Use of arylsulphonic acids for histochemical demonstration of cholesterol and its esters.

A histochemical technique is being described using for the demonstration of cholesterol and its esters a reagent composed of sulphuric acid and 2,5-dimethylbenzene sulphonic acid or p-toluene sulphonic acid dissolved in glacial acetic acid. The results of this method have been compared with other methods for cholesterol identification and verified by extraction and chromatographic techniques. The main advantage of the presented procedure is a better preservation of sections, due to the use of smaller amounts of sulphuric acid, permitting thus a more exact histological localization of cholesterol.

Immunohistochemical staining using coupled immunofluorescence and immunocomplex technique.

An immunomicroscopic method combining the use of fluorescent antibodies and enzyme immunocomplex technique is presented. The basic steps of this procedure are: 1. Rabbit antiserum (IgG) to the antigen to be determined, 2. FITC-conjugated antirabbit globulin, 3. Peroxidase + antiperoxidase complex, 4. 3,3-diaminobenzidine + H2O2. The method renders it possible to study simultaneously in the same section antigens and antibodies that are to be localized, and that both by immunofluorescent and immunoenzymatic techniques, it provides means for further elimination of some of the deficiencies of immunofluorescent technique, and in suitable cases it also makes electronoptic examination available.

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.

[The importance of pulmonary angiography in the surgical treatment of congenital heart defects with a left-right shunt and pulmonary hypertension]. / Význam pulmoangiografie pre indikáciu na chirurgickú liecbu vrodených srdcových chýb s l'avo-pravým skratom a pl'úcnou hypertenziou.

The studied series consisted of 14 patients with ventricular septal defect and pulmonary hypertension aged from 3 months to 15 years. Three examination methods were compared: (1) Invasive hemodynamic examination; (2) Pulmoangiographic examination by means of wedge peripheral pulmoangiography; (3) Histological examination of bioptic samples. Direct correlation was found to exist between mean PA pressure, PAR/m2, TPR/SR and pulmoangiographic records, in which the following parameters were evaluated: (a) length of the narrowing of the peripheral branch of the PA, (b) background opacity, (c) circulation time. The histological findings obtained in the bioptic samples corresponded practically in all cases with the hemodynamic and PAG findings. Only in two patients did the histological picture display a less severe degree of affection than found by PAG and hemodynamic examination. The obtained results suggest that in light of surgical indications, PAG can be considered a suitable and valuable supplementary method in assessing borderline findings of pulmonary hypertension in children with congenital heart defects and left-to-right shunt.

[Morphology of ferritin-induced shock in sensitized guinea pigs]. / K morfológii soku vyvolaného feritínom u senzibilizovaných morciat.

With the aid of morphological methoda, differences have been described between the active ferritin shock and the ferritin-antiferritin shock in guinea-pigs, which clinically appear as identical. The two processes can be differentiated from each other on the light microscopical level, but with particular clarity on the electron microscopical level. In contrast to the active ferritin shock, the guinea pigs in whom the ferritin-antiferritin shock had been produced showed agglomerates of platelets and ferritin-antiferritin complexes either present free in the dirculation or ingested in macrophages or granulocytes.

[Experimental administration of a fatal dose of antiglomerular antibodies]. / Experimentálna aplikácia letálnej dávky protiglomerulärnych protilátok.

The administration of a sufficient dose of heterologous antibodies against renal glomeruli in rats produces symptoms of respiratory insufficiency and pulmonary oedema causing death of the animals. Under these experimental conditions the basic pathogenic mechanism appears to be binding of the injected antibodies on the basement membrane structures of pulmonary capillaries resulting in their damage. Such changes are followed by widespread haemorrhagic exsudative changes in the lungs. Localization of the injected antiglomerular antibodies has been studied with the aid of immunohistochemical methods and in addition, by a method using antibodies labelled with radioactive isotopes of iodine, the so-called "paired administration technique". After the administration of a lethal dose of antiglomerular antibodies, they later appeared to be predominantly localized in the lungs. This fact correlates well with the idea of the effect of the antibodies upon pulmonary parenchyma in the present experiment. A remarkable finding was a rather high localization of the injected antibodies in the liver parenchyma which could be explained either by a cross reaction between the antibodies and the antigens of hepatic vascular spaces, or by a shock reaction associated with heart failure and circulatory disturbances.

[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.