Monoclonal antibodies against chordin. Use in structural and immunohistochemical studies.

Eight MAbs have been developed against chordin and designated as At2-At9. It is shown that all antibodies are directed against identical, spatially overlapping or closely positioned epitopes of chordin. The chordin molecule has repetitive sites wherein epitopes for the eight MAbs are located. This site lies within a proteinase-resistant fragment of chordin, presumably a glycopeptide, of molecular mass between 2 and 10 kDa. Fluorescence staining of cryostat sections from stellate sturgeon with the use of At5 (indirect Coons' method) has revealed a positive reaction with notochord cells and sheath and with the spinal cord. No significant reaction with cartilage, muscle and kidney was detected.

Relative distribution of fibronectin and type I, III, IV, V collagens in normal and atherosclerotic intima of human arteries.

Spatial distribution of fibronectin and type I, III, IV and V collagen has been investigated in normal arterial intima, fatty streaks, and atherosclerotic plaques by indirect immunofluorescence on transverse sections. Two distinct types of extracellular matrix were revealed in atherosclerotic lesions. The fibrous plaques consisted mostly of interstitial collagen types I and III, contained moderate amounts of type V and none of type IV collagen or fibronectin. In the extracellular matrix of the fatty streaks and in some areas of the fibrous plaques containing large amounts of subendothelial cells, some interstitial collagen was revealed, an increased amount of type IV, some type V collagen and a lot of fibronectin. Similarities of the extracellular matrix in atherosclerotic lesions and granulation tissues are discussed.

Confocal and conventional immunofluorescent and immunogold electron microscopic localization of collagen types III and IV in human placenta.

Confocal and conventional indirect immunofluorescence and immunogold electron microscopic methods were applied to examine the distribution of extracellular matrix constituents (collagens types III and IV) in the villi of immature and term human placentae. The immunofluorescence study revealed that collagen type III is more distinct in the villous stroma of term placenta as compared with that of the first trimester. Collagen type IV was detected mainly in endothelial and epithelial basement membranes and interestingly also to a certain extent in the stroma. Results obtained using immunoelectron microscopy support the proposal that collagen types III and IV are characteristic of stromal and basement membranes, respectively. Stromal collagen type IV is apparently localized in association with the interstitial types of collagen (I and III), in the villous stroma of term placenta.

Participation of collagen types I, III, IV, V, and fibronectin in the formation of villi fibrosis in human term placenta.

The indirect immunofluorescence method was used to study the human term placenta in pathological pregnancy for the distribution of collagen types I, III, IV, V, and fibronectin in fibrosis stromatis villi. All collagen types and fibronectin were shown to participate in fibrosis villorum formation. Fibronectin was also detected in the fibrinoid that surrounded villi at stroma. The presence of free cytotrophoblast cells in the fibrinoid was accompanied by a noticeable increase in fibronectin fluorescence. A significant amount of collagen types IV and V and a less amount of collagen types I and III were identified.

[Immunofluorescence study of the extracellular matrix of the human placenta]. / Immunofluorestsentnoe izuchenie éktratselliuliarnogo matriksa platsenty cheloveka.

Distribution of collagen types I, III, IV, V and fibronectin in human placental villi has been studied by indirect immunofluorescence. During 9-12 weeks of pregnancy the extracellular matrix of villi represents a network of filaments organized in bundles and aggregates that contain collagen types I and III and finer filaments of collagen types IV and V. Collagen type IV is regularly detected in basal membrane of capillaries and particularly in villous epithelium, collagen type V and fibronectin are occasionally detected there. Marked immunofluorescent reaction on collagen types IV and V and fibronectin, and weak reaction on collagen type III is observed in cellular islets around cytotrophoblasts. In the fetus born in term placental villi have uniform immunofluorescence in thick basal membranes of fetal capillaries and of chorionic epithelium. The immunofluorescent reaction specific for all collagen types is uniform in villous stroma. Distribution of different collagen types and fibronectin, including the unusual localization of membrane collagen type IV, in villous stroma and cellular islets of early and mature placenta is discussed.

