In vivo antitumor effect of proteoglycan fraction from Agaricus brasiliensis does not depend on the production of antitumor antibodies / O efeito antitumoral in vivo da fração proteoglicana de Agaricus brasiliensis não depende da produção de anticorpos antitumorais

Polysaccharides isolated from the edible mushroom Agaricus brasiliensis were previously shown to have antitumor, antiproliferative, and immunomodulatory activity. Here, we evaluated the in vivo effect of the acid-treated fraction from A. brasiliensis (ATF) on the subcutaneous growth of Ehrlich tumor cells (EHR) and on the production of tumor-specific antibodies. Mice (n=10) were inoculated with 2 × 106 EHR and injected subcutaneously in the tumor inoculum region with 0.1 mL ATF or saline. Control (tumor-free) group received ATF or saline. Treatments were carried out for 7, 14, or 30 days, with three consecutive doses and an interval of 4 days between the first and last doses, being repeated until the end of each experimental period.  Histopathological analysis shows the infiltration of mononuclear and polymorphonuclear cells into the tumor site of all tumor-bearing mice. Tumor stimulated the influx of polymorphonuclear cells in the early stages, especially at 7 days, while the influx of mononuclear cells was higher in the final stages, at 14 and 30 days in all animals, independently of the treatment with ATF. Treatment of animals for 30 days reduced the tumor weight in 30% but we did not find a correlation with the antitumor antibody production since both treated and untreated mice were able to produce them

Evaluation of in vitro Cytotoxic Effects of Especifico Pessoa Phytotherapic Tincture on Ehrlich Tumor Cells and Mice Spleen Cells

Especifico Pessoa (EP) is traditionally used for the treatment of snakebite envenoming. The traditional use of EP and its properties have been reported. In this study, we evaluated the in vitro cytotoxic effects of EP on Ehrlich tumor and mice spleen cells. Cytotoxicity assay was carried out by using Trypan blue exclusion method. Spleen cell suspension was prepared (n=2) with RPMI medium and tumor cell suspension was prepared from ascitic fluid of Ehrlich tumor-bearing mice (n=1); both the suspensions contained 4 x 106 cells mL-1. Pure EP or EP diluted in RPMI (1:2; 1:4) was used. The results were expressed as percentage of cell viability and demonstrate that EP is toxic to Ehrlich cells at all concentrations (Control: 96.42 ± 3.40; Pure: 1.55 ± 2.91; 1:2: 4.85 ± 5.04; 1:4: 13.39 ± 5.08), but nontoxic to spleen cells in at the lowest dilution (Control: 72.86 ± 13.79; Pure: 13.52 ± 6,36; 1:2: 41.36 ± 13.51; 1:4: 56.59 ± 8.62). Therefore, the results demonstrate that EP has cytotoxic effects, depending on the dose and the cell line evaluated.Especifico Pessoa (EP) é utilizado no envenenamento por serpentes. Estudos comprovam seu uso tradicional e descrevem outras propriedades, como antitumoral. O objetivo deste estudo foi avaliar seu efeito citotóxico em células do Tumor de Ehrlich e esplênicas de camundongos. O ensaio foi realizado pelo método de exclusão do Azul Tripan. A suspensão de células esplênicas foi obtida (n = 2) em RPMI e a tumoral a partir do fluido ascitico de camundongos portadores do tumor (n = 1), ambos a 4 x 106 células mL-1. O EP foi usado puro ou diluído em RPMI (1:2; 1:4). Os resultados são expressos como porcentagem de viabilidade celular e demonstraram que o EP é tóxico para células tumorais, em todas as concentrações (Controle: 96,42 ± 3,40; Puro: 1,55 ± 2,91; 1:2: 4,85 ± 5,04; 1:4: 13,39 ± 5,08), mas atóxico para os esplenócitos na menor diluição (Controle: 72,86 ± 13,79; Puro: 13,52 ± 6,36; 1:2: 41,36 ± 13,51; 1:4: 56,59 ± 8,62). Portanto, os resultados demonstram que o EP tem efeito citotóxico, dependendo da dose e linhagem celular utilizada

