilirrubina total e aumento significativo do nível de albumina quando comparado ao G2. Os níveis séricos de ureia, creatinina, íons potássio e íons cloreto aumentaram significativamente, enquanto os íons sódio diminuíram significativamente no G2 quando comparado ao G1. Além disso, houve um aumento do nível de MDA para o grupo tratado com etoposídeo com os ratos controle correspondentes. No entanto, houve uma notável diminuição nos níveis de SOD, GPX e CAT no G2 em relação ao G1. Houve aumento significativo dos níveis séricos de peróxido de hidrogênio (H2O2) e óxido nítrico (NO) no grupo tratado com etoposídeo quando comparado ao grupo controle. Foi perceptível que a administração de alecrim isoladamente ou com etoposídeo não teve efeito sobre os níveis de H2O2 e NO. O nível sérico de T3 e T4 foi significativamente aumentado em ratos administrados com etoposídeo em comparação com o G1. A administração de alecrim, isoladamente ou com etoposídeo, aumentou significativamente os níveis séricos de T3 e T4 quando comparada aos ratos controle. A análise da expressão gênica mostrou desregulação significante da SOD e GPx hepática em G2 quando comparado com o G1. O tratamento com extrato de alecrim produziu aumento significativo das enzimas antioxidantes mRNA SOD e GPx. O gene MDA estava aumentado em G2 quando contrastado com o G1. O tratamento dos ratos induzidos por etoposídeo com extrato de alecrim proporcionou diminuição significativa na expressão do gene MDA quando comparado ao grupo etoposídeo. Ratos tratados com etoposídeo apresentaram declínio significativo na expressão da proteína Nrf2 hepática quando comparados ao G1. Enquanto isso, a suplementação de ratos administrados com etoposídeo e alecrim produziu uma elevação significativa nos níveis de proteína hepática Nrf2. Além disso, a estrutura histológica do fígado apresentou degeneração perceptível e infiltração celular nas células hepáticas. É possível inferir que o alecrim tem um papel potencial e que deve ser pesquisado como um componente natural para proteção da toxicidade induzida por etoposídeos.

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An evaluation was made of the larvicidal efficacy of lotilaner (Credeli®) in the treatment of dogs naturally infested with Dermatobia hominis larvae. A total of 12 dogs presenting at least three live D. hominis larvae were medicated. The animals were medicated orally with a single dose of no less than 20 mg/kg lotilaner. After drug administration, the animals remained at their homes, and observations were made to verify the larvicidal effect 6 hours after treatment. Live larvae were considered any parasite that exhibited motility after removal. For each animal was using the formula: 100 x [(total of live larvae before treatment − total live larvae after treatment) /total of live larvae before treatment] as criteria for evaluating lotilaner efficacy. A total of 98 larvae were counted in 12 dogs, with an average of 8.1 larvae per animal. The effectiveness of lotilaner was 80.6%. Nineteen larvae were found alive, albeit presenting hypomobility and lethargic behavior. However, note that the evaluation was performed just six hours after administration of the drug. Lotilaner administered orally in a single dose of 20 mg/kg showed 80.6% efficacy six hours after treating dogs naturally infested with D. hominis.(AU)

Foi avaliada a eficácia larvicida do lotilaner (Credeli®) no tratamento de cães naturalmente infestados com larvas de Dermatobia hominis. Foram medicados 12 cães, apresentando pelo menos três larvas vivas de D. hominis. Os animais foram medicados por via oral com dose única não inferior a 20 mg/kg de lotilaner. Após a administração desse medicamento, os animais permaneceram em seus domicílios, e foram feitas observações para verificar o efeito larvicida 6 horas após o tratamento. Larvas vivas foram consideradas quaisquer parasitas que exibissem motilidade após a remoção. Para cada animal foi utilizada a fórmula: 100 x [(total de larvas vivas antes do tratamento - total de larvas vivas após o tratamento) /total de larvas vivas antes do tratamento] como critério para avaliar a eficácia do lotilaner. Foram contadas 98 larvas em 12 cães, com média de 8,1 larvas por animal. A eficácia do lotilaner foi de 80,6%. Dezenove larvas foram encontradas vivas, porém apresentando hipomobilidade e comportamento letárgico. No entanto, vale ressaltar que a avaliação foi realizada apenas 6 horas após a administração do medicamento. Lotilaner, administrado por via oral em dose única de 20 mg/kg, apresentou eficácia de 80,6%, seis horas após o tratamento de cães naturalmente infestados por D. hominis.(AU)

