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1.
Mitochondrial DNA B Resour ; 6(3): 1007-1008, 2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33796718

RESUMO

Halocynthia aurantium (Stolidobranchia: Pyuridae) is a species of tunicate of commercial value that is commonly found in the northern Pacific Ocean and in the Bering Sea. Here, we determined the complete mitogenome of sea peach H. aurantium using 150 PE high-throughput sequencing. The assembled mitogenome is 14,979 bp in length (overall A + T contents 56.2%), and contains 13 protein-coding genes, 21 transfer RNAs, two ribosomal RNAs. Phylogenetic analysis of the mitogenome sequence of H. aurantium fully resolved it in a clade with H. roretzi. These data and results will be useful for future studies on the evolution of the Halocynthia and the Pyuridae.

2.
J Immunol Res ; 2020: 1731457, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33299895

RESUMO

Tumor microenvironment components dictate the growth and progression of various cancers. Tumor-associated macrophages are the most predominant cells in TME and play a major role in cancer invasiveness. Gastric cancer is one of the most common cancers in Asia, and recently, various cases of resistance to fluorouracil treatment have been reported. In this study, we investigated the role of alternatively activated macrophages in the resistance of AGS gastric cancer cells to fluorouracil. THP-1 cells were polarized using recombinant human IL-4, then were cocultured with AGS cells treated with fluorouracil. Cell viability, Western blot, immunofluorescence, and cell invasion were performed for this investigation. Our results demonstrated that polarized macrophages initiated the survival of treated AGS cells and induced the resistance in AGS by upregulating the expression of integrin ß3, focal adhesion protein (FAK), and cofilin proteins. These results reveal that integrin ß3, focal adhesion protein (FAK), and cofilin proteins are potential targets for the improvement of fluorouracil efficacy in gastric cancer treatment.


Assuntos
Fatores de Despolimerização de Actina/genética , Resistencia a Medicamentos Antineoplásicos , Fluoruracila/farmacologia , Proteína-Tirosina Quinases de Adesão Focal/genética , Integrina beta3/genética , Macrófagos/imunologia , Macrófagos/metabolismo , Fatores de Despolimerização de Actina/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Polaridade Celular , Resistencia a Medicamentos Antineoplásicos/genética , Resistencia a Medicamentos Antineoplásicos/imunologia , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Integrina beta3/metabolismo , Ativação de Macrófagos/imunologia , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias Gástricas/etiologia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia
3.
Liver Cancer ; 9(2): 182-192, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32399432

RESUMO

Background/Aim: Uniform treatment of hepatocellular carcinoma (HCC) with molecular targeted drugs (e.g., sorafenib) results in a poor overall tumor response when tumor subtyping is absent. Patient stratification based on actionable gene expression is a method that can potentially improve the effectiveness of these drugs. Here we aimed to identify the clinical application of actionable genes in predicting response to sorafenib. Methods: Through quantitative real-time reverse transcription PCR, we analyzed the expression levels of seven actionable genes (VEGFR2, PDGFRB, c-KIT, c-RAF, EGFR, mTOR, and FGFR1) in tumors versus noncancerous tissues from 220 HCC patients treated with sorafenib. Our analysis found that 9 responders did not have unique clinical features compared to nonresponders. A receiver operating characteristic curve evaluated the predictive performance of the treatment benefit score (TBS) calculated from the actionable genes. Results: The responders had significantly higher TBS values than the nonresponders. With an area under the curve of 0.779, a TBS combining mTOR with VEGFR2, c-KIT, and c-RAF was the most significant predictor of response to sorafenib. When used alone, sorafenib had a 0.7-3% response rate among HCC patients, but when stratifying the patients with actionable genes, the tumor response rate rose to 15.6%. Furthermore, actionable gene expression is significantly correlated with tumor response. Conclusions: Our findings on patient stratification based on actionable molecular subtyping potentially provide a therapeutic strategy for improving sorafenib's effectiveness in treating HCC.

