Cytologic detection of cervical and endometrial carcinoma with other genital tract involvement.

OBJECTIVE: To investigate the possibility of a correct cytologic diagnosis of cervical and endometrial carcinoma with other genital organ involvement. STUDY DESIGN: From uteri removed during hysterectomy due to cervical (33 cases) and endometrial (44 cases) cancer, samples were taken by cytobrush or spatula from the ectocervix, endocervix and endometrium of uteri opened longitudinally. Smears and cytosediments were stained by the Papanicolaou polychrome method. Moreover, acid beta-galactosidase activity was demonstrated in serial cytosediments by the indigogenic method of Lojda. From quenched tissue samples taken from the same sites as those for cytology, a series of cryostat sections was prepared and stained by hematoxylin and eosin or azure A, or subjected to the reaction for acid beta-galactosidase. RESULTS: In 17 of 33 patients with cervical cancer, the same type of cancer was also found in smears of the endocervix and endometrium. In six patients the type of cancer was different. Of 44 patients with endometrial cancer, 16 had an endocervical malignancy of the same type. In seven cases the type of cancer was different. The reaction for acid beta-galactosidase helped in the differentiation between squamous (negative reaction in cancer cells) and cylindrocellular (positive reaction) cancer in cytologic preparations. CONCLUSION: Before treatment, it is necessary to determine if there is involvement of the endocervix in endometrial cancer and of the endometrium in cervical cancer. Routine cytologic examination supplemented by the reaction for acid beta-galactosidase proved to be useful for this purpose.

Are intestinal tumours in Apc+/-mice a suitable model of colorectal carcinoma in man?

In snap frozen sections of the duodenum, jejunum, ileum, the right and left colon of APC+/-mice mucosubstances, activities of brush border glycosidases and proteases, immunoreactivity of sucrase and activities of some enzymes of pericellular proteolysis were studied. Multiple adenomas (tubular or tubulovillous) the numbers of which decreased in the aboral direction occurred in the small intestine. Two tubulovillous adenomas with dysplastic nuclei but with no invasion were found in the right colon. The morphological and histochemical findings resembled those of human colorectal tumours. Activities of brush border enzymes and sucrase immunoreactivity were decreased to various extent or were not present at all. The findings fluctuated even within the same section. Activities of enzymes of pericellular proteolysis were slightly increased in comparison with non affected mucosa. This model is suitable and deserves further studies.

[The effect of exposure to silicon dioxide on the occurrence of porphyria tarda]. / Vliv expozice oxidu kremicitému na výskyt porfyria cutanea tarda.

The incidence of porphyria cutanea tarda was studied in two groups of silicon dioxide risk workers and compared to that in a control group. In the group with higher exposure to SiO2 the illness occurred in 12 out of 440 workers, in the group with lower exposure in 12 out of 1000 workers. In the control group porphyria was found in 18 out of 12,100 individuals examined. The difference in the incidence of the illness between the SiO2 risk individuals and the control group is statistically significant (p less than 0.01), while the difference between the group with variously high SiO2 exposure is on the level of statistically significance (p less than 0.05). The causes of the higher incidence rate of porphyria cutanea tarda in free SiO2 risk workers are discussed.

[The effect of exposure to silicon oxides on the occurrence of porphyria cutanea tarda]. / Vliv expozice oxidu kremicitému na výskyt porfyria cutanea tarda.

The incidence of porphyria cutanea tarda was studied in two groups of silicon dioxide risk workers and compared to that in a control group. In the group with higher exposure to SiO2 the illness occurred in 12 out of 440 workers, in the group with lower exposure in 12 out of 1000 workers. In the control group porphyria was found in 18 out of 12,100 individuals examined. The difference in the incidence of the illness between the SiO2 risk individuals and the control group is statistically significant (p less than 0.01), while the difference between the group with variously high SiO2 exposure is on the level of statistical significance (p less than 0.05). The causes of the higher incidence rate of porphyria cutanea tarda in free SiO2 risk workers are discussed.

[Occurrence of symptomatic liver porphyria in workers at risk for exposure to free SiO2 during subway construction]. / Výskyt symptomatické jaterní porfyrie u pracovníku z rizika volného SiO2 pri výstavbe metra.

Symptomatic hepatic porphyria was followed up in workers exposed to the risk of free SiO2. This disease was found in 12 out of 998 workers engaged in the construction of the underground and exposed to SiO2 (group I), and in 12 at-risk persons out of 440 patients from different workplaces who were hospitalized for SiO2 exposure (group II). The average age of the workers of group I was 40.6 +/- 7.7, that of the group II 54.9 +/- 13.3. In both these high-risk groups, porphyria was found to be significantly more frequent than in the control group not exposed to free SiO2. The authors discuss the long-lasting effects of free SiO2 on the occurrence of symptomatic hepatic porphyria.

Uneven distribution of alkaline phosphatase in individual layers of rabbit and ox cornea. Histochemical and biochemical study.

In the rabbit and bovine cornea the activity of alkaline phosphatase using histochemical as well as biochemical methods was investigated. Biochemically the enzyme activity was studied in separated corneal layers. In the histochemical investigation the best results were obtained in cryostat sections using the azocoupling method with naphthol AS-MX phosphate and Variamine Blue RT Salt. The enzyme activity was found not only in the epithelium and endothelium (as was described previously) but even in keratocytes. The mutual relation of activities in the epithelium and in keratocytes differed in both species. The overall activity found by histochemical methods is in good agreement with the biochemical determination of alkaline phosphatase (p-nitrophenyl phosphate as the substrate). Besides the histochemical approach shows an uneven distribution of alkaline phosphatase activity in individual cells which cannot be assessed by the biochemical determination.