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Abstract The present research was made to determine the micronuclei and cytotoxic capacity of the antidepressant venlafaxine in an in vivo acute and subchronic assays in mouse. In the first study, we administered once 5, 50, and 250 mg/kg of the drug, and included a negative and a daunorubicin treated group. Observations were daily made during four days. The subchronic assay lasted 5 weeks with daily administration of venlafaxine (1, 5, and 10 mg/kg) plus a negative and an imipramine administered groups. Observations were made each week. In the first assay results showed no micronucleated polychromatic erythrocytes (MNPE) increase, except with the high dose at 72 h. The strongest cytotoxic effect was found with 250 mg/kg at 72 h (a 51% cytotoxic effect in comparison with the mean control level). In the subchronic assay no MNPE increase was found; however, with the highest dose a significant increase of micronucleated normochromatic erythrocytes was observed in the last three weeks (a mean of 51% respect to the mean control value). A cytotoxic effect with the two high doses in the last two weeks was observed (a polychromatic erythrocyte mean decrease of 52% respect to the mean control value). Results suggest caution with venlafaxine.

Resumo A presente pesquisa foi feita para determinar a capacidade micronuclei e citotóxica do antidepressivo venlafaxina em ensaios agudos e subcrônicos in vivo em camundongos. No primeiro estudo, administramos uma vez 5, 50 e 250 mg/kg do medicamento e incluímos um grupo negativo e um grupo tratado com daunorubicina. As observações foram feitas diariamente durante quatro dias. O ensaio subcrônico durou cinco semanas com administração diária de venlafaxina (1, 5, e 10 mg/kg) mais um grupo negativo e um grupo administrado de imipramina. As observações foram feitas a cada semana. No primeiro ensaio, os resultados não mostraram aumento de eritrócitos policromáticos micronucleados (MNPE), exceto com a dose elevada a 72 h. O efeito citotóxico mais forte foi encontrado com 250 mg/kg a 72 h (um efeito citotóxico de 51% em comparação com o nível médio de controle). No ensaio subcrônico não foi encontrado aumento de MNPE; entretanto, com a dose mais alta, um aumento significativo de eritrócitos normocromáticos micronucleados foi observado nas últimas três semanas (média de 51% em relação ao valor médio de controle). Foi observado um efeito citotóxico com as duas altas doses nas últimas duas semanas (uma diminuição média de 52% em relação ao valor médio de controle dos eritrócitos policromáticos). Os resultados sugerem cautela com a venlafaxina.

Resumo

Hepatitis C virus infection (HCV) is the foremost reason of progressive hepatic fibrosis and cirrhosis, with an elevated risk of hepatocellular carcinoma (HCC) development. Medicinal plants have been used for human health benefits for several years, but their therapeutic potential needs to be explored. The main objective of this study was to figure out the in vitro antiviral and anticancer characteristics of total crude protein of Iberis gibraltarica against HCV and HCC. Total crude protein of Iberis gibraltarica was isolated and quantified. The level of cytotoxicity was measured against the HepG2 cell line and it shows no significant cytotoxicity at the concentration of 504µg/ml. The anti-HCV effect was determined by absolute quantification via real time RT-PCR method and viral titer was reduced up to 66% in a dose dependent manner against the total protein of Iberis gibraltarica. The anticancer potential of Iberis gibraltarica was also examined through mRNA expression studies of AFP and GPC3 genes against the total protein of Iberis gibraltarica-treated HepG2 cells. The results show up to 90% of the down-regulation expression of AFP and GPC3. The obtained results indicate the therapeutic potential of total protein of Iberis gibraltarica against HCV and hepatocellular carcinoma in vitro.

A infecção pelo vírus da hepatite C (HCV) é a principal causa de fibrose hepática progressiva e cirrose, com risco elevado de desenvolvimento de carcinoma hepatocelular (HCC). As plantas medicinais vêm sendo utilizadas para benefícios à saúde humana há vários anos, mas seu potencial terapêutico precisa ser explorado. O principal objetivo deste estudo foi descobrir as características antivirais e anticancerígenas in vitro da proteína bruta total de Iberis gibraltarica contra HCV e HCC. A proteína bruta total de Iberis gibraltarica foi isolada e quantificada. O nível de citotoxicidade foi medido contra a linha celular HepG2 e não apresenta citotoxicidade significativa na concentração de 504µg/ml. O efeito anti-HCV foi determinado por quantificação absoluta através do método RT-PCR em tempo real e o título viral foi reduzido em até 66% de forma dose-dependente contra a proteína total de Iberis gibraltarica. O potencial anticancerígeno de Iberis gibraltarica também foi examinado através de estudos de expressão de mRNA dos genes AFP e GPC3 contra a proteína total de células HepG2 tratadas com Iberis gibraltarica. Os resultados mostram até 90% da expressão de regulação negativa de AFP e GPC3. Os resultados obtidos indicam o potencial terapêutico da proteína total de Iberis gibraltarica contra HCV e carcinoma hepatocelular in vitro.