4.
Cancers (Basel) ; 12(4)2020 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-32225122

RESUMO

Preoperative chemoradiotherapy (PCRT) and subsequent surgery is the standard multimodal treatment for locally advanced rectal cancer (LARC), albeit PCRT response varies among the individuals. This creates a dire necessity to identify a predictive model to forecast treatment response outcomes and identify patients who would benefit from PCRT. In this study, we performed a gene expression study using formalin-fixed paraffin-embedded (FFPE) tumor biopsy samples from 156 LARC patients (training cohort n = 60; validation cohort n = 96); we identified the nine-gene signature (FGFR3, GNA11, H3F3A, IL12A, IL1R1, IL2RB, NKD1, SGK2, and SPRY2) that distinctively differentiated responders from non-responders in the training cohort (accuracy = 86.9%, specificity = 84.8%, sensitivity = 81.5%) as well as in an independent validation cohort (accuracy = 81.0%, specificity = 79.4%, sensitivity = 82.3%). The signature was independent of all pathological and clinical features and was robust in predicting PCRT response. It is readily applicable to the clinical setting using FFPE samples and Food and Drug Administration (FDA) approved hardware and reagents. Predicting the response to PCRT may aid in tailored therapies for respective responders to PCRT and improve the oncologic outcomes for LARC patients.

5.
Mitochondrial DNA B Resour ; 5(3): 3538-3539, 2020 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-33458232

RESUMO

The complete mitochondrial genome of pitted stingray, Bathytoshia brevicaudata (Myliobatiformes: Dasyatoidea) was investigated by next-generation sequencing. The analyzed mitochondrial genome was 17,640 nucleotides in length and had 59.2% for AT contents. This genome contains 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes. and 1 putative control region. Five protein-coding genes (ATPase6, COII, ND2, ND3, ND4) including incomplete stop codons and four tRNAs have atypical codons. The phylogenetic inference including 13 species of the same family revealed a close relationship with Pteroplatytrygon violacea. This is the first mitochondrial genome report from genus Bathytoshia.

6.
Colloids Surf B Biointerfaces ; 183: 110455, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31493630

RESUMO

Green chemistry is beneficial for the production of eco-friendly and stable nanoparticles using biological agents. The present study was performed to explore the potential of the marine bacterium Paracoccus haeundaensis BC74171T for the extracellular synthesis of gold nanoparticles (AuNPs). Cell-free supernatant-mediated AuNPs were characterized by different techniques and analyzed for their antioxidant activity and antiproliferative effect on normal and cancer cells. Visual observations indicated the formation of AuNPs by the development of a ruby red color and were confirmed by a UV-vis absorbance peak at about 535 nm. The synthesized AuNPs were spherical in shape and had an average size of 20.93 ± 3.46 nm, as determined by transmission electron microscopy and a dynamic light scattering particle size analyzer, respectively. From Fourier transform infrared spectroscopy, the interaction of functional groups was determined and the presence of biomaterial on the AuNP surface was confirmed. Concentration-dependent antioxidant activity of AuNPs was observed by the 2,2-diphenyl-1-picrylhydrazyl method. The AuNPs synthesized do not show growth inhibition on HaCaT and HEK293 normal cells, while they show concentration-dependent growth inhibition in the case of A549 and AGS cancer cells. Thereby, this study proves that AuNP synthesis using P. haeundaensis is a facile method and that the AuNPs synthesized are non-toxic to human cells, which indicates that they can be useful in biomedical applications.


Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Proliferação de Células/efeitos dos fármacos , Ouro/química , Nanopartículas Metálicas/química , Paracoccus/metabolismo , Células A549 , Antibacterianos/química , Antioxidantes/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Espaço Extracelular/metabolismo , Células HEK293 , Humanos , Testes de Sensibilidade Microbiana , Água do Mar/microbiologia
7.
Drug Dev Res ; 80(4): 504-512, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30860609

RESUMO

The fungus Chaetomium sp. is a causative agent of infections in humans and is ubiquitous in the natural environment. The secondary metabolites of this genus exhibit many biological activities, including antifungal activity and toxicity in mitochondria. In this study, we isolated cristazine from the fungus C. cristatum, which has the potential to inhibit the growth of human epidermoid carcinoma (A431) cells in a dose- and time-dependent manner. Its inhibitory activity was examined using a cell viability assay and cell death was elucidated by western blot analysis. Cristazine triggered apoptotic cell death via the Type I death receptor pathway including the activation of caspases and other target proteins. However, cristazine did not have any effect on mitochondrial apoptotic proteins such as Bid, cytochrome c, and apoptosis-inducing factor. Cristazine inhibited the cell cycle progression by arresting the G1 /S phase and up-regulating the inhibitory proteins of cyclin-dependent kinases. Thus, cristazine has great potential for inducing apoptosis in A431 cells via both cell cycle arrest and the inhibition of cell growth.