[Prion disease and brain amyloidosis]. / Prionovye bolezni i amiloidoz golovnogo mozga.

Human prion disorders include Kuru, Creutzfeld-Jakob disease (CJD), Gerstman-Straussler-Scheinkler syndrome (GSS), fatal familial insomnia (FFI) and prion protein cerebral amyloid angiopathy (PrPCAA). Prion diseases manifest as infections, genetic and sporadic disorders. In these diseases an abnormal form of the host's protein, prion protein protease-resistant (PrPres), is essential for pathogenic process. Host protein, prion protein protease-sensitive (PrPsen) in humans is encoded by a single copy gene (PRNP) located in the short arm of chromosome 20. To date, 19 different mutations in PRNP have been found that cause inherited prion disease. In these diseases PrPsen undergoes conformational changes involving a shift from alpha-helix to beta-sheet structures. This conversion is important for PrP-amyloidogenesis which occurs to the highest degree in GSS, while it is less frequently seen in other prion diseases. Pathomorphologically, amyloidogenesis in the brain is characterized by formation of PrPres conglomerates, diffuse homogeneous deposits and pleomorphic fibrillar amyloid plaques. The neurotoxic activity of PrPres and its fragments supports the causal relationship between PrPres deposits and neuropathological events in prion diseases. Congo-red and certain sulfated glycans potently inhibit PrPres formation. This raises the potential of therapeutic strategies for the treatment of these diseases.

[Immunomorphological method of study with a peroxidase antibody label]. / Immunomorfologicheskii metod issledovaniia s metkoi antitel peroksidazoi.

Among different variants of immunoperoxidase method preference is given to its indirect variant with labeling of pure antibody with peroxidase by a two-step procedure. Examples of the use of this method for the detection of serum proteins, tissue antigens, and autoantibody are presented. The best results are achieved in treatment of sections with pure antibody instead of antisera.

[Characteristics of the extracellular matrix in the formation of sclerosis of the chorionic villi (immunofluorescence studies)]. / Osobennosti ékstratselliuliarnogo matriksa v formirovanii skleroza vorsin platsenty (immunofliuorestsentnoe issledovanie).

Distribution of the eight components of extracellular matrix in small villi of human placenta was studied with indirect immunofluorescence. Normal full-term pregnancy was found to present rough fibers and type I collagen conglomerates accumulated in the stromal center, while type III and V collagens occurred all over the villous surface. Stromal localization of type IV collagen was recorded as well as its presence in basal membranes of the epithelium and vessels. Fibronectin showed diffuse distribution concentrating along the vessels. Laminin, entactin and in lesser degree heparan sulphate proteoglycan were registered in epithelial and vascular basal membranes. Isolated sclerosis of the villi in pathological pregnancy involved stromal accumulation of type I, III collagens, small fragments of collagens type IV, V and fibronectin. Multiple sclerosis of adherent villi demonstrated hyalin-like conglomerates of various collagens and fibronectin filling up the stroma. Laminin, heparan sulphate proteoglycan and entactin were not observed at these sites.

[Obtaining pure antibodies with the aid of an immunosorbent based on sephadex]. / Poluchenie chistykh antitel pri pomoshchi immunosorbenta na osnove sefadeksa.

A method for production of pure antibodies by means of an immunosorbent based on sephadex according to the technique of Wilson and Nakane modified by the authors is described. The method requires no special equipment, is simple and may be used in any laboratory for immunomorphological studies.

[Myofibroblasts in desmoids]. / Miofibroblasty v desmoidakh.