Natural products: the contribution of research developed by a university in Sinop, Brazil

Bioactive compounds present in plants, microorganisms, and animals are the subject of scientific research, mainly focusing on the discovery of novel pharmacologically active molecules for application in human and veterinary medicine, as well as agriculture. Brazil is the country with the greatest biodiversity globally, accounting for 15–20% of global diversity. Therefore, the objective of the present study was to carry out a bibliographic survey on studies involving natural compounds conducted by staff of the Federal University of Mato Grosso, Sinop campus. The review included scientific articles published in specialized journals by the staff from the moment they were hired, according to information in their Lattes Curriculum. During the survey period, the campus had 314 staff, and 311 scientific articles were published. Among the 311 articles, 122 studies met the inclusion criteria and were selected for this literature review. According to the findings, UFMT – Sinop has contributed to research in the field of natural products, carrying out studies on the biological effects of compounds present in extracts obtained from plants, microorganisms, and animal secretions

Evaluation of Euphorbia tirucalli L. Immunomodulatory and Antitumor Activity

Euphorbia tirucalli L. is commonly used to treat various pathologies, including cancer. The objective of this study was to evaluate the effect of a crude extract of E. tirucalli on the development of Ehrlich solid tumors and immune function. We prepared an extract by macerating the aerial parts of the plant with 80% ethanol (1:10 g/v) for 15 days. Its effects on tumor development were assessed in male Swiss mice (n=10). The mice were injected subcutaneously with 10⁶ tumor cells and then treated by gavage daily with the extract (33.33, 67, and 133.34 mg kg-1) or saline (0.5% ethanol) for 30 days. The treatment had no toxic effect and did not reduce tumor growth. However, the weight of tumor mass was lowest in the mice treated with extract at 67 mg kg-1. Immunomodulatory activity was evaluated in mice (n=8), with and without tumors, treated with the extract (67 mg kg-1), under the same conditions as described above for 7 and 14 days. Lymphoproliferation, NK activity and IL-12, TNF-a, IFN-y, and IL-10 levels did not differ significantly between the groups. However, IL-4 levels were reduced in the E. tirucalli treated groups at 7 and 14 days when compared to the controls. We concluded that the extract is not toxic and cannot inhibit Ehrlich solid tumor development. Its immunomodulatory activity involves its ability to modulate IL-4 levels.Euphorbia tirucalli L. é empregado popularmente para o tratamento de diversas patologias, inclusive o câncer. O objetivo deste trabalho foi avaliar o efeito do extrato bruto da planta sobre o desenvolvimento do tumor sólido de Ehrlich (EHR) e sobre o sistema imunológico. O extrato foi preparado com as partes aéreas da planta maceradas com etanol 80% (1:10 g/v, 15 dias). A análise do desenvolvimento tumoral, em camundongos Swiss (n=10; machos) inoculados subcutaneamente com 10⁶ células EHR e tratados (via intragástrica, diariamente) com o extrato (33,33, 67 ou 133,34 mg kg-1) ou solução salina (0,5% etanol) durante 30 dias, demonstrou que o tratamento não apresentou efeito tóxico e não reduziu o crescimento do tumor, sendo o extrato de 67 mg kg-1 o que apresentou o menor valor da massa tumoral. A atividade imunomoduladora foi avaliada em camundongos swiss (n = 8; machos), portadores do tumor ou não, e tratados com 67 mg kg-1 do extrato, nas mesmas condições, durante 7 e 14 dias. A linfoproliferação, atividade NK e a produção de IL-12, TNF-a, IFN-y e IL-10 não apresentaram diferença significativa entre os grupos, mas a produção de IL-4 foi reduzida no grupo Aveloz e Aveloz + EHR, aos 7 e 14 dias, em comparação aos controles. Concluímos que o extrato não é tóxico, porém não foi capaz de inibir o desenvolvimento tumoral, e a sua ação imunomoduladora foi concentrada na modulação da IL-4

