Localization of specific carbohydrate configurations in the hemophilic synovial membrane

Fluorescein isothiocyanate-conjugated lectins include: concanavalin A (Con-A), Dolichos biflorus agglutinin (DBA), Griffonia simplicifolia-I (GS-I), Griffonia simplicifolia-II (GS-II), Arachis hypogaea agglutinin (PNA), Maclura pomifera agglutinin (MPA), Ricinus communis agglutinin-I (RCA-I), Glycine max agglutinin (SBA), Ulex europaeus agglutinin-I (UEA-I), and wheat germ agglutinin (WGA). These lectins were used to histochemically demonstrate lectin bindings on hemophilic synovial membrane. GS-I (galactose and galactosamine specific) and SBA (galactosamine specific) was shown to bind strongly in the cytoplasm of the lining cells. Lymphocytes and mast cells were largely bound by Con-A (mannose and glucose specific) and GS-II (glucosamine specific) positive. UEA-1 (fucose specific) was shown to bind specifically to synovial vascular endothelial cells. Lectin histochemistry is a useful method for classification of the cells on the synovial membrane.     

老年性骨关节炎与类风湿关节炎滑膜细胞转化特性的比较研究

目的与骨关节炎比较,探讨类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)胞外信号活化蛋白激酶(ERK)的活化状态和P53蛋白的异常表达。方法原代培养RA FLS和OA FLS,应用western blot检测二者ERK活化状态的差异;应用抗P53抗体(DO-1和PAb240)与抗PCNA抗体对RA和OA的滑膜组织和细胞进行免疫组织化学染色;并进一步应用western blots确定RA FLS P53的异常表达。结果与OA相比,RA滑膜衬里层P53蛋白表达增多,主要为与抗突变型P53抗体阳性反应细胞积聚,并进一步证实RA衬里层成纤维样滑膜细胞P53表达异常;而且RA FLS在低血清和非贴壁情况下ERK活化状态均明显高于骨关节炎。结论类风湿关节炎衬里层成纤维样滑膜细胞P53蛋白异常表达,而且ERK处于持续活化状态,这为RAFLS的转化特性提供了新的佐证。     

Lectin affinity of the seminiferous epithelium in healthy and cryptorchid post-pubertal boars

The present study describes the sugar content of the seminiferous epithelium, using lectin histochemistry, in healthy boars and in boars with unilateral and bilateral abdominal cryptorchidism. In healthy boars the apical cytoplasm of Sertoli cells exhibited abundant glucosyl (Con A and WGA lectins), galactosyl (HPA, DBA, SBA and PNA lectins), and fucosyl (AAA lectin) residues. Spermatogonia and spermatocytes contained abundant glucosyl (Con A and WGA lectins) and fucosyl (AAA lectin) residues. In spermatids, galactosyl (SBA and PNA lectins) and glucosyl (Con A and WGA lectins) residues increased progressively throughout spermiogenesis, and fucosyl (AAA lectin) residues decreased. As compared with healthy boars, the scrotal testis of unilateral cryptorchid boars showed decreased amounts of fucosyl (AAA lectin) and galactosyl (HPA and DBA lectins) residues on the Sertoli cell apical cytoplasm; spermatocytes exhibited higher content of glucosyl (Con A lectin) residues and spermatids showed altered nature of glucosyl (Con A and WGA lectins) and galactosyl (SBA and PNA lectins) complexes. In abdominal testes of unilateral and bilateral cryptorchid boars, immature Sertoli cells and spermatogonia showed decreased fucosyl (AAA lectin), and increased glucosyl (Con A and WGA lectins) and galactosyl (SBA and PNA lectins) contents. These results suggest that the seminiferous epithelium of healthy boars has polarized activity with the apical compartment implicated in germ cell-Sertoli cell adhesion and interaction, in transport of ions, substrates and fluids, and in acrosomal differentiation. In scrotal testes, unilateral abdominal cryptorchidism could lead to defective germ cell-Sertoli cell adhesion, impaired acrosomal differentiation and increased ionic transport in the apical compartment of the seminiferous epithelium. Unilateral and bilateral cryptorchidism could induce increased ionic transport and membrane permeability in the seminiferous epithelium of abdominal testes.     