Assuntos
Alcaloides/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Piperazinas/farmacologia , Receptores de Morte Celular/metabolismo , Alcaloides/isolamento & purificação , Antineoplásicos/isolamento & purificação , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Técnicas de Cultura de Células , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Chaetomium/química , Relação Dose-Resposta a Droga , Humanos , Estrutura Molecular , Piperazinas/isolamento & purificação , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Fatores de Tempo
8.
Mar Drugs ; 17(2)2019 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-30795639

RESUMO

ß-thymosin is known for having 43 amino acids, being water-soluble, having a light molecular weight and ubiquitous polypeptide. The biological activities of ß-thymosin are diverse and include the promotion of wound healing, reduction of inflammation, differentiation of T cells and inhibition of apoptosis. Our previous studies showed that oyster ß-thymosin originated from the mantle of the Pacific oyster, Crassostrea gigas and had antimicrobial activity. In this study, we investigated the anti-inflammatory effects of oyster ß-thymosin in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells using human ß-thymosin as a control. Oyster ß-thymosin inhibited the nitric oxide (NO) production as much as human ß-thymosin in LPS-induced RAW264.7 cells. It also showed that oyster ß-thymosin suppressed the expression of prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, oyster ß-thymosin reduced inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6). Oyster ß-thymosin also suppressed the nuclear translocation of phosphorylated nuclear factor-κB (NF-κB) and degradation of inhibitory κB (IκB) in LPS-induced RAW264.7 cells. These results suggest that oyster ß-thymosin, which is derived from the mantle of the Pacific oyster, has as much anti-inflammatory effects as human ß-thymosin. Additionally, oyster ß-thymosin suppressed NO production, PGE2 production and inflammatory cytokines expression via NF-κB in LPS-induced RAW264.7 cells.


Assuntos
Anti-Inflamatórios/farmacologia , Crassostrea/química , Dinoprostona/biossíntese , Macrófagos/efeitos dos fármacos , Óxido Nítrico/biossíntese , Timosina/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Sobrevivência Celular , Células Cultivadas , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Queratinócitos/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Células RAW 264.7 , Alinhamento de Sequência , Transdução de Sinais/efeitos dos fármacos , Timosina/isolamento & purificação
9.
Mitochondrial DNA B Resour ; 4(2): 3179-3181, 2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-33365908

RESUMO

The complete mitochondrial genome of Pseudotolithus elongatus (Perciformes: Sciaenidae) is determined based on NGS technology. The assembled mitogenome is a 16,497 bp in length containing a typical set of the 13 protein-coding genes, 22 tRNAs, 2 rRNA genes, and the 1 putative control region. The overall base composition is A (27.8%), T (25.3%), G (16.1%), and C (30.8%) with an A-T content of 53.1%. The phylogenetic analysis of 36 mitogenomes from the GenBank indicated that P. elongatus is closely related to the Aplodinotus grunniens. This mitogenome information of the P. elongatus would be useful to understand evolutionary and phylogenetic analysis of the family Sciaenidae fishes.

10.
Int J Oncol ; 53(5): 2300-2308, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30226597

RESUMO

In recent decades, various bioactive compounds from plants have been investigated for their potential use in the treatment of diseases in humans. Aster incisus extract (AIE) is the extract of a common plant that is mostly found in Asia. It has traditionally been used for medicinal purposes in South Korea. In this study, we evaluated the potential anticancer effects of a methanolic extract of Aster incisus in a normal human cell line (HaCaT keratinocytes) and in 4 different types of human cancer cell lines (A549, lung cancer; Hep3B, liver cancer; MDA­MB­231, breast cancer; and AGS, gastric cancer). The HaCaT, A549, Hep3B, MDA­MB­231 and AGS cells were treated with various concentrations of AIE and following treatment, cell survival was evaluated. Additional analyses, such as WST-1 assay, western blot analysis, DAPI staining, flow cytometry, immunofluorescence staining and wound healing assay were performed to elucidate the mechanisms and pathways involved in the cell death induced by AIE. Treatment with AIE induced morphological changes and considerably reduced the viability of the both normal and cancer cell lines. Further analysis of the AGS gastric cancer cells revealed that AIE led to the induction of apoptosis and a high accumulation of cells in the G1 cell phase following treatment with AIE in a dose-dependent manner. The results also revealed that AIE successfully suppressed the migration of the AIE-treated AGS cells. The results of western blot analysis indicated that AIE increased the expression of pro-apoptotic proteins, particularly Bid, Bad, Bak, cytochrome c, apoptosis inducing factor (AIF), cleaved caspase­3, -8 and -9 and cleaved poly(ADP-ribose) polymerase (PARP). Additionally, AIE decreased the expression of the anti-apoptotic proteins, Bcl-2 and Bcl-xL. On the whole, the findings of this study demonstrate that AIE induces apoptosis through the activation of the caspase­dependent pathway mediated by the mitochondrial pathway and by arresting the cell cycle in AGS cells.


