RESUMO
The time of average polypeptide chain synthesis (tc), distribution of synthesized polypeptides according to their molecular masses, the ratio of translating and nontranslating ribosomes and polyribosomes of different size have been analyzed for diploid and aneuploid strains of fibroblasts. The magnitude of tc as well as the size of polypeptide chains synthesized were found to be similar for both kinds of fibroblasts. The relative cellular content of the translating ribosomes has been shown to decrease during the transition of cells from both strains to the stationary growth phase. The relative content of heavy polyribosomes is lower in aneuploid cells as compared with that in diploid cells. The process of translation in aneuploid fibroblasts is concluded to have no essential deviations from normal.
Assuntos
Aneuploidia , Diploide , Biossíntese de Proteínas , Células Cultivadas , Embrião de Mamíferos , Fibroblastos/metabolismo , Fibroblastos/ultraestrutura , Humanos , Biossíntese Peptídica , Polirribossomos/análiseRESUMO
The degradation rate of long-lived and short-lived proteins was determined in diploid fibroblasts and fibroblasts with trisomy 7 derived from human embryos. Two fractions of proteins were detected in the exponentially growing diploid fibroblasts with half-lives (T 1/2) 37 and 19 hours. The rate of protein degradation increases in diploid fibroblasts as they approach confluence and protein fractions with T 1/2 30, 18 and 12 hours appear. The rate of protein degradation in trisomic fibroblasts does not change for the long-lived and short-lived proteins and is the same in both exponential (T 1/2 31 and 14 hours) and stationary phase (T 1/2 33 and 17 hours). The relative amount of the short-lived proteins in trisomic fibroblasts in the stationary phase decreased as compared with the one in diploid fibroblasts. It is apparent that a mechanism of regulation of protein catabolism in trisomic fibroblasts is impaired.
Assuntos
Diploide , Proteínas/metabolismo , Trissomia , Células Cultivadas , Embrião de Mamíferos , Fibroblastos/metabolismo , Fibroblastos/ultraestrutura , Humanos , Hidrólise , CinéticaRESUMO
The electrophoretical analysis and CNBr-peptide mapping of the collagens, isolated from the costal cartilage of 30 patients with non-classified and syndromal forms of pex excavatum (funnel chest) (27 patients) and pex carinatum (3 patients) was carried out. In case of one patient with the nonclassified form of funnel chest the electrophoretical mobility of CB 9.7-peptide was found to be decreased. The electrophoretical mobilities of other peptides are not markedly changed. The data obtained allow one to suggest the mutation causing the defect in the region about 160 amino acid residues distant from the C-end of alpha 1 (II) chain of type II collagen of the patient.
Assuntos
Colágeno/análise , Tórax em Funil/genética , Adolescente , Sequência de Aminoácidos , Criança , Pré-Escolar , Colágeno/genética , Brometo de Cianogênio , Tórax em Funil/metabolismo , Humanos , MutaçãoRESUMO
Imbalance between the synthesis and degradation of collagen is a common sign of tissues of hypertrophic and cheloid scars. The fact that the collagen of abnormal scars of the both types have a polyfunctional linkage with a structure of pyridinol and high amounts of a polyfunctional linkage with a structure of pyrrole (Ehlich's chromogen) in the collagen of cheloid scars support the assumption that the collagen synthesis-degradation imbalance may be due to the enhanced resistance of structurally unchanged collagen to collagenolytic enzymes. The comparative studies of the ratio of type 1 collagen homotrimers to heterotrimers, that of type 1 collagen dimers to monomers, and that of types 1 to type 3 collagen in the pepsin extracts of abnormal scars of various age and patients' intact skin suggest that there are certain differences in the magnitude and scar age-related changes in the expression of collagen genes and posttranslational collagen modifications in the tissue of hypertrophic and cheloid scars. It is suggested that age-specific time course of collagen metabolic and structural changes in the abnormal scars may result from their alterations of the population composition of fibroblasts with a different phenotype.
Assuntos
Cicatriz Hipertrófica/metabolismo , Colágeno/metabolismo , Queloide/metabolismo , Cicatriz Hipertrófica/etiologia , Fibroblastos/metabolismo , Humanos , Queloide/etiologia , Cicatrização/fisiologiaRESUMO
The publications on the structural and functional features of the main molecular shaperon of li-chain were updated. In the 1990s the determination of molecular shaperon as a group of mutually unrelated protein molecules taking part in the assembly of other polypeptides was worked out. In humans the main isoform of li-chain is the protein with a mol. wt. of 31-33 kD. A great variety of functions of this shaperon is linked with definite amino acid sequences. In particular, the molecular shaperon of the molecules of the main histocompatibility complex-II has functions connected with the presentation of antigen and the differentiation of B lymphocytes.
