RESUMO
Modular and tuneable genetic tools for Metabolic Engineering fuels the development of chassis for the efficient production of biocompounds at industrial scale. We have constructed an autoinduction device for gene expression in Bacillus subtilis based on the LuxR/I quorum sensing system [1]. Here, we present raw and processed data regarding to B. subtilis growth measured as OD600, performed in three different scales: microcultivation on 96-well plates (200 µL), test tubes (12 mL), and Erlenmeyer flasks (50 mL). We also present raw and processed data on gene expression measured as GFP fluorescence (485/535 nm), luminescence and riboflavin production. Measurements were performed on a microplate reader Tecan 200 PRO (iControl software) and on spectrophotometer (Thermo Fisher Scientific GENESYS 10S UV-Vis). Processed data are presented as product/OD600, maximum and minimum promoter activity, fold of induction, and the induction OD600.
RESUMO
Intense synthesis of proteins and chemicals in engineered microbes impose metabolic burden, frequently leading to reduced growth and heterogeneous cell population. Thus, the correct balance between growth and production is important. Such balance can be engineered through dynamic control of pathways, but few broadly applicable tools are available to achieve this. We present an autonomous control of gene expression mediated by quorum sensing in Bacillus subtilis, able to self-monitor and induce expression without human supervision. Two variations of the induction module and seven of the response module were engineered generating a range of induction folds and strengths for gene expression control. Our strongest response promoter is 2.5 and 3.2 times stronger than the well-characterized promoters PsrfA and Pveg, respectively. We applied our strongest autoinduction device for the production of the vitamin B2. This study presents a toolbox of autoinduction modules for B. subtilis that is modular and tunable.
Assuntos
Bacillus subtilis , Regulação Bacteriana da Expressão Gênica , Engenharia Metabólica , Percepção de Quorum , Bacillus subtilis/genética , Bacillus subtilis/metabolismoRESUMO
A Split-hand/foot Malformation, muitas vezes aceita como sinônimo de ectrodactilia, é uma malformação com diferentes padrões de hereditariedade que pode se apresentar isoladamente ou como parte de síndromes de maior expressão clínica. Discutimos as peculiaridades do seu diagnóstico e das manifestações associadas ao quadro. Descrevemos o caso esporádico de um paciente com ectrodactilia que desenvolveu uma Síndrome Mielodisplásica associada a manifestações reumatológicas e a Trombose Venosa Profunda. Consideramos o paciente como portador da forma isolada da Split-hand/foot Malformation e as suas outras manifestações como consequências atípicas da Síndrome Mielodisplásica.
The Split-hand/foot Malformation often accepted as a synonym for ectrodactyly is a malformation with different patterns of heredity that can present it individually or as part of syndromes with most clinical significance. We discussed the peculiarities of their diagnosis and clinical manifestations associated with the condition presented. We describe a sporadic case of a patient with ectrodactyly who developed a myelodysplastic syndrome associated with rheumatic manifestations and Deep VeinThrombosis. We considered the patient as suffering from an isolated form of Split-hand/foot Malformation and its other manifestations as atypical consequences of myelodysplastic syndrome.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Anormalidades Musculoesqueléticas/genética , Deformidades Congênitas do Pé , Deformidades Congênitas da Mão , Síndromes Mielodisplásicas/diagnóstico , Trombose VenosaRESUMO
Justificativa e objetivos: Histoplasmose disseminada aguda (HDA) é considerada uma doença definidora da síndrome da imunodeficiência adquirida (Aids). Este relato se destina a demonstrar um caso de HDA associado à Aids, sua dificuldade diagnóstica e alta letalidade. Relato do caso: Paciente do sexo masculino, 26 anos, sem patologias prévias, iniciou quadro agudo de febre diária associado à afecção pulmonar e do trato gastrointestinal. Foi realizado anti-HIV, que detectou positividade. Durante a internação apresentou piora clínica, sendo então tratado empiricamente para as principais doenças oportunistas desta imunodeficiência, porém sem resposta eficaz. Evoluiu para o óbito, sendo obtido tardiamente o diagnóstico de HDA através da necrópsia. Conclusão: HDA deve ser aventada em todo e qualquer portador de HIV com síndrome febril e CD4 baixo, independente de sua procedência, ocupação ou história epidemiológica de exposição ao Histoplasma capsulatum. A associação da Aids e HDA pode ter um curso rápido e fatal...
Assuntos
Humanos , Masculino , Adulto Jovem , Febre , Histoplasmose , Síndrome da Imunodeficiência AdquiridaRESUMO
OBJECTIVE: The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes. METHODS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors. RESULTS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells. CONCLUSIONS: Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors.
Assuntos
Células da Medula Óssea/metabolismo , Quinase 2 de Adesão Focal/metabolismo , Síndromes Mielodisplásicas/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 11/análise , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Quinase 2 de Adesão Focal/análise , Proteína-Tirosina Quinases de Adesão Focal/análise , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/genética , Reação em Cadeia da Polimerase , Prognóstico , Proteína Tirosina Fosfatase não Receptora Tipo 11/metabolismo , Fatores de Risco , Estatísticas não Paramétricas , Adulto Jovem , Domínios de Homologia de src/fisiologiaRESUMO
OBJECTIVE: The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes. METHODS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors. RESULTS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells. CONCLUSIONS: Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors. .