Assuntos
Adenocarcinoma/tratamento farmacológico , Aster/química , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Medicina Tradicional Coreana/métodos , Extratos Vegetais/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Adenocarcinoma/epidemiologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Incidência , Metanol/química , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , República da Coreia/epidemiologia , Transdução de Sinais/efeitos dos fármacos , Neoplasias Gástricas/epidemiologia
11.
J Microbiol Biotechnol ; 28(10): 1645-1653, 2018 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-30176712

RESUMO

The genus Acer contains several species with various bioactivities including antioxidant, antitumor and anti-inflammatory properties. However, Acer okamotoanum Nakai, one species within this genus has not been fully studied yet. Therefore, in this study, we investigated the anti-adipogenic activities of leaf extract from A. okamotoanum Nakai (LEAO) on 3T3-L1 preadipocytes. Adipogenesis is one of the cell differentiation processes, which converts preadipocytes into mature adipocytes. Nowadays, inhibition of adipogenesis is considered as an effective strategy in the field of anti-obesity research. In this study, we observed that LEAO decreased the accumulation of lipid droplets during adipogenesis and down-regulated the expression of key adipogenic transcription factors such as peroxisome proliferator-activated receptor γ (PPAR γ) and CCAAT/enhancer binding protein α (C/EBP α). In addition, LEAO inactivated PI3K/Akt signaling and its downstream factors that promote adipogenesis by inducing the expression of PPAR γ. LEAO also activated ß-catenin signaling, which prevents the adipogenic program by suppressing the expression of PPAR γ. Therefore, we found that treatment with LEAO is effective for attenuating adipogenesis in 3T3-L1 cells. Consequently, these findings suggest that LEAO has the potential to be used as a therapeutic agent for preventing obesity.


Assuntos
Acer/química , Adipogenia/efeitos dos fármacos , Proteínas Estimuladoras de Ligação a CCAAT/genética , Regulação da Expressão Gênica/efeitos dos fármacos , PPAR gama/genética , Extratos Vegetais/farmacologia , Células 3T3-L1 , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Animais , Sobrevivência Celular , Metabolismo dos Lipídeos/efeitos dos fármacos , Camundongos , Obesidade/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , beta Catenina/metabolismo
12.
Microb Pathog ; 116: 84-90, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29339306

RESUMO

Facile, eco-friendly synthesis of metal nanoparticles has been proposed as a cost effective method. In the present study, we propose the facile synthesis of silver-silver chloride (Ag-AgCl) nanoparticles (NPs) using the medicinally important Agrimonia pilosa plant extract without addition of capping or stabilizing agents. The Ag-AgCl NPs synthesis was observed at 40 °C after 10 min incubation; the synthesis of Ag-AgCl NPs was indicated by color change and confirmed by UV-vis spectroscopic peak at 454 nm. TEM analysis confirmed Ag-AgCl NPs were 10-20 nm in size and spherical, and oval in shape. Elemental composition was determined by energy dispersive X-ray analysis, and crystalline structure was confirmed by X-ray diffraction spectroscopy. Different phytocomponents present in the plant extract were analyzed by Gas Chromatography-Mass spectrometry, and the interaction of biomolecules in reduction process was analyzed by Fourier transform infrared spectroscopy studies. The synthesized Ag-AgCl NPs showed significant antibacterial efficiency, analyzed by well diffusion assay against pathogenic bacteria including Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Staphylococcus saprophyticus, Escherichia coli, Pseudomonas putida. Minimum inhibitory concentration and minimum bactericidal concentration were evaluated by microbroth dilution, and spread plate method, respectively. The possible mechanism of bacterial growth inhibition is due to changes in bacterial cell wall morphology that was studied by FE-SEM analysis.


