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Biomedical subjects

Z Lojda

Publications and source records attributed to Z Lojda.

At least 55 records · Page 3Linked to original sources

Histochemistry of the gastric and duodenal mucosa under cimetidine and carbenoxolone treatment.

Histological and histochemical patterns of the gastric and duodenal mucosa was investigated in nine subjects with duodenal ulcer or ulcerlike syndrome after 4 to 5 weeks treatment with cimetidine or carbenoxolone. No spectacular changes were found after carbenoxolone except for a slight increase in goblet cells in the duodenal bulb. In the patients treated with cimetidine, findings suggesting an influence of this drug on the biochemical cytology of parietal cells and some interference in the differentiation of enterocytes were established.

Adult

[The ratio of enterocytes and goblet cells in the mucosa of the small intestine in experimental infection of piglets with the coccidium Isospora suis].

The proportions of cup-shaped cells and enterocytes were studied in piglets infected the first and fifth day after birth with 200,000 oocysts of Isospora suis. The greatest imbalance was found in the ratio of the cells of the intestinal mucous membrane in the region of middle and lower jejunum (ratio of 10:1.75)--this was recorded the third to fourth day after infection when the ratio made 10:0.62, whereas in the control animals it was 10:5.7. On the eighth to ninth day after infection the number of cup-shaped cells began to increase, the ratio reaching the values of 10:2.62. Even the 13th day after infection the ratio remained far from normal (10:2.50).

Animals

Enzyme histochemistry in the developing suprarenal gland of human embryos.

The localization of the activities of some selected phosphatases, peptidases and dehydrogenases were studied in cryostat sections of the developing anlage of the suprarenal gland of human embryos from 8 to 20 weeks of the intra-uterine life. In the youngest fetuses under our notice (weeks 8-12), the activity of alkaline phosphatase (AP) on the cellular membranes of the fetal cortex was very low. In contrast, the activity of acid phosphatase (AcP) was comparatively high. Peak activity was found in the cells of the central zone of the fetal cortex. Compared to the activity of the latter, the activity of non-specific esterase (ANE) was somewhat lower. Both its localization and the gradient were identical with those of acid phosphatase. Of the peptidases studied, only dipeptidyl peptidase IV (DPP IV) exhibited slight activity in deeper layers of the primitive fetal cortex after week 8. The other peptidases exhibited only traces of activity. As early as in the first stages followed, the activity of glycero-3-phosphate-dehydrogenase (alpha-GPDH) was very high in all cells of the differentiating fetal cortex. The intensity of the activity of succinate dehydrogenase (SDH) was markedly lower. In older fetuses (weeks 13-20) there was a gradual increase in the activities of most enzymes, seen, after week 15 of the intrauterine life, also in the cells of the so-called definitive cortex. Most pronounced were the increases in the activities of acid phosphatase and non-specific esterase. The relatively low activities of the enzymes under study point to a relatively low degree of cell differentiation of both the primitive and, after week 15, the definitive cortex. Pronounced morphological and functional changes occur after the 20th week of the intrauterine life.

Adrenal Glands

[Comparison of tolerance to hydrophilic contact lenses made of Hema (37% H2O) and Hema-Degma (55%, 65% H2O) in the rabbit eye. I. Changes in corneal transparency due to disturbed hydration].

The authors compared on the rabbit eye the tolerance of hydrophilic contact lenses with equal parameters (0.2 mm central and peripheral thickness, 7.4 radius, 15 mm diameter) with a different degree of hydration (37% H2O-Hema), (55% and 65% H2O-Hema-Degma) during continuous wear for a period of two weeks (1, 2, 3, 4, 7 and 14 days). Special attention was devoted to changes in the transparency of the cornea. Changes of the transparency due to wearing of contact lenses were due to changes of corneal hydration. The cause of increased corneal hydration were metabolic and later also morphological disorders in the corneal endothelium. The activity of Na+-K+-dependent adenosine triphosphatase and gamma-glutamyl transferase were reduced, followed by a change in the shape and size of endothelial cells. Later the activities of both enzymes were reduced also in the epithelium. Keratocytes had reduced alkaline phosphatase and gamma-glutamyl transferase activities. The staining properties of glycosaminoglycans in the stroma remained, however, unaltered, similarly as the activity of acid glycosidases and other investigated lysosomal enzymes. The onset of increased corneal hydration caused by a disorder of the active water ion transport and of metabolites in the cornea depended on the percentage of water in hydrophilic contact lenses. It was observed latest after application of contact lenses with 65% water.

Animals

[Comparison of tolerance to hydrophilic contact lenses made of Hema (37% H2O) and Hema-Degma (55%, 65% H2O) in the rabbit eye. II. Changes in corneal transparency due profound degenerative processes].