Lectins in diagnosis of bladder carcinoma.

With the purpose of studying changes in the expression of glycoconjugate structures in nonmalignant and cancerous lesions of urothelium the lectins ConA, TKA, PNA, DBA, STA, LFA, UEA, MPA, RCA, LCA, GSA1, SBA, GSA2, WGA, PHA and Lot were tested in formalin-fixed, paraffin-embedded tissue sections of (1) cold biopsies from normal urothelium and bladder cancer of different grades (G1-G3) in humans, (2) normal transitional epithelium and N-butyl-N(4-hydroxybutyl)nitrosamine (BBN)-induced bladder cancer in animal experiments (Wistar rat), and (3) human transitional cancer cell line HT 1376. In human urothelium TKA and SBA were positive markers demonstrating positive staining reactions in all tumor grades without binding to normal epithelium. They stained also the human transitional carcinoma cell line HT 1376 (G3). In Wistar rats DBA, ConA, LCA, SBA, GSA2 and WGA had a specific affinity to BBN-induced carcinoma. Findings of positive lectin marker in transitional cell cancer may offer progress in diagnostics and therapy.     

Lectins expression of parathyroid glands with primary hyperparathyroidism

The binding sites of lectins in parathyroid glands were determined by an immunohistochemical method in normal parathyroid gland, hyperplasia, adenoma and carcinoma, the used lectins were commercially available Glycine max (SBA), Concanavalin enciformis (Con A), Triticum vulgaris (WGA), Richinus communis (RCA), Banderiaea simplicifolia II (BSA II) and Arachis hypogaea (PNA). For normal parathyroid glands (2 cases) and hyperplasia (2 cases), WGA and BSA II were stained in cytoplasma and cell membrane. For carcinoma (1 cases), all lectins but BSA II were positively stained. In particular, SBA revealed more stronger stain than any other hystological types. From the staining patterns of lectins, it was suggested that adenomas (22 cases) be divided into one group similar to carcinoma and the others to normal parathyroid gland and hyperplasia. But there was no difference in clinical data of patients between the two groups.     

Capacitated and acrosome reacted spermatozoa of goat (Capra indicus): a fluorescence and electron microscopic study

Summary. Plasma membrane alterations accompanying in vitro capacitation and acrosome reaction of goat spermatozoa were studied using lectin labelling, scanning electron microscopy, and freeze-fracture methods. Fluorescein isothi-ocyanate linked lectins namely; Canavalia ensiformis (ConA), Maclura pomifera (MPA), Arachis hypogaea (PNA), Glycine max (SBA) and Triticum vulgaris (WGA) agglutinin were used to examine the distribution of surface carbohydrates during these two events. The head and the sperm tail reveal altered lectin labelling features after capacitation and acrosome reaction. After capacitation the surface coat components for MPA, SBA, and WGA are shed from the spermatozoon head. ConA receptors on the head are retained after capacitation but are partially shed in the acrosome reacted spermatozoa. SBA receptor sites appear on the sperm tail of the capacitated spermatozoa. Unusual morphological changes attending capacitation involve the sperm tail-end which develops a novel entity, which we have termed 'spatula'. The 'spatula' shows strong binding with ConA and WGA only. In the acrosome reacted spermatozoa the spatulated tail-end unwinds with a concomitant loss of lectin labelling. Highly ordered membrane particles, 'ladders' of the middle piece of the epididymal sperm tail, disappear and IMP clearings appear on the middle piece and in the spatulated ends of the capacitated spermatozoa. But in the acrosome reacted sperm IMPs reappear and are randomly disposed on the middle-piece and are clustered in small patches on the principal-piece. IMP free areas appear on the plasma membrane covering the acrosome and the outer acrosomal membrane (OAM) of the capacitated spermatozoa. The plasma membrane and OAM fuse at multiple foci and appear as acrosomal 'ghosts' which remain associated with the sperm head even after acrosome reaction.     

