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Peptidylarginine deiminase (PAD) is an enzyme known to be involved in the pathogenesis of rheumatoid arthritis (RA). Since many of the molecular events present in the joints in RA also take place in the injured liver, we postulated in this study that PAD may be involved in liver fibrosis. The objectives of this study therefore were to find out if PAD could be demonstrated immunohistochemically in liver biopsies of patients with chronic hepatitis and if it is associated with METAVIR activity and fibrosis scores. Liver biopsies were obtained from 100 patients with chronic liver diseases between September 2006 and 2007. The biopsies were scored by two histopathologists according to the METAVIR activity and fibrosis scores after histological preparation. Immunohistochemistry for PAD was performed on the biopsies using a monoclonal antibody against PAD. PAD could not be demonstrated in normal liver biopsies but was found in the hepatocytes of patients with chronic hepatitis. PAD labeling could distinguish patients with no fibrosis from either F1 or F2 or F3 or F4 fibrosis. Similarly, PAD labeling could separate patients with no inflammatory activity from those with mild or moderate or severe activity. We concluded that PAD could be demonstrated immunohistochemically in liver biopsies of patients with chronic hepatitis and that its immunodetection was significantly associated with Metavir activity and fibrosis scores., (Copyright © 2009 Elsevier GmbH. All rights reserved.)

A case of storage disease followed up during 12 a was studied by morphological, histochemical, immunological, and biochemical techniques. Data were analysed in an attempt to differentiate an acquired storage by excessive cell degradation from a storage of genetic origin. If, as it is our belief, a conclusion of acquired storage can be made, this observation in which the same cholesterol metabolism abnormalities as in Niemann Pick type C were seen, leads to a rediscussion of their diagnostic significance.

Within the uterine glands, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the activities of G-6-PDH, 6-PGDH, and cytochrome oxidase increase within secreting cells during the 2nd half of pregnancy. The activities of the other enzymes remained almost unchanged during the period of investigation. The description of our results distinguishes between gland neck, middle, and distal part of the secretory unit, respectively. In general, the enzyme activities are similar within the middle and distal gland segments, but lower in the epithelia of the neck region. The activity of dehydrogenases was medium to intensive within the middle and distal gland segments, but only low to medium within the neck portion. Of the hydrolases, the acid phosphatase, ATPase, leucine aminopeptidase, and beta-galactosidase demonstrated an intensive activity within activity secreting cells. The enzyme activities of the gland epithelia are compared with these of the uterine surface epithelia and the histochemical results are discussed in context with their significance in histiotrophic nutrition.

Activity changes of thiamine pyrophosphatase (TPPase), acid phosphatase (aP), non-specific esterase (nE), acid beta-galactosidase (a beta Gal), beta-glucuronidase (beta-Gluc), and beta-D-N-acetylglucosaminidase (NAG) in follicles during atresia were investigated in the ovaries of mice, rats, Mongolian gerbils, hamsters, guines pigs, rabbits, cats, and pigs. Changes of hydrolase activity were highly enzyme dependent, species-specific and mostly confined to the granulosa. Decrease of TPPase activity and increase of lysosomal enzyme activities during atresia appeared to be true for all mammals. The start of activity changes in the time course of atresia depended on the occurrence of the enzyme in the growing granulosa. Continuous increase of lysosomal enzyme activity appeared in follicles where these enzymes could also be found in the growing granulosa. In contrast, when lysosomal enzyme activity was low or could not be detected in the growing granulosa, increased enzyme activity could only be observed at a time when degenerative processes have already progressed considerably. This distribution pattern suggests that hydrolytic enzymes in the granulosa cells as well as hydrolases of invading macrophages participate in this degenerative process. In some mammals, enzyme activity changes appeared in the cumulus oophorus for the first time in advanced stages of degeneration. In some mammals enzyme activity changes were dependent on the developmental stage of the follicle. This stage dependency argues for an interrelationship between activation of lysosomal enzymes and androgen metabolism.