Resumo

Canine leishmaniasis has a wide variety of clinical signs, and, depending on the stage of the disease, the kidneys are the organs most affected. To stage the disease and carry out treatment, kidney assessment is of great importance, along with drug interactions and the deposition of immune complexes. In this study, we evaluated the renal morphology and function by means of B-mode ultrasonography and vascular Doppler, biochemical, urinalysis, and blood pressure tests, correlating the findings in dogs positive for leishmaniasis and treated with miltefosine. For this, 38 dogs were used, 12 healthy (G1) ones and 26 naturally infected with Leishmania sp.; of these, 12 animals were not treated (G2), and 14 were treated with miltefosine (G3). Evaluations were performed twice, with an interval of 30 days, before and after treatment with miltefosine. The average values of blood pressure as well as biochemical and urinary parameters were within the normal ranges for the species. In the volumetric Doppler measurement, no statistical differences were observed for systolic velocity, diastolic velocity, and resistivity index between the kidneys and the treated and untreated groups. According to the results obtained, treatment with miltefosine does not influence the renal parameters evaluated.(AU)

A leishmaniose canina possui uma ampla variedade de sinais clínicos, tendo os rins como os órgãos mais afetados. Para o estadiamento da doença e realização do tratamento, a avaliação renal é de grande importância, além das interações medicamentosa podem ocasionar com a deposição de imunocomplexos. O presente trabalho, teve como objetivo avaliar a morfologia e função renal por meio da ultrassonografia modo B e Doppler vascular, exames bioquímicos, urinálise e de pressão arterial, correlacionando seus achados de cães positivos para leishmaniose e tratados com miltefosina. Para tanto, foram utilizados 38 cães, 12 hígidos (G1), 26 naturalmente infectados por Leishmania sp, destes, 12 animais que não foram tratados (G2) e 14 tratados com miltefosina (G3). As avaliações foram feitas em dois momentos, com intervalo 30 dias, para contemplar as avaliações antes e após o tratamento com miltefosina. Os valores médios de pressão arterial, parâmetros bioquímicos e urinários, encontravam-se dentro da normalidade para a espécie, na mensuração Doppler volumétrica não foi observado diferença estatística entre as variáveis de velocidade sistólica, velocidade diastólica e índice de resistividade entre os rins e os grupos tratados e não tratados. De acordo com os resultados obtidos, o tratamento com miltefosina não influencia nos parâmetros renais avaliados.(AU)

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Abstract This study was carried out to evaluate the effect of Glutamine, as a dipeptide or a free amino acid form, on the progression of burn injuries in rats. Thirty male Wistar rats were burned with a comb metal plate heated in boiling water (98 °C) for three minutes, creating four rectangular full-thickness burn areas separated by three unburned interspaces (zone of stasis) in both dorsum sides. The animals were randomized into three groups (n=10): saline solution (G1-Control) and treated groups that orally received Glutamine as dipeptide (G2-Dip) or free amino acid (G3-FreeAA). Two and seven days after burn injury, lesions were photographed for unburned interspaces necrosis evolution assessment. Seven days after injury, glutathione seric was measured and histopathological analysis was performed. By photographs, there was a significant reduction in necrosis progression in G3-Free-AA between days two and seven. Histopathological analysis at day 7 showed a significantly higher stasis zone without necrosis and a higher number of fibroblasts in G2-Dip and G3-FreeAA compared with G1-Control. Also, glutathione serum dosage was higher in G2-Dip. The plasmatic glutathione levels were higher in the G2-Dip than the G1-Control, and there was a trend to higher levels in G3-FreeAA. The reduction in histological lesions, greater production of fibroblasts, and greater amounts of glutathione may have benefited the evolution of burn necrosis, which showed greater preservation of interspaces.

Resumo Este estudo foi realizado para avaliar o efeito da Glutamina, como um dipeptídeo ou forma de aminoácido livre, na progressão de queimaduras em ratos. Trinta ratos Wistar machos foram queimados com um pente de metal aquecido em água fervente (98 °C) por três minutos, criando quatro áreas retangulares queimadas separadas por três interesespaços não queimados (zona de estase) em ambos os lados do dorso. Os animais foram randomizados em três grupos (n = 10): solução salina (G1-Controle) e grupos tratados que receberam glutamina via oral como dipeptídeo (G2-Dip) ou aminoácido livre (G3-FreeAA). Dois e sete dias após a queimadura, as lesões foram fotografadas para avaliação da evolução da necrose entre os espaços não queimados. Sete dias após a lesão, foi dosada a glutationa sérica e realizada análise histopatológica. Pelas fotografias, houve uma redução significativa na progressão da necrose no G3-Free-AA entre os dias dois e sete. A análise histopatológica no dia 7 mostrou uma zona de estase significativamente maior sem necrose e número mais elevado de fibroblastos em G2-Dip e G3-FreeAA em comparação com G1-Controle. Os níveis plasmáticos de glutationa foram maiores no G2-Dip em relação ao G1-Controle, e houve tendência a níveis mais elevados no G3-FreeAA. A redução das lesões histológicas, maior produção de fibroblastos, maior quantidade de glutationa podem ter beneficiado a evolução da necrose da queimadura, que mostrou maior preservação dos interespaços.

