RESUMO
Protein malnutrition causes anemia and leukopenia as it reduces hematopoietic precursors and impairs the production of mediators that regulate hematopoiesis. Hematopoiesis occurs in distinct bone marrow niches that modulate the processes of differentiation, proliferation and self-renewal of hematopoietic stem cells (HSCs). Mesenchymal stem cells (MSCs) contribute to the biochemical composition of bone marrow niches by the secretion of several growth factors and cytokines, and they play an important role in the regulation of HSCs and hematopoietic progenitors. In this study, we investigated the effect of protein malnutrition on the hematopoietic regulatory function of MSCs. C57BL/6NTaq mice were divided into control and protein malnutrition groups, which received, respectively, a normal protein diet (12% casein) and a low protein diet (2% casein). The results showed that protein malnutrition altered the synthesis of SCF, TFG-ß, Angpt-1, CXCL-12, and G-CSF by MSCs. Additionally, MSCs from the protein malnutrition group were not able to maintain the lymphoid, granulocytic and megakaryocytic-erythroid differentiation capacity compared to the MSCs of the control group. In this way, the comprehension of the role of MSCs on the regulation of the hematopoietic cells, in protein malnutrition states, is for the first time showed. Therefore, we infer that hematopoietic alterations caused by protein malnutrition are due to multifactorial alterations and, at least in part, the MSCs' contribution to hematological impairment.
Assuntos
Células da Medula Óssea/efeitos dos fármacos , Proteínas Alimentares/administração & dosagem , Hematopoese/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Deficiência de Proteína/metabolismo , Animais , Células da Medula Óssea/fisiologia , Técnicas de Cocultura , Meios de Cultivo Condicionados , Hematopoese/fisiologia , Leucócitos Mononucleares/fisiologia , Camundongos , Proteínas Proto-Oncogênicas c-kit/metabolismo , RNA/efeitos dos fármacos , RNA/genética , RNA/metabolismoRESUMO
Introducción: el seguimiento virológico de los pacientes con hepatitis C se realiza mediante la determinación cuantitativa del ácido ribonucleico viral por técnicas de biología molecular. Objetivos: evaluar el comportamiento virológico de los pacientes con hepatitis crónica C tratados con antivirales cubanos. Materiales y métodos: se realizó un estudio descriptivo, prospectivo en 45 pacientes con hepatitis crónica C atendidos en Consulta de Hepatología del Hospital Universitario Faustino Pérez, de Matanzas, en el período comprendido entre enero 2014 a diciembre 2017, tratados durante 48 semanas con PEG-heberon y ribavirina. De ellos se analizaron las características basales así como los diferentes tipos de respuesta al tratamiento según resultados virológicos. Resultados: predominaron los pacientes del sexo femenino, menores de 45 años, vírgenes de tratamiento y con cargas virales basales altas. Se alcanzó la respuesta virológica rápida en el 31,1%, la temprana total en el 19,4 %, al final del tratamiento en el 77,1% y la respuesta virológica sostenida en el 59,3%. Entre los respondedores predominaron los rápidos con respuesta virológica sostenida y entre los no respondedores, los nulos. Conclusiones: los estudios cuantitativos de ácido ribonucleico viral son esenciales para el seguimiento de los pacientes con hepatitis C ya que a través de sus determinaciones basales, durante el tratamiento y posterior a este, puede evaluarse la respuesta al tratamiento (AU).
