Determining acid phosphatase (AcP[E.C. 3.1.3.2.]) and adenosine triphosphatase (ATPase [E.C. 3.6.1.3.]) activity in the lungs of rats following a single administration of ashes from industrial high-heat facilities

There was examined pulmonary tissue of white rats, which had been administered intratrachealy a single dose of the respirable fraction of ashes sample from 6 different power stations elektrohasting plants and hasting plants in Poland (0.2 ml suspension; 50 mg of the examined sample in 0.6 cm3 of NaCl solution). 9 months after the application of the ashes, biopsies of the left lung were taken and there was determined the activity of acid phosphatase (AcP) and adenosinetriphosphatase (ATP-ase) histoenzymatically. There was found sensitivity of these hydrolases and changes of their activity connected with chemical composition of the examined ashes.     

Underfeeding and exposure to short photoperiod alters rat pineal and Harderian gland lysosomal enzyme activities

Harderian gland (HG) weight and lysosomal enzyme activity were evaluated after 21-day-old female rats were singly caged in a long (LP; 14:10 LD) or short (SP; 8:16 LD) photoperiod and fed on one of two dietary regimens (fed ad libitum or 50% underfed) for 50 days; an additional fed and an underfed group of animals in LP were injected every afternoon with 100 micrograms melatonin. Absolute HG weights were significantly lower in all underfed groups compared to their respective fed controls or to the LP fed control group. Absolute HG weights of underfed rats in SP were significantly lower than the underfed rats in LP. Relative HG weights (mg/100 g body wt) were significantly higher in the underfed saline or melatonin-treated groups compared to their respective fed controls; however, HG of the underfed SP group were not different from SP-fed controls. No significant differences in HG acid phosphatase, hexosaminidase, and beta-glucuronidase activities were observed in any of the treatment groups maintained in LP. Acid phosphatase, hexosaminidase, and beta-glucuronidase activities were significantly elevated in HG of underfed animals maintained in SP compared to their respective fed controls or to the LP-underfed group. Both the underfed control and the underfed-melatonin treated groups had lower pineal protein values than their respective fed groups; underfed animals in 8:16 LD had similar pineal protein values compared to those of the fed control group in SP. Significant effects of photoperiod and underfeeding with no interaction between these variables were observed on pineal acid phosphatase. The fed group maintained in 8:16 LD had significantly higher acid phosphatase activity than the fed group kept in 14:10 LD. In conclusion, underfeeding resulted in severely reduced body weights and absolute Harderian gland weights. Increased activity in certain lysosomal enzymes occurred in both the pineal and Harderian gland and in some instances this was dependent upon the light cycle and dietary regimen to which the animals were exposed.     

The influence of coal ashes and soil dust on the activity of two selected hydrolytic enzymes in the liver of laboratory animals

The effect of a single intratracheal application of the electroenergetic ashes and the soil dust on the liver cell was studied by means of the histoenzymatic methods for the determination of the activities of chosen hydrolases (AcP, E.C. 3.1.3.2. and ATP, E.C. 3.6.1.). The degree of the histochemical changes in hepatocytes depended on the kind of the applied dust and the trace elements' content in it. The activity's increase of the acid phosphatase and ATP-ase in each experimental group was observed, especially in the preparations collected from the animals were given the electroenergetic ashes.     

Lysosomal enzymes in the rat harderian gland are altered by either bromocriptine treatment or hypophysectomy and hormone replacement therapy

Four groups of adult male hypophysectomized rats were injected subcutaneously twice daily between 0800-0900 hr and 1600-1700 hr with either saline diluent, 150 micrograms sheep prolactin and/or growth hormone (GH); intact rats received either saline or 150 micrograms bromocriptine twice daily. After 4 days of treatment, lysosomal enzyme assays revealed significant elevations in both acid phosphatase and alpha-mannosidase enzyme activities in the Harderian glands of saline-injected hypophysectomized rats compared to those in intact controls. beta-Glucuronidase levels were depressed and hexosaminidase activity unaffected by hypophysectomy treatment alone compared to intact controls. Lysosomal enzyme activities in hypophysectomized animals treated with prolactin were not different from the hypophysectomized control animals. However, treatment with GH alone or in combination with prolactin had a significant inhibitory effect on beta-glucuronidase, hexosaminidase, and alpha-mannosidase enzyme activities in the Harderian gland of hypophysectomized animals. Bromocriptine treatment in intact rats only elevated acid phosphatase activity. In summary, the patterns of responses did not reveal a role for prolactin in the control of Harderian gland lysosomal enzyme activities by the pituitary. However, some of the influence on this target system may be exerted by growth hormone.     