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The aim of this study was to determine the allergenic activity of components present in crude extracts of Pterobothrium crassicolle plerocerci (CPE) and blastocysts (CBE) obtained from Micropogonias furnieri in a murine model. Two groups of seven animals each received 50 µg of CPE or CBE on days 1, 35 and 120. Serum samples were tested by ELISA and Immunoblotting. Specific IgG and IgE levels were detected by ELISA, showing specific humoral responses for the primary immunization for both immunoglobulins and continuously growing titers for IgE. Positive Passive Cutaneous Anaphylaxis tests in rats sensitized with anti-CBE sera and tested by CBE, showed biologically, the allergenic activity of the extracts. The CPE and CBE showed some different recognition regions but both experimental groups recognized all regions of the extracts when tested for cross reactions, showing that CPE and CBE could share antigenic recognition sites.(AU)

O objetivo deste estudo foi determinar a atividade alergênica de componentes presentes em extratos crus de plerocercos (CPE) e de blastocistos de Pterobothrium crassicolle (CBE), obtidos de Micropogonias furnieri, em modelo murino. Dois grupos de sete animais receberam cada um 50 µg de CPE ou CBE nos dias 1, 35 e 120. As amostras de soro foram testadas por ELISA e Imunoblot. Os níveis específicos de IgG e IgE foram detectados por ELISA, mostrando respostas humorais específicas para a imunização primária para ambas as imunoglobulinas e títulos crescentes de IgE. Testes positivos de Anafilaxia Cutânea Passiva em ratos sensibilizados com soros anti-CBE e testados por CBE, demonstraram biologicamente, a atividade alergênica dos extratos. O CPE e o CBE evidenciaram algumas regiões de reconhecimento diferentes, mas ambos os grupos experimentais reconheceram todas as regiões dos extratos, quando testados para reações cruzadas, mostrando que o CPE e o CBE poderiam compartilhar locais de reconhecimento antigênico.(AU)

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Acute liver injury (ALI) is an important medical problem that requires effective therapy. Astaxanthin (AST) is a carotenoid, and the beneficial effects of astaxanthin, including anti-oxidative, anti-inflammatory and anti-tumour activities, have been identified. The present study was designed to elucidate the protective effects of astaxanthin against ALI and their underlying mechanisms. RAW264.7 macrophages were treated with dimethyl sulfoxide combined with different doses of astaxanthin for 12 h. Mice were fed with or without astaxanthin for up to 7 days. LPS was administered to induce inflammation. We assessed histopathology, oxidative stress, inflammation and apoptosis .The results indicated that astaxanthin attenuated LPS-induced oxidative stress, inflammation and cell apoptosis both in vivo and in vitro. In vivo and in vitro experiments showed that astaxanthin down regulated the nuclear factor-kappa beta (NF-κB), nuclear factor erythroid 2-related factor 2 (Nrf2) and NLR family pyrin domain containing 3 (NLRP3) signalling pathways, inhibiting the LPS-induced inflammatory response, oxidative stress and cell apoptosis, and alleviating LPS-induced ALI in mice.

A lesão hepática aguda (ALI) é um problema médico importante que requer terapia eficaz. A astaxantina (AST) é um carotenóide, e os efeitos benéficos da astaxantina, incluindo atividades antioxidantes, anti-inflamatórias e antitumorais, foram identificados. O presente estudo foi desenhado para elucidar os efeitos protetores da astaxantina contra ALI e seus mecanismos subjacentes. Macrófagos RAW264.7 foram tratados com dimetil sulfóxido combinado com diferentes doses de astaxantina por 12 h. Os camundongos foram alimentados com ou sem astaxantina por até sete dias. O LPS foi administrado para induzir a inflamação. Histopatologia, estresse oxidativo, inflamação e apoptose foram avaliados. Os resultados indicaram que a astaxantina atenuou o estresse oxidativo induzido por LPS, inflamação e apoptose celular in vivo e in vitro. Experimentos in vivo e in vitro mostraram que a astaxantina regulou negativamente as vias de sinalização do fator nuclear-kappa beta (NF-κB), fator nuclear 2 relacionado ao eritróide 2 (Nrf2) e domínio de pirina da família NLR contendo 3 (NLRP3), inibindo o LPS- resposta inflamatória induzida, estresse oxidativo e apoptose celular e alívio da ALI induzida por LPS em camundongos.

