RESUMO
The presence of TNF-alpha mRNA and protein in circulating human blood monocytes isolated by continuous Percoll gradient fractionation was studied. The technique of RNA isolation from the blood samples was used to study TNF-alpha mRNA expression. It was shown that human blood monocytes of healthy donors contained no presynthesized pool of TNF-alpha mRNA as well as no TNF-alpha protein.
Assuntos
Leucócitos Mononucleares/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Northern Blotting , Células Cultivadas , Fracionamento Químico , Humanos , RNA Mensageiro/isolamento & purificaçãoRESUMO
It has been shown that patients with nonspecific aortoarteritis undergoing acute and subacute stages of the process demonstrate sensitization to collagen, types I and III. The activity of inflammation was discovered to affect the disease progression and involvement of new arterial areas. Marked suppression of leukocyte migration in response to collagen, types I and III, in patients with nonspecific aortoarteritis points to a high probability of further disease progression.
Assuntos
Aortite/etiologia , Arterite/etiologia , Adulto , Anticorpos/análise , Aorta Abdominal , Aorta Torácica , Aortite/imunologia , Arterite/imunologia , Tronco Braquiocefálico , Inibição de Migração Celular , Colágeno/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Artéria RenalRESUMO
The interaction of type III collagen (CIII) with washed human platelets was studied, using a CIII preparation from human placenta. CIII was labeled with 125I, and the monomeric and fibrillar forms of 125I-CIII (125I-CIIIm and 125I-CIIIf, respectively) were incubated with the platelets at room temperature. The platelet-associated and free labels were separated by centrifugation through 20% sucrose. The binding of 125I-CIIIf was unsaturable, linearly dependent on the label concentration and made up to 28 +/- 3% of the added protein. In comparison with CIIIf, the binding of 125I-CIIIm was minimal, i.e., only 0.9 +/- 0.2% of the added protein; so it did not significantly increase the background level (label sedimented through 20% sucrose in the absence of platelets). Although the level of 125I-CIIIm was very low, the binding was also unsaturable and linearly dependent on the concentration of the labeled protein. Platelet activation did not influence the level of CIIIf binding, nor did it stimulate the binding of CIIIm. The binding of 125I-CIIIf was not inhibited by the unlabeled CIIIm. The data obtained testify to the absence of high affinity platelet collagen receptors and support the hypothesis on multiple low affinity interactions between collagen fibrils and platelet surface. The binding of CIIIf to platelets was characterized by very fast kinetics; the level of binding reached a plateau within the range of 1 min and was similar in the presence of Ca2+/Mg2+ and EDTA.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Plaquetas/metabolismo , Colágeno/metabolismo , Plaquetas/efeitos dos fármacos , Ácido Edético/farmacologia , Epoprostenol/farmacologia , Humanos , Técnicas In Vitro , Radioisótopos do Iodo , Cinética , Ativação Plaquetária , Conformação Proteica , Trombina/farmacologiaRESUMO
Immunofluorescence was employed to study distribution of collagen type I, III, IV and V, and fibronectin in skin biopsy specimens from patients with Ehlers-Danlos syndrome (EDS) and cutis laxa. Abnormal distribution of collagen type V in the skin biopsies was found out in EDS type VII. A high level of dermal fibronectin occurred in EDS type III. Tissue fibronectin was absent in the skin extracellular matrix in EDS type X. Defective distribution of collagen, predominantly of type III and V and fibronectin absence were registered in the skin connective tissue in cutis laxa.
Assuntos
Colágeno/análise , Cútis Laxa/metabolismo , Síndrome de Ehlers-Danlos/metabolismo , Fibronectinas/análise , Pele/análise , Biópsia , Cútis Laxa/patologia , Síndrome de Ehlers-Danlos/patologia , Matriz Extracelular/análise , Imunofluorescência , Humanos , Pele/patologiaRESUMO
Enzymatic activity of cells, antigenic cellular markers and extracellular matrix of the hyperplastic intima of the aorta and carotid arteries was investigated in non-specific aorto-arteritis by immunomorphological and histochemical techniques. The cells of subendothelial layer of thickened arterial intima contained smooth muscle cell myosin, gave positive reactions to myosin ATP-ase and revealed high activity of thiamine pyrophosphatase. Fibronectin and type IV and V collagen were located in close proximity to these cells. The data obtained make it possible to consider these cells as modified smooth muscle cells. Type III collagen was the prevalent type of extracellular matrix of the thickened intima. A great number of blood vessels of the capillary and precapillary types have been found to penetrate into the intima from the adventitia. A possible role of pericytes surrounding newly formed capillaries as the precursors of subendothelial cell population in the hyperplastic intima is discussed.