Overexpression of active TGF-beta-1 in the murine knee joint: evidence for synovial-layer-dependent chondro-osteophyte formation

OBJECTIVE: To investigate the impact of a prolonged and constant active TGF-beta expression by the synovial lining cells on cartilage and ligamentous joint structures in vivo. DESIGN: An adenoviral vector (AdTGF-beta1(223,225)) was used for the overexpression of active TGF-beta1 in knee joints of C57Bl/6 mice. RESULTS: It was found that physiological relevant levels of active TGF-beta1 produced by the synovial lining layer resulted in histopathological changes: hyperplasia of synovium and chondro-osteophyte formation at the so-called chondro-synovial junctions. No histological changes were seen after intra-articular injection of an empty control vector (AdDL70-3) or by overexpression of latent TGF-beta1 (AdTGF-beta1). The predominant site of TGF-beta production in osteoarthritis (OA) and rheumatoid arthritis (RA) is the synovial lining layer. To address the question whether the TGF-beta-induced changes were related to the expression site in the synovial lining, the synovial lining layer was depleted by local treatment with liposomes encapsulating clodronate. Depletion of the lining resulted in a dramatic change of TGF-beta1-induced pathology: markedly reduced chondro-osteophyte formation and increased accumulation of extracellular matrix in the synovium. CONCLUSION: This study shows that overexpression of active TGF-beta1 in the knee joint results in OA-like changes and suggests the synovial lining cells contribute to the chondro-osteophyte formation.     

Characterizing the glycocalyx of poultry spermatozoa: I. Identification and distribution of carbohydrate residues using flow cytometry and epifluorescence microscopy

The aim of the present work was to use a battery of lectins to 1) delineate the carbohydrate content of sperm glycocalyx in the turkey and chicken using flow cytometry analysis, and 2) evaluate the distribution of existing sugars over the sperm plasma membrane surface with epifluorescent microscopy. Carbohydrate groups (corresponding lectins) that were investigated included galactose (GS-I, Jacalin, RCA-I, PNA), glucose and/or mannose (Con A, PSA, GNA), N-acetyl-glucosamine (GS-II, s-WGA, STA), N-acetyl-galactosamine (SBA, WFA), fucose (Lotus, UEA-I), sialic acid (LFA, LPA), and N-acetyl-lactosamine (ECA). Spermatozoa were assessed before and after treatment with neuraminidase to remove sialic acid. Mean fluorescence intensity (MnFI) was used as indicator of lectin binding for flow cytometry analysis. Nontreated spermatozoa from both species showed high MnFI when incubated with RCA-I, Con A, LFA, and LPA, as did chicken spermatozoa incubated with s-WGA. Neuraminidase treatment increased the MnFI for most lectins except LFA and LPA, as expected. Differences in MnFI between species included higher values for s-WGA and ECA in chicken spermatozoa and for WFA in turkey spermatozoa. Microscopy revealed segregation of some sugar residues into membrane-specific domains; however, the 2 staining techniques (cell suspension vs fixed preparation) differed in identifying lectin binding patterns, with fixed preparations yielding a high degree of nonspecific binding. We conclude that 1) the glycocalyx of turkey and chicken spermatozoa contains a diversity of carbohydrate groups, 2) these residues are extensively masked by sialic acid, 3) the glycocalyx composition is species-specific, and 4) some glycoconjugates appear to be segregated into membrane-specific domains. Characterization of the poultry sperm glycocalyx is the first step in identifying the physiological impact of semen storage on sperm function.     

Distribution of glycoconjugates in normal rat ventral prostate and their use as markers of androgen-controlled secretory function in culture

The distribution of glycoconjugates in uncultured and cultured rat ventral prostate was studied by using eight fluorescent lectins. The prostate pieces were cultured in defined medium with or without testosterone for 1-14 days. Each lectin revealed a characteristic binding pattern. Con A, LCA, WGA, and RCA I stained both epithelial and interstitial components. SBA and PNA were specific for the epithelium: SBA stained the region of the Golgi complex; PNA showed the brightest fluorescence in the apical part of the cells representing the region of secretory granules. In culture without testosterone the epithelial cells gradually lost their fluorescence, whereas the stromal fluorescence increased. The basement membrane was disorganized. With testosterone the integrity of the epithelium and stroma was maintained, and the staining pattern of the lectins was in the main similar as in vivo. However, at 14 days a change in the staining pattern of apical cytoplasm with PNA was noted, indicating that in long-term cultures, in addition to testosterone, other hormones and growth factors are necessary to complete especially the last stages of the secretory process in the epithelial cells.     

