[Tumors caused by methylcholanthrene and lapachol. Follow-up of development with cytology]. / Tumores provocados por metilcolantreno y lapachol. Seguimiento del desarrollo mediante citología.

This present work was carried out in order to study the effect of Lapachol (LAP) administrated to rats simultaneously with a chemical carcinogen 20-Methylcholanthrene (MCA). Animals were divided in 4 groups: A-Group treated with 80 mg of MCA I(n = 11 rats), B-Group treated with 80 mg of MCA+LAP 100 mg/kg weight/day, (n = 15 rats), C-Group treated with LAP 100 mg/kg weight/day (n = 12 rats), D-Group control-no treatment (n = 13 rats). Cytological studies as well as cytochemical techniques allowed the recognition of benign and malignant conditions at the time of the tumor appearance. Histopathological evaluation posteriorly confirmed the development of tumors in 53% of the animals in group B. Morphologically consistent with poorly differentiated adenocarcinomas of the salivary glands and fibroadenomas of the breast in 18.2% (2/11) of the rats in group A. Besides the presence of one or several supra-hepatic nodules in vecinity of the suspensory ligament corresponding to nodular hyperplasia were observed in 33% (4/12) of group C and in 13.3% (12/15) of group B. No nodules were observed in groups A and D. Tubular dilatation of kidneys were noted in 60% (9/15) and 83.3/ (10/12) of the rats in group E and C respectively. From the original salivary gland tumor a series of transplantables tumors were developed and followed by cytological evaluations. The importance of the cytological and histopathological diagnosis for the pharmacological effects studies of certain drugs like Lapachol that are widely used as antitumoral agents without exact knowledge of the adverse effects is emphasized.

Glycosidases in the rat uterus during the oestrus cycle.

Seven acid glycosidases activities have been measured in normal rat uterus during the oestrus cycle. It has been observed that alpha-fucosidase, alpha-mannosidase, beta-galactosidase, alpha-galactosidase, aryl sulfatase, acid phosphatase and hexosaminidase activities changed cyclically during the oestrus cycle. From onward oestrus to metaoestrus the enzyme activities are in their highest level, but then decline slightly towards the resting one, as the cycle progresses. It is possible that changes in glycosidases content bear a lysosomal relationship, since it is known the increase in the lysosome content of the uterus during the oestrus and metaoestrus. The increased enzyme content could be related to uterus glycoproteins secretion and degradation during the normal oestrus cycle.

Glycosidases from the rat uterus.

Six glycosidase activities were detected in homogenates of the rat uterus by using the p-Nitrophenyl-glycoside as substrate. The enzyme activities were separated by Sephadex G-200 column chromatography. The uterus contained alpha-galactosidase, beta-galactosidase and N-acetylglucosaminidase activity which had a similar pH optima of 4.4, whereas the pH optimum for beta-fucosidase was found to be of 4.4 - 4.6. The alpha-mannosidase activity showed two peaks of maximal activity at pH 5.0 and pH 6.0. The uterus contained two forms of the alpha-L-fucosidase activity separable by Sephadex G-200 which had a pH optima of 4.4 - 4.6 (form I) and pH 5.0 (form II). The glycosidases differed in heat stability, Km values and other several characteristics, which suggests that each enzyme apparently did not show any cross-contamination.

Glycosidases in the rat uterus during the oestrus cycle.

Seven acid glycosidases activities have been measured in normal rat uterus during the oestrus cycle. It has been observed that alpha-fucosidase, alpha-mannosidase, beta-galactosidase, alpha-galactosidase, aryl sulfatase, acid phosphatase and hexosaminidase activities changed cyclically during the oestrus cycle. From onward oestrus to metaoestrus the enzyme activities are in their highest level, but then decline slightly towards the resting one, as the cycle progresses. It is possible that changes in glycosidases content bear a lysosomal relationship, since it is known the increase in the lysosome content of the uterus during the oestrus and metaoestrus. The increased enzyme content could be related to uterus glycoproteins secretion and degradation during the normal oestrus cycle.

Glycosidases from the rat uterus.

Six glycosidase activities were detected in homogenates of the rat uterus by using the p-Nitrophenyl-glycoside as substrate. The enzyme activities were separated by Sephadex G-200 column chromatography. The uterus contained alpha-galactosidase, beta-galactosidase and N-acetylglucosaminidase activity which had a similar pH optima of 4.4, whereas the pH optimum for beta-fucosidase was found to be of 4.4 - 4.6. The alpha-mannosidase activity showed two peaks of maximal activity at pH 5.0 and pH 6.0. The uterus contained two forms of the alpha-L-fucosidase activity separable by Sephadex G-200 which had a pH optima of 4.4 - 4.6 (form I) and pH 5.0 (form II). The glycosidases differed in heat stability, Km values and other several characteristics, which suggests that each enzyme apparently did not show any cross-contamination.

Hypophysis effect on the topographical distribution of toad oviduct mucoprotein components.