Assuntos
Antígenos de Diferenciação de Linfócitos B/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Chaperonas Moleculares/imunologia , Sequência de Aminoácidos , Animais , Apresentação de Antígeno , Antígenos de Diferenciação de Linfócitos B/química , Linfócitos B/imunologia , Diferenciação Celular , Membrana Celular/química , Citoplasma/química , Antígenos de Histocompatibilidade Classe II/química , Humanos , Chaperonas Moleculares/química , Dados de Sequência Molecular , Peso Molecular , Isoformas de Proteínas/química , Isoformas de Proteínas/imunologiaRESUMO
Publications on C-lectin receptors, multifunctional surface molecules of antigen presenting cells are updated. The study of C-lectin receptors is stipulated by their role in antimicrobial protection. The importance of C-lectin receptors of dendrite cells in the system of the recognition of microbial carbohydrate as well as their properties of the targeted supply of tumor cell and other antigens to dendrite cells with the purpose of the formation of immunity against faintly immunogenic antigens is considered. Suggestion is made on the potential of their use as experimental preparations for immunotherapy.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Cálcio , Células Dendríticas/imunologia , Lectinas Tipo C/fisiologia , Animais , Apresentação de Antígeno , Células Apresentadoras de Antígenos/metabolismo , Antígenos de Neoplasias/imunologia , Antígenos de Neoplasias/metabolismo , Diferenciação Celular , Movimento Celular , Células Dendríticas/metabolismo , Humanos , Imunoterapia , Lectinas Tipo C/uso terapêutico , Polissacarídeos Bacterianos/imunologia , Polissacarídeos Bacterianos/metabolismo , Receptores Imunológicos/fisiologia , Receptores Imunológicos/uso terapêuticoRESUMO
High-titer antidiphtheria antitoxic rabbit serum has been obtained, and on the basis of this serum a coagglutinating diagnosticum has been developed. The sensitivity of the test has been found to depend on the content of antitoxic antibodies in the serum and on its purity. Diagnostica prepared from native serum containing 500 I. U./ml (a titer of 1:51, 200 in the passive hemagglutination test) permit the detection of 0.02-0.03 Lf/ml of diphtheria toxin. A decrease in antibody titer to 5-25 I. U./ml leads to a drop in sensitivity to 0.2-2 Lf/ml. The use of LgG fraction and pure antibodies increases the sensitivity of the test to 0.002-0.003 Lf/ml. The possibility of detecting toxin in Corynebacterium diphtheriae strains is shown.
Assuntos
Testes de Aglutinação/métodos , Toxina Diftérica/análise , Animais , Anticorpos Antibacterianos/análise , Corynebacterium diphtheriae/imunologia , Toxina Diftérica/imunologia , Toxoide Diftérico/imunologia , Soros Imunes/imunologia , Soros Imunes/isolamento & purificação , Imunização , Imunoglobulina G/imunologia , Coelhos , Proteína Estafilocócica A/imunologiaRESUMO
The immunogenic potency, toxicity, homologous and heterologous protective activity of lipopolysaccharide preparations obtained from serogroup A N. meningitidis (LPS A) were studied in animal experiments. These preparations were shown to possess very high protective activity. The alkaline treatment of native LPS A decreased the toxicity of the preparation almost 20 times and did not affect its immunogenic potency. Detoxified LPS A was capable of protecting mice from fatal meningococcemia resulting from infection with N. meningitidis strains, serogroups A, B and C; the adsorption of the preparation on aluminium hydroxide did not affect its protective activity. In view of the properties of detoxified LPS A revealed in this investigation, it may be regarded as a possible vaccinal preparation.
Assuntos
Vacinas Bacterianas/imunologia , Lipopolissacarídeos/imunologia , Neisseria meningitidis/imunologia , Vacinas Atenuadas/imunologia , Adsorção , Hidróxido de Alumínio , Animais , Vacinas Bacterianas/uso terapêutico , Vacinas Bacterianas/toxicidade , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Dose Letal Mediana , Lipopolissacarídeos/uso terapêutico , Lipopolissacarídeos/toxicidade , Infecções Meningocócicas/prevenção & controle , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Sepse/prevenção & controle , Vacinas Atenuadas/uso terapêutico , Vacinas Atenuadas/toxicidadeRESUMO
The protective activity of the sera of mice immunized with the preparations of native and detoxified N. meningitidis lipopolysaccharide (LPS), group A, as well as with monoclonal antibodies to N. meningitidis antigens, groups A and B, was studied on the mucin model of meningococcal infection. The study showed that the maximum level of anti-LPS antibodies in mice was observed on day 7 after the injection of LPS. Immune sera obtained from mice were capable of protecting the animals from fetal meningococcemia induced by N. meningitidis strains of homologous and heterologous groups. As shown by the results of this study, the alkaline treatment of N. meningitidis native LPS did not decrease the protective properties of antibodies. The monoclonal antibodies under study were found to possess high preventive activity in mice challenged with N. meningitidis, groups A and B. Anti-LPS monoclonal antibodies showed greater protective activity than antipolysaccharide monoclonal antibodies.