Lichtmikroskopischer Nachweis von Lektin-Rezeptoren an der Oberfläche humaner Spermatozoen/Demonstration of Lectin-receptors on the Surface of Human Spermatozoa by Light-microscopy

The topography of lectin binding sites on human spermatozoa has been examined by using particular series of peroxydase labeled lectin-konjugates (GS-1, GS-2, WGA, PNA, SBA, DBA, BPA, MPA, UEA-1, LPA, Con-A) which demonstrated typical patterns of lectin-receptor distributions concerning acrosomal and equatorial as well as postacrosomal areas.     

Expression of large tenascin-C splice variants in synovial fluid of patients with rheumatoid arthritis.

Tenascin-C (TN-C) is a hexameric glycoprotein component of extracellular matrix, and alternative RNA splicing creates two major TN-C size variants (the small and large variants). The large TN-C variants play key roles in many pathologic conditions in adults, including tumorigenesis, regeneration, and inflammation. This cross-sectional study compared levels of large TN-C variants in synovial fluid of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Synovial fluid samples were obtained from knees of 26 patients with advanced RA and 79 with advanced OA. Expression of TN-C splice variants was examined using Western blotting. The levels of large TN-C variants in synovial fluid were determined by an enzyme-linked immunosorbent assay. Synovium were analyzed for TN-C by immunohistochemistry. Immunoblotting showed the presence of large TN-C variants in synovial fluid from patients with RA and OA. However, levels of large TN-C variants were fourfold higher in RA samples compared with OA samples (p < 0.01). Synovial fluid levels of TN-C in RA did not correlate with C-reactive protein levels. Immunohistochemistry of the synovium showed stronger reactivity in RA samples than in OA samples. These results indicate that local synthesis of TN-C is increased during rheumatic disease.     

Characterizing the glycocalyx of poultry spermatozoa: II. In vitro storage of Turkey semen and mobility phenotype affects the carbohydrate component of sperm membrane glycoconjugates

The turkey sperm glycocalyx is known to contain residues of sialic acid, alpha-mannose/alpha-glucose, alpha- and beta-galactose, alpha-fucose, alpha- and beta-N-acetyl-galactosamine, monomers and dimers of N-acetyl-glucosamine, and N-acetyl-lactosamine. Potential changes in these carbohydrates during in vitro semen storage at 4 degrees C were evaluated using males of both high- and low-sperm-mobility phenotypes. Changes in carbohydrate residues were quantified by flow cytometry analysis using a battery of 14 fluorescein isothiocyanate-labeled lectins in combination with control (sialylated) or neuraminidase-treated (nonsialylated) sperm. Sperm were evaluated at 0, 2, 4, 8, 12, and 24 hours of storage. For control sperm, 4 different patterns of lectin binding were observed over time: 1) increased mean fluorescence intensity (MnFI) at 2 hours (Griffonia simplicifolia lectin-I [GS-I]) and 8 hours (Ricinus communis lectin-I [RCA-I]) that remained elevated during storage; 2) increased MnFI at specific time points (Limax flavus lectin [LFA], 2 hours; Artocarpus integrifolia lectin [jacalin] and succinyl Triticum vulgare lectin [sWGA], 8 hours; Galanthus nivalis lectin [GNA], 12 hours) followed by decreasing MnFI during the remainder of the 24-hour storage period; 3) increased MnFI only at the 24-hour time point (Lotus tetragonolobus lectin [lotus] and Arachis hypogaea lectin [PNA]); and 4) no changes in MnFI during the 24-hour storage period (Erythrina cristagalli lectin [ECA], GS-II, Pisum sativum lectin [PSA], Glycine max lectin [SBA], and Wisteria floribunda lectin [WFA]). For nonsialylated sperm, increased binding of ECA, GS-II, SBA, and WFA was observed at variable time points; only Canavalia ensiformis lectin (Con A) and PSA remained unchanged during storage. Differences between mobility phenotypes existed for lectins Con A, GS-II, LFA, PSA, SBA, and sWGA, with sperm from low-mobility males exhibiting higher MnFI than high-mobility males throughout 24 hours of storage. We concluded that the observed increases in lectin binding during semen storage indicate an augmentation of nonsialylated terminal residues, which could alter sperm antigenicity and negatively impact fertility. Further, spermatozoa from low-mobility males may have higher antigenicity even before semen storage. Other possible functional implications are discussed.     