Ten desmoids are examined electron-microscopically and by the fluorescent antibody method. The cells with the ultrastructural signs of myofibroblasts are found in tumours. Immunomorphologically these cells were found to contain cytoplasmic myosin occurring in the cells of both muscular and non-muscular origin. The structural myosin of smooth muscle cells was not detected in these elements. The above cells are considered to be the modification of fibroblasts which have no relation to the smooth muscle cells. Non-homogeneity of myofibroblast population is noted.

[Distribution of collagen types I, III, IV and V in the walls of human arteries]. / Raspredelenie kollagena I, III, IV, V tipov v stenke arterii cheloveka.

Using immunofluorescence the localization of I, II, IV, V type collagen in different layers of the artery wall was established. The adventitia was shown to contain only I and III type collagen. All collagen types studied were identified in the media. The structural organization and quantity of different types of collagen were found to depend on the artery caliber. The distribution of I, III, IV, V type collagen in the aortal intima is described. The localization of athrombogenic collagen of type IV and V in the area of the endothelial basal membrane is highlighted. The authors revealed the presence of I and III type collagen in the subendothelium and the age-related increase in interstitial collagen levels in the intima which is important for predicting thrombosis in deendothelization . Employing immunoelectron microscopy, previously undescribed forms of I, III, IV, V type collagen organization into microgranular structures were ascertained.

[Immunocytochemical study of the myoepithelial cells in fibrocystic disease and ductal carcinoma of the breast]. / Immunotsitokhimicheskoe izuchenie mioepitelial'nykh kletok pri fibrozno-kistoznoi mastopatii i protokovom rake molochnoi zhelezy.

Myoepithelial cells (MC) were identified and the pattern of their distribution in dysplasia and duct carcinoma of the mammary gland was studied using a monospecific antiserum to smooth-muscle myosin of the human uterus, by the indirect Coons' method under luminescent microscope. Samples of the operation material from 10 patients with fibrous-cystic mastopathy and 26 patients with duct carcinoma were examined. In solid (7 observations) and comedo (4 observations) carcinomas with the structure of cancer in situ, MC were found intact in the periphery. In invasive growth of these tumors as well as in scirrhus (5 observations) and adenocarcinoma (10 observations) MC disappeared from the duct structures and were absent in carcinoma proliferates. Positive immunofluorescence in the conventional location of MC in the alveolar duct system of the mammary gland allows sufficiently valid identification and study of the distribution and changes in these cells. The immunocytochemical verification of MC in dysplasia and duct carcinoma indicates the possibility of using MC as markers of duct carcinoma in situ; MC disappearance from the duct structures characterizes the beginning of the infiltrative tumor growth.

[Fibronectin content of the intima of normal and atherosclerotic arteries]. / Soderzhanie fibronektina v intime arterii v norme i pri ateroskleroze.

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.

[Immunomorphologic characteristics of the distribution of collagen types I, III, IV and V in normal intima and in atherosclerosis of the major arteries and aorta in man]. / Immunomorfologicheskaia kharakteristika raspredeleniia kollagena I, III, IV, V tipov v normal'noi intime i pri ateroskleroze krupnykh arterii i aorty cheloveka.

The distribution of collagen of the above types in normal intima, sites of its physiological thickening, lipid streaks, atherosclerotic plaques is studied by the immunoluminescent method. All collagen types at various proportions are found to be located in the subendothelial areas of intima. Besides that the IV and V collagen types are detected in the zone of endothelial basal membrane. The essential property of atherosclerotic plaques is the prevalence of rough fibrillar structures from interstitial collagen which do not occur normally.

[Pathohistology of costal cartilage and immunomorphologic characteristics of collagen in funnel chest]. / Patogistologiia rebernogo khriashcha i immunomorfologicheskaia kharakteristika kollagena pri voronkoobraznoi grudi.