Peptidases, phosphatases, glycosidases, non-specific esterases, succinate dehydrogenase, cholinesterases, and carbohydrate components were studied in bioptic material of the normal and diseased human stomach using well established older, modified older, and new qualitative histochemical methods. For the first time, an enzyme pattern is reported for all regions of the human mucosa. Local and regional enzyme histochemical differences existed between the cardiac, fundic, body, and pyloric mucosa. Differences were absent, however, in the same region, and no differences were found between the anterior and posterior wall and the large and small curvature of the stomach. In cases of histologically less severe gastritis as a rule, enzyme histochemical changes were not found. They were numerous, however, in biopsies of patients with severe gastritis; only amino-peptidases A and M were unchanged. Dipeptidyl peptidase IV was absent; gamma-glutamyl transpeptidase exhibited individual differences. Alkaline phosphatase occurred in the pericapillary stroma and adenosine phosphates were not hydrolysed in atrophic glandular epithelia. Activity increases of lysosomal dipeptidyl peptidase I and beta-D-glucuronidase were typical for inflammatory infiltration processes of the gastric mucosa. Severe atrophy was accompanied by an activity decrease of glandular non-specific esterases, dipeptidyl peptidase II, and beta-N-acetyl-D-glucosaminidase and an activity decrease of the stromal peptidases and glycosidases. Enzyme activity was absent in the gastric glands proper in cases of total atrophy. An increase in macrophage number was primarily linked with an increase in acid phosphatase activity. Alkaline phosphatase, aminopeptidase M and gamma-glutamyl transpeptidase activities were enhanced in malignant neoplasms. High activities of all peptidases and alkaline phosphatase were found in the brush border of surface epithelial cells in cases of intestinal metaplasia. Except for dipeptidyl peptidase I and II, the enzyme pattern corresponds to that of small intestinal enterocytes. Compared with histological routine procedures for gastric diagnosis and assessment of the course enzyme histochemical methods deliver additional information; practically, however, the enzyme histochemical analysis of gastric biopsies are only useful in special cases.

Proteases and gamma-glutamyl transferase linked to plasma membranes, lysosomes and secretion granules were investigated cytochemically together with phosphatases, glycosidases and non-specific esterases in the mature guinea-pig and marmoset yolk sac. Species-independently, the yolk sac epithelial cells showed high activities of lysosomal proteases, glycosidases and non-specific esterases and phosphatases, whereas species-dependent patterns were found for plasma membrane proteases, gamma-glutamyl transferase and phosphatases. Furthermore, the guinea-pig yolk sac epithelium showed marked regional differences and that of both species intercellular differences, which allowed the subdivision of yolk sac epithelial cells in various types. The high activities of all lysosomal enzymes suggests their general importance for the yolk sac function in guinea-pigs and marmosets. The different equipment of the plasma membrane with proteases, gamma-glutamyl transferase and phosphatases indicates a more species-specific physiological role of this part of the yolk sac epithelial cells.

For human mammary and ovarian carcinomas and transplantation tumours of rats clear hints found for an enzymatic-lytic effects of infiltrating tumour cells as a premise of the invasive tumour growth. In contrast in malignant melanomas there is no functional sign for a specific enzymatic-lytic effect of the living tumour cells to the surrounding tissue. In the mamma carinomas and ovarian carcinomas infiltratively growing tumour districts were characterized by an increased activity of the NADH-D, LDH, G-6-P-DH, IDH, MDH, SPase, UE, LAP and beta-GD. The transplantation tumours showed a high number of tumour cells with a high leucine amino-peptidase- and beta-glucuronidase-activity in a middle zone that was localized under the tumour edge district. The increased activity of the LAP and beta-GD found in the infiltration zone of the tumours is considered as an demonstration of a strong proteolytic activity of the tumour cells. The findings are discussed in the aspect of the invasive tumour growth.

As a model alkaline phosphatase and non-specific esterase were investigated with indoxyl substrates and 11 tetrazolium salts of different reducible substances in livers and kidneys of rat. Besides their localizing qualities, the reducibility of the used tetrazolium salts is the most important parameter for their use in histochemistry of hydrolases. The composition of the reaction products depends on the reducibility of the indicator and the pH of the reaction. The of phenazine methosulphate improves the indoxyl-tetrazolium techniques above all in the neutral and acid range.