一氧化氮在骨性关节炎发病机制中的作用

目的 研究一氧化氮与骨性关节炎发病的关系。方法 抽取骨性关节炎和类风湿性关节类患者的关节积液和血清,及关节镜下清理术中切除的骨性关节炎滑膜和退变软骨作为标本,用硝酸还原酶法测定其中的一氧化氮含量并进行比较。结果 骨性关节炎患者的软骨及滑膜中一氧化氮含量明显高于其血清及关节液,且软骨中一氧化氮含量明显高于滑膜。类风湿性关节炎患者关节液中一氧化氮浓度明显高于其血清及骨性关节炎的关节液。结论 骨性关节炎     

Matrix metalloproteinase protein expression profiles cannot distinguish between normal and early osteoarthritic synovial fluid

ABSTRACT: BACKGROUND: Osteoarthritis (OA) and Rheumatoid arthritis (RA) are diseases which result in the degeneration of the joint surface articular cartilage. Matrix Metalloproteinases (MMPs) are enzymes that aid in the natural remodelling of tissues throughout the body including cartilage. However, some MMPs have been implicated in the progression of OA and RA as their expression levels and activation states can change dramatically with the onset of disease. Yet, it remains unknown if normal and arthritic joints demonstrate unique MMPs expression profiles, and if so, can the MMP expression profile be used to identify patients with early OA. In this study, the synovial fluid protein expression levels for MMPs 1, 2, 3, 7, 8, 9, 12 & 13, as well as those for the Tissue Inhibitors of MMPs (TIMPs) 1, 2, 3, & 4 were examined in highly characterized normal knee joints, and knee joints with clinically diagnosed OA (early and advanced) or RA. The purpose of this study was to determine if normal, OA, and RA patients exhibit unique expression profiles for a sub-set of MMPs, and if early OA patients have a unique MMP expression profile that could be used as an early diagnostic marker. METHODS: Synovial fluid was aspirated from stringently characterized normal knee joints, and in joints diagnosed with either OA (early and advanced) or RA. Multiplexing technology was employed to quantify protein expression levels for 8 MMPs and 4 TIMPs in the synovial fluid of 12 patients with early OA, 17 patients diagnosed with advanced OA, 15 with RA and 25 normal knee joints. Principle component analysis (PCA) was used to reveal which MMPs were most influential in the distinction between treatment groups. K - means clustering was used to verify the visual grouping of subjects via PCA. RESULTS: Significant differences in the expression levels of MMPs and TIMPs were observed between normal and arthritic synovial fluids (with the exception of MMP 12). PCA demonstrated that MMPs 2, 8 & 9 can be used to effectively separate individuals diagnosed with advanced arthritis from early osteoarthritic and normal individuals, however, these MMP profiles do not separate early OA from normal synovial fluid. An apparent separation between advanced OA and RA subjects was also revealed through PCA. K-means clustering verified the presence of 3 clusters: normal joints clustered with early OA, and separate clusters of advanced OA or RA. CONCLUSIONS: This study demonstrates that unique MMP and TIMP expression profiles are present within normal, advanced OA and RA synovial fluid. These MMP profiles can be used to distinguish advanced OA & RA synovial fluid from early OA & normal synovial fluid, and even between synovial fluid samples from OA and RA joints. Although this methodology cannot be used for the diagnosis of early OA, high throughput multiplex technology of MMPs and TIMPs in synovial fluid may prove useful in determining the severity of the disease state, and/or quantifying the response of individuals to disease interventions.     

