RESUMO
OBJECTIVE AND DESIGN: To evaluate the potency of RORγt blockade for treatment of Inflammatory Bowel Disease (IBD), the efficacy of TAK-828F, a novel RORγt inverse agonist, in anti-TNF-α mAb non-responsive mouse colitis model and effect of TAK-828F on IL-17 production in peripheral mononuclear blood cells (PBMCs) of anti-TNF-α naive and treatment-failure patients of IBD was investigated. METHODS AND RESULTS: The colitis model showed Th17-dependent pathogenicity and response to anti-IL-12/23p40 monoclonal antibody (mAb), but no response to anti-TNF-α mAb. In the model, TAK-828F, at oral dosages of 1 and 3 mg/kg, inhibited progression of colitis and reduced the immune reaction that characterize Th17 cells. Anti-IL-17A mAb showed neither efficacy nor change in the T cell population and colonic gene expression in the model. In the normal mouse, a 4-week treatment of TAK-828F at 30 mg/kg did not severely reduce lymphocyte cell counts in peripheral and intestinal mucosa, which was observed in RORγ-/- mice. TAK-828F strongly inhibited IL-17 gene expression with IC50 values from 21.4 to 34.4 nmol/L in PBMCs from anti-TNF mAb naive and treatment-failure patients of IBD. CONCLUSIONS: These results indicate that RORγt blockade would provide an effective approach for treating refractory patients with IBD by blocking IL-23/Th17 pathway.
Assuntos
Acetatos/farmacologia , Acetatos/uso terapêutico , Doenças Inflamatórias Intestinais/tratamento farmacológico , Leucócitos Mononucleares/efeitos dos fármacos , Naftiridinas/farmacologia , Naftiridinas/uso terapêutico , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/imunologia , Adolescente , Adulto , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Células Cultivadas , Colo/efeitos dos fármacos , Colo/imunologia , Colo/patologia , Modelos Animais de Doenças , Feminino , Humanos , Doenças Inflamatórias Intestinais/genética , Doenças Inflamatórias Intestinais/imunologia , Interleucina-17/antagonistas & inibidores , Interleucina-17/genética , Interleucina-17/imunologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Leucócitos Mononucleares/imunologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Knockout , Camundongos SCID , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto JovemRESUMO
A series of tetrahydroisoquinoline derivatives were designed, synthesized, and evaluated for their potential as novel orally efficacious retinoic acid receptor-related orphan receptor-gamma t (RORγt) inverse agonists for the treatment of Th17-driven autoimmune diseases. We carried out cyclization of the phenylglycinamide core by structure-based drug design and successfully identified a tetrahydroisoquinoline carboxylic acid derivative 14 with good biochemical binding and cellular reporter activity. Interestingly, the combination of a carboxylic acid tether and a central fused bicyclic ring was crucial for optimizing PK properties, and the compound 14 showed significantly improved PK profile. Successive optimization of the carboxylate tether led to the discovery of compound 15 with increased inverse agonistic activity and an excellent PK profile. Oral treatment of mice with compound 15 robustly and dose-dependently inhibited IL-17A production in an IL23-induced gene expression assay.
Assuntos
Descoberta de Drogas , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/agonistas , Tetra-Hidroisoquinolinas/farmacologia , Administração Oral , Animais , Cristalografia por Raios X , Citocinas/biossíntese , Relação Dose-Resposta a Droga , Humanos , Injeções Intradérmicas , Interleucina-23/administração & dosagem , Interleucina-23/farmacologia , Células Jurkat , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Modelos Moleculares , Estrutura Molecular , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Relação Estrutura-Atividade , Tetra-Hidroisoquinolinas/administração & dosagem , Tetra-Hidroisoquinolinas/químicaRESUMO
A series of novel phenylglycinamides as retinoic acid receptor-related orphan receptor-gamma t (RORγt) inverse agonists were discovered through optimization of a high-throughput screen hit 1. (R)-N-(2-((3,5-Difluoro-4-(trimethylsilyl)phenyl) amino)-1-(4-methoxyphenyl)-2-oxoethyl)-3-hydroxy-N-methylisoxazole-5-carboxamide (22) was identified as one of the best of these compounds. It displayed higher subtype selectivity and specificity over other nuclear receptors and demonstrated in vivo potency to suppress the transcriptional activity of RORγt in a mouse PD (pharmacodynamic) model upon oral administration.