Assuntos
Anticorpos Antibacterianos/uso terapêutico , Anticorpos Monoclonais/uso terapêutico , Lipopolissacarídeos/imunologia , Neisseria meningitidis/imunologia , Animais , Avaliação Pré-Clínica de Medicamentos , Imunização , Imunização Passiva , Meningite Meningocócica/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Sepse/prevenção & controleRESUMO
The tumorigenic capacity of mouse B-cell hybridomas in both cloned and primary cultures was studied. The cells were selected for inoculation from 24-well plates and introduced into the spleen of syngeneic mice. The cells took in 50% of the animals. The cells of hybridoma tumors induced as the result of intrasplenic inoculation, when subcultured in the second passage following the standard scheme, i.e. inoculated intraperitoneally in a dose of 1 X 10(7) cells into mice previously treated with vaseline oil or pristane, produced tumors in 100% of the animals.
Assuntos
Linfócitos B/citologia , Hibridomas/citologia , Baço/citologia , Animais , Anticorpos Monoclonais/análise , Técnicas Citológicas , Relação Dose-Resposta Imunológica , Hibridomas/imunologia , Hibridomas/transplante , Imunização/métodos , Camundongos , Camundongos Endogâmicos A , Camundongos Endogâmicos BALB C , Mieloma Múltiplo/patologia , Transplante de Neoplasias , Neoplasias Experimentais/etiologia , Neoplasias Experimentais/imunologiaRESUMO
Synthesis of nonribosomal RNA was increased 2-2.5-fold in nuclei of rat liver cells at early steps of regeneration /2 days after partial hepatectomy/under conditions of protein synthesis inhibition by means of cycloheximide within 3 hrs. Amount of mRNA, transferred from nuclei into cytoplasm, was also 2-fold higher under these conditions as compared with control cells. The mRNA, transferred from nuclei into cytoplasm under the conditions of cycloheximide treatment of liver cells at early steps of regeneration, did penetrate the polyribosomes although the protein synthesis did not occur on the polyribosomes formed. At the later steps of regeneration /within 5 days after partial hepatectomy/ under the conditions of protein synthesis inhibition by cycloheximide within 3 hrs, formation of nonribosomal RNA was decreased by 40% in liver cell nuclei; amount of mRNA, transferred into cytoplasm, was also decreased by 40%. When the cycloheximide treatment was carried out during 1 hr, synthesis of nonribosomal RNA was increased 1.5-fold in liver cell nuclei at the later steps of regeneration although the amount of mRNA, transferred from nuclei into cytoplasm, was decreased by 30% as compared with the control cells. The data obtained suggest that during rat liver tissue regeneration a change occurred in the molecular mechanisms linking translation, biosynthesis and nuclear-cytoplasmic transport of mRNA of "tumoral" type/at early steps of regeneration/ or "normal type"/ at later steps of regeneration.
Assuntos
Cicloeximida/farmacologia , Regeneração Hepática , Fígado/metabolismo , Biossíntese de Proteínas , RNA Mensageiro/metabolismo , Animais , Transporte Biológico , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Hepatectomia , Polirribossomos/metabolismo , Ratos , Ratos EndogâmicosRESUMO
The ratio of collagens I/III was studied in human diploid, trisomic and triploid fibroblast strains. The following material was used in experiments: 5 strains obtained from human spontaneous abortuses (trisomy 7, trisomy 9, triploidy), 6 diploid embryonic strains, 4 strains from patients with Down syndrome, 3 strains from healthy children, 4 strains from healthy adult donors and 3 strains from very old persons (81-107 years old). Various types of collagen were separated by means of electrophoresis. Distinct ontogenetic alterations were not observed in content of collagen III in diploid and aneuploid human cultivated fibroblasts. However, considerable alterations in the ratio of collagens I/III correlated with genetic pathologies only in the patients examined.