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Type-2 diabetes mellitus (T2DM) is characterized by defects in insulin secretion and combined peripheral resistance to the hormone. Several non-human primates (NHP) species develop T2DM, mainly captive animals with reduced physical activity and incorrect feeding. This case report describes the T2DM treatment of a black-eared marmoset (Callithrix penicillata) by diet reformulation and metformin oral administration. An adult female was diagnosed with T2DM after hyperglycemia and high serum fructosamine associated with glycosuria and obesity. Metformin hydrochloride (125mg/animal, orally, q24h) associated with feeding intervention was started. After 26 days, a significant reduction in weight, glycemia, and serum fructosamine could be observed, showing satisfactory results for the adopted therapy. Metformin is considered a safe drug for T2DM treatment due to its low hypoglycemia risk. The new diet consisted of sweet potato, squash, and varied fruits offered twice daily. In addition, thawed-mice newborns, egg whites, and small portions of pelleted primate food. In the present report, metformin use, associated with a low glycemic index diet, was effective in treating this particular marmoset and may present a potential for T2DM treatment in other NHPs.

Diabetes mellitus tipo 2 (DM2) caracteriza-se por uma combinação de defeitos na secreção de insulina e resistência periférica ao hormônio. Diversas espécies de primatas não humanos (PNH) desenvolvem DM2, sobretudo animais cativos com atividade física reduzida e alimentados incorretamente. Este trabalho descreve o tratamento de DM2 em sagui-de-tufo-preto (Callithrix penicillata), através da reformulação da dieta e administração oral de metformina. Uma fêmea adulta foi diagnosticada com DM2 após apresentar hiperglicemia e frutosaminemia elevadas associadas à glicosúria e à obesidade. Iniciou-se o uso do cloridrato de metformina (125mg/animal, VO, SID) associado ao controle de consumo alimentar com ajustes da dieta. Após 26 dias pode-se observar redução significativa de peso, adequação da glicemia e frutosaminemia, constatando resultado satisfatório da terapêutica adotada. A metformina é considerada um medicamento seguro para o tratamento de DM2, devido ao baixo risco de hipoglicemia. A base da nova dieta era batata-doce, abóbora e frutas variadas oferecidas duas vezes ao dia. Além disso, camundongos recém-nascidos descongelados, clara de ovo e pequenas porções de ração primata peletizada. No presente relato, a metformina associada a uma dieta com baixo índice glicêmico, foi eficaz para tratamento de DM2 podendo apresentar potencial terapêutico de DM2 em outros PNH.

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Purpose: To evaluate the chemotherapeutic activity of temozolomide counter to mammary carcinoma. Methods: In-vitro anticancer activity has been conducted on MCF7 cells, and mammary carcinoma has been induced in Wistar rats by introduction of 7, 12-Dimethylbenz(a)anthracene (DMBA), which was sustained for 24 weeks. Histopathology, immunohistochemistry, cell proliferation study and apoptosis assay via TUNEL method was conducted to evaluate an antineoplastic activity of temozolomide in rat breast tissue. Results: IC50 value of temozolomide in MCF7 cell has been obtained as 103 µM, which demonstrated an initiation of apoptosis. The temozolomide treatment facilitated cell cycle arrest in G2/M and S phase dose dependently. The treatment with temozolomide suggested decrease of the hyperplastic abrasions and renovation of the typical histological features of mammary tissue. Moreover, temozolomide therapy caused the downregulation of epidermal growth factor receptor, extracellular signal-regulated kinase, and metalloproteinase-1 expression and upstream of p53 and caspase-3 proliferation to indicate an initiation of apoptotic events. Conclusions: The occurrence of mammary carcinoma has been significantly decreased by activation of apoptotic pathway and abrogation of cellular propagation that allowable for developing a suitable mechanistic pathway of temozolomide in order to facilitate chemotherapeutic approach.

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Background: Parkinson's disease (PD) is the second most prevalent neurodegenerative disease. There is no effective treatment for neurodegenerative diseases. Snake venoms are a cocktail of proteins and peptides with great therapeutic potential and might be useful in the treatment of neurodegenerative diseases. Crotapotin is the acid chain of crotoxin, the major component of Crotalus durissus collilineatus venom. PD is characterized by low levels of neurotrophins, and synaptic and axonal degeneration; therefore, neurotrophic compounds might delay the progression of PD. The neurotrophic potential of crotapotin has not been studied yet. Methods: We evaluated the neurotrophic potential of crotapotin in untreated PC12 cells, by assessing the induction of neurite outgrowth. The activation of the NGF signaling pathway was investigated through pharmacological inhibition of its main modulators. Additionally, its neuroprotective and neurorestorative effects were evaluated by assessing neurite outgrowth and cell viability in PC12 cells treated with the dopaminergic neurotoxin MPP+ (1-methyl-4-phenylpyridinium), known to induce Parkinsonism in humans and animal models. Results: Crotapotin induced neuritogenesis in PC12 cells through the NGF-signaling pathway, more specifically, by activating the NGF-selective receptor trkA, and the PI3K/Akt and the MAPK/ERK cascades, which are involved in neuronal survival and differentiation. In addition, crotapotin had no cytotoxic effect and protected PC12 cells against the inhibitory effects of MPP+ on cell viability and differentiation. Conclusion: These findings show, for the first time, that crotapotin has neurotrophic/neuroprotective/neurorestorative potential and might be beneficial in Parkinson's disease. Additional studies are necessary to evaluate the toxicity of crotapotin in other cell models.(AU)