Altered reabsorption of protein by the renal cortex in rats treated with hypertonic saline or mannitol.

The reabsorption of horseradish peroxidase (HRP) by the proximal tubule cells of rat kidneys was investigated by measuring the concentration of HRP in total particulate fractions of the cortex 1/4 and 1 hr after intravenous injection, and by correlated cytochemical observations. When compared to the corresponding values of the control animals, the concentration of HRP 1 hr after injection was decreased approximately 10-fold in the renal cortex of rats which had received an intravenous injection of hypertonic saline or two subcutaneous injections of mannitol. The plasma clearance and the urinary excretion of HRP were not altered significantly after injection of hypertonic saline, but the plasma clearance was decreased and the urinary excretion increased after injection of mannitol. When the dose of injected HRP was varied, the reabsorption of HRP by the renal cortex was proportional to the dose in the experimental and the control animals. Cytochemical staining for peroxidase activity also showed that the phagosomes and phagolysosomes of the proximal tubule cells contained much less peroxidase in the experimental rats than in the control rats. After injection of mannitol, large vacuoles appeared in the proximal tubule cells. The vacuoles often contained peroxidase-positive granules (phagosomes) which varied in diameter from the limit of microscopic visibility up to several microns. Most of the vacuoles did not react for acid phosphatase activity, but lysosomes were often aggregated around the vacuoles and seemed to release acid phosphatase into the cytoplasm. Certain analogies between the reabsorption of protein and that of water by the proximal tubule cells are discussed.     

Changes of enzyme activity in the rat liver at different age in circadian cycle

Most of the biological processes in the living organisms of both animals and man are known to be of rhythmical nature. Variability of enzymatic activity in circadian cycle depends on many factors among other on age, sexual maturity, diet as well as medication. The results obtained in our studies indicate, that the activity changes of acid phosphatase and ATP-ase Mg++ dependent in the liver of all the examined age groups were of 24 hour circadian rythm. As to the acid phosphatase activity the results of this experiments showed that in circadian cycle in all examined age groups there is only one peak of elevated activity. ATP-ase Mg++ dependent showed two activity peaks appearing at the same hour both in 30 and 60 days old animals. It should be noticed that the activities of ATP-ase Mg++ dependent in 100 day old animals were two times higher than in 30 and 60 days old rats.     

沙利度胺对实验性矽肺血管生成的干预研究

目的观察实验性矽肺形成过程中肺组织局部血管生成的动态变化,以及沙利度胺对矽肺肺纤维化血管生成的干预作用。方法SD大鼠54只,随机分为3组,矽肺组和沙利度胺组气管内注入二氧化硅混旋液复制实验性大鼠矽肺模型,对照组在相同条件下给予生理盐水。第二天起沙利度胺组给予沙利度胺饲料喂养,其余各组在相同条件下给予普通饲料喂养。采用HE染色、羟脯氨酸含量测定、免疫组织化学染色等方法,观察实验性大鼠矽肺的发病过程,肺组织中p-Akt蛋白和局部血管生成的动态变化及其与肺胶原蛋白含量的关系。结果矽肺组第7天新生血管明显增多,第30天较第7天有所减少,到第60天肺组织正常结构基本消失,取代为广泛结节性纤维化,少见血管;沙利度胺组血管生成在第7天轻于矽肺组,但仍高于对照组。免疫组化显示p-Akt蛋白在矽肺组第7天明显达到高峰,第30天时较第7天有所减弱,第60天时明显减弱。沙利度胺组p-Akt蛋白和羟脯氨酸含量均低于矽肺组,但高于对照组。结论矽肺早期血管生成显著,血管生成在矽肺肺纤维化发生早期可能起重要作用;沙利度胺能在一定程度上抑制过度的血管生成,对实验性矽肺具有一定的阻抑作用。     