Assuntos
Agrimonia/metabolismo , Antibacterianos/metabolismo , Bactérias/citologia , Bactérias/efeitos dos fármacos , Nanopartículas Metálicas , Prata/metabolismo , Bacillus cereus , Contagem de Colônia Microbiana , Escherichia coli , Cromatografia Gasosa-Espectrometria de Massas , Listeria monocytogenes , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Transmissão , Extratos Vegetais/metabolismo , Pseudomonas putida , Prata/química , Espectrometria por Raios X , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de Fourier , Staphylococcus aureus , Staphylococcus saprophyticus , Temperatura , Difração de Raios X
13.
Mediators Inflamm ; 2018: 4675204, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30622433

RESUMO

Aster incisus is a common flower found in almost all regions of South Korea. In the current study, we investigated the potential antioxidant and anti-inflammatory properties of the Aster incisus methanol extract in LPS-stimulated RAW 264.7 cells. We analyzed the phytochemicals contained in the extract by GC-MS. GC-MS results showed that the Aster incisus extract contains 9 known compounds. Later on, DPPH assay, WST-1 assay, nitric oxide (NO) assay, Western blot, and RT-PCR were conducted to investigate the anti-inflammatory effects of the extract. Our WST-1 assay results revealed that Aster incisus did not affect the viability of all tested cell lines up to a concentration of 200 µg/ml; therefore, lower concentrations (50 µg/ml and 150 µg/ml) were used for further assays. Aster incisus scavenged DPPH and inhibited the production of NO. Aster incisus also reduced significantly the production of inflammation-related enzymes (iNOS, Cox-2) and cytokines (TNFα, IL-1ß, and IL-6) and the gene expression of the proinflammatory cytokines. Additionally, further Western blot results indicated that Aster incisus inhibited the expression of p-PI3K, p-IκBα, p-p65 NF-κB, p-ERK1/2, p-SAPK/JNK, and p-Akt. Our results demonstrated that Aster incisus suppressed the expression of the inflammation mediators through the regulation of NF-κB, MAPK, and Akt pathways.


Assuntos
Aster/química , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Camundongos , Células RAW 264.7
14.
Int J Mol Med ; 41(2): 1103-1109, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29207042

RESUMO

Lovastatin is a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor that is clinically used for the prevention of cardiovascular diseases. Although it has been reported that lovastatin has anti-inflammatory properties in several studies, how lovastatin regulates the inflammation is still unclear. To evaluate the effect of lovastatin on nitric oxide production (NO) in RAW264.7 macrophages, NO production assay was performed. Also, cell viability was measured to confirm cytotoxicity. Level of tumor necrosis factor-α (TNF-α) transcription was measured by reverse transcription polymerase chain reaction (RT-PCR) from total RNA in RAW264.7 cells. Western blot analysis and immunofluorescence staining were used to investigate the regulation of lovastatin on the expression, phosphorylation, and nuclear translocation of cellular proteins. The results of the present study revealed that lovastatin reduced nitric oxide production via the reduction of inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. The mRNA level of TNF-α was reduced in presence of lovastatin. In addition, lovastatin downregulated histone deacetylase 1 (HDAC1), resulting in the accumulation of acetylated histone H3 and heat shock protein 70. Furthermore, the expression of phosphoinositide 3-kinase catalytic subunits α and ß was reduced under lovastatin treatment, and the phosphorylation of Akt and mammalian target of rapamycin was consequently inhibited. Lovastatin also inhibited the phosphorylation of inhibitor of nuclear factor (NF)-κBα and the translocation of NF-κB into the nucleus. Therefore, the present study demonstrates that lovastatin inhibits the expression of pro-inflammatory mediators, including iNOS and TNF-α, through the suppression of HDAC1 expression, PI3K/Akt phosphorylation and NF-κB translocation in LPS-stimulated RAW264.7 macrophage cells.


Assuntos
Anti-Inflamatórios/administração & dosagem , Histona Desacetilase 1/genética , Inflamação/tratamento farmacológico , Lovastatina/administração & dosagem , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Histona Desacetilase 1/antagonistas & inibidores , Humanos , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/patologia , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Macrófagos/patologia , Camundongos , NF-kappa B/genética , Óxido Nítrico Sintase Tipo II/genética , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Células RAW 264.7 , Serina-Treonina Quinases TOR/genética , Fator de Necrose Tumoral alfa/genética
15.
Genome Announc ; 5(36)2017 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-28883126

RESUMO

We report the draft genome sequences of a novel member of the Picornavirales isolated from Pacific abalone (Haliotis discus hannai). The full length of the assembled draft genome sequences, obtained by use of a next-generation sequencing technique, were 8,019 nucleotides, including an RNA-dependent RNA polymerase gene (5,088 nucleotides) and a capsid protein gene (2,553 nucleotides). This genome sequence will be useful for understanding viral disease of Pacific abalone.