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Mammary gland tumors are the most common neoplasms in female intact dogs and share some biological and histopathological aspects with those in women with breast cancer, making them a good model in comparative oncology. Resveratrol is a polyphenol found in several plants, and some studies have indicated that it acts in the neoplastic process as an anticancer drug. Viscum album is a hemiparasitic plant widely used as an adjuvant treatment for cancer in some countries. Thus, this study aimed to evaluate the antitumor potential of resveratrol and homeopathic Viscum album together and separately using two previously characterized canine mammary tumor cell lines (UNESP-CM9 and UNESP-CM60). The cell viability test (MTT) was performed, which revealed an IC50 of 3.11 µl/100 ml for UNESP-CM9 and 2.993 µl/100 ml for UNESP-CM60 for Viscum album, and for resveratrol, the IC50 was 281.6 µM for UNESP-CM9 and 105.5 µM for UNESP-CM60. The combination of both natural compounds led to tumor cell death at a lower IC50. The cell migration assay demonstrated an increase in cell migration time with both treatments. UNESP-CM9 closed 35.66% of the wounds in the control group and 15.51% of the wounds in the viscum group, while UNESP-CM60 closed 39.46% of the wounds in the control group and 19.95% of the wounds in the viscum group and 2.41% of the wounds in the resveratrol group. Thus, these two compounds have antitumor potential, making them possible alternatives to conventional treatments.(AU)

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Cyathostomins are the largest group of parasites in horses that can be controlled by ivermectin (IVM). This study aimed to run a four-dose titration trial of IVM in 28 naturally infected Thoroughbred yearlings. The local Strongyle population had been recorded to be resistant to IVM (200 µg/kg). The parasite fecal egg count (FEC) was performed to investigate the egg reappearance period (ERP) of two and five weeks (w2pt and w5pt) after IVM treatment. FEC was > 1000 on day zero for all groups. Although 100% FEC reduction was reported at w2pt for all concentrations, the FEC at w5pt revealed < 83% efficacy. This study reports the reduction of ERP using the label dose as well as 300, and 400 µg/kg (double dose) of IVM. The protocol allowed IVM to significantly suppress FEC w2pt although not eliminating adult worms, failing to guarantee an extension of its protection period over 8 weeks. Moreover, the FEC at w5pt possibly means the infection was not cleared, and worms reestablished egg laying. We raised the possibility of withdrawing IVM of control programs when the drug has less than 80% FEC reduction at w5pt.(AU)

Ciatostomíneos são o maior grupo de parasitos de equinos, controlados com ivermectina (IVM). Este estudo teve o objetivo de realizar um teste com quatro doses de IVM, em 28 potros Puro-Sangue Inglês. Os estrôngilos eram resistentes a IVM (200 µg/kg). A contagem de ovos por grama de fezes (OPG) foi realizada após tratamento (pt), para determinar o período de reaparecimento de ovos (PRO) após duas e cinco semanas (s2pt e s5pt). A OPG foi >1000 para todos os grupos. Observou-se 100% de redução na OPG na s2pt para todas as concentrações de IVM. A OPG na s5pt revelou < 83% de redução. Este estudo relata uma redução considerável do PRO utilizando a dose de bula, e as doses de 300 e 400 µg/kg (dobro da dose) de IVM contra ciatostomíneos em equinos. O protocolo permitiu observar a redução da postura de ovos na s2pt com IVM, falhando na garantia da extensão de proteção da droga. O reaparecimento da OPG significa que a infecção não foi eliminada. Foi levantada a possibilidade de restringir o uso de IVM, quando o medicamento atingir menos de 80% de redução da OPG na s5pt.(AU)