In the present work the effects of the hypophysis hormones on oviduct mucoprotein components distribution patterns were studied. Remarkable changes after treating the toad with hypophysis injections were apparent. The distribution pattern for hexose, sialic acid, hexosamine and phosphate from 18 hours hypophysis treated toads were found to be identical with those obtained from preovulatory period animals. On the other hand, the levels for mucoprotein components from hypophysis treated animals were found to be approximately one-half or more higher than those obtained from postovulatory period toads. Otherwise, hypophysis treatment of the toads in preovulatory period had not effect on the levels and distribution patterns of mucoprotein components. These results suggest that hypophysis hormones are involved in the increase of the oviduct secretory activity.

Hypophysis effect on the topographical distribution of toad oviduct mucoprotein components.

In the present work the effects of the hypophysis hormones on oviduct mucoprotein components distribution patterns were studied. Remarkable changes after treating the toad with hypophysis injections were apparent. The distribution pattern for hexose, sialic acid, hexosamine and phosphate from 18 hours hypophysis treated toads were found to be identical with those obtained from preovulatory period animals. On the other hand, the levels for mucoprotein components from hypophysis treated animals were found to be approximately one-half or more higher than those obtained from postovulatory period toads. Otherwise, hypophysis treatment of the toads in preovulatory period had not effect on the levels and distribution patterns of mucoprotein components. These results suggest that hypophysis hormones are involved in the increase of the oviduct secretory activity.

Carbohydrate utilization by the avian oviduct during the laying cycle.

In the present work the carbohydrate utilization by the avian oviduct during the egg formation was studied. The pattern obtained for total hexose, fucose, hexosamines, sialic acid, fructose and sulphate demonstrated that in the domestic fowl's oviduct the isthmus and the shell gland secrete the precursors for the protein carbohydrate substance of the mammillae and shell matrix and may contribute during the shell formation with the substrates necessary for the metabolic energy of the organ. The infundibulum only participates as a donor for the metabolic energy.

Carbohydrate utilization by the avian oviduct during the laying cycle

In the present work the carbohydrate utilization by the avian oviduct during the egg formation was studied. The pattern obtained for total hexose, fucose, hexosamines, sialic acid, fructose and sulphate demonstrated that in the domestic fowl’s oviduct the isthmus and the shell gland secrete the precursors for the protein carbohydrate substance of the mammillae and shell matrix and may contribute during the shell formation with the substrates necessary for the metabolic energy of the organ. The infundibulum only participates as a donor for the metabolic energy.

Carbohydrate utilization by the avian oviduct during the laying cycle.

In the present work the carbohydrate utilization by the avian oviduct during the egg formation was studied. The pattern obtained for total hexose, fucose, hexosamines, sialic acid, fructose and sulphate demonstrated that in the domestic fowls oviduct the isthmus and the shell gland secrete the precursors for the protein carbohydrate substance of the mammillae and shell matrix and may contribute during the shell formation with the substrates necessary for the metabolic energy of the organ. The infundibulum only participates as a donor for the metabolic energy.

Dialyzable sialic acid of toad oviduct.

1. Small quantities of dialyzable sialic acid in toad oviduct were found. 2. About one-third of the dialyzable sialic acid reacts directly in the thiobarbituric acid procedure, a finding which suggests that the other two-thirds of the dialyzable sialic acid were probably derived from low molecular weight compounds containing sialic acid. 3. When a soluble fraction was chromatographed on Dowex 1-X4 column, three fractions (I, II and III) were isolated. Peak I was a material which shows identical to authentic NANA, colorimetrically as well as chromatographically. Several studies have indicated that both peaks, II and III, could be sialic acid-bound compounds of low molecular weight.

Dialyzable sialic acid of toad oviduct

1. Small quantities of dialyzable sialic acid in toad oviduct were found. 2. About one-third of the dialyzable sialic acid reacts directly in the thiobarbituric acid procedure, a finding which suggests that the other two-thirds of the dialyzable sialic acid were probably derived from low molecular weight compounds containing sialic acid. 3. When a soluble fraction was chromatographed on Dowex 1-X4 column, three fractions (I, II and III) were isolated. Peak I was a material which shows identical to authentic NANA, colorimetrically as well as chromatographically. Several studies have indicated that both peaks, II and III, could be sialic acid-bound compounds of low molecular weight.

Dialyzable sialic acid of toad oviduct.

1. Small quantities of dialyzable sialic acid in toad oviduct were found. 2. About one-third of the dialyzable sialic acid reacts directly in the thiobarbituric acid procedure, a finding which suggests that the other two-thirds of the dialyzable sialic acid were probably derived from low molecular weight compounds containing sialic acid. 3. When a soluble fraction was chromatographed on Dowex 1-X4 column, three fractions (I, II and III) were isolated. Peak I was a material which shows identical to authentic NANA, colorimetrically as well as chromatographically. Several studies have indicated that both peaks, II and III, could be sialic acid-bound compounds of low molecular weight.