Assuntos
Descoberta de Drogas , Glicina/análogos & derivados , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/agonistas , Administração Oral , Animais , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Glicina/administração & dosagem , Glicina/química , Glicina/farmacologia , Humanos , Células Jurkat , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Modelos Moleculares , Estrutura Molecular , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Relação Estrutura-AtividadeRESUMO
The application of char to agricultural land is recognized as a potential way to sequester atmospheric carbon (C) assimilated by plants in soil, thus decelerating global warming. Such a process would also be expected to improve plant growth and the physical and chemical properties of soil. However, field investigations of the effects of continuous char application have not been reported. In the present study, the effects of repetitive bamboo char application on CO2, CH4, and N2O flux from soil, soil C content, and crop yield were investigated at two upland fields over five crop seasons. Three treatments: chemical fertilizer (CF) applied plots (Control plot); cattle manure (CM) (10 t ha(-1)) and CF applied plot (CM plot); and bamboo char (20 t ha(-1)), cattle manure (10 t ha(-1)), and CF applied plot (Char/CM plot), were arranged in each field. After three crop seasons, the fourth treatment with char was applied without CF (Char plot) was given to one of the fields. CM and/or char were applied every crop season. Gas fluxes were measured using the static chamber method. Seasonal variations in CO2 flux and total CO2 emissions were consistently similar between the CM and Char/CM plots and between the Char and Control plots. As such, the decomposition rate of bamboo char was quite small, and the positive or negative effect of char on CM decomposition was not significant in the fields. Soil C analysis provided confirmation of this. CM application enhanced N2O emission mainly in the summer crop season. The differences in total N2O emission between the Char/CM and CM plots as well as between the Char and Control plots were insignificant in most cases. Total CH4 flux was negligibly small in all cases. Although the yield of winter crop (broccoli) in the Char/CM plots was twice observed to be higher than that in the Control and CM plots at one of the fields, in general, the char application had no effect on overall crop yield. Thus, the repeated application of bamboo char had no significant influence on greenhouse gas emissions and crop yields, but a high C accumulating function was found.
Assuntos
Agricultura/métodos , Poluentes Atmosféricos/metabolismo , Carvão Vegetal/metabolismo , Produtos Agrícolas/crescimento & desenvolvimento , Solo/química , Animais , Dióxido de Carbono/metabolismo , Bovinos , Carvão Vegetal/administração & dosagem , Fertilizantes/análise , Gases/metabolismo , Efeito Estufa , Japão , Esterco/análise , Metano/metabolismo , Óxido Nitroso/metabolismo , Distribuição Aleatória , Estações do AnoRESUMO
The Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study) was established in the late 1980s to evaluate the risk impact of lifestyle factors and levels of serum components on human health. During the 20-year follow-up period, the results of the study have been published in almost 200 original articles in peer-reviewed English-language journals. However, continued follow-up of the study subjects became difficult because of the retirements of principal researchers, city mergers throughout Japan in the year 2000, and reduced funding. Thus, we decided to terminate the JACC Study follow-up at the end of 2009. As a final point of interest, we reviewed the population registry information of survivors. A total of 207 (0.19%) subjects were ineligible, leaving 110 585 eligible participants (46 395 men and 64 190 women). Moreover, errors in coding date of birth and sex were found in 356 (0.32%) and 59 (0.05%) cases, respectively, during routine follow-up and final review. Although such errors were unexpected, their impact is believed to be negligible because of the small numbers relative to the large total study population. Here, we describe the final cohort profile at the end of the JACC Study along with selected characteristics of the participants and their status at the final follow-up. Although follow-up of the JACC Study participants is finished, we will continue to analyze and publish study results.