Bioaccumulation of metals from a nickel smelter waste in P and F1 generations of exposed animals. II. Modulation of immune processes

A group of female Chinchilla rabbits was exposed through inhalation to the metal aerosol derived from dumped waste of a nickel smelter. The experiments were carried out in a field exposure station. Increased levels of tissue immune complexes were found in the myocardium and lungs of P females, whereas F1 rabbits (exposed both prenatally and 6 weeks postnatally) from the same group of P females had significantly elevated serum circulating immune complexes as compared to controls. In P rabbits, nonspecific serum tumoricidal activity was increased by 8.2%, while in F1 animals the increase was by 14%. Transplantation immunity was examined in a group of inbred Lewis rats following the transplantation of a skin allograft from the ear of inbred Berlin-Druckrey rats. The mean time of allograft survival in animals following i.v. administration metal dust suspension 2 days prior to transplantation, was prolonged as compared to controls. On day 22 after allograft transplantation, lactate dehydrogenase activity was found to be reduced in peripheral lymphocytes, and the liver and spleen weight proved to be diminished. These findings suggest a modulating effect of the metal dust from a nickel smelter regarding nonspecific serum tumoricidal activity and transplantation immunity as well as immune complex formation.     

The effect of testosterone propionate, 5 alpha-dihydrotestosterone and 5 alpha-androstan-3 beta, 17 beta-diol on acid phosphatase activity in the prostate of castrated rats

The acid phosphatase is studied in the prostate gland of rats. It is shown that 10 days after gonadectomy the activity of acid phosphatase lowered considerably. Administration of testosterone propionate or 5 alpha-dihydrotestosterone to castrated animals restored the enzyme activity whereas 5 alpha-androstan-3 beta,17-diol was not effective. Administration of testosterone propionate with one of its metabolites increased the activity of acid phosphatase in the prostate gland, however to a less extent than with the use of the hormone itself. The lowest activity is detected with the simultaneous application of three androgens.     

Effects of acetylphenylhydrazine (APHZ) on the stromal cells of the bone marrow in rats (in vitro)

The study has been carried out on Wistar rats. The animals were divided into two groups: the control rats received 0.9% saline solution intraperitoneally in there consecutive days. The experimental animals were administered 1% solution of acetylphenylhydrazine (APHZ) intraperitoneally also in three days. Prior to the experiments samples of blood have been collected from tail veins and reticulocyte counts made: the counts had also been determined before the culture of tissue was started. On the fifth day of the primary culture, the morphology and enzymatic activity of the stromal cells of the bone marrow were studied. The cells grown in vitro were cultured by the use of the Eagle's medium enriched with 20% calf serum and L-glutamate. The type of the cells, their number and rate of growth were noted. In the cells the activity of specific esterase as well as acid and alkaline phosphatase was determined. Statistically significant differences between the control and experimental animals were found both in the numbers of stromal cells and in the differences in score values of activity of particular enzymes in various types of cells.     

Effect of fasting and refeeding on duodenal alkaline phosphatase activity in monosodium glutamate obese rats

In the present work the effects of fasting and refeeding on fat pad weight and alkaline phosphatase activity in the brush border of individual duodenal enterocytes have been evaluated in male Wistar rats with obesity induced by monosodium glutamate (MSG) treatment during the early postnatal period. Neonatal rats were treated subcutaneously with MSG (2 mg/g b.w.) or saline (controls) for 4 days after birth. At 4 months of age, two types of experiments were performed. In the first experiment rats, were submitted to 3 or 6 days lasting food deprivation. In the second experiment the rats were refed for 3 or 6 days ad libitum or restrictedly (60% of pre-fasting intake) after a 6 day-fasting period. Fasting and refeeding influenced the body fat and function of the duodenum in MSG-treated rats differently as compared to the controls. However, alkaline phosphatase activity and the weight of epididymal and retroperitoneal fat depots were significantly increased in MSG obese rats (P<0.001) during all the periods examined. While 3 days of food deprivation resulted in both groups in a similar loss of adipose tissue weight and alkaline phosphatase activity, the decrements of these parameters after 6 days of fasting were lower in obese rats suggesting that their capacity to spare body fat stores was enhanced. After 3 days of ad libitum refeeding, a more marked adaptational increase of food consumption and also a significantly increased alkaline phosphatase activity above the pre-fasting level (P<0.01) was observed in the MSG-treated rats. Consequently, a more rapid body fat restoration was demonstrated in these animals. Refeeding of rats at 60% of the pre-fasting intake level resulted in a significant increase of alkaline phosphatase activity in both the MSG and control group; moreover, as food restriction continued, MSG-treated rats tended to further increase the enzyme activity. Our results revealed that MSG treatment of neonatal rats may significantly change the intestinal functions. Permanently increased alkaline phosphatase activity observed in MSG obese rats during all investigated periods suggests that this functional alteration is probably not a consequence of actual nutritional variation but could be a component of regulatory mechanisms maintaining their obesity at critical values.     