Analysis of cartilage-derived retinoic-acid-sensitive protein (CD-RAP) in synovial fluid from patients with osteoarthritis and rheumatoid arthritis

We have measured the concentration of cartilage-derived retinoic-acid-sensitive protein (CD-RAP) in synovial fluid (SF) from the knees of 49 patients with osteoarthritis (OA) and 79 with rheumatoid arthritis (RA) in order to investigate the correlation between the type of joint disease and level of CD-RAP. The mean concentration of CD-RAP in synovial fluid was significantly higher in OA than in RA. The level of CD-RAP in the group of patients with mild OA was significantly higher than in the moderate or severe groups and that in the group with mild RA was also significantly higher than in the other RA groups and decreased with progression of the disease. Immunohistochemical studies showed expression of CD-RAP in the cytoplasm of chondrocytes in newly-formed fibrocartilage. Since CD-RAP is mainly produced in young and proliferating chondrocytes, our results suggest that the level of CD-RAP in synovial fluid reflects remodelling of articular cartilage and may be used as a marker to estimate objectively the restorative reaction of chondrocytes.     

Distinct serum and synovial fluid interleukin (IL)-33 levels in rheumatoid arthritis, psoriatic arthritis and osteoarthritis

ObjectivesRecent evidence suggests a role for interleukin (IL)-33 and its receptor ST2 in arthritis. In this study, we quantified IL-33 and soluble (s)ST2 levels in serum and synovial fluid (SF), and assessed synovial IL-33 expression levels and pattern in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), or osteoarthritis (OA).MethodsSerum and SF IL-33 and sST2 levels were assessed by ELISA. IL-33 mRNA was quantified by RT-qPCR. Synovial IL-33 protein expression pattern was examined by immunohistochemistry.ResultsSerum and SF IL-33 levels tended to be higher in RA than in OA patients. In contrast to RA, IL-33 was not detectable in PsA serum and SF. Serum sST2 levels were higher in RA than in OA. There was a wide variation of synovial tissue IL-33 mRNA expression within each disease group and IL-33 mRNA levels were not significantly different between the groups. A similar IL-33 protein expression pattern was observed in RA, PsA and OA synovium, with strong nuclear expression of IL-33 in endothelial cells and, in a subset of RA, PsA and OA patients, in cells morphologically consistent with synovial fibroblasts.Discussion/ConclusionsThis study confirms increased circulating IL-33 levels in RA. In addition, we report that IL-33 is undetectable in the serum or SF of PsA patients. Local expression of IL-33 in the synovium was observed at similar variable levels in RA, PsA and OA, suggesting that inflamed joints do not represent the primary source of elevated serum and SF levels of IL-33 in RA.     

Distribution of Lectin Binding in Rat Testis and Epididymis

Summary: Seven lectins (PNA, RCA I, SBA, Con A, WGA, UEA I, DBA) conjugated with rhodamine were employed to analyse the staining pattern of glycoproteins with varying sugar residues in the testis and epididymis of adult Wistar rats. Some lectins (UEA I, SBA, DBA) gave rather specific staining of the mature acrosome, while others (PNA, RCA I) showed affinity for the early stages of acrosome formation or had a wide affinity for germinal and non-germinal cells and structures (Con A, WGA). In the epididymis the sperm mass had a homogenous staining reaction with some lectins (PNA, RCA I, Con A, WGA, DBA) which also showed a rather strong reaction on the epithelial surface. It was concluded that this reaction is at least partially due to the secretory products synthetized by principal, apical, narrow and light cells of the epididymal epithelium. Some differences in the staining pattern of these cells were recorded indicating specialization of the cells for the production of distinct glycoproteins. The staining pattern of the interstitial and intertubular compartment of the testis and epididymis was also recorded. Zusammenfassung: Die Verteilung der Lektin-Bindung im Hoden und Nebenhoden der Ratte Sieben Lektine (PNA, RCA I, SBA, Con A, WGA, UEA I, DBA) wurden verbunden mit Rhodamin verwendet, um das Darstellungsmuster von Glykoproteinen mit verschiedenen Zuckerresten im Hoden und Nebenhoden erwachsener Wistar-Ratten zu analysieren. Einige Lektine (UEA I, SBA, DBA) gaben eine recht spezifische Darstellung des reifen Akrosoms, wohingegen andere (PNA, RCA I) eine Affinität zu den frühen Stadien der Akrosomformation zeigten oder eine breite Affinität zu Germinal- und Nichtgerminalzellen und -strukturen aufwiesen (Con A, WGA). Im Nebenhoden zeigte die Spermatozoenmasse ein homogenes Darstellungsmuster mit einigen Lektinen (PNA, RCA I, Con A, WGA, DBA), welche auf der epithelialen Oberfläche auch eine recht starke Reaktion aufwiesen. Es wurde daraus geschlossen, daβ diese Reaktion zumindest teilweise auf die sekretorischen Produkte, die in den Haupt-, Apikal-, Schmal- und Hellzellen des Nebenhodens synthetisiert werden, zurückzuführen ist. Einige Unterschiede im Darstellungsmuster dieser Zellen wurden aufgezeichnet, die die Spezialisierung der Zellen für die Produktion bestimmter Glykoproteine anzeigen. Das Darstellungsmuster des interstitiellen und intertubulären Anteils des Hodens und Nebenhodens wurde ebenfalls aufgezeichnet.     

