Human hyaline cartilage--biochemical and immunochemical study.

Hyaline cartilage from the knee or rib of newborns was divided in neutral salts into a soluble and insoluble part. The insoluble part was treated with collagenase and was separated into 2 peaks by gel chromatography on Sephadex G-200. The 1st peak revealed a low concentration of proteins, no hydroxyproline, and very high levels or uronates. In the 2nd peak, very high values of hydroxyproline and very low values of uronate were found. Thus the 1st peak contained only proteoglycans and no collagen, while the entire collagen was found in the 2nd peak. The individual fractions were inoculated into rabbits to obtain antibodies. Compared to the fraction containing macromolecular proteoglycans, antisera proved remarkably suitable particularly for analysis of human spleen, yet also of other organs. These anti-cartilage antisera visualized primarily spongy structures of pericapillary sheaths and circumferential reticulum of the periarterial lymphatic sheaths, but to a lesser extent also other extracellular structures of the spleen and of other organs.

The use of anti-cartilage antisera in the study of human spleen.

Antisera against human hyaline cartilage, particularly antisera against collagenase-treated residue obtained after extraction of cartilage homogenate with 3.4 mol dm-3 NaCl and extraction with 0.05 mol dm-3 sodium citrate buffer at pH = 3.2, reacted with different fibrillary and membraneous structures of the human spleen. Very intensive immunofluorescence was observed in special spongiform structures at the end of the arterial system, referred to as spongy arterial terminals. Fibrillary and membraneous structures situated perivascularly, in lymphatic sheaths and the periphery of follicles displayed similar reactions. The discontinuous walls of sinuses and reticular fibrils in pulp cords reacted less intensively. The given antisera are used to advantage as immunohistologic reagents since they clearly visualize the basic structure, and particularly the spongy arterial terminals, otherwise inconspicuous. In the hyaline cartilage of the rib and knee, the cartilage channels yielded the most intensive reaction. Of the other organs studied the marked reaction in the mesangium of the renal glomerulus was the most remarkable.

The use of lectins identified with specific antibodies in lectin histochemistry of NZB/W F1 mouse kidney.

The affinity of Helix pomatia, peanut, Pisum sativum, soy bean, and wheat germ agglutinins to various nephron parts of NZB/W F1 mice was different and is assumed to be age dependent. The affinity of Pisum sativum agglutinin to basal membranes of small renal vessels increased with the age of NZB/W F1 mice. The wheat germ agglutinin bound to structures with alkaline phosphatase activity.

Immunofluorescence and enzyme cytochemical analysis of arterial terminals in the human spleen.

The simultaneous application of immunofluorescent and enzyme cytochemical methods made it possible to analyze the components of the walls of the arterial branches and arterial capillaries, and particularly the spongy arterial terminals in the red pulp of human spleen. The endothelium was identified by alkaline phosphatase demonstration, the basement membranes (BM) by means of antisera to glomerular BM, the media by using the same antisera, and the adventitia by means by antisera to hyaline cartilage, which are presumed to react particularly with adventitial structures. In the spongy arterial terminals visualized by means of anti-hyaline-cartilage antisera, the endothelium and mostly also BM could be detected, yet the enzyme cytochemical reaction to acid phosphatase and alpha-naphthyl acetate esterase and the immunofluorescent reaction to lysozyme yielded negative reactions. The findings suggest that neither macrophages nor other cells containing the given enzymes in detectable form are accumulated in these structures.

[Antibodies against hyaline cartilage. Antigen isolation and their characterization]. / Protilátky proti hyalínovej chrupke. Izolácia antigénov a ich charakterizácia.

Extracts from human hyaline cartilage were analyzed quantitatively. The extracts were obtained by the action of salts, enzymatic processing and chromatography on Sephadex G-200 columns. The obtained fractions were characterized by thin layer chromatographic methods, electrophoretically, and immunochemically by means of polyclonal antibodies to individual components. Qualitative analysis of amino acids and electrophoretic separation showed that the 1st fraction contained mainly proteins and glycosaminoglycans after separation on Sephadex G-200, while the 2nd fraction exhibited mainly amino acids characteristic for collagen.

[Lectin histochemical analysis of the epithelial lining of the appendix in rabbits]. / Lektínová histochemická analýza epitelovej výstelky králicieho apendixu.

The binding of PSA, PNA, HPA, WGA, Con A, LCL, RCA, SBA, and PHA lectins to epithelial structures of the normal rabbit appendix was studied. Differences were observed in the affinity of some lectins to the epithelium of the intercryptal lining of the rabbit appendix, to the epithelium hemming the glandules and crypts of the domes of Peyer's patches. The obtained results document the difference in the affinity of individual batches of anti-WGA antibodies to this lectin after its binding to the epithelial structures studied. This implies the possibility that differently reacting antibodies may develop to different commercially available batches of WGA. Precipitation of WGA at sites of alkaline phosphatase occurrence demonstrates the relationship of this enzyme to WGA binding sites in tissues.

[Structural basis of appendiceal function in rabbits]. / Struktúrne základy funkcie králicieho apendixu.