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Tilapia masculinization can be induced by oral administration of α-methyltestosterone (MT), which is commonly dissolved in ethanol to be added to the feed. However, there are many benefits in using alternative vehicles, such as oil. The incorporation time, vehicles quantity, safety for handlers, fish and the environment are favorable factors. In fry fed for 35 days under temperature control, we found that masculinization rate was similar in both incorporation vehicles of MT (oil or ethanol) in the concentrations studied (30 and 60 mg MT kg-1 feed). In an experiment, using hormone oil dissolution and oral administration at 30 mg MT kg-1 feed, it was observed that the longer the administration time, the lower the coefficient of variation in the masculinization rate. Therefore, administration for 32 days showed the lowest variability in the masculinization rate (99.8 ± 0.5 %), compared to 24 (98.5 ± 3.0 %), 16 (97.0 ± 6.0 %) and 8 (89.0 ± 8.8 %) days. The field experiment confirmed the results obtained in the lab. We concluded that the oil can be used as MT vehicle and we recommend to dispense it at the lowest hormonal concentration (30 mg MT kg-1 feed) for 32 days for tilapia masculinization.(AU)

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The aim of this study is to investigate the effect of Raphanus raphanistrum (radish) on chronic kidney disease damage by reactive oxygen species or free radicals in animal model rats. Total of 18 rats were used in this study, divided into 3 groups and each group consist of 6 rats. Group 1 control (C), group 2 model (M) and group 3 test (T). Model and test group were treated with alcohol to produce chronic kidney disease by reactive oxygen spices for 9 weeks a dose of 1 ml. After that test group was treated with Raphanus raphanistrum juice for 4 weeks 80mg/kg body weight to determine it effect. Raphanus raphanistrum juice effect on behavior of rats through increases the locomotor activity and anxiety. The serum creatinine and uric acid level were significantly improved in T group. The reactive oxygen enzyme test shows that Super Oxide Dismutase (SOD) and Glutathione Peroxidase (GPx) was increase in T group. The Glutathione S-Transferases (GST) and Catalase (CAT) enzyme level was nearly same in C and T groups. This study concludes that compound 1,1-diphenyl-2-picrylhydrazyl found in Raphanus raphanistrum juice and possess strong antioxidant activity on Chronic kidney disease induce by ethanol through reactive oxygen species. There is need of more researches to determine the use of natural compound to treat different disease.

O objetivo deste estudo é investigar o efeito de Raphanus raphanistrum (rabanete) nos danos da doença renal crônica por espécies reativas de oxigênio ou radicais livres em ratos modelo animal. Um total de 18 taxas foi utilizado neste estudo, divididas em 3 grupos e cada grupo consistia em 6 ratos. Grupo 1 controle (C), grupo 2 modelo (M) e grupo 3 teste (T). O modelo e o grupo de teste foram tratados com álcool para produzir doença renal crônica por meio de especiarias reativas de oxigênio por 9 semanas, uma dose de 1 ml. Após esse teste, o grupo foi tratado com suco de Raphanus raphanistrum por 4 semanas 80 mg / kg de peso corporal para determinar seu efeito. Efeito do suco de Raphanus raphanistrum no comportamento de ratos por meio do aumento da atividade locomotora e ansiedade. A creatinina sérica e o nível de ácido úrico melhoraram significativamente no grupo T. O teste da enzima oxigênio reativa mostra que a Super Óxido Dismutase (SOD) e a Glutationa Peroxidase (GPx) aumentaram no grupo T. O nível das enzimas Glutationa S-Transferases (GST) e Catalase (CAT) foi quase o mesmo nos grupos C e T. Este estudo conclui que o composto 1,1-difenil-2-picrilhidrazil encontrado no suco de Raphanus raphanistrum possui forte atividade antioxidante na doença renal crônica induzida pelo etanol por meio de espécies reativas de oxigênio. Há necessidade de mais pesquisas para determinar o uso de compostos naturais no tratamento de diferentes doenças.

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Abstract Bulbine natalensis and Chorophytum comosum are potential medicinal source for the treatment of cancers. Chronic myeloid leukaemia is a hematopoietic stem cells disorder treated by tyrosine kinase inhibitors but often cause recurrence of the leukaemia after cessation of therapy, hence require alternative treatment. This study determines the anti-cancer effect of leaf, root and bulb methanolic and aqueous extracts of B. natalensis and C. comosum in chronic human myelogenous leukaemia (K562) cell line by MTT, Hoechst bis-benzimide nuclear and annexin V stain assays. The root methanolic extract of B. natalensis and C. comosum showed a high cytotoxicity of 8.6% and 16.7% respectively on the K562 cell line at 1,000 μg/ml concentration. Morphological loss of cell membrane integrity causing degradation of the cell and fragmentation were observed in the root methanolic extract of both plants. A high apoptosis (p < 0.0001) was induced in the K562 cells by both leaf and root extracts of the C. comosum compared to the B. natalensis. This study shows both plants possess apoptotic effect against in vitro myelogenous leukaemia which contributes to the overall anti-cancer properties of B. natalensis and C. comosum to justify future therapeutic applications against chronic myelogenous leukaemia blood cancer.