16.
J Microbiol Biotechnol ; 25(11): 1801-9, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26215267

RESUMO

A phytoene synthase gene, crtB, was isolated from Kocuria gwangalliensis. The crtB with 1,092 bp full-length has a coding sequence of 948 bp and encodes a 316-amino-acids protein. The deduced amino acid sequence showed a 70.9% identity with a putative phytoene synthase from K. rhizophila. An expression plasmid, pCcrtB, containing the crtB gene was constructed, and E. coli cells containing this plasmid produced the recombinant protein of approximately 34 kDa , corresponding to the molecular mass of phytoene synthase. Biosynthesis of lycopene was confirmed when the plasmid pCcrtB was co-transformed into E. coli containing pRScrtEI carrying the crtE and crtI genes encoding lycopene biosynthetic pathway enzymes. The results obtained from this study will provide a base of knowledge about the phytoene synthase of K. gwangalliensis and can be applied to the production of carotenoids in a non-carotenoidproducing host.


Assuntos
Escherichia coli/metabolismo , Geranil-Geranildifosfato Geranil-Geraniltransferase/biossíntese , Micrococcaceae/enzimologia , Carotenoides/biossíntese , Clonagem Molecular , Escherichia coli/genética , Expressão Gênica , Geranil-Geranildifosfato Geranil-Geraniltransferase/química , Geranil-Geranildifosfato Geranil-Geraniltransferase/genética , Licopeno , Micrococcaceae/genética , Dados de Sequência Molecular , Peso Molecular , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Análise de Sequência de DNA
17.
J Pharm Pharmacol ; 67(9): 1297-305, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25904113

RESUMO

OBJECTIVES: The purpose of this study is to investigate anti-inflammatory effects of toluhydroquinone in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. METHODS: Toluhydroquinone was purified from a fungal strain, Aspergillus sp. We investigated that levels of nitric oxide (NO) using Griess reagent, production of prostaglandin E2 (PGE2 ) and pro-inflammatory cytokines using ELISA assay. We conducted Western blot analysis to investigate regulatory effects of toluhydroquinone on expression of inducible nitric oxide synthase (iNOS), cyclooxyganse-2 (COX-2), nuclear factor-κB (NF-κB), Akt and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW264.7 cells. The translocation of NF-κB was detected by immunofluorescence staining. KEY FINDINGS: Toluhydroquinone inhibited production of NO and PGE2 via suppressing protein expression of iNOS and COX-2, respectively. Secretion and expression of inflammatory cytokines were down-regulated by toluhydroquinone as well. Toluhydroquinone reduced phosphorylation of Akt, NF-κB and MAPKs. Moreover, toluhydroquinone inhibited translocation of NF-κB from the cytosol into the nucleus. CONCLUSIONS: We revealed that inhibitory effects of toluhydroquinone on expression of inflammatory mediators are induced through inactivation of Akt, NF-κB and MAPKs. Thus, our results suggest that toluhydroquinone may be used for a potential anti-inflammatory reagent.


Assuntos
Anti-Inflamatórios/farmacologia , Aspergillus/metabolismo , Benzoquinonas/farmacologia , Inflamação/tratamento farmacológico , Lipopolissacarídeos/farmacologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Animais , Linhagem Celular , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Células HEK293 , Humanos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
18.
Biomed Pharmacother ; 70: 129-39, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25776491

RESUMO

Angiogenesis, the growth of new blood vessels from the existing ones, occurs during embryo development and wound healing. However, most malignant tumors require angiogenesis for their growth and metastasis as well. Therefore, inhibition of angiogenesis has been focused as a new strategy of cancer therapies. To treat cancer, there are marine microorganism-derived secondary metabolites developed as chemotherapeutic agents. In this study, we used toluhydroquinone (2-methyl-1,4-hydroquinone), one of the secondary metabolites isolated from marine algae symbiotic fungus, Aspergillus sp. We examined the effects of toluhydroquinone on angiogenesis using HUVECs. We identified that toluhydroquinone inhibited the activity of ß-catenin and down-regulated Ras/Raf/MEK/ERK signaling which are crucial components during angiogenesis. In addition, the expression and activity of MMPs are reduced by the treatment of toluhydroquinone. In conclusion, we confirmed that toluhydroquinone has inhibitory effects on angiogenic behaviors of human endothelial cells, HUVECs. Our findings suggest that toluhydroquinone can be proposed as a potent anti-angiogenesis drug candidate to treat cancers.


