RESUMO
Morphological properties and the size of microvesicles were assessed using atomic force microscopy, electron microscopy, and granulometric analysis. As these methods require significant numbers of microvesicles, we chose microvesicles derived from cell lines for our research.
Assuntos
Membrana Celular/ultraestrutura , Micropartículas Derivadas de Células/ultraestrutura , Células Endoteliais/ultraestrutura , Células Matadoras Naturais/ultraestrutura , Trofoblastos/ultraestrutura , Linhagem Celular , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Células THP-1RESUMO
The study involving 48 patients with the preliminary diagnosis of acute coronary syndrome aged 54-78 years included analysis of clinical, laboratory and instrumental data for subgroups with different definitive diagnosis (myocardial infarction and unstable angina in 28 and 22 patients respectively). On the whole, the mean content of HSP-70 at admission averaged 2.1 ± 0.3 ng/ml and decreased to 1.6 ± 0.4 ng/ml (p < 0.05) after therapy. In the subgroup with myocardial infarction, the HSP-70 level (1.6 ± 0.4 ng/ml) was significantly lower than in patients with unstable angina (2.1 ± 0.3 ng/ml); p < 0.05. The reduced HSP-70 level associated with a severe myocardial lesion gives evidence against the hypothesis of these proteins as markers of myocardial dysfunction whereas a rise in their content at the onset of ischemic process suggests their protective role.
Assuntos
Angina Instável , Proteínas de Choque Térmico HSP70/sangue , Infarto do Miocárdio , Miocárdio , Idoso , Angina Instável/diagnóstico , Angina Instável/metabolismo , Angina Instável/terapia , Biomarcadores/sangue , Diagnóstico Diferencial , Tratamento de Emergência , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/terapia , Miocárdio/metabolismo , Miocárdio/patologia , Fatores de Risco , Estatística como AssuntoRESUMO
For the first time, the biodistribution of recombinant heat shock protein in rhHsp70 rats with grafted intracranial C6 glioma was evaluated. It was assessed using the fluorescent antibody accumulation chaperone rhHsp70 conjugated with fluorochrome Alexa Fluor 555 in tumor cells by intratumoral or intravenous administration. Assessment of the distribution and accumulation of labeled protein was carried out on the model of subcutaneous B16/F10 melanoma in C57BL/6 mice with the use of single-photon emission computer tomography. After 60 minutes after intravenous administration rhHsp70-I123 (20 MBq, 5 mg chaperone) accumulation of the drug mainly in the liver and tumor tissue was showed. The coefficient of the differential accumulation of the labeled protein KDN(tumor/background) was 3.14. It was turned out that comparing the level of fixation of rhHsp70-I123 in the liver and the tumor KDN(tumor/ liver) = 0.76. After 24 hours from the time of injection of rhHsp70-I123 it was observed increase the level of fixation of the labeled protein in the liver and melanoma: KDN(tumor/background) = 3.43; KDN(tumor/liver = 0.78.
Assuntos
Neoplasias Encefálicas/metabolismo , Glioma/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Fígado/metabolismo , Melanoma Experimental/metabolismo , Neoplasias Cutâneas/metabolismo , Animais , Corantes Fluorescentes , Proteínas de Choque Térmico HSP70/administração & dosagem , Proteínas de Choque Térmico HSP70/farmacocinética , Injeções Intralesionais , Injeções Intravenosas , Radioisótopos do Iodo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais/metabolismo , Ratos , Ratos Wistar , Fatores de Tempo , Distribuição Tecidual , Tomografia Computadorizada de Emissão de Fóton ÚnicoRESUMO
Kinetics of the chaperone activity of proteins Hsp70 and Hdj1 were analyzed in human U-937 promonocytes during their response to heat shock or to treatment with the echinochrome triacetyl glucoside derivative U-133. To measure the chaperone activity of both proteins, a special test was developed for their recognition and binding of a denatured protein. Using this test, the chaperone activity could be concurrently estimated in large numbers of cellular or tissue extracts. We also estimated the contents of both chaperones in cells by immunoblotting. The values for contents of Hsp70 and Hdj1 obtained by two independent test systems coincided, and this suggested that the substrate-binding activity could change proportionally to the chaperone content in the protein mixture. Therefore, the test developed by us can be employed for high throughput screening of drugs activating cellular chaperones. The analysis of quantity and activity of two cellular chaperones during the cell response to heat stress or to the drug-like substance U-133 showed that both factors caused the accumulation of chaperones with similar kinetics. We conclude that the efficiency of drug preconditioning could be close to the efficiency of hyperthermia and that the high activity of chaperones could be retained in human cells for no less than 1.5 days.