Assuntos
Neoplasias/epidemiologia , Adulto , Distribuição por Idade , Idoso , Causas de Morte , Estudos de Coortes , Feminino , Seguimentos , Humanos , Incidência , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Sistema de Registros , Distribuição por SexoRESUMO
BACKGROUND: We investigated the association of baseline body mass index (BMI) and weight change since age 20 years with liver cancer mortality among Japanese. METHODS: The data were obtained from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study). A total of 31 018 Japanese men and 41 455 Japanese women aged 40 to 79 years who had no history of cancer were followed from 1988 through 2009. RESULTS: During a median 19-year follow-up, 527 deaths from liver cancer (338 men, 189 women) were documented. There was no association between baseline BMI and liver cancer mortality among men or men with history of liver disease. Men without history of liver disease had multivariable hazard ratios (HR) of 1.95 (95%CI, 1.07-3.54) for BMI less than 18.5 kg/m(2) and 1.65 (1.05-2.60) for BMI of 25 kg/m(2) or higher, as compared with a BMI of 21.0 to 22.9 kg/m(2). BMI was positively associated with liver cancer mortality among women and women with history of liver disease. Weight change since age 20 years was positively associated with liver cancer mortality among women regardless of history of liver disease. Women with history of liver disease had a multivariable HRs of 1.96 (1.05-3.66) for weight gain of 5.0 to 9.9 kg and 2.31 (1.18-4.49) for weight gain of 10 kg or more, as compared with weight change of -4.9 to 4.9 kg. CONCLUSIONS: Both underweight (BMI <18.5 kg/m(2)) and overweight (BMI ≥25 kg/m(2)) among men without history of liver disease, and weight gain after age 20 (weight change ≥5 kg) among women with history of liver disease, were associated with increased mortality from liver cancer.
Assuntos
Índice de Massa Corporal , Neoplasias Hepáticas/mortalidade , Aumento de Peso , Redução de Peso , Adulto , Idoso , Feminino , Seguimentos , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Adulto JovemRESUMO
Aspergillus lacticoffeatus WU-2020 is a citric acid hyperproducer that is suitable for solid culture. Here, we present a high-quality draft of its genome sequence (35.9 Mb), which consists of 11 scaffolds and contains 11,490 genes. We also present the mitochondrial genome, which is 31.3 kb in length.
RESUMO
BACKGROUND: Phosphatidylcholine hydroperoxide (PCOOH) is a primary oxidation product of PC, and is markedly accumulated in blood plasma and arterial walls in atherosclerotic animals and humans. The role of PCOOH in the induction of angiogenesis is unknown. METHODS: In this study, we investigated whether PCOOH stimulated angiogenic responses (e.g., vascular endothelial growth factor (VEGF)-induced cell proliferation, migration, and tube formation, and angiogenesis-related gene/protein expression) in human umbilical vein endothelial cells (HUVEC) and in an ex vivo rat aorta model. RESULTS: VEGF induced proliferation, migration, and tube formation of HUVEC, and these angiogenic responses were all enhanced by PCOOH but not by native (nonoxidized) PC. The angiogenic effects of PCOOH are considered to be mediated via generation of reactive oxygen species and activation of both PI3K/AKT and MAPK pathways. The angiogenic activities of PCOOH were also confirmed by the rat aortic ring assay. CONCLUSIONS: These results indicate that PCOOH can elicit several angiogenic responses. GENERAL SIGNIFICANCE: The present study implies an important role of PCOOH in atherosclerosis progression and plaque instability.
Assuntos
Aorta/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Fosfatidilcolinas/farmacologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Animais , Sequência de Bases , Western Blotting , Células Cultivadas , Primers do DNA , Endotélio Vascular/enzimologia , Endotélio Vascular/fisiologia , Humanos , Técnicas In Vitro , Sistema de Sinalização das MAP Quinases , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RatosRESUMO
Supplementation to an AIN93G-based diet of tocotrienol (T3) for 13 weeks administered to Fischer 344/slc rats showed a safety profile with no side effects. Dose-dependent T3 levels were detected in many tissues. Under the present experimental conditions, a continuous intake of the T3 concentrate would be safe in the rats as long as the T3 content was less than 0.20% of the dietary intake.