Assuntos
Inibidores da Angiogênese/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Hidroquinonas/metabolismo , Rodófitas/metabolismo , Simbiose/efeitos dos fármacos , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Aspergillus/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Humanos , Hidroquinonas/química , Hidroquinonas/farmacologia , Simbiose/fisiologia
19.
Int J Mol Med ; 35(1): 202-10, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25352364

RESUMO

In the present study, we investigated regulatory effects of veratric acid on the production of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. NO production was significantly decreased by veratric acid in the LPS-stimulated RAW264.7 cells in a dose-dependent manner. The reduction in nitric oxide production was induced by the downregulation of inducible NO synthase (iNOS) expression. Veratric acid suppressed the LPS-induced effects on the regulatory and catalytic subunits of phosphoinositide 3-kinase (PI3K), comprised of p85, p110α, p110ß and Akt. The acetylation of p300 and the phosphorylation of activating transcription factor 2 (ATF-2) induced by LPS were downregulated following treatment with veratric acid; similar effects were observed following treatment with LY294002, a specific inhibitor of PI3K/Akt. The LPS-induced expression of histone deacetylase (HDAC)3 decreased to basal levels following treatment with veratric acid, and its expression was also downregulated by LY294002. In the measurement of histone acetylation levels, the LPS-stimulated acetylation of histone H4 was significantly attenuated by veratric acid, and was also reduced following the inhibition of PI3K/Akt with LY294002. From our data, it can be concluded that veratric acid exerts a regulatory effect on LPS-induced iNOS expression. Our results suggest that veratric acid impedes the PI3K/Akt-mediated histone acetyl-transferase (HAT) activation and HDAC expression induced by LPS, thereby abrogating iNOS expression.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Histonas/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Fosfatidilinositol 3-Quinases/metabolismo , Ácido Vanílico/análogos & derivados , Acetilação/efeitos dos fármacos , Animais , Linhagem Celular , Ativação Enzimática/efeitos dos fármacos , Feminino , Histona Desacetilases/genética , Histona Desacetilases/metabolismo , Lipopolissacarídeos , Camundongos , Óxido Nítrico , Óxido Nítrico Sintase Tipo II/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ácido Vanílico/farmacologia
20.
Int J Mol Med ; 34(4): 1101-9, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25109657

RESUMO

The aim of the present study was to identify the anti-inflammatory and anti-oxidative effects of peat moss aqueous extract (PME) on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. To demonstrate the anti-inflammatory and antioxidant effects of PME, the levels of nitric oxide (NO) and cytokines were measured using Griess reagent and cytokine ELISA kits, respectively. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analysis were conducted to evaluate the expression of genes and proteins. Immunofluorescence was used to measure the expression and translocation of transcription factors. Pre-treatment with PME inhibited the production of prostaglandin E(2) and NO by suppressing the gene expression of cyclooxygenase-2 and inducible NO synthase, respectively. The LPS-stimulated gene expression and the production of tumor necrosis factor-α and interleukin-1ß were significantly reduced by PME. In the LPS-stimulated RAW 264.7 cells, nuclear factor­κB (NF-κB) translocated from the cytosol to the nucleus, while pre-treatment with PME induced the sequestration of NF-κB in the cytosol through the inhibition of IκBα degradation. In the same manner, PME contributed to the inhibition of the activation of mitogen-activated protein kinases. In addition, the PME-treated RAW 264.7 cells facilitated the activation of nuclear factor-like 2 (Nrf2) , and in turn, enhanced heme oxygenase-1 (HO-1) expression. These results indicate that PME exerts anti-inflammatory and antioxidant effects, and suggest that PME may neutralize inflammation and prevent cellular damage by oxidative stress.


Assuntos
Anti-Inflamatórios/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Extratos Vegetais/farmacologia , Sphagnopsida/química , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Dinoprostona/metabolismo , Ativação Enzimática/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/enzimologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo
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