Biopsy samples from the costal cartilage tissue were studied for 68 children with funnel deformity and from 20 children with normally formed chest. The authors present general morphologic features characteristic of the costal cartilage structure in norm and in case of funnel chest. These features include vast acellular sites, map-like areas, unmasked chondrin fibers and "marrow" cavities. In funnel chest they develop, however, 6-7 years earlier, than normally, and are consequent stages of the accelerated costal cartilage involution. Costal cartilage matrix was found to have an increased content of fibronectin and V collagen type in case of funnel chest. Besides III and IV procollagen types were noted in the chondrocytes. The accelerated growth of costal cartilages is involved in the formation of funnel chest.

[Types of myoepithelial cell proliferation in dyshormonal breast dysplasias and benign breast tumors]. / Tipy proliferatsii mioépitelial'nykh kletok pri disgormonal'nykh displaziiakh i dobrokachestvennykh opukholiakh molochnoi zhelezy.

Myoepithelial cells (MC) were identified and types and forms of their hyperplasia in dysplasias and bening mammary gland tumors in dog and man were studied by indirect Coons' method using highly purified monospecific antiserum to smooth muscle myosin and by performing alkaline phosphatase test. Operation material from 75 patients and 12 dogs was studied by immunohistochemical method and from 26 persons and 12 dogs by histochemical method. Comparative analysis of immunohistochemical and histochemical identification of MC revealed differences in the results of staining in 7 out of 38 observations due to negative test for alkaline phosphatase in the presence of fluorescence. A high degree of coincidence of positive tests in immunohistochemical and histochemical methods of the study suggests that the test for alkaline phosphatase is a sufficiently reliable marker of MC. The principal similarity of types and forms of MC hyperplasia in canine and human mammary gland tissue indicates that dogs may be used as an adequate model for the study of various diseases of this organ. In addition to the known centripetal and centrifugal types, a uniformly concentric and smooth-muscle proliferations of MC were distinguished in parallel immunohistochemical and histochemical studies on variants of MC proliferation.

[Role of increased vascular permeability in the formation of dimorphic amyloid in mouse myocardium]. / Rol' povyshennoi sosudistoi pronitsaemosti v obrazovanii dimorfnogo amiloida v miokarda myshi

Electron-microscopic investigation carried out following the intravenous administration of ferritin revealed an increase in permeability of the myocardium capillary wall in mice in experimental casein amyloidosis. Additionally, a complex electron-microscopic, immunohistochemical and histological investigation of the development of amyloidosis in the myocardium of mice was conducted. It was established that deposits of typical amyloid in the myocardium of mice consisted of two ultrastructural components--granular material and fibrils. It was shown that formation of amyloid with characteristic tinctorial properties was preceded by depositions of specific antigen fibrils of granular ultrastructure.

[The viral and immunological characteristics of cardiomyopathies and myocarditis]. / Virusoimmunologicheskaia kharakteristika kardiomiopatii i miokarditov.

As many as 97 patients with myocardial lesions: congestive and hypertrophic cardiomyopathy (CMP), postmyocarditis CMP (PM CMP), myocarditis (MC), alcoholic heart injury (AHI), coronary heart disease (CHD), vegetodysovarian myocardiodystrophy were examined by means of a complex of the virological tests (for Coxsackie B, Epstein-Barr and hepatitis B viruses) and immunoassays (for antibodies to different components of the myocardium, leukocyte migration inhibition test, antibody-dependent cellular cytotoxicity test, measurements of T and B lymphocytes and their subpopulations, and so forth). Virus infection was shown to be of a role for the onset of acute MC (usually reversible) and congestive CMP. At the same time the autoimmune mechanisms of the lesions were conclusively ascertained in MC associated with heart failure and in PM CMP. In patients with congestive CMP and AHI coupled with heart failure, antibodies to nerve fibers of the myocardium could be demonstrated in the presence of T-lymphocyte deficiency and high titers of antibodies to Epstein-Barr virus. This does not allow excluding myocardial denervation leading to refractory heart failure. Some immunological parameters made use of in the study provide an opportunity of an objective evaluation of the effect glucocorticoid treatment produces on patients suffering from MC and PM CMP.