Assuntos
Aortite/enzimologia , Artérias/enzimologia , Arterite/enzimologia , Adenosina Trifosfatases/metabolismo , Aorta/enzimologia , Artérias Carótidas/enzimologia , Imunofluorescência , Histocitoquímica , Humanos , Tiamina Pirofosfatase/metabolismoRESUMO
Fibronectin and collagen I, III, IV, V distribution in preexisting and newly formed connective tissue of fibroadenomas and leaf-like human breast tumour was studied by the immunofluorescent method. Extracellular matrix of a newly formed connective tissue was characterized by the predominance of collagen type III, the presence of collagen type IV, a high content of fibronectin in a leaf-like tumour. Such characteristics of the extracellular matrix structure in benign tumours are interpreted as a manifestation of desmoplasia.
Assuntos
Adenofibroma/metabolismo , Neoplasias da Mama/metabolismo , Colágeno/metabolismo , Fibronectinas/metabolismo , Feminino , Imunofluorescência , HumanosRESUMO
Fibronectin and collagen, types I, III, IV and V, in the granulation tissue replacing the myocardial infarction, in the focal and diffuse cardiosclerosis were studied by means of the immunofluorescent method. The extracellular matrix in the granulation tissue contained fibronectin and collagen of the above types. Intracellular fibronectin was also found in the necrotized cardiomyocytes. Fibronectin and collagen of type IV were not detected in the postinfarction scars. The extracellular matrix consisted mainly of collagen, types III and V. Fibronectin and collagen, types I, III and V, are observed in the connective tissue in diffuse cardiosclerosis in the presence of the coronary arteries stenosis. No prevalence of the collagen of any type was found.
Assuntos
Colágeno/imunologia , Fibronectinas/imunologia , Miocárdio/imunologia , Idoso , Especificidade de Anticorpos , Doença das Coronárias/imunologia , Doença das Coronárias/patologia , Imunofluorescência , Tecido de Granulação/imunologia , Tecido de Granulação/patologia , Humanos , Pessoa de Meia-Idade , Infarto do Miocárdio/imunologia , Infarto do Miocárdio/patologia , Miocárdio/patologia , Recidiva , Esclerose , Fatores de TempoRESUMO
An immunofluorescent study demonstrated the localization of fibronectin and collagen of types 1, 3, 4 and 5 in normal arterial intima and atherosclerotic patches. In the atherosclerotic patches, fibronectin is mostly grouped among cells of smooth-muscle origin together with collagen of type 4, while the fibrous tissue of the patches contains little fibronectin. It is assumed that changed extracellular matrix composition reflects the development of atherosclerotic patches, and fibronectin can be regarded as a marker of early stages of atherosclerosis.
Assuntos
Arteriosclerose/metabolismo , Fibronectinas/análise , Adulto , Idoso , Colágeno/análise , Imunofluorescência , Humanos , Pessoa de Meia-IdadeRESUMO
By means of immunoperoxidase and immunoferritin techniques collagen of the I, III, IV and V types has been revealed in cryostat sections of the popliteal artery and in the musculus quadriceps of the femur. Areas of the vascular wall without any macroscopical signs of lesions have been investigated. They have been obtained from amputated extremities of young persons (17-22 years old), and muscle pieces have been taken during operations performed in the knee joint. After certain immunocytochemical procedures the cryostat slices are embedded in mixture of epon 812 and araldit, non-contrasted ultrathin slices are examined in the electron microscope JEM 100CX. Collagen of the I and III types is revealed in fibrills 20-80 nm thick either with or without cross striation, as well as in microfibrills. Collagen of the III type in the intercellular substance of the arterial wall occurs in nonfibrillar form. Collagen of the IV type is revealed in basal membranes of the smooth muscle cells of the arterial wall, of the muscle fibers and of endothelium of blood capillaries of the skeletal muscle. Collagen of the V type is found as accumulations having various size and form; they localize in many places of the intercellular substance of the arterial wall. A tight contact is revealed between the formations including collagen of the V type with drops of elastin and elastic fibers. A suggestion is made that collagen of the V type participates in formation of elastic fibers.