The effect of anoxia on cerebral acid hydrolases in the five-day-old rat

1. Five-day-old anaesthetized rats subjected to slow, prolonged asphyxia (50-55 min) were either allowed to die or resuscitated when at the point of death. Activities of various cerebral acid hydrolases known to be associated with lysosomes were determined in these animals and in littermate controls. 2. Asphyxia to death resulted in a significant increase in the activities of acid phosphatase, cathepsin (pH5.0) and beta-glucuronidase in whole-brain homogenates. 3. The effect of asphyxia on beta-glucuronidase activity was not apparent when the assay was performed in the presence of Triton X-100 (0.1%, v/v). 4. In resuscitated animals whole-brain-homogenate beta-glucuronidase activity showed the greatest increase (31%) 15 min after recovery. After a 60 min recovery period differences between control and asphyxiated animals were no longer apparent. 5. In animals anoxiated to death activities of acid phosphatase and beta-N-acetylglucosaminidase in brain high-speed supernatants were significantly higher than in controls. Acid phosphatase activity was similarly increased in asphyxiated animals resuscitated for 5 or 60 min. 6. It is suggested that the response of the immature rat brain to asphyxia involves a disruption or increased fragility of lysosomal particles.     

Activities of digestive enzymes in rats kept on standard or excessive breast feeding and on low-protein diet at once after weaning

Activities of digestive enzymes (maltase, alkaline phosphatase, aminopeptidase M, and glycyl-L-leucine dipeptidase) in small and large intestine, liver, and kidney were studied in rats of different ages kept at the period of lactation under conditions of the standard (8 individuals per litter) and low (3 individuals) number of pups per litter. The low-protein diet for 10 days at once after weaning was found to change the mass of organs and their digestive enzyme activities in all studied rat groups. The revealed changes were more prominent in rats kept under conditions of excessive breast feeding. In adult animals of this group, distribution of the alkaline phosphatase activity along the small intestine differed from that in control rats. The obtained results seem to confirm that any disturbance of the nutrition quality in early ontogenesis leads to disturbance of the «metabolic programming of enzyme systems» of digestive organs.     

Characterization and localization of alkaline phosphatase activity in rat testes

Alkaline phosphatase activity in extracts of testes of sexually immature (13 days old) and sexually mature rats has been characterized by its heat sensitivity, the extent of inhibition by homoarginine and phenylalanine, and by polyacrylamide gel electrophoresis. The testicular enzyme appears to be a liver-bone-kidney-type alkaline phosphatase. There are no significant differences in the properties of the enzyme from animals of these two ages. Spermatocytes and early spermatids contain very little alkaline phosphatase activity; the specific activity of a nonflagellate germinal cell suspension is only 1/20th that of the whole testis. Since the constant level of activity in immature and mature animals is not consistent with the enzyme activity being present only in late spermatids, we conclude that the majority of the testicular enzyme is present in nongerminal cells. The presence of alkaline phosphatase in plasma membrane purified from testes of adult rats was demonstrated.     