Assuntos
Suplementos Nutricionais , Tocotrienóis/farmacocinética , Vitamina E/farmacocinética , Animais , Peso Corporal/efeitos dos fármacos , Dieta , Esquema de Medicação , Masculino , Ratos , Ratos Endogâmicos F344 , Distribuição Tecidual , Tocotrienóis/administração & dosagem , Vitamina E/administração & dosagemRESUMO
Prolyl-tRNA synthetase (PRS) is a clinically validated antimalarial target. Screening of a set of PRS ATP-site binders, initially designed for human indications, led to identification of 1-(pyridin-4-yl)pyrrolidin-2-one derivatives representing a novel antimalarial scaffold. Evidence designates cytoplasmic PRS as the drug target. The frontrunner 1 and its active enantiomer 1-S exhibited low-double-digit nanomolar activity against resistant Plasmodium falciparum (Pf) laboratory strains and development of liver schizonts. No cross-resistance with strains resistant to other known antimalarials was noted. In addition, a similar level of growth inhibition was observed against clinical field isolates of Pf and P. vivax. The slow killing profile and the relative high propensity to develop resistance in vitro (minimum inoculum resistance of 8 × 105 parasites at a selection pressure of 3 × IC50) constitute unfavorable features for treatment of malaria. However, potent blood stage and antischizontal activity are compelling for causal prophylaxis which does not require fast onset of action. Achieving sufficient on-target selectivity appears to be particularly challenging and should be the primary focus during the next steps of optimization of this chemical series. Encouraging preliminary off-target profile and oral efficacy in a humanized murine model of Pf malaria allowed us to conclude that 1-(pyridin-4-yl)pyrrolidin-2-one derivatives represent a promising starting point for the identification of novel antimalarial prophylactic agents that selectively target Plasmodium PRS.
Assuntos
Aminoacil-tRNA Sintetases , Antimaláricos , Malária Falciparum , Malária , Animais , Antimaláricos/farmacologia , Humanos , Malária/tratamento farmacológico , Malária Falciparum/tratamento farmacológico , Camundongos , Plasmodium falciparumRESUMO
Recent studies have demonstrated that tocotrienol (T3) is superior to tocopherol (Toc) for cancer chemoprevention. However, there is little information on whether Toc influences the anticancer property of T3. In this study, we investigated the influence of Toc on the cytotoxic effects of delta-T3 in DLD-1 human colorectal adenocarcinoma cells. Toc, especially alpha-Toc, attenuated delta-T3-induced cytotoxicity and apoptosis in DLD-1 cells, whereas Toc alone did not exhibit any cytotoxic effect. delta-T3-induced cell cycle arrest and proapoptotic gene/protein expression (e.g., p21, p27, and caspases) were abrogated by alpha-Toc. Furthermore, coadministration of alpha-Toc decreased delta-T3 uptake into DLD-1 cells in a dose-dependent manner. These results indicate that alpha-Toc is not only less cytotoxic to cancer cells, but it also reduces the cytotoxicity of delta-T3 by inhibiting its cellular uptake.
Assuntos
Adenocarcinoma/metabolismo , Neoplasias Colorretais/metabolismo , Vitamina E/análogos & derivados , alfa-Tocoferol/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Vitamina E/antagonistas & inibidores , Vitamina E/metabolismo , Vitamina E/farmacologiaRESUMO
Several lines of evidence support the beneficial effect of tocotrienol (T3; an unsaturated vitamin E) on inhibition of tumor development. Many factors, including decrease in oxidative stress and modulation of cell signaling pathways in tumor and endothelial cells, have been implicated in such anticancer action of T3, while the in vivo potency and exact intracellular mechanisms for the anticancer properties of T3 remain not fully understood. We have hypothesized that the inhibitory effect of T3 on cancer may be attributable to the antiangiogenic activity of T3, and we found that T3 acts as a potent regulator of growth-factor-dependent signaling in endothelial cells and as an antiangiogenic agent minimizing tumor growth. In this work, we review the history and biological action (i.e., anticancer) of vitamin E and describe current research on the antiangiogenic effects of T3 and its mechanisms.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Anticarcinógenos/uso terapêutico , Antineoplásicos/uso terapêutico , Tocotrienóis/uso terapêutico , Apoptose/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Humanos , Neoplasias/patologia , Neoplasias/prevenção & controle , Receptores de Fatores de Crescimento do Endotélio Vascular/efeitos dos fármacos , Receptores de Fatores de Crescimento do Endotélio Vascular/fisiologiaRESUMO
We investigated the potency of TAK-828F, a RORγt inverse agonist, in murine experimental autoimmune encephalomyelitis (EAE) model. TAK-828F inhibited the differentiation of Th17 and Th1/17 cells in inguinal lymph node. Increase of these cells in central nervous system (CNS) was also inhibited by TAK-828F. Prophylactic and therapeutic treatments of TAK-828F were efficacious in the model. Plasma concentration of TAK-828F was higher than that in CNS. These results indicate that TAK-828F mainly acts at peripheral and results in the reduction of Th17- and Th1/17-dependent inflammation in CNS. Blocking RORγt may be a promising strategy for treatment of multiple sclerosis.
