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1.
BMC Microbiol ; 16(1): 290, 2016 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-27955621

RESUMO

BACKGROUND: Pharmaceutical industry demands innovation for developing new molecules to improve effectiveness and safety of therapeutic medicines. Preclinical assays are the first tests performed to evaluate new therapeutic molecules using animal models. Currently, there are several models for evaluation of treatments, for dermal oedema or infection. However, the most common or usual way is to induce the inflammation with chemical substances instead of infectious agents. On the other hand, this kind of models require the implementation of histological techniques and the interpretation of pathologies to verify the effectiveness of the therapy under assessment. This work was focused on developing a quantitative model of infection and oedema in mouse pinna. The infection was achieved with a strain of Streptococcus pyogenes that was inoculated in an injury induced at the auricle of BALB/c mice, the induced oedema was recorded by measuring the ear thickness with a digital micrometer and histopathological analysis was performed to verify the damage. The presence of S. pyogenes at the infection site was determined every day by culture. RESULTS: Our results showed that S. pyogenes can infect the mouse pinna and that it can be recovered at least for up to 4 days from the infected site; we also found that S. pyogenes can induce a bigger oedema than the PBS-treated control for at least 7 days; our results were validated with an antibacterial and anti-inflammatory formulation made with ciprofloxacin and hydrocortisone. CONCLUSIONS: The model we developed led us to emulate a dermal infection and allowed us to objectively evaluate the increase or decrease of the oedema by measuring the thickness of the ear pinna, and to determine the presence of the pathogen in the infection site. We consider that the model could be useful for assessment of new anti-inflammatory or antibacterial therapies for dermal infections.


Assuntos
Modelos Animais de Doenças , Pavilhão Auricular/efeitos dos fármacos , Pavilhão Auricular/microbiologia , Edema/tratamento farmacológico , Dermatopatias Bacterianas/tratamento farmacológico , Infecções Estreptocócicas/tratamento farmacológico , Streptococcus pyogenes/fisiologia , Animais , Antibacterianos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Avaliação Pré-Clínica de Medicamentos/métodos , Pavilhão Auricular/patologia , Edema/microbiologia , Edema/patologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Dermatopatias Bacterianas/microbiologia , Dermatopatias Bacterianas/patologia , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/patologia
2.
Tumour Biol ; 32(3): 561-8, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21225484

RESUMO

The aim of this study was to identify the gene expression profile in biopsies of patients with cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3, and microinvasive cancer by suppression subtractive hybridization and Southern blotting. After analyzing 1,800 cDNA clones, we found 198 upregulated genes, 166 downregulated, and no significant change of gene expression in 86 clones (p = 0.005). These results were validated by Northern blot analysis (p = 0.0001) in the identification of 28 overexpressed and 7 downregulated transcripts. We observed a set of genes related to the Notch signaling pathway that may be involved in the transformation of cervical cells and in the development to malignancy. The differentially expressed genes may provide useful information about the molecular mechanisms involved in human cervical carcinoma and as diagnostic markers.


Assuntos
Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Neoplasias do Colo do Útero/genética , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/fisiologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/fisiologia , Feminino , Papillomavirus Humano 16/isolamento & purificação , Humanos , México , Proteína Proto-Oncogênica N-Myc , Proteínas Nucleares/genética , Proteínas Oncogênicas/genética , Receptor Notch3 , Receptores Notch/genética , Receptores Notch/fisiologia , Transcrição Gênica , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia
3.
Mol Biotechnol ; 42(1): 61-7, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19058034

RESUMO

Analysis of the Thermoplasma acidophilum DSM 1728 genome identified two putative alcohol dehydrogenase (ADH) open reading frames showing 50.4% identity against each other. The corresponding genes Ta0841 and Ta1316 encode proteins of 336 and 328 amino acids with molecular masses of 36.48 and 36.01 kDa, respectively. The genes were expressed in Escherichia coli and the recombinant enzymes were functionally assessed for activity. Throughout the study only Ta1316 ADH resulted active in the oxidative reaction in the pH range 2-8 (optimal pH 5.0) and temperatures from 25 to 90 degrees C (optimal 75 degrees C). This ADH catalyzes the oxidation of several alcohols such as ethanol, methanol, 2-propanol, butanol, and pentanol during the reduction of the cofactor NAD(+). The highest activity was found in the presence of ethanol producing optically pure acetaldehyde. The specific enzyme activity of the purified Ta1316 ADH with ethanol as a substrate in the optimal conditions was 628.7 U/mg.


Assuntos
Álcool Desidrogenase/química , Álcool Desidrogenase/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Thermoplasma/enzimologia , Álcool Desidrogenase/genética , Álcoois/metabolismo , Sequência de Aminoácidos , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Cinética , Dados de Sequência Molecular , Proteínas Recombinantes/genética , Análise de Sequência de DNA , Especificidade por Substrato , Temperatura
4.
Salud Publica Mex ; 48(2): 155-65, 2006.
Artigo em Espanhol | MEDLINE | ID: mdl-16619872

RESUMO

The Notch signaling pathway plays a crucial role at different stages of cell development, such as proliferation, growth, differentiation, and apoptosis. Recent studies demonstrate that depending on the expression level and cellular context, the Notch receptors play a role in apoptosis resistance in malignant cells. These findings suggest that Notch signaling components may be a potential target in the development of new cancer therapies. This review describes the function of the Notch pathway and new strategies in the modulation of its signal.


Assuntos
Neoplasias/tratamento farmacológico , Neoplasias/etiologia , Receptores Notch/antagonistas & inibidores , Receptores Notch/fisiologia , Transdução de Sinais , Apoptose , Humanos
5.
Salud pública Méx ; 48(2): 155-165, mar.-abr. 2006.
Artigo em Espanhol | LILACS | ID: lil-429954

RESUMO

La vía de señalización Notch desempeña un papel fundamental en las diferentes etapas del desarrollo celular como la proliferación, crecimiento, diferenciación y apoptosis. Estudios recientes han demostrado que, dependiendo del nivel de expresión y del contexto celular, los receptores de membrana Notch contribuyen en la resistencia a apoptosis en células tumorales. Estos descubrimientos sugieren que componentes de la vía de señalización Notch son un blanco potencial para el desarrollo de terapias más efectivas contra el cáncer. Esta revisión describe la función de la vía Notch y nuevas estrategias utilizadas en la modulación de su señal.


Assuntos
Humanos , Neoplasias/tratamento farmacológico , Neoplasias/etiologia , Receptores Notch/antagonistas & inibidores , Receptores Notch/fisiologia , Transdução de Sinais , Apoptose
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