EFFECT OF THYROID DEFICIENCY ON THE LEVELS OF SEVERAL ENZYMES IN RAT BRAIN

—Activities of acid phosphatase, alkaline phosphatase and β-glucuronidase have been estimated in the brain tissues, using various subcellular particles, in growing thyroidectomized rats and also using cytoplasmic extracts free from debris and nuclear fraction in young hypothyroid animals. Hepatic glucose-6-phosphate dehydrogenase activity was markedly reduced after thyroidectomy but the enzyme was brought back to normal levels by thyroxine treatment. There was no change, however, in the activity of neural glucose-6-phosphate dehydrogenase after thyroidectomy. In the thyroidectomized animals an increase only in the free acid phosphatase activity in the neural synaptosomes was found and this increase in activity was not counteracted by administration of thyroxine. In the hypothyroid young animal β-glucuronidase, acid phosphatase and alkaline phosphatase activities were found to be affected during development.     

Transported Enzymes in Sciatic Nerve and Sensory Ganglia of Rats Exposed to Maternal Antibodies Against Nerve Growth Factor

Abstract: The accumulations by axoplasmic transport of selected enzyme activities proximal and distal to a ligature placed on the sciatic nerve were monitored in rats exposed in utero to maternal antibodies to nerve growth factor (NGF) and in control rats. Littermates of the animals exposed to anti-NGF were shown elsewhere to have had a 70% reduction in the number of sensory neurons in dorsal root ganglia and a 90% reduction in number of neurons in superior cervical (sympathetic) ganglion. The accumulation of F^--sensitive acid phosphatase activity was depressed 75% both proximal and distal to the tie. Accumulation of F^--resistant acid phosphatase activity was depressed nearly 50% proximal to the tie. Distal accumulation of this activity did not occur in either group of rats. Accumulation of acetylcholinesterase activity was not affected. Proximal accumulation of glutamic dehydrogenase activity was depressed 30%. Distal accumulation of the activities of β-glucuronidase and hexokinase was depressed 50%. In the lumbar dorsal root ganglia, dry weight was reduced 40%, and the activities of peroxide-sensitive, F^--resistant acid phosphatase and of the mitochondrial enzymes hexokinase, glutamic dehydrogenase, glutamic-oxalacetic transaminase, and NAD-dependent isocitric dehydrogenase were all reduced a little more, 45–50% per ganglion. However, the activities of the lysosomal enzymes, F^--sensitive acid phosphatase and β-glucuronidase, of the peroxide-resistant, F^--resistant acid phosphatase, and of the mitochondrial enzyme glutaminase were all reduced about 60% per ganglion. The results of these measurements were interpreted to suggest that much, and perhaps all, of the F^--sensitive acid phosphatase activity in motion in peripheral nerve in rat is confined to sensory axons.     

Histochemical studies of rat lungs after the short-term influence of soil dust

There was examined the biological activity of soil dusts using histochemical methods. Intratracheal administration of dusts used in this study is a common method for testing activity of industrial dusts. The used soil dusts were characterized by high content of free silicon dioxide, 3 times higher than its content in dusts from power stations released in the process of burning coal and approximately to the amount in graphite dust. Both coal dusts and graphite dusts absorbed by the lungs cause silicoanthracosis. The investigations have been shown that soil dusts caused stimulation of the mitochondrial metabolism giving an increase of the activity of succinic dehydrogenase, lactic dehydrogenase, and ATPase induced by Mg++ ions. This was additionally confirmed by an increase of NADP activity which is an enzyme binding a chain or reactions regulating the hydrocarbonic metabolism. There was also observed an increased activity of the hydrolytic enzyme acid phosphatase. High activity occurred in the epithelium of bronchi and bronchioli and focally in pulmonary parenchyma.     

Influence of gender on tamoxifen-induced biochemical changes in serum of rats

Tamoxifen, the widely prescribed drug in the prevention and therapy of breast cancer, may cause side effects which may be influenced by gender. The present study was undertaken to investigate the impact of gender on tamoxifen-induced toxic and biochemical changes following oral administration of tamoxifen at high dose level of 20 mg/kg once daily for a 2-week period in both male and female rats. The results showed marked increases in serum activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in female rats. In contrast, treatment with tamoxifen in male animals significantly decreased the activity of ALT, with a tendency for a decrease in serum AST levels. In female rats, a significant reduction in the serum activity of acid phosphatase (ACP) was noted, compared with a non-significant decrease in males. Non-significant changes in serum levels of alkaline phosphatase (ALP) were seen in both sexes. Tamoxifen lowered serum contents of total lipid and total cholesterol in both male and female rats. Serum levels of triglycerides were reduced in female rats as compared to a non-significant decrease in male animals. The serum albumin concentration was decreased in both male and female rats, while total protein was decreased only in female animals. Tamoxifen markedly increased serum levels of creatinine in female rats, compared with a non-significant rise in males. Total serum contents of calcium were similarly reduced in both males and females. This is the first study which points to gender-related differences in tamoxifen-induced toxic and metabolic changes in rats. The results indicated that females are more susceptible than males to tamoxifen toxicity, probably due to the ability of tamoxifen to antagonize the action of estrogen in females.     