Assuntos
Acetatos/farmacologia , Encefalomielite Autoimune Experimental/imunologia , Naftiridinas/farmacologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/antagonistas & inibidores , Animais , Diferenciação Celular/efeitos dos fármacos , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th17/efeitos dos fármacos , Células Th17/imunologiaRESUMO
IL-17-producing Th17 cells and IFN-γ and IL-17 double-producing Th1/17 cells have been identified as the pathogenic cells in inflammatory bowel disease (IBD). Retinoic acid-related orphan receptor γt (RORγt) is a master regulator for the differentiation and activation of Th17 and Th1/17 cells. We discovered a novel orally available TAK-828F, a strong and selective RORγt inverse agonist. To assess the potential of RORγt blockade in the therapy for IBD, the efficacy of TAK-828F in activated T cell transfer mouse colitis model was investigated. This model was highly sensitive to the prophylactic treatment of anti-TNF-α monoclonal antibody but partially susceptible to sulfasalazine, tacrolimus, and prednisolone. Oral administration of TAK-828F, at doses of 1 and 3 mg/kg, b.i.d, strongly protected the progression of colitis. TAK-828F decreased the population of Th17 and Th1/17 cells in a dose-dependent manner in the mesenteric lymph node. Moreover, expression of mRNA that are characteristic of the Th17 signature, such as IL-17A and IL-17F in the colon, were inhibited by TAK-828F, while the expression of IL-10, an anti-inflammatory cytokine, was increased. In the therapeutic treatment, TAK-828F lessened disease severity compared to the vehicle control mice. Interestingly, gene expression of zonula occludens-1 (ZO-1) and mucin 2 (Muc2), which play an important role in barrier function of the intestinal mucosa, was recovered by TAK-828F. These results indicate that blocking RORγt has promising pharmacological profile in the colitis model. RORγt blockade may provide a novel therapeutic paradigm for treatment of IBD with unique mechanism by which improves imbalance of the immune system.
Assuntos
Acetatos/farmacologia , Colite/tratamento farmacológico , Agonismo Inverso de Drogas , Naftiridinas/farmacologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/antagonistas & inibidores , Animais , Diferenciação Celular , Modelos Animais de Doenças , Mucosa Intestinal/metabolismo , Linfonodos/citologia , Camundongos , Células Th17/citologia , Células Th17/efeitos dos fármacosRESUMO
Retinoic-acid-related orphan receptor γt (RORγt) inverse agonists could be used for the treatment of autoimmune diseases. Previously, we reported a novel quinazolinedione 1 a with a flexible linear linker as a novel RORγt inverse agonist. A U-shaped conformation in the complex structure of 1 a with RORγt protein was confirmed. Further improvement of the pharmacokinetic (PK) profiles was required because of the low drug exposure in mice upon oral administration (mouse AUC of 1 a: 27â ng â h â mL-1 at 1â mg â kg-1 , p.o.). To improve the PK profiles, conformationally constrained U-shaped scaffolds were investigated. As a result, morpholine analogues with improved PK profiles and high potency were successfully identified. The substituent at the N1 position of the quinazoline moiety was also modified, leading to an enhancement of reporter activity. Consequently, compound 43 (N2 -(3-chloro-4-cyanophenyl)-N4 -(3-(cyclopropylmethyl)-1-isopropyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-yl)morpholine-2,4-dicarboxamide) exhibited improved drug exposure (mouse AUC: 1289â ng â h â mL-1 at 1â mg â kg-1 , p.o.). In addition, suppression of IL-17A gene expression by IL-23 stimulation in a mouse pharmacodynamics model was observed for 43. The conformation of 43 with RORγt protein was also confirmed as U-shape by X-ray co-crystal structure analysis. The key interaction that boosts potency is also discussed.