The authors investigated on the rabbit eye the effect of long-term continuous wearing (14, 21, 28 days) hydrophilic contact lenses (h. c. l.) with varying degrees of hydration (37%, 55%, 65% water). Long-term wearing of the lenses caused disorders in all corneal layers. These changes are identical in all types of contact lenses, only their onset is earlier and the extent is more marked in contact lenses with a low hydration (37% water). Contrary to epithelial changes in the corneal stroma, the changes are qualitatively different depending on the degree of hydration of the contact lens. Prolonged application of contact lenses with 37% water leads to infiltration of the stroma by inflammatory cells which carry the activities of destructive lysosomal enzymes. The latter are gradually released into tissues and local degenerative processes develop which in some instances culminate by ulceration. Other corneas are vascularized. The described changes in stroma were not found in hydrophilic contact lenses with 55% and 65% water. After prolonged application of highly hydrated contact lenses (65% water) the authors, however, observed uptake of liquid from the upper third of the stroma due to the contact lens which dried on the eye and acted on the cornea as a hypertonic solution. In this subepithelial zone the staining of glycosaminoglycans was reduced, similarly as the enzymatic activities in keratocytes which suggests serious degenerative processes, i. e. disorders in the synthesis of components of the ground substance of the corneal stroma.

Animals

[Mucus synthesis in the goblet cells of the small intestine in experimental infection with the coccidium Isospora suis in piglets].

The state of mucus synthesis in the goblet cells of the small intestine was studied in conventional piglets infected with a dose of 200,000 oocytes of the coccidium Isospora suis the first and fifth day after parturition. The synthesis of mucus and its chemical characteristics undergo significant changes during the third and fourth day after infection. The activity of acid and neutral mucous substances declines; their level and the physiological synthetic function of goblet cells begin to return to the normal during the period starting on the eight to tenth day after infection. However, there were no fully functioning goblet cells in the broken numerical ratio even at the end of the period of investigation, i.e. the 13th day after infection. The thin surface layer of mucus remained almost unchanged within the whole extent of the small intestine parts studied.

Animals

[Alkaline phosphatase activity levels during infection with the coccidia Isospora suis in piglets].

The activity of alkaline phosphatase (EC 3.1.3.1.) was studied in the small intestines of piglets infected with the coccidia Isospora suis on the first and fifth day of age. Alkaline phosphatase activity is reduced significantly the third to fourth day after infection (DAI) throughout the small intestine in piglets infected on the fifth day of age. Similar results were obtained in the piglets killed the eighth to tenth DAI which had also been infected on the fifth day of their age. Piglets infected the first day of age and killed on the 8th to 13th DAI exhibited no larger differences from the above-mentioned groups, but alkaline phosphatase activity was distributed more regularly all over the surface of the villi. The normal values, comparable with the controls, were not obtained within 13 days after infection (piglet age 14 days).

Alkaline Phosphatase

[Activity of selected dehydrogenases and monoamine oxidases in the small intestine of gnotobiotic piglets infected with the coccidium Isospora suis].

In the small intestine of 16 gnotobiotic piglets infected a day post partum (DPP) by Isospora suis coccidia the activities were studied of selected dehydrogenases and monoaminoxidase (O2 oxidoreductase, MAOx, EC 1.4.3.4.). The following dehydrogenases were investigated: succinate dehydrogenase (SDH, EC 1.3.99.1.), glycerol-3-phosphate dehydrogenase (glycerol-3-phosphate:menadion oxidoreductase, GPOX, EC 1.1.99.5.) and tetrazolium oxidoreductase (NADH, ES 1.6.99.3.). The activities of NADH and GPOX were found to decrease, a decrease being somewhat milder in MAOx, at a high infection dose of I. suis oocysts (750,000 oocysts), in comparison with the control, already on the first day after infection (DAI). The SDH levels did not change. In piglets infected by a low infection dose of I. suis oocysts a double marked decrease (negative to slightly positive finding) was recorded in the period from the third to the eighth day after infection (DAI). A similar pattern with a longer time interval between the decreases was observed in GPOX (4th to 11th day after infection). The findings of SDH and MAOx activities were different. The SDH activity is maintained at the same level (++) for the whole period of investigation and there occurs a decrease (+) only on the 9th day after infection, persisting until 11 DAI. The MAOx activity and its change correspond to the SDH activity; the difference being that in the second group the starting level is high ( ) and on the eleventh day after infection it is low or medium (0-++), in comparison with the standard. This variability is discernible from 8th DAI.

Animals

Disturbances in the rabbit cornea after short-term and long-term wear of hydrogel contact lenses. Usefulness of histochemical methods.

The influence of soft contact lenses (SCL) with low (37%, L) and high (65%, H) water content on rabbit corneas was investigated. The lenses were worn continuously for 1, 2, 4, 7, 10, 14, 21 or 28 days. The changes in corneal transparency, hydration and enzyme activities were studied. A slight change in corneal transparency due to higher hydration caused by a decreased activity of Na+-K+-dependent adenosine triphosphatase (Na+-K+-ATPase) in the corneal endothelium is followed by a decrease in the activity of gamma-glutamyl transferase (GGT). Slight morphological disturbances appear within 4 days in animals wearing SCL (L). SCL (H) produce similar changes one week later. Subsequently, the corneal epithelium becomes thinner and changes in the size of corneal endothelial cells are obvious. Disturbances of enzyme activities in cells of all corneal layers are present. In the epithelium highly increased activities of acid glycosidases, acid phosphatase, and dipeptidyl peptidase I and II, in keratocytes decreased activities of alkaline phosphatase and GGT, and in the endothelium decreased activity of Na+-K+-ATPase and GGT were found. These changes are more severe after SCL (L). In this case, inflammatory cells displaying high activities of lysosomal hydrolases appear in the anterior part of the stroma during the 3rd and 4th weeks and local degradation of glycosaminoglycans and proteins takes place. In contrast, after SCL (H) a remarkable thinning of the corneas was observed during extended wear, accompanied by decreased stainability of stromal glycosaminoglycans and highly decreased enzyme activities in keratocytes. The histochemical methods proved very useful in the assessment of lesions caused by a continuous wear of SCL.