Introduction: the virological follow-up of patients with hepatitis C is made through the quantitative determination of the viral ribonucleic acid using techniques of molecular biology. Objectives: to assess the virological behavior of patients with hepatitis C treated with Cuban antivirals. Materials and methods: a prospective, descriptive study was carried out in 45 patients with hepatitis C who attended the Consultation of Hepatology of the University Hospital "Faustino Pérez", of Matanzas, in the period from January 2014 to December 2017, treated with PEG-eberon and ribavirin for 48 weeks. Their basal characteristics were analyzed and also the different kinds of answer to the treatment according to the virological results. Results: female sex, patients aged less than 45 years old, non-treated before and with high viral loads. The fast virological answer was reached in 31 % of the patients, and the total early answer in 19.4 %; at the end of the treatment in 77.1 %, and the sustained virological answer in 59.3 % of the patients. Among the answering ones predominated the fast with sustained virological answer, and among the non-answering predominated the null ones. Conclusions: quantitative studies of viral ribonucleic acid are essential for the follow-up of patients with hepatitis C, because through their basal determinations, during and after the treatment, the answer to the treatment can be evaluated (AU).
Assuntos
Humanos , Masculino , Feminino , Antivirais/uso terapêutico , Hepatite C/virologia , Pacientes , RNA/efeitos dos fármacos , RNA/uso terapêutico , RNA/farmacologia , Resultado do TratamentoRESUMO
OBJECTIVE: The aim of the study was to evaluate the predictive value of clinical and molecular risk factors, including peripheral blood mononuclear cell (PBMC) mitochondrial DNA (mtDNA) and mitochondrial RNA (mtRNA), for the development of lactic acidosis (LA) and symptomatic hyperlactataemia (SHL). METHODS: In a substudy of a large multicentre, randomized trial of three antiretroviral regimens, all containing didanosine (ddI) and stavudine (d4T), in antiretroviralnaïve, HIV-1-infected patients, patients with LA/SHL ('cases') were compared with those without LA/SHL in a univariate analysis, with significant parameters analysed in a multivariate model. In a molecular substudy, PBMC mtDNA and mtRNA from cases and matched controls at baseline and time of event were examined. RESULTS: In 911 subjects followed for a median of 192 weeks, 24 cases were identified (14 SHL and 10 LA). In univariate analysis, cases were more likely to be female (P=0.05) and to have a high body mass index (BMI) (P=0.02). In multivariate analyses, only BMI remained an independent predictor of the development of LA/SHL (P=0.03). Between cases and controls there was no significant difference in mtDNA copy number at baseline (389 vs. 411 copies/cell, respectively; P=0.60) or at time of event (329 vs. 474 copies/cell, respectively; P=0.21), in the change in mtDNA copy number from baseline to event (-65 vs. +113 copies/cell, respectively; P=0.12), in mtRNA expression at baseline or time of event, or in the change in mtRNA expression from baseline to event. CONCLUSION: The development of LA/SHL was associated with increased BMI, but PBMC mtDNA and mtRNA did not predict LA/SHL. This demonstrates the ineffectiveness of routine measurement of PBMC mtDNA in patients on ddI and d4T as a means of predicting development of LA/SHL.
Assuntos
Acidose Láctica/etiologia , Índice de Massa Corporal , DNA Mitocondrial/metabolismo , Infecções por HIV/complicações , HIV-1 , Leucócitos Mononucleares/metabolismo , RNA/metabolismo , Acidose Láctica/induzido quimicamente , Acidose Láctica/epidemiologia , Acidose Láctica/genética , Adulto , Fármacos Anti-HIV/administração & dosagem , Fármacos Anti-HIV/efeitos adversos , Australásia/epidemiologia , DNA Mitocondrial/efeitos dos fármacos , DNA Viral/efeitos dos fármacos , DNA Viral/metabolismo , Didanosina/administração & dosagem , Didanosina/efeitos adversos , Europa (Continente)/epidemiologia , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/epidemiologia , Infecções por HIV/genética , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Estudos Multicêntricos como Assunto , Análise Multivariada , América do Norte/epidemiologia , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , RNA/efeitos dos fármacos , RNA Mitocondrial , RNA Viral/efeitos dos fármacos , RNA Viral/metabolismo , Ensaios Clínicos Controlados Aleatórios como Assunto , Fatores de Risco , Fatores Sexuais , América do Sul/epidemiologia , Estavudina/administração & dosagem , Estavudina/efeitos adversosRESUMO