The association of lysosomal enzymes with nuclei during enzyme induction in rat liver.

Male rats of the Holtzman strain were fasted for 3 days and refed a diet high in carbohydrate (68.9%). The induction of liver glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase was monitored for up to 48 h after refeeding. Induction occurred by 24 h, and by 48 h, 4.2- and 1.5-fold increases were observed for glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, respectively, compared with that of livers of pellet-fed rats. After refeeding, lysosomes increased in fragility as judged by an increased release of acid phosphatase activity during standard homogenization. Fragility was greatest 3 h after refeeding, but normal fragility was observed 24 h after refeeding. Nuclei were isolated from the liver samples before and after refeeding. Those isolated just before refeeding revealed small latent acid phosphatase activity (4–6%). However, after refeeding the carbohydrate-rich diet, a transient and significant (P < 0.01) increase in the latent activity occurred that was maximal (20%) at 1 h, returning to normal by 24 h. Cross-mixing the 800g nuclear pellet from livers of animals starved for 3 days with the 800g supernatant fraction from livers of animals refed the carbohydrate-containing diet did not alter the nuclear lysosomal-free (overt) or latent (detergent-released) enzyme activity. Similarly, mixing the 800g nuclear pellet from livers of animals refed for 1 h with the 800g supernatant fraction from livers of animals starved for 3 days, but not refed, did not change the nuclear lysosomalfree or latent enzyme activity. Purified nuclei, further washed in hypotonic buffer, lost acid phosphatase activity, but those isolated from livers of rats refed for 1 h retained 10% of the enzyme latency, whereas all latency was lost from those isolated from uninduced rats. A second lysosomal enzyme, β-galactosidase, became associated with the nuclei with the same temporal pattern as for acid phosphatase. However, no variation in nuclear content of cytosolic lactate dehydrogenase occurred as a result of feeding the high-carbohydrate diet to starved rats. When similarly starved rats were refed a diet high in lipid and carbohydrate-free, no induction of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase was observed. Lysosomes were not temporarily fragile and purified nuclei did not exhibit increased latency of acid phosphatase activity. Though the evidence presented does not establish a direct correlation between lysosome migration to nuclei as a required function in enzyme induction, the temporal and specific nature of the phenomenon support the hypothesis that liver lysosomal enzymes participate in early signals in the induction of enzymes of lipogenesis.     

Morphochemical investigations of the liver and biochemical studies of the blood of guinea pigs in anaphylactic shock

The experimental investigations have been carried out on 116 guinea pigs divided in two groups: the experimental and control group. The animals of the experimental group were sensitized with a 25% egg white suspension in 0.86% conc. of NaCl applied subcutaneously. After 21 days the same animals were exposed to the action of the same antigen in aerosol according to the method of Gerszanowicz, [16]. It has been shown, that in anaphylactic shock (acute and chronic) the damage of the lysosomal membranes in hepatocytes appeared which may be the cause of liberation among others also of acid phosphatase from the liver into the blood. Histochemically it was found a low phosphorylase and glucose-6-phosphatase activity, which was the basis of the assumption, that in anaphylactic shock we have to do with an enzymatic block--phosphorylase kinase--phosphorylase and inhibition of the enzymatic activity of glucose-6-phosphatase in the liver of guinea pigs. The comparison of the histochemical and biochemical results concerned with the amount of lipids, glycogen and nucleic acids in the liver revealed that the increasing amount of lipids is paralleled by decrease of glycogen. Among nucleic acids a growing level of ribonucleic acid was found while the level of the desoxyribonucleic acid remained stable.