Animals

Are Z-Arg-Gly-Phe-Phe-Leu-MNA and Z-Arg-Gly-Phe-Phe-Pro-MNA suitable substrates for the demonstration of cathepsin D activity?

The suitability of Z-Arg-Gly-Phe-Phe-Leu-MNA and Z-Arg-Gly-Phe-Phe-Pro-MNA for the assessment of cathepsin D activity was tested in biochemical and histochemical experiments. Substrates were dissolved in dimethylformamide and used at 0.1-0.5 mM in various buffers over a pH range of 3.5-7.4. Homogenates of various rat organs and isolated purified enzymes [cathepsin D from bovine spleen, dipeptidyl peptidase (DPP) IV from porcine kidney and rat lung] were used as enzyme sources. Pepstatin, di-isopropylfluorophosphate (DFP), p-chloromercuribenzoate, o-phenanthroline and a series of DPP IV inhibitors were used in inhibitor experiments. At pH 3.5 and 5.0, substrates were used in a two-step postcoupling procedure with aminopeptidase M and dipeptidyl peptidase IV as auxiliary enzymes and Fast Blue BB as coupling agent. Results were compared with those obtained with haemoglobin. Above pH 5.0 substrates were used in a one-step postcoupling procedure. Cryostat sections of snap-frozen or cold aldehyde-fixed tissue pieces of various rat organs and biopsies of human jejunal mucosa were used in histochemical experiments. As in biochemical tests a two-step procedure was used in the pH range 3.5-5.0, but Fast Blue B was used in the second step for the simultaneous coupling. Above pH 5.0 a one-step simultaneous azo coupling procedure was used with Fast Blue B as coupling agent. At pH 3.5 the hydrolysis rate of both synthetic substrates was about 100x lower than that of haemoglobin when cathepsin D from bovine spleen was used.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Naphthylamine

Histochemistry of proteases in ependyma, choroid plexus and leptomeninges.

Aminopeptidase M (APM), aminopeptidase A (APA), dipeptidyl peptidase IV (DPP IV) and gamma-glutamyl transferase (GGT) were demonstrated histochemically in cryostat sections of the rat brain to show the reaction pattern of ependyma, choroid plexus and leptomeninges. GGT was only demonstrable in the cell membranes of ependymal cells and in the leptomeninges; however, APA, APM and DAP IV showed a variable degree of activity in the capillary endothelium of the choroid plexus as well as in the leptomeninges. On the basis of these results, it is postulated that peptides in the cerebrospinal fluid can be cleaved extraventricularly by the enzymes demonstrated in the leptomeninges.

Animals

Dipeptidyl peptidases of human lymphocytes.

In human lymphocytes three dipeptidyl peptidases were discovered in our laboratory. For a correct demonstration of activities of these enzymes discriminating substrates must be used. Dipeptidyl peptidase IV (DPP IV) is revealed with Gly-Pro-4-methoxy-2-naphthylamide (Gly-Pro-MNA) and Fast Blue B (FBB). It is present in the surface membrane of about 40% lymphocytes of the peripheral blood. Only T-lymphocytes bear the reaction. Reacting lymphocytes belong predominantly to OKT4+ subset. Some OKT8+ lymphocytes also react. With more sensitive substrates (Lys-Pro-MNA, Phe-Pro-MNA and Ala-Pro-MNA) a co-reaction of DPP II was demonstrated "in situ" and in zymograms. In haemoblastoses a positive reaction in cells indicates their derivation from the T-lineage of lymphocytes. A negative reaction does not exclude a T-cell malignancy, however. A decreased number of DPP IV positive lymphocytes in the peripheral blood indicates a diminished immunocompetent potential of T-cells, e.g. immunodeficiency in patients with malignant lymphoma, gastric and colocrectal carcinoma, AIDS, etc. DPP II demonstrated with Lys-Ala-MNA occurs in about 60% of lymphocytes belonging to T and B subsets. It is localized in lysosomes. Although Lys-Pro-MNA is a more sensitive substrate a co-reaction of DPP IV must always be considered. Patients with chronic B-lymphocytic leukaemia displaying a high number of DPP II+ cells usually have a worse prognosis. DPP I assessed with Gly-Pro-MNA and nitrosalicylaldehyde occurs in about 20% of T and B lymphocytes. The number of positively reacting cells increases after corticosteroid therapy. The influence of the treatment on the activity can be shown very well in histograms of DPP I activity measured by computer-assisted microfluorometry.

Cathepsin C