OBJECTIVE: Parenteral diets are often administered to critically ill patients. To study one of the effects of commercially available parenteral lipid diets, rich in triacylglycerol esters of omega-6 polyunsaturated fatty acids or omega-9 monounsaturated fatty acids, on the immune system of such patients, we evaluated the cytotoxicity of oleic and linoleic acids on Raji cells that had been derived from human B-lymphocytes. METHODS: Cell death intensity and type were investigated by flow cytometry by quantitation of cell volume, granularity, DNA fragmentation, mitochondrial depolarization, and lipid accumulation. Fluorescence microscopy was used to determine chromatin condensation and type of cell death (acridine orange/ethidium bromide assay). Gene expression of BCL-XL, BCL-XS, C-MYC, and P53 was studied by reverse transcriptase polymerase chain reaction. RESULTS: Oleic acid was less toxic than linoleic acid to Raji cells. Both fatty acids promote apoptosis and necrosis of these cells. The mechanism of cell death induced by these fatty acids seemed to involve mitochondrial depolarization, lipid accumulation, and overexpression of C-MYC and P53. CONCLUSION: Oleic acid may offer a less harmful alternative to linoleic acid in parenteral diets with respect to patient B-lymphocyte-mediated immunologic activity.
Assuntos
Linfócitos B/efeitos dos fármacos , Ácido Linoleico/toxicidade , Ácido Oleico/toxicidade , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Células Cultivadas , Cromatina/efeitos dos fármacos , Relação Dose-Resposta a Droga , Citometria de Fluxo/métodos , Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas In Vitro , Microscopia de Fluorescência/métodos , RNA/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fatores de TempoRESUMO
3-Aminobenzamide (3AB) is an inhibitor of poly (ADP-ribose) polymerase (PARP), an enzyme implicated in the maintenance of genomic integrity, which is activated in response to radiation-induced DNA strand breaks. cDNA macroarray membranes containing 1536 clones were used to characterize the gene expression profiles displayed by mouse BALB/3T3 fibroblasts (A31 cell line) in response to ionizing irradiation alone or in combination with 3AB. A31 cells in exponential growth were pre-treated with 3AB 4mM 1h before gamma-irradiation (4Gy), remaining in culture during 6h until harvesting time. A31 cells treated with 3AB alone presented a down-regulation in genes involved in protein processing and cell cycle control, while an up-regulation of genes involved in apoptosis and related to DNA/RNA synthesis and repair was verified. A31 cells irradiated with 4Gy displayed 41 genes differentially expressed, being detected a down-regulation of genes involved in protein processing and apoptosis, and genes controlling the cell cycle. Concomitantly, another set of genes for protein processing and related to DNA/RNA synthesis and repair were found to be up-regulated. A positive or negative interaction effect between 3AB and radiation was verified for 29 known genes. While the combined treatment induced a synergistic effect on the expression of LCK proto-oncogene and several genes related to protein synthesis/processing, a negative interaction effect was found for the expression of genes related to cytoskeleton and extracellular matrix assembly (SATB1 and Anexin III), cell cycle control (tyrosine kinase), and genes participating in DNA/RNA synthesis and repair (RNA helicase, FLAP endonuclease-1, DNA-3 glycosylase methyladenine, splicing factor SC35 and Soh1). The present data open the possibility to investigate the direct participation of specific genes, or gene products acting in concert in the mechanism underlying the cell response to radiation-induced DNA damage under the influence of PARP inhibitor.