Enzymo-, immuno- and lactin-histochemical methods were used to study the structure of the rabbit appendix wall. The value of some structural components in the function of this part of the intestine is discussed. Some findings were documented electronmicroscopically. In addition to its resorptive function, the rabbit appendix is equipped with a potent defense mechanism against adverse environmental effects of the appendix content. Individual structures of this defensive barrier are closely characterized with regard to cellular equipment and possibilities of its morphological identification.

[Structural basis of the spleen in rats]. / Struktúrne základy funkcie sleziny potkana.

The basic function of the spleen in the rat, similarly as in man, is to cleanse the blood of damaged old particles of the body itself, but also of foreign particles. To fulfill this function, the spleen is equipped with the white and red pulp with a specific structure of blood circulation. In an open system of circulation, blood from the arterial terminals opens into the cords of the red pulp, where it is adequately processed. From the cords the blood gets through the walls of sinuses, acting as the last filtration barrier, into the lumen of sinuses and then into the venous circulation. Unlike the human spleen, this organ in the rat has a marked marginal sinus and channel systems bridging the marginal zone. The channels mediate the circulation of lymphocytes between the red white pulp.

[Morphology of systemic anaphylactic reaction in rabbits to bovine serum albumin and ferritin. I. Manifestations of the reaction after antigen administration: laboratory, macroscopic and optical microscopic findings]. / K morfológii systémovej anafylaktickej reakcie králikov na hovädzí sérový albumín a feritín. I. Niektoré prejavy reakcie organizmu na podaný antigén, laboratórne, makroskopické a opticko-mikroskopické nálezy.

A report is presented on systemic anaphylactic reaction [SAR] of rabbits to bovine serum albumin and ferritin. Active anaphylactic sensibilization with a high yield of sensibilized animals was achieved by administering antigens to rabbits aged a few hours up to 3 months. Tre rabbits showed no presence of precipitins in the circulation, but there were reagins against bovine serum albumin or ferritin. There were various types of SAR activity. Macroscopic findings showed acute emphysema and signs of right heart overload. Apart from acute pulmonary emphysema, histology showed occasional bronchospasm and certain manifestations of RES activation as there was evidence of oedematous infiltration of the peribronchial and perivascular spaces. The above morphological findings cannot as yet be regarded as pathognomonic for SAR.

[Reactivity of antibodies to collagen types I to IV and antibodies to chondroitin sulfate in the spleen]. / Reaktivita protilátok proti kolagénu I.-IV. typu a protilátok proti chondroitínsulfátu v slezine.

Antibodies to collagen type I and III reacted negatively, antibodies to collagen type IV positively with reticulin, trabeculae and circumferent reticulum of lymphatic sheaths, poorly positively with capsula, strongly positively with subcapsular zone. Antibodies to collagen type II reacted positively with capsula, poorly with subcapsular zone, strongly with sinus wall and poorly with trabeculae. They did not react with circumferent reticulum of periarterial lymphoid sheaths. Antibodies to collagen type II and IV reacted positively with central arteries. Antibodies to chondroitinsulphate C reacted poorly and antibodies to chondroitinsulphate B strongly positively with sinus walls and oval cells spread in the white and red pulpa. Antibodies to chondroitin sulphate A reacted similarly as antibodies to chondroitinsulphate B.

[Dermoid cyst of the spleen]. / Dermoidná cysta sleziny.

The case report of a 14-year old girl is given. In her spleen of the weight of 1580 g an irregular cyst of glossy inner surface and with striking trabeculation developed. It was filled with clear greenish liquid. The wall was formed by hyalinised collagen tissue which was covered by epidermoid and cuboidal epithelium on the inner surface. Vessel conglomerates were frequent, some bleeding or mononuclear infiltrates, foci of giant cell granulation tissue, lymphoid or fatty tissue were present.

A comparative study of the morphology of the systemic anaphylactic reaction (SAR) and the shock reaction induced by antigen-antibody complexes in rabbits.

Differences in the morphological picture of the systemic anaphylactic reaction (SAR), and of the shock reaction induced by antigen-antibody (ag-ab) complexes in rabbits, are described. In rabbits SAR is characterized by acute distension of the lungs, with oedematous swelling in the vicinity of bronchi and veins, intravascular stasis of basophils without degranulation or visible fixation of the inducing antigen, small clusters of blood platelets, leukostasis, minimal decomposition of neutrophil granulocytes and absence of circulating or fixed ag-ab complexes. The shock reaction induced by ag-ab complexes is characterized by acute distension of the lungs without bronchospasm, swelling of the vicinity of bronchi and veins, hyaline thrombi particularly in the pulmonary microcirculation, leukostasis, formation of antigen-antibody complexes and thrombocyte clusters in the microcirculation, and by phagocytosis of these complexes and thrombocytes by various cells.

On morphology of anaphylactic reaction elicited by ferritin.

The differences between the systemic anaphylactic reaction (SAR) of guinea-pigs actively sensitized with ferritin and those guinea-pigs who were administered with ferritin-antiferritin complexes, which were of a clinically and macroscopically identical result, are described by means of methods of the morphologic examination. Except some details, SAR of guinea-pigs and rabbits elicited by ferritin possesses a similar morphological picture.