Assuntos
Aberrações Cromossômicas , Colágeno/metabolismo , Diploide , Células Cultivadas , Fibroblastos/metabolismo , Fibroblastos/ultraestrutura , HumanosRESUMO
Effect of cAMP on the activity of polynucleotide phosphorylase (PNPase) was studied in polyribosome fraction of rat liver tissue. Intraperitonel administration of cAMP or of theophilline into rats distinctly decreased the PNPase activity in the polyribosome fraction. The cAMP (1-10(-4) M) inhibited the enzymatic activity only by 8% in polyribosome fraction in vitro, as it was estimated by the reaction of phosphorolysis of endogenous RNA and polyA added. Any attempts were proved to be uncucessful to reveal cAMP, ATP-dependent proteinkinase in rat liver, responsible for the decrease in the PNPase activity in the polyribosome fraction. The cAMP inhibited the increase in the PNPase activity, coupled with protein biosynthesis in polyribosomes. Moreover, cAMP caused a decrease in the PNPase activity in reaction of polyA phosphorolysis and did not affect the rate of endogenous RNA phosphorolysis in polyribosome fraction, isolated from postmitochondrial fraction after incubation for 15 min at 30 degrees. The 3',5'-cyclo AMP (2-10(-6)-2-10(-4) M) stimulated incorporation of 14C-leucine into acid-insoluble material, when postmitochondrial fraction was incubated under the same conditions. The data obtained suggest that cAMP either inhibits specifically the PNPase synthesis or represses the coupled with protein biosynthesis formation of active "heavy" type of PNPase from less active "light" type.
Assuntos
AMP Cíclico/farmacologia , Fígado/enzimologia , Polirribonucleotídeo Nucleotidiltransferase/metabolismo , Polirribossomos/enzimologia , Animais , Leucina/metabolismo , Fígado/ultraestrutura , Masculino , Ratos , Teofilina/farmacologiaRESUMO
Relative synthesis of collagen was studied in skin fibroblasts of children with funnel chest and of corresponding age children (control) in presence of ascorbic acid. In presence of ascorbic acid the rate of collagen synthesis was 2-2.4-fold lower that in corresponding controls both in proliferating and stationary cultures. At the same time, relative synthesis of collagen was quite similar both in the patient and control fibroblasts in presence of ascorbic acid. Estimation of the 14C-Hypro/14C-Pro ratio in collagens isolated from cultural media showed that there was no difference in the hydroxylation rate of collagens in control and patient fibroblasts. These data suggest that funnel chest is one of the forms of systemic connective tissue diseases, which impaired mainly the cartilages.
Assuntos
Ácido Ascórbico/farmacologia , Colágeno/biossíntese , Tórax em Funil/metabolismo , Pele/metabolismo , Células Cultivadas , Criança , Síndrome de Ehlers-Danlos/complicações , Síndrome de Ehlers-Danlos/metabolismo , Fibroblastos/metabolismo , Humanos , Síndrome de Marfan/complicações , Síndrome de Marfan/metabolismoRESUMO
Age-dependent alterations in excretion of hydroxyproline were studied in 62 healthy persons (3-78 years old) by means of analysis of the morning portion of urine after gel filtration on Sephadex G-10. With ageing the following patterns were decreased: content of total hydroxyproline (r = -0.78; P less than 0.001), the ratio hydroxyproline/creatinine (r = -0.80, P less than 0.001) and content of hydroxyproline in peptide fraction with MW greater than 700-1000 (r = -0.50; P less than 0.001). The reverse correlation was found between the contents of total and free hydroxyproline (r = -0.45; P less than 0.001) but total content of hydroxyproline correlated directly with its content in the peptide fraction (r = 0.92; P less than 0.001). At the same time, reverse correlation was observed between the contents of free hydroxyproline and of the peptide-bound amino acid (r = -0.58; P less than 0.001). Approximate ranges of normal excretion of hydroxyproline with urine were calculated in various age groups.
Assuntos
Hidroxiprolina/urina , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de ReferênciaRESUMO
In a group of women, which had children with hereditary isolated and syndromal chest deformations, acute respiratory diseases, tonsillitis or influenza occurred during pregnancy in 42.9 +/- 6.6% and 53.8 +/- 13.8% of cases, respectively, whereas in the control group only 17.8 +/- +/- 7.2% of the women were impaired with identical infectious diseases during pregnancy (P less than 0.02 and P less than 0.05, respectively). Excretion of hydroxyproline was distinctly dissimilar in 22 children with isolated and in 13 children with syndromal chest deformations depending on presence or absence of the above-mentioned infections during pregnancy. Under conditions of these infections the higher level of total hydroxyproline excretion was noted as well as relatively lower content of bound hydroxyproline was detected in those peptides, which appear to be responsible for the content of newly synthesized collagen. The data obtained suggest that acute respiratory diseases, influenza, chronic and acute tonsillitis may impair collagen metabolism in children with hereditary chest deformations as well as that these infectious diseases occurred during pregnancy may increase the risk of the pathology development in the children.