Osteopontin expression identifies a subset of recruited macrophages distinct from Kupffer cells in the fatty liver

Metabolic-associated fatty liver disease (MAFLD) represents a spectrum of disease states ranging from simple steatosis to non-alcoholic steatohepatitis (NASH). Hepatic macrophages, specifically Kupffer cells (KCs), are suggested to play important roles in the pathogenesis of MAFLD through their activation, although the exact roles played by these cells remain unclear. Here, we demonstrated that KCs were reduced in MAFLD being replaced by macrophages originating from the bone marrow. Recruited macrophages existed in two subsets with distinct activation states, either closely resembling homeostatic KCs or lipid-associated macrophages (LAMs) from obese adipose tissue. Hepatic LAMs expressed Osteopontin, a biomarker for patients with NASH, linked with the development of fibrosis. Fitting with this, LAMs were found in regions of the liver with reduced numbers of KCs, characterized by increased Desmin expression. Together, our data highlight considerable heterogeneity within the macrophage pool and suggest a need for more specific macrophage targeting strategies in MAFLD

Desenvolvimento de melanoma em animais selecionados pela intensidade de resposta inflamatória aguda (Airmax/Airmin): caracterização das células Natural Killer, NKT, Treg e perfil de citocinas

Animais selecionados de acordo com a intensidade da sua resposta inflamatória aguda (AIR) à poliacrilamida, foram inoculados s.c. com células de melanoma B16F10 ou S91. Aos 7, 14 ou 30 dias, os animais foram sacrificados para obtenção do baço e avaliação do número de células T, Treg, NK e NKT, por citometria de fluxo, e do perfil de algumas citocinas (ELISpot). Os resultados demonstraram que os animais AIRmax apresentaram maior incidência do melanoma B16F10, associado ao aumento do número de células Treg, e também incidência elevada do melanoma S91, associada ao aumento da produção IL-10 e redução de IFN-g, em relação aos animais AIRmin. Estes resultados permitem inferir que a sensibilidade ao desenvolvimento de melanomas pode ser resultado de alterações nos mecanismos regulatórios da resposta imune, que neste caso, inibiram o desenvolvimento de uma resposta eficiente nos animais AIRmax.Not available.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Enhanced natural killer activity and production of pro-inflammatory cytokines in mice selected for high acute inflammatory response (AIRmax)

Strains of mice with maximal and minimal acute inflammatory responsiveness (AIRmax and AIRmin, respectively) were developed through selective breeding based on their high- or low-acute inflammatory responsiveness. Previous reports have shown that AIRmax mice are more resistant to the development of a variety of tumours than AIRmin mice, including spontaneous metastasis of murine melanoma. Natural killer activity is involved in immunosurveillance against tumour development, so we analysed the number and activity of natural killer cells (CD49b+), T-lymphocyte subsets and in vitro cytokine production by spleen cells of normal AIRmax and AIRmin mice. Analysis of lymphocyte subsets by flow cytometry showed that AIRmax mice had a higher relative number of CD49b+ cells than AIRmin mice, as well as cytolytic activity against Yac.1 target cells. The number of CD3+ CD8+ cells was also higher in AIRmax mice. These findings were associated with the ability of spleen cells from AIRmax mice in vitro to produce higher levels of the pro-inflammatory cytokines tumour necrosis factor-α, interleukin-12p40 and interferon-γ but not the anti-inflammatory interleukin-10. Taken together, our data suggest that the selective breeding to achieve the AIRmax and AIRmin strains was able to polarize the genes associated with cytotoxic activity, which can be responsible for the antitumour resistance observed in AIRmax mice

First phytochemical study and biological activity of the leaves ethanolic extract from Cissus spinosa Cambess