Distribution of acid phosphatase, beta-glucuronidase, n-acetyl-beta-d-glucosaminidase and beta-galactosidase in cornea of albino rabbit.

Activities of acid phosphatase, beta-glucuronidase, N-acethyl-beta-D-glucosaminidase and acid beta-galactosidase were investigated histochemically in rabbit corneas. Frozen sections after block fixation in cold 4% formaldehyde with 1% CaCl2 followed by washing in cold physiological saline as well as cold microtome sections of corneas quenched in petroleter chilled with acetone-dry ice mixture, transferred to nonprecooled slides or semipermeable membranes were used. Standard aqueous media were employed in the case of free-floating frozen sections of fixed corneas as well as of cold mictrotome sections (postfixed in cold 4% formaldehyde). Agar media were used in connection with the technic of semipermeable membranes. Gomori method (in the case of acid phosphatase), simultaneous azocoupling methods (substrates derivated of naphthol-AS-BI with hexazonium-p-rosanilin) in the case of acid phosphatase, beta-glucuronidase and N-acetyl-beta-D-glucosaminidase and the indigogenic method in the case of acid beta-galactosidase were applied. Enzyme activities in sections of fixed corneas were minimal in comparison with those in cold microtome sections of unfixed material revealed particularly with the technic of semipermeable membranes which is to be preferred. This technic is recommended in studies concerned with lysosomal enzymes in the cornea, particularly in keratocytes. All enzymes investigated were present in corneal epithelium, keratocytes and endothelium. Acid phosphatase displayed the highest activity followed by beta-glucuronidase and acetyl-beta-D-glucosaminidase. The activity of beta-galactosidase was the lowest. For the demonstration of activities in keratocytes sections parallel to the surface are very suitable. In these sections enzyme activities were demonstrated in small granules (apparently lysosomes) present in the central part of their cytoplasm as well as in projections. Diffuse staining was also seen, being the highest in the case of acid phosphatase.

Alkali burns of the rabbit cornea. I. A histochemical study of beta-glucuronidase, beta-galactosidase and N-acetyl-beta-D-glucosaminidase.

In alkali burned rabbit corneas activities of beta-glucuronidase, N-acetyl-beta-D-glucosaminidase and acid beta-galactosidase were studied histochemically in various time intervals after the traumatization. The technic with semipermeable membranes was employed. Within four days after the injury enzyme activities in the traumatized area were almost lacking. The corresponding activities in the unaffected part of the cornea were within the norm. On the 7th day enzyme activities were on an increase (but still subnormal) in the traumatized area. This area was surrounded by a zone of keratocytes with high levels of enzyme activities. This was particularly remarkable in keratocytes subjacent to the epithelium. The activation of all enzymes studied was present in the basal layer of the epithelium and in the endothelium as well. On the 14th day enzyme activities in the traumatized area were nearly restored and on the 32nd day they could not be distinguished from the normal cornea. Beta-galactosidase displayed a relatively maximal increase in the activity of all enzymes investigated.

Alkali burns of the rabbit cornea. II. A histochemical study of glycosaminoglycans.

In alkali burned rabbit cornea the stainability of glycosaminoglycans in cold microtome setions was investigated. Staining by Alcian blue in 3% acetic acid, Alcian blue in various MgCl2 concentration and toluidine blue (pH 4.5) was employed. From the 1st to the 4th experimental day the intensity of reactions was decreased. This is most probably due to an increased hydration of the corneal stroma. On the 7th day hydration was markedly suppressed and reached nearly the normal level. In this time interval a decreased stainability of glycosaminoglycans was seen accompanied by a complete loss of staining in the marginal zone. On the 14th day the stainability in the traumatized area began to restore and in the marginal zone appeared. On the 32nd day the staining intensity of both areas was normalised, however when lower concentrations of MgCl2 were used; in the presence of higher concentrations of MgCl2 the decreased staining intensity persisted and points to a lower sulfatation of glycosaminoglycans. This was particularly remarkable in the area bordering the injured zone. This decrease runs parallel to the increased activities of acid glycosidases (especially of acid beta-galactosidase) which were reported previously.

The histochemical demonstration of aminopeptidase with bromoindolyl leucinamide.

The indigogenic method for aminopeptidase of Pearson et al. (1963) was critically evaluated. The localization obtained with it is not correct due to diffusion artifacts. Ferricyanide cannot be used as an oxidation agent. Based on experiments with other oxidation agents (phenazonium methosulfate, nitro BT, tetranitro BT) a new method was devised. The recommended incubation medium contains 0.9 mM L-N-(5-bromoindol-3-yl) leucinamide hydrobromide (chloride), 0.73 mM tetranitro BT, 0.27 mM phenazonium methosulfate and 0.67 M phosphate buffer pH 7.4. The enzyme activity is indicated by the deposition of tetranitro BT formazan. Results with this method in rat kidney, jejunum, liver, lung, and submaxillary gland, in monkey kidney and jejunum, and in human jejunal biosies are almost identical with those obtained with L-leucyl-4-methoxy-beta-naphthylamide applied in a simultaneous azocoupling procedure. The given principle of the demonstration of aminopeptidase activity with an indolylamine substrate deserves a further exploration in the demonstration of peptidases "in situ" both on optical as well as electronmicroscopical levels.