Assuntos
Células 3T3/efeitos da radiação , Benzamidas/farmacologia , Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica/efeitos da radiação , Ribonucleoproteínas , Células 3T3/efeitos dos fármacos , Células 3T3/fisiologia , Animais , Anexina A3/efeitos dos fármacos , Anexina A3/genética , Anexina A3/efeitos da radiação , Dano ao DNA/genética , Dano ao DNA/efeitos da radiação , Endodesoxirribonucleases/efeitos dos fármacos , Endodesoxirribonucleases/genética , Endodesoxirribonucleases/efeitos da radiação , Endonucleases Flap , Raios gama , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/efeitos dos fármacos , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/efeitos da radiação , Proteínas de Ligação à Região de Interação com a Matriz/efeitos dos fármacos , Proteínas de Ligação à Região de Interação com a Matriz/genética , Proteínas de Ligação à Região de Interação com a Matriz/efeitos da radiação , Camundongos , Proteínas Nucleares/efeitos dos fármacos , Proteínas Nucleares/genética , Proteínas Nucleares/efeitos da radiação , Análise de Sequência com Séries de Oligonucleotídeos , Inibidores de Poli(ADP-Ribose) Polimerases , Proto-Oncogene Mas , RNA/biossíntese , RNA/efeitos dos fármacos , RNA/efeitos da radiação , RNA Helicases/efeitos dos fármacos , RNA Helicases/genética , RNA Helicases/efeitos da radiação , Fatores de Processamento de Serina-ArgininaRESUMO
Xanthorrizol (1) and 4-(1',5'-dimethylhex-4'-enyl)-2-methylphenol (2) were identified as the principal antimicrobial components of a CH(2)Cl(2)-MeOH (1:1) extract derived from Iostephane heterophylla. Compound 2 is a new natural product, but has been synthesized. Both compounds exhibited low level activity (MICs of 16-32 microg/mL) against methicillin-resistant staphylococci and vancomycin-resistant enterococci. They were either inactive or poorly active against Gram-negative bacteria and yeast. Mechanistic studies performed in Escherichia coli imp suggested nonspecific inhibition of DNA, RNA, and protein synthesis by both of these compounds. Compound 1 was tested in an in vivo model; it did not provide protection to mice infected with Staphylococcus aureus.
Assuntos
Anti-Infecciosos/isolamento & purificação , Antifúngicos/isolamento & purificação , Asteraceae/química , Fenóis/isolamento & purificação , Plantas Medicinais/química , Animais , Antibacterianos , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Antifúngicos/química , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , DNA/biossíntese , DNA/efeitos dos fármacos , Modelos Animais de Doenças , Resistência Microbiana a Medicamentos , Enterococcus faecium/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Feminino , Espectroscopia de Ressonância Magnética , México , Camundongos , Camundongos Endogâmicos , Testes de Sensibilidade Microbiana , Fenóis/química , Fenóis/farmacologia , Raízes de Plantas/química , Biossíntese de Proteínas , Proteínas/efeitos dos fármacos , RNA/biossíntese , RNA/efeitos dos fármacos , Espectrofotometria Ultravioleta , Staphylococcus aureus/efeitos dos fármacos , Vancomicina/farmacologiaRESUMO
It is now well documented that changes in gene expression take place during cell isolation and culture. Here, we report the change in the expression of the mRNAs for alpha(1)-adrenoceptor subtypes, during dissociation of guinea pig liver cells with collagenase. Using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) assays, it was observed that during the isolation procedure, the mRNA for the alpha(1A)-adrenoceptor, normally expressed in whole liver, was degraded and the mRNA for alpha(1D) subtype, barely expressed in whole liver, increased in an actinomycin D-sensitive manner. When the isolation procedure was performed in the presence of cycloheximide, the mRNA for the alpha(1A)-adrenoceptor did not diminish and the induction of the alpha(1D)-adrenoceptor mRNA was even more evident. Our data indicate that cell isolation alters alpha(1)-adrenoceptor mRNA expression.