AIMS: The objective of this study was to identify the phytochemical profile and to evaluate the biological effects of the crude ethanolic extract (EE) and the ethanolic fraction (EF) of leaves of the species Cissus spinosa Cambess, after oxidative stress induced by cyclophosphamide (CP) in mice.METHODS: Phytochemical profile was performed detecting functional groups and, analysis of total flavonoids and phenols concentration, as well as the antiradical activity in EE and EF. The phytochemical characterization was done for the identification of flavonoids present in the leaves of the plant. In the biochemical tests, hematological parameters, glucose and total cholesterol dosages in plasma, enzymatic and non-enzymatic antioxidants and lipid damage marker were evaluated in different tissues (liver, kidney and heart), besides genotoxic and immunological analyzes. The animals received 15 days of treatment, via gavage, with EE (50 mg kg-1) or EF (50 mg kg-1) and on the 15th day, an intraperitoneal injection of CP (100 mg kg-1) or saline (0.9%). After 24 h the last treatment, the animals were anesthetized for blood withdrawal, sacrificed and removal of the organs.RESULTS: In the phytochemical analyzes, the presence of alkaloids, flavonoids and phenols was identified, the latter presented a higher concentration for EF. Eight flavonoids were identified - Rutin, Quercetin-3-β-D-glucoside, Quercitrin, Taxifolin, Quercetin, Canferol, Luteolin and Apigenin. In the biochemical analyzes, in general, EE showed a better antioxidant action against oxidative damages, hypoglycemic and antitilipemic action when comparing with EF, probably due to the synergism caused by flavonoids. It was observed the reduction and an increase of micronucleated polychromatic erythrocytes, due to the action of antioxidant compounds and alkaloids present in the plant, also considering the question of the seasonal period that directly interferes in the production of these compounds. In the immunological analysis, the extracts did not stimulate the spontaneous production of oxygen peroxide (H2O2) and nitric oxide (NO•). CONCLUSIONS: Other studies, such as the variation of the chemical composition of the plant by local seasonality, hypoglycemic and antilipemic action, should be carried out to better delineate the biological action present in this plant.Objetivos: O objetivo deste estudo foi identificar o perfil fitoquímico e avaliar os efeitos biológicos do extrato etanólico bruto (EE) e da fração etanólica (EF) das folhas da espécie Cissus spinosa Cambess, após estresse oxidativo induzido pela ciclofosfamida. (CP) em camundongos.Métodos: O perfil fitoquímico foi realizado detectando grupos funcionais e, análise da concentração de flavo-noides e fenóis totais, bem como a atividade antirradical em EE e EF. A caracterização fitoquímica foi realizada para a identificação de flavonoides presentes nas fo-lhas da planta. Nos testes bioquímicos, os parâmetros hematológicos, as dosagens de glicose, colesterol total, creatinina, fosfatase alcalina e aminotransferases no plasma, os antioxidantes enzimáticos e não enzimá-ticos e o marcador de dano lipídico foram avaliados em diferentes tecidos (fígado, rim e coração), além de análises genotóxicas e imunológicas. Os animais receberam 15 dias de tratamento, via gavagem, com EE (50 mg kg-1) ou EF (50 mg kg-1) e no 15º dia, injeção intraperitoneal de CP (100 mg kg-1) ou soro fisiológico (0,9%) . Após 24 h do último tratamento, os animais fo-ram anestesiados para retirada do sangue, sacrificados e após a retirada dos órgãos.Resultados: Nas análises fitoquímicas, identificou-se a presença de alcaloides, flavonoides e fenóis, estes últimos apresentaram maior concentração para EF. Oito flavonoides foram identificados - rutina, quercetina-3-β--D-glicosídeo, quercitrina, taxifolina, quercetina, can-ferol, luteolina e apigenina. Nas análises bioquímicas, em geral, o EE apresentou melhor ação antioxidante contra os danos oxidativos, ação hipoglicemiante e antitilipêmica quando comparada à EF, provavelmente devido ao sinergismo causado pelos flavonoides. Observou-se a redução e um aumento de eritrócitos policromáticos micronucleados, devido à ação de compostos antioxidantes e alcaloides presentes na planta, considerando também a questão do período sazonal que interfere diretamente na produção desses compostos. Na análise imunológica, os extratos não estimularam a produção espontânea de peróxido de oxigênio (H2O2) e óxido nítrico (NO•).Conclusões: Outros estudos, como a variação da composição química da planta por sazonalidade local, ação hipoglicemiante e antilipêmica, devem ser rea-lizados para melhor delineamento da ação biológica presente nesta planta