Assuntos
Tórax em Funil/etiologia , Hidroxiprolina/urina , Complicações Infecciosas na Gravidez , Adolescente , Criança , Pré-Escolar , Feminino , Tórax em Funil/urina , Humanos , Influenza Humana , Gravidez , Infecções RespiratóriasRESUMO
Content of collagen and relative content of unextractable collagen were increased by 35% and 50%, respectively, while the content of immobilized water was decreased in costal cartilage of children with the isolated form of funnel chest or with the deformation accompanied by Ehlers-Danlos syndrome. These anomalies may be considered as indicators of the cartilage premature ageing. In skin of the children with isolated form of funnel chest the ratio of collagen extracted with acetic acid was increased 2-fold.
Assuntos
Cartilagem/análise , Colágeno/isolamento & purificação , Tórax em Funil/metabolismo , Costelas/análise , Pele/análise , Criança , Pré-Escolar , Humanos , Solubilidade , Ácidos Urônicos/análiseRESUMO
Relationship between the state of natural cytotoxicity and collagen catabolism were studied in patients with focal sclerodermia. In the impairment activity of natural killer cells (NK cells) was markedly altered depending on such clinical parameters as duration, severity, activity and distribution of the disease as well as it was related to definite transformations in catabolism of collagen proteins. Low rate of collagen production and intensive collagenolytic processes correlated with the high level of hydrolytic activity of NK cells at the early steps of systemic morphea guttata accompanied by highly progressive course of the disease. Under conditions of subacute forms of focal morphea guttata activity of NK cells was distinctly decreased down to 50% of that of normal state and was accompanied by alterations in the spectrum of collagenolytic enzymes as well as by considerable variations in collagen production and in posttranslational modification of collagen fibrils. During the chronic step of sclerodermia activity of NK cells was normalized and correlated with the lowered rate of collagen production as well as with the subsequent alterations in collagen proteins catabolism. Alterations in the level of natural cytotoxicity appear to correlate with the state of collagen metabolism in focal sclerodermia.
Assuntos
Colágeno/metabolismo , Citotoxicidade Imunológica , Células Matadoras Naturais/imunologia , Esclerodermia Localizada/imunologia , Adulto , Idoso , Feminino , Humanos , Hidroxiprolina/urina , Masculino , Pessoa de Meia-Idade , Peso Molecular , Esclerodermia Localizada/metabolismoRESUMO
Catabolism of collagen was studied in 25 children with keloid and hypertrophic scars by means of evaluation of total and polypeptide-bound hydroxyproline excreted with urine. Formation of postburn pathological scars led to an increase in excretion of collagen degradation products, the rate of the increase depended on area and age of scar. In hypertrophic cicatrization rate of collagen metabolites excretion correlated with the scar age reversely, demonstrating the tendency to decrease of the scar collagen metabolites with its ageing. Excretion of collagen metabolites with urine in patients with keloid scars depended on area of the scar and only slightly on its age. Besides, in patients with keloids rather high, statistically distinct correlation (r = 0.855) was found between the ratio macrophages/neutrophils under studies using "skin window" as well as between the ratio of hydroxyproline content in the polypeptide fractions with molecular mass above 1,500 and in the fraction containing oligopeptides and free hydroxyproline. This correlation was not found in patients with hypertrophic scars. Interaction of macrophages, neutrophils and, probably, eosinophiles appears to have a definite importance in regulation of collagen degradation, impairment of which may be substantial for scars formation.
Assuntos
Queimaduras/urina , Cicatriz/urina , Hidroxiprolina/urina , Queloide/urina , Queimaduras/patologia , Criança , Cicatriz/patologia , Colágeno/metabolismo , Humanos , Hipertrofia , Queloide/patologia , Técnica de Janela CutâneaRESUMO
Collagen production was studied in human embryonal fibroblasts. Synthesis of collagen in the cells with trisomy 7 was decreased 2-fold as compared with diploid cells in spite of 50% increase of the collagen genes localized on the chromosome 7. Diploid and aneuploid cells synthesized collagens of the I and III types at similar weight ratios. At the same time, an increase in synthesis of type I collagen trimer was found in culture of cells with anomalous caryotype. A decrease of collagen production in cells with anomalous caryotype appears to be one of the reasons responsible for spontaneous abortion.