Assuntos
Ciclopentanos/farmacologia , Desenho de Fármacos , Furanos/farmacologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/agonistas , Administração Oral , Animais , Cristalografia por Raios X , Ciclopentanos/administração & dosagem , Ciclopentanos/síntese química , Transferência Ressonante de Energia de Fluorescência , Furanos/administração & dosagem , Furanos/síntese química , Camundongos , Modelos Moleculares , Conformação Molecular , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismoRESUMO
Both conjugated linoleic acid (CLA), which contains conjugated double bonds, and eicosapentaenoic acid (EPA), an n-3 polyunsaturated fatty acid, have antitumor effects. Hence, we hypothesized that a combination of conjugated double bonds and an n-3 highly unsaturated fatty acid may produce a stronger antitumor effect, and we have previously shown that conjugated EPA (CEPA), prepared by alkaline treatment of EPA, induces strong and selective apoptosis in vitro and in vivo, with the mechanism proceeding via lipid peroxidation. In this study, we examined CEPA-induced gene expression in DLD-1 colorectal adenocarcinoma human cells carrying a mutant p53, in order to understand the details of CEPA-induced apoptosis via lipid peroxidation. DNA microarray analysis of 9970 genes was performed by comparison of CEPA-treated DLD-1 cells with untreated DLD-1 cells, thereby allowing determination of the differential gene expression profile induced by CEPA in these cells. CEPA treatment caused up-regulation of expression of genes induced by p53 and activation of the mitochondrial apoptosis pathway via Bax and the death pathway via TRAIL, leading to apoptosis of DLD-1 cells. In addition, activation of the mutant p53 was also induced by CEPA, and these effects showed lipid-peroxidation dependency. This is the first such gene expression analysis of the effects of CEPA, and our results confirm that CEPA induces lipid peroxidation, activates mutant p53, and causes p53-dependent apoptosis in DLD-1 cells.
Assuntos
Adenocarcinoma/metabolismo , Apoptose/efeitos dos fármacos , Neoplasias Colorretais/metabolismo , Ácido Eicosapentaenoico/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Mutação , Proteína Supressora de Tumor p53/metabolismo , Adenocarcinoma/genética , Linhagem Celular Tumoral , Neoplasias Colorretais/genética , Ácido Eicosapentaenoico/análogos & derivados , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
Tocotrienol (T3), unsaturated vitamin E, has recently gained considerable attention as a potent antiangiogenic agent minimizing tumor growth, the exact intracellular mechanisms of which remain poorly understood. Because hypoxia-inducible factor-1alpha (HIF-1alpha), its downstream target vascular endothelial growth factor (VEGF), and other angiogenic factors such as interleukin-8 (IL-8) and cyclooxygenase 2 (COX-2) play critical roles in neovascularization, we tested the hypothesis that the inhibitory effect of T3 on tumor angiogenesis is via regulation of these angiogenic factors. We used 2 cancer cell lines, human colorectal adenocarcinoma cells (DLD-1) and human hepatoma cells (HepG2). T3 isomers (2 micromol/L) inhibited hypoxia-induced VEGF secretion from DLD-1, with delta-T3 showing potent inhibition. Delta-T3 suppressed hypoxia-induced VEGF and IL-8 expression in DLD-1 at both mRNA and protein levels, and we found the inhibitory mechanism of delta-T3 by reducing HIF-1alpha protein expression or increasing HIF-1alpha degradation. Also, delta-T3 (2 micromol/L) did not affect hypoxia-induced COX-2 mRNA expression; however, delta-T3 tended to suppress (P = 0.044) hypoxia-induced COX-2 protein expression, implying a possible post-transcriptional mechanism by delta-T3. Overall, our results confirmed that T3 has an inhibitory effect on angiogenic factor secretion from cancer cells and revealed the possible mechanisms, providing new information about the antiangiogenic effects of T3.