Natural killer activity of the spleen cells of Ehrlich tumor-bearing mice treated with Copaifera multijuga extract

AIMS: Copaifera multijuga Hayne oleoresin is commonly used in traditional medicine owing to its anti-inflammatory, antiseptic, antitumor, and antibacterial properties. However, little is known about the effect of the compounds from the bark of this plant. In this study, the immunomodulatory effect of the ethanolic extract of C. multijuga bark via natural killer activity of non-adherent spleen cells of Ehrlich tumor-bearing mice was evaluated.METHODS: Male Swiss mice were inoculated subcutaneously with 1×106 Ehrlich tumor cells (Ehrlich and Ehrlich/C. multijuga group) or phosphate buffered saline solution (control group and C. multijuga group) and treated orally daily with C. multijuga extract (200 mg kg-1, 0.1 mL per mouse, for the Ehrlich/C. multijuga and C. multijuga groups) or phosphate buffered saline solution (control group and Ehrlich group). The four experimental groups consisted in eight mice each and were organized in two sets, one treated for seven days and another treated for 14 days, totalizing 64 mice throughout the experiment. Twenty-four hours after the last oral administration, the mice were euthanized and the spleen tissue was isolated to prepare a non-adherent spleen cell suspension and to evaluate natural killer activity. Data are presented as the cell lysis percentage of Yac.1 target cells by non-adherent spleen cells.RESULTS: Treatment for seven days increased natural killer activity in the Ehrlich/C. multijuga group (21.20±8.89, p0.05).CONCLUSIONS: Treatment for seven days with an ethanolic extract of C. multijuga bark enhanced the natural killer activity of non-adherent spleen cells from Ehrlich tumor-bearing mice.***Atividade natural killer das células esplênicas de camundongos portadores do tumor de Ehrlich tratados com extrato de Copaifera multijuga***OBJETIVOS: O óleo-resina de Copaifera multijuga Hayne é popularmente utilizado na medicina tradicional por suas propriedades anti-inflamatória, antisséptica, antitumoral e antibacteriana. Entretanto, há poucos estudos sobre o efeito dos compostos obtidos da casca da planta. Assim, o objetivo deste trabalho foi avaliar o efeito imunomodulador do extrato etanólico da casca da C. multijuga na atividade natural killer das células esplênicas não aderentes de animais portadores do tumor de Ehrlich.MÉTODOS: Camundongos Swiss machos foram inoculados subcutaneamente com 1 x 106 células do tumor de Ehrlich (grupo Ehrlich e Ehrlich/C. multijuga) ou com solução salina tamponada (grupo controle e C. multijuga) e tratados diariamente (gavagem) com extrato de C. multijuga (200 mg kg-1, 0,1mL cada um, para os grupos Ehrlich/C. multijuga e C. multijuga) ou com solução salina tamponada (grupo controle e grupo Ehrlich). Os quatro grupos experimentais consistiram de oito animais cada e foram organizados em dois conjuntos, um tratado por sete dias e outro tratado por 14 dias, totalizando 64 animais durante todo o experimento. Após 24 horas do término do tratamento, os animais foram eutanasiados para obtenção da suspensão de células esplênicas não aderentes e avaliação da atividade natural killer. Os resultados são apresentados como porcentagem da lise celular das células alvo Yac.1 pelas células esplênicas não aderentes.RESULTADOS: A análise dos resultados demonstrou que aos sete dias de tratamento a C. multijuga aumentou a atividade natural killer no grupo Ehrlich/C. multijuga (21,20±8,89, p0,05).CONCLUSÕES: Os resultados demonstraram que o tratamento com o extrato etanólico da casca de C. multijuga favoreceu a atividade natural killer das células esplênicas não aderentes dos animais portadores do tumor de Ehrlich apenas aos sete dias de tratamento