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1.
Appl Microbiol Biotechnol ; 103(1): 211-223, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30343427

RESUMO

Lycopene is a red carotenoid pigment with strong antioxidant activity. Saccharomyces cerevisiae is considered a promising host to produce lycopene, but lycopene toxicity is one of the limiting factors for high-level production. In this study, we used heterologous lycopene biosynthesis genes crtE and crtI from Xanthophyllomyces dendrorhous and crtB from Pantoea agglomerans for lycopene production in S. cerevisiae. The crtE, crtB, and crtI genes were integrated into the genome of S. cerevisiae CEN.PK2-1C strain, while deleting DPP1 and LPP1 genes to inhibit a competing pathway producing farnesol. Lycopene production was further improved by inhibiting ergosterol production via downregulation of ERG9 expression and by deleting ROX1 or MOT3 genes encoding transcriptional repressors for mevalonate and sterol biosynthetic pathways. To further increase lycopene production, CrtE and CrtB mutants with improved activities were isolated by directed evolution, and subsequently, the mutated genes were randomly integrated into the engineered lycopene-producing strains via delta-integration. To relieve lycopene toxicity by increasing unsaturated fatty acid content in cell membranes, the OLE1 gene encoding stearoyl-CoA 9-desaturase was overexpressed. In combination with the overexpression of STB5 gene encoding a transcription factor involved in NADPH production, the final strain produced up to 41.8 mg/gDCW of lycopene, which is approximately 74.6-fold higher than that produced in the initial strain.


Assuntos
Licopeno/metabolismo , Microrganismos Geneticamente Modificados , NADP/biossíntese , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Basidiomycota/genética , Membrana Celular/metabolismo , Evolução Molecular Direcionada , Farneseno Álcool/metabolismo , Farnesil-Difosfato Farnesiltransferase/genética , Farnesil-Difosfato Farnesiltransferase/metabolismo , Ácidos Graxos Insaturados/metabolismo , Regulação Fúngica da Expressão Gênica , Pantoea/genética , Engenharia de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Estearoil-CoA Dessaturase/genética , Estearoil-CoA Dessaturase/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
2.
Viruses ; 7(9): 5133-44, 2015 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-26402693

RESUMO

The antiviral effect of a catalytic RNA-hydrolyzing antibody, 3D8 scFv, for intranasal administration against avian influenza virus (H1N1) was described. The recombinant 3D8 scFv protein prevented BALB/c mice against H1N1 influenza virus infection by degradation of the viral RNA genome through its intrinsic RNA-hydrolyzing activity. Intranasal administration of 3D8 scFv (50 µg/day) for five days prior to infection demonstrated an antiviral activity (70% survival) against H1N1 infection. The antiviral ability of 3D8 scFv to penetrate into epithelial cells from bronchial cavity via the respiratory mucosal layer was confirmed by immunohistochemistry, qRT-PCR, and histopathological examination. The antiviral activity of 3D8 scFv against H1N1 virus infection was not due to host immune cytokines or chemokines, but rather to direct antiviral RNA-hydrolyzing activity of 3D8 scFv against the viral RNA genome. Taken together, our results suggest that the RNase activity of 3D8 scFv, coupled with its ability to penetrate epithelial cells through the respiratory mucosal layer, directly prevents H1N1 virus infection in a mouse model system.


Assuntos
Anticorpos Catalíticos/administração & dosagem , Antivirais/administração & dosagem , Células Epiteliais/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Ribonucleases/administração & dosagem , Anticorpos de Cadeia Única/administração & dosagem , Administração Intranasal , Animais , Antivirais/farmacocinética , Hidrólise , Camundongos Endogâmicos BALB C , RNA Viral/metabolismo , Anticorpos de Cadeia Única/farmacocinética , Resultado do Tratamento
3.
J Med Syst ; 32(6): 471-80, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19058651

RESUMO

The advanced technology of computing system was followed by the rapid improvement of medical instrumentation and patient record management system. The typical examples are hospital information system (HIS) and picture archiving and communication system (PACS), which computerized the management procedure of medical records and images in hospital. Because these systems were built and used in hospitals, doctors out of hospital have problems to access them immediately on emergent cases. To solve these problems, this paper addressed the realization of system that could transmit the images acquired by medical imaging systems in hospital to the remote doctors' handheld PDA's using CDMA cellular phone network. The system consists of server and PDA. The server was developed to manage the accounts of doctors and patients and allocate the patient images to each doctor. The PDA was developed to display patient images through remote server connection. To authenticate the personal user, remote data access (RDA) method was used in PDA accessing the server database and file transfer protocol (FTP) was used to download patient images from the remove server. In laboratory experiments, it was calculated to take ninety seconds to transmit thirty images with 832 x 488 resolution and 24 bit depth and 0.37 Mb size. This result showed that the developed system has no problems for remote doctors to receive and review the patient images immediately on emergent cases.


Assuntos
Telefone Celular , Computadores de Mão , Sistemas de Informação/instrumentação , Segurança Computacional , Compressão de Dados/métodos , Atenção à Saúde/métodos , Desenho de Equipamento , Humanos , Processamento de Imagem Assistida por Computador/métodos , Software
4.
Conf Proc IEEE Eng Med Biol Soc ; 2005: 3749-52, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-17281044

RESUMO

Diabetes is an incurable chronic disease with the prevalence high in developed countries. The number of patients in Korea is also on a steady increase. Patients have to self-manage their blood glucose level by daily test and insulin injection. Therefore, it is very important to assist them in self-management procedure. In this study, PDA(personal digital assistant) based PDMS (personal diabetes management system) was developed in the integrated environment of Visual Studio .NET 2003, which consisted of four main menus to manage personal information, diet, exercise, and blood glucose. It gives quantitative health indices, such as BMI (body mass index) and diabetes index, based on personal physical information, pedigree, and living habits. In addition, it is capable of predicting change of the body weight, which may remind patients of the importance of the disease management. The present system can be used as a mobile device within the tele-healthcare system integrated with medical center through Internet.

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