Assuntos
Proteínas Angiogênicas/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Tocotrienóis/farmacologia , Proteínas Angiogênicas/genética , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Neoplasias Colorretais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Two incubation experiments were conducted under controlled moisture and temperature conditions to determine the effects of soil amendment treatments based on pruning waste biochar and oyster shell, on N2O and CO2 emissions from an orchard soil. In experiment 1, four treatments were tested including, control (CK), pruning waste biochar at 2% (B2%), at 10% (B10%), and oyster shell (OS), mixed with soil from two different depths, namely, from the 0-5 cm and the 0-10 cm layers. In experiment 2, only the 0-10 cm soil layer was used to study the effect of surface application of pruning waste biochar (B2% and B10%) on soil N2O and CO2 emissions. The results showed that soil pH, total C and C: N ratio increased with biochar amendment treatments. Significant reduction in soil NO3- content was observed for the B10% treatment. Although OS application increased soil pH, no effect was observed on soil mineral N content, total C or C: N ratio. The rate of N2O emissions from the 0-5 cm soil layer after B2% and B10% addition, significantly declined by 12.5% and 26.3%, respectively. However, only the B10% treatment caused significant reduction in N2O emissions from the 0-10 cm soil layer and from surface soil, by 15.1% and 13.8%, respectively. Oyster shell application had no effect on either soil N2O or CO2 emissions from either soil layer tested. Our results suggest that the addition of pruning waste biochar at a high rate has the potential to mitigate N2O emissions from orchard soils; while, oyster shell can be used for liming without altering soil N2O nor CO2 emissions.
RESUMO
A laboratory study was conducted to study the effects of liming and different biochar amendments on N2O and CO2 emissions from acidic tea field soil. The first experiment was done with three different rates of N treatment; N 300 (300 kg N ha-1), N 600 (600 kg N ha-1) and N 900 (900 kg N ha-1) and four different rates of bamboo biochar amendment; 0%, 0.5%, 1% and 2% biochar. The second experiment was done with three different biochars at a rate of 2% (rice husk, sawdust, and bamboo) and a control and lime treatment (dolomite) and control at two moisture levels (50% and 90% water filled pore space (WFPS)). The results showed that dolomite and biochar amendment significantly increased soil pH. However, only biochar amendment showed a significant increase in total carbon (C), C/N (the ratio of total carbon and total nitrogen), and C/IN ratio (the ratio of total carbon and inorganic nitrogen) at the end of incubation. Reduction in soil NO3--N concentration was observed under different biochar amendments. Bamboo biochar with the rates of 0.5, 1 and 2% reduced cumulative N2O emission by 38%, 48% and 61%, respectively, compare to the control soil in experiment 1. Dolomite and biochar, either alone or combined significantly reduced cumulative N2O emission by 4.6% to 32.7% in experiment 2. Reduction in N2O production under biochar amendment was due to increases in soil pH and decreases in the magnitude of mineral-N in soil. Although, both dolomite and biochar increased cumulative CO2 emission, only biochar amendment had a significant effect. The present study suggests that application of dolomite and biochar to acidic tea field soil can mitigate N2O emissions.
Assuntos
Carbonato de Cálcio/química , Carvão Vegetal/química , Magnésio/química , Óxido Nitroso/química , Solo/química , Ácidos/químicaRESUMO
Retinoic acid-related orphan receptor γt (RORγt) is a key master regulator of the differentiation and activation of IL-17 producing CD4+ Th17, CD8+ Tc17 and IL-17/IFN-γ co-producing cells (Th1/17 cells). These cells play critical roles in the pathogenesis of autoimmune diseases such as inflammatory bowel disease and multiple sclerosis. Thus, RORγt is an attractive target for the treatment of these diseases. We discovered TAK-828F, an orally available potent and selective RORγt inverse agonist. The inhibitory effect on the activation and differentiation of Th17 cells by TAK-828F was evaluated in mouse and human primary cells. TAK-828F inhibited IL-17 production from mouse splenocytes and human peripheral blood mononuclear cells dose-dependently at concentrations of 0.01-10⯵M without affecting the production of IFN-γ. Additionally, TAK-828F strongly inhibited Th17, Tc17 and Th1/17 cells' differentiation from naive T cells and memory CD4+ T cells at 100â¯nM without affecting Th1 cells' differentiation. In addition, TAK-828F improved Th17/Treg cells' population ratio by inhibiting Th17 cells' differentiation and up-regulating Treg cells. Furthermore, TAK-828F, at 100â¯nM, reduced the production of Th17-related cytokines (IL-17, IL-17F and IL-22) without affecting IFN-γ production in whole blood. These results demonstrate that TAK-828F has the potent and selective inhibitory activity against RORγt both in mouse and human cells. Additionally, oral administration of TAK-828F showed promising efficacy in naive T cell transfer mouse colitis model. TAK-828F may provide a novel therapeutic option to treat immune diseases by inhibiting Th17 and Th1/17 cells' differentiation and improving imbalance between Th17 and Treg cells.