Changes in the lectin binding sites on the testicular, epididymal, vas, and ejaculated spermatozoon surface of dog

Summary. Changes in the localization of sperm surface glycocomponents of testicular, epididymal, vas deferens, and ejaculated spermatozoa of dog ( Canis domesticus ) were studied employing fluorescein isothiocyanate conjugated lectins viz., Concanavalin A (ConA), Triticum vulgaris (WGA), Maclura pomifera (MPA), and Arachis hypogaea (PNA) agglutinins. The plasma membrane clothing the acrosome of the testicular, epididymal, and vas deferens spermatozoa shows reactivity with all the lectins used. However, in the ejaculated spermatozoa, the entire sperm surface shows reactive sites for ConA, WGA, and PNA. Variation in the labelling of the cytoplasmic droplet in different stages of spermatozoon transit in the epididymis has been discussed.     

Fluorescence study of renal cell carcinoma with antibodies to renal tubular antigens, intermediate filaments, and lectins

A fluorescence study was performed in 16 renal cell carcinomas using antibodies to renal tubular antigens (RTA), two intermediate filaments, cytokeratin and vimentin, and two lectins, soybean agglutinin (SBA) and peanut agglutinin (PNA). We observed the presence of RTA, cytokeratin, and vimentin in all of our specimens. The expression of vimentin, the cytoskeletal protein of mesenchymal cells, was considered to be very interesting feature of the tumor. Binding sites of SBA, normally present in glomeruli, proximal and distal tubules, were detectable in the neoplastic cells in only 37.5% of our specimens. PNA did not react with the tumor except for the small area of 2 specimens. Lectins may be useful for estimating the characteristics or renal cell carcinoma including its malignant potentials, and antibodies to RTA and intermediate filaments seem to be available for the diagnosis of the tumor in metastatic lesions.     

Osteoarthritis synovial fluid activates pro-inflammatory cytokines in primary human chondrocytes

Purpose

Two of the most common joint diseases are rheumatoid arthritis (RA) and osteoarthritis (OA). Cartilage degradation and erosions are important pathogenetic mechanisms in both joint diseases and have presently gained increasing interest. The aim of the present study was to investigate the effects of the synovial fluid environment of OA patients in comparison with synovial fluids of RA patients on human chondrocytes in vitro.

Methods

Primary human chondrocytes were incubated in synovial fluids gained from patients with OA or RA. The detection of vital cell numbers was determined by histology and by using the Casy Cell Counter System. Cytokine and chemokine secretion was determined by a multiplex suspension array.

Results

Microscopic analysis showed altered cell morphology and cell shrinkage following incubation with synovial fluid of RA patients. Detection of vital cells showed a highly significant decrease of vital chondrocyte when treated with RA synovial fluids in comparison with OA synovial fluids. An active secretion of cytokines such as vascular endothelial growth factor (VEGF) of chondrocytes treated with OA synovial fluids was observed.

Conclusions

Significantly increased levels of various cytokines in synovial fluids of RA, and surprisingly of OA, patients were shown. Activation of pro-inflammatory cytokines of human chondrocytes by synovial fluids of OA patient supports a pro-inflammatory process in the pathogenesis of OA.