Assuntos
Fígado/metabolismo , RNA Mensageiro/metabolismo , Receptores Adrenérgicos alfa 1/genética , Animais , Cicloeximida/farmacologia , Dactinomicina/farmacologia , Regulação da Expressão Gênica , Cobaias , Fígado/citologia , Fígado/efeitos dos fármacos , Masculino , Inibidores da Síntese de Proteínas/farmacologia , RNA/efeitos dos fármacos , RNA/genética , RNA/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaAssuntos
Estradiol/farmacologia , Chumbo/efeitos adversos , Receptores de Estrogênio/efeitos dos fármacos , Útero/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Exposição Ambiental , Eosinófilos/efeitos dos fármacos , Feminino , Chumbo/farmacologia , Mitose/efeitos dos fármacos , RNA/biossíntese , RNA/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Útero/fisiologiaRESUMO
The ability of C8-substituted guanine (Gua) ribonucleosides to induce B cell proliferation has been well documented in the literature. These compounds are analogues of adducts formed from free radical attack on ribonucleosides and RNA. Here we examined the proliferative properties of two of these radical adducts, 8-methylguanosine (8-MeG) and 8-oxo-7,8-dihydroguanosine (8-OxoG) and compared them with those of the well studied B cell mitogen, 8-bromoguanosine (8-BrG). 8-MeG and 8-OxoG were synthesized in the considerable yields of 28, and 55%, respectively, and were characterized by UV, NMR and CG-MS. Their effects upon [3H]thymidine uptake into DNA by Swiss mouse splenocytes, mouse embryo 3T3 fibroblasts (A31) and mouse B16F10 melanoma were examined. Both guanosine (G) radical adducts were shown to increase [3H]thymidine uptake by mouse splenocytes but displayed selectivity in respect to continuous cell lines. 8-MeG acted upon 3T3 fibroblasts whereas 8-OxoG acted upon B16F10 melanoma. The non-physiological analogue 8-BrG acted upon all tested cells. Parallel experiments of cell counting, cytotoxicity,and cell sorting indicated that DNA synthesis induced by the C8-substituted G's reflected cell growth. It is proposed that the compounds act intracellularly because their proliferative effects were blocked in the presence of a nucleoside transport inhibitor but were not inhibited by an antagonist of the A2 purine receptor. The present results, taken together with data from the literature, suggest that in the case of 3T3 fibroblasts and mouse splenocytes the proliferative effects of the compounds are not dependent on metabolism through purine salvage pathways. In the case of melanoma, however, the compounds are likely to become part of the purine nucleoside pool. The demonstration that adducts produced by free radical attack on ribonucleosides and RNA are able to induce cell proliferation opens new perspectives for the understanding of free radical mediated carcinogenesis.
Assuntos
Guanosina/análogos & derivados , RNA/efeitos dos fármacos , Ribonucleosídeos/metabolismo , Células 3T3/citologia , Células 3T3/efeitos dos fármacos , Células 3T3/metabolismo , Adjuvantes Imunológicos/síntese química , Adjuvantes Imunológicos/farmacologia , Animais , Divisão Celular/efeitos dos fármacos , DNA/biossíntese , DNA de Neoplasias/biossíntese , Radicais Livres/metabolismo , Radicais Livres/farmacologia , Guanosina/síntese química , Guanosina/farmacologia , Masculino , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos BALB C , Mitógenos/síntese química , Mitógenos/farmacologia , RNA/metabolismo , Baço/citologia , Baço/efeitos dos fármacos , Baço/metabolismoRESUMO
Modern recombinant biotechnology has made possible the production of large amount of interferons and their use as immunotherapeutic agents. Most of the biological, physical and chemical characteristics of interferons has been established, including their classification, genetic structure, chemical composition and possible mechanisms of action. Interferons have been utilized in clinical studies with human and experimental animals against bacterial, mycotic, parasitic and viral infections. Success has been reported mainly when administered prophylactically against acute infections. Favorable results have been obtained, both prophylactic and therapeutically, in some chronic diseases and in those in which the microorganism has an intracellular phase during its life cycle. Moreover, a promising future has been suggested for the combined use of interferon with other antimicrobial drugs.