Assuntos
Artérias/metabolismo , Colágeno/metabolismo , Músculos/metabolismo , Adolescente , Adulto , Artérias/ultraestrutura , Humanos , Técnicas Imunoenzimáticas , Microscopia Eletrônica , Pessoa de Meia-Idade , Músculos/ultraestrutura , Artéria Poplítea/metabolismoRESUMO
Results of an immuno-morphological analysis of the I, II, IV and V collagen types in 7 synovial sarcomas of various location are presented. A fibrillar matrix of synovial sarcomas is represented by a collagen of type I and collagen of type V which is co-distributed with the former. Collagen of type III is found in a smaller quantity and in areas of fibrosarcoma-like structure being included in the largest fibrillar structures. Collagen of type IV is presented in all structural areas of tumours. The greatest amount of the IV type collagen is observed in areas of the epithelioid and hemangiopericytoma-like structure, less in areas of fibrosarcoma-like structure. These results may be used in differential diagnostics. In the walls of fissure-like spaces, only the IV type collagen was found as distinct from vascular tumours. On the basis of the data obtained and analysis of the literature a mesodermal origin of synovial sarcoma is suggested.
Assuntos
Colágeno/metabolismo , Sarcoma Sinovial/metabolismo , Adolescente , Adulto , Criança , Diagnóstico Diferencial , Feminino , Imunofluorescência , Histocitoquímica , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Sarcoma Sinovial/diagnóstico , Sarcoma Sinovial/patologiaRESUMO
The results of immunomorphological analysis of 4 collagen types in desmoids of different localization and behaviour are presented. Biopsies of 12 nodes (6 cases of abdominal and 6 cases of extraabdominal localization) were studied. Two groups of desmoids were defined depending on the character of different collagen types distribution. Desmoids of the first group were characterized by a high content of type I and III collagen, both types being found at approximately equal proportions. Clinically predominant abdominal localization and primary character of processes were typical for this group. Collagen of type III clearly predominated in the desmoids of the second group; extraabdominal localization, recurrences, and a young age of patients were characteristic features of this group.
Assuntos
Neoplasias Abdominais/patologia , Colágeno/imunologia , Fibroma/patologia , Neoplasias Abdominais/imunologia , Adulto , Nádegas , Feminino , Fibroma/imunologia , Imunofluorescência , Humanos , Perna (Membro) , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/imunologia , Recidiva Local de Neoplasia/patologia , OmbroRESUMO
Fibrosis is the most important manifestation of human vascular atherosclerosis. Primary culture of human aortic cells makes it possible to study some manifestations of fibrosis in vitro. Indirect immunofluorescence has shown that primary culture cells contain intracellular antigens to all the antibodies used but that only type I collagen forms extracellular matrix in culture. Modulation of the cultivation conditions does not lead to the appearance on the cell surface of other type collagens. It is assumed that in primary culture, human aortic cells synthesize the same collagen types as those synthesized in vitro. This circumstance might enable using such a culture as model for studying the mechanisms of fibrosis during atherosclerosis.
Assuntos
Aorta/análise , Colágeno/análise , Imunofluorescência , Humanos , Técnicas In VitroRESUMO
The distribution of activity of the elongation factors EF-1 and EF-2 among the components of rabbit reticulocyte lysate separated by sucrose density gradient centrifugation was studied. At low ionic strength (0.01 M KCl) about 30% of the EF-1 activity was found in polyribosomes. At moderate ionic strength (0.1 M KCl) the EF-1 activity was absent in the polyribosomes. An addition of RNA excess to the lysate prior to centrifugation at low ionic strength resulted in elimination of the EF-1 activity from the polyribosomes. This indicates that EF-1 is reversibly bound to the polyribosomes and that EF-1 may be retained on them due to interaction with RNA of polysomes mediated by its RNA-binding site. After dissociation of polyribosomes containing EF-1 in the presence of EDTA and subsequent fractionation of the dissociation products at low ionic strength (0.01 M KCl) the EF-1 activity was revealed in the ribosomal subparticles (predominantly in 60S). At 0.1 M KCl EF-1 mainly sedimented in the zone of distribution of polyribosomal informosomes. The elongation factor EF-2 was not revealed in polyribosomes during lysate centrifugation even at low ionic strength which corresponds to its lower affinity for RNA.