Resumo Bulbine natalensis Baker e Chorophytum comosum (Thunb.) Jacques são potenciais fontes medicinais para o tratamento de cânceres. A Leucemia Mieloide Crônica (LMC) é um distúrbio das células-tronco hematopoiéticas que é tratado com inibidores da tirosina quinase, mas frequentemente, causa recorrência da leucemia após a interrupção da terapia, portanto, requer um tratamento alternativo. Este estudo determinou o efeito anticancerígeno de extratos metanólicos e aquosos de folha, raiz e bulbo de B. natalensis e C. comosum na linhagem celular de leucemia mieloide humana crônica (K562) por ensaios de MTT, Hoechst bis-benzimida nuclear e anexina V. O extrato metanólico da raiz de B. natalensis e C. comosum apresentou alta citotoxidade de 8,6% e 16,7% respectivamente, na linhagem celular K562 com a concentração de 1,000 μg / ml. Perda morfológica da integridade da membrana celular causando degradação dos núcleos, citoplasma e encolhimento celular foi observada no extrato metanólico da raiz de ambas as plantas. Uma alta apoptose (p <0,0001) foi induzida nas células K562 por extratos de folhas e raízes de C. comosum em comparação com B. natalensis. Este estudo mostrou que ambas as plantas possuem efeito apoptótico contra leucemia mieloide in vitro que contribui para as propriedades anticâncer gerais de B. natalensis e C. comosum para justificar futuras aplicações terapêuticas contra câncer de sangue de LMC.

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Aerobic vaginitis (AV) is a recently defined vaginal recurring infection, which is treated with antibiotics. However, excessive and prolonged use of antibiotics disrupts healthy vaginal microflora and leads to the emergence of antibiotic resistance among pathogens. This situation has directed researchers to explore alternative antimicrobials. The current study describes in vitro and in vivo antimicrobial efficacy and pharmaceutical interactions between plant essential oils (EOs) and five lactic acid bacteria (LABs), isolated from the healthy vagina, against E. faecalis, one of the major etiological agents of AV. In vitro experiments confirm good antimicrobial activity of both plant EOs and cell free supernatant (CFS) from LABs. Based on high antimicrobial efficacy, Moringa essential oil (MO) was selected to determine its nature of interaction with CFS of five LAB strains. Synergism was recorded between MO and CFS of L. reuteri (MT180537). To validate in vitro findings, prophylactic responses of individual and synergistic application of MO and L. reuteri (MT180537) were evaluated in an E. faecalis (MW051601) induced AV murine model. The prophylactic efficacy was evidenced by a reduction in intensity of clinical symptoms, E. faecalis (MW051601) count per vaginal tissue along with a reduction in AV associated changes in histological markers of infection in animals receiving Moringa essential oil and L. reuteri (MT180537) alone or in combination. However, significant synergism between Moringa essential oil and L. reuteri (MT180537) could not be observed. Our data confirms the importance of in vivo experiments in deducing pharmacological interactions.

Vaginite aeróbica (VA) é uma infecção vaginal recorrente definida recentemente, que é tratada com antibióticos. No entanto, o uso excessivo e prolongado de antibióticos perturba a microflora vaginal saudável e leva ao surgimento de resistência aos antibióticos entre os patógenos. Esta situação levou os pesquisadores a explorar antimicrobianos alternativos. O presente estudo descreve a eficácia antimicrobiana in vitro e in vivo e as interações farmacêuticas entre óleos essenciais vegetais (OE) e cinco bactérias lácticas (BAL), isoladas de vagina sã, contra E. faecalis, um dos principais agentes etiológicos da AV. Os experimentos in vitro confirmam a boa atividade antimicrobiana de ambos os EOs de plantas e sobrenadante livre de células (CFS) de LABs. Com base na alta eficácia antimicrobiana, o óleo essencial de Moringa (MO) foi selecionado para determinar sua natureza de interação com o sobrenadante livre de células (CFS) de cinco cepas de LAB. Sinergismo foi registrado entre MO e CFS de L. reuteri (MT180537). Para validar os resultados in vitro, as respostas profiláticas da aplicação individual e sinérgica de MO e L. reuteri (MT180537) foram avaliadas em um modelo murino AV induzido por E. faecalis (MW051601). A eficácia profilática foi evidenciada por uma redução na intensidade dos sintomas clínicos, contagem de E. faecalis (MW051601) por tecido vaginal, juntamente com uma redução nas alterações associadas a AV nos marcadores histológicos de infecção em animais que receberam óleo essencial de Moringa e L. reuteri (MT180537) sozinho ou em combinação. No entanto, não foi possível observar sinergismo significativo entre o óleo essencial de Moringa e L. reuteri (MT180537). Nossos dados confirmam a importância dos experimentos in vivo na dedução de interações farmacológicas.