The influence of changes in food availability on the activities of key degradative and metabolic enzymes in the liver and epaxial muscle of the golden perch

This study investigated the influence of feeding frequency on the activities of important degradative enzymes and potentially rate-limiting enzymes in glycolysis and gluconeogenesis in the liver and white epaxial muscle of Macquaria ambigua . Adult animals were either fed daily to satiety (fed), deprived of food for up to 180 days (starved), or starved for 150 days then fed daily to satiety for 30 days (starved/fed). The activities of lipolytic, glycogenolytic and glycolytic enzymes in the livers of starved fish were maintained as long as liver energy stores were available, but became significantly reduced following their exhaustion indicating a decline in metabolism in response to prolonged starvation. The response of epaxial muscle metabolism to changes in food availability was different to that of the liver, as no significant change in the activities of muscle lipolytic or glycogenolytic enzymes were observed in response to starvation. Muscle tissue metabolism was reduced after 60–90 days of starvation, but then returned to prestarvation levels.     

Morphometric studies of pancreas islets of rats after subphrenic vagotomy]

Analysis of the morphometric data showed that 7 days after vagotomy significant changes ocurred in the pancreatic islets, mainly as a result of variation in B cells. At later periods (45 and 90 days after surgery) normalization of some morphofunctional parameters is noted, however, absolute restoration is not achieved.     

Biochemical changes in progressive muscular dystrophy. XII. Cyclic nucleotides in lymphoid and nonlymphoid organs of dystrophic mice

Concentrations of cAMP and cGMP were measured (per milligram DNA) in the lymphoid (thymus, spleen) and nonlymphoid organs (liver, brain, kidney, lungs, heart, pancreas, skeletal muscle, lens) of normal (+/+) and dystrophic (dy/dy) 129 ReJ mice aged 30, 60, and 90 days. The cAMP concentrations in the thymus did not reveal any significant differences at 30 and 60 days of dystrophy, but were considerably higher (2-fold) at 90 days. cGMP concentrations were decreased in the thymus at 30 days (0.20-fold) and markedly elevated at 60 (2-fold) and 90 days (3-fold) of the disease. The [cAMP]/[cGMP] ratio was increased (1.30-fold) at 30 days of dystrophy, and this was followed by a sharp decline at 60 days (2-fold), with a lesser decrease at 90 days (0.34-fold). In the spleen, the cAMP concentrations were augmented significantly in all stages of dystrophy (1.5- to 2.6-fold). cGMP (per milligram DNA) did not show any significant variation at 30 and 60 days of the disease but was increased (3-fold) at 90 days. The [cAMP]/[cGMP] ratio, which was enhanced in the spleen at 30 (2-fold) and 60 days (1.5-fold), demonstrated no change at 90 days of dystrophy. These results indicated significant differences in the concentration of cyclic nucleotides and their ratios in the thymus and spleen of 129 ReJ dy/dy mice. The modifications were not limited to lymphoid organs alone, having been noted in the nonlymphoid organs as well. These changes could, in turn, influence immune responsiveness and could cause immunodepression in dystrophic mice.     

Long-term fatty liver-induced insulin resistance in orotic acid-induced nonalcoholic fatty liver rats

We investigated whether fatty liver preceded insulin resistance or vice versa using a long-term orotic acid (OA)-induced nonalcoholic fatty liver disease (NAFLD) model without the confounding effects of obesity and hyperlipidemia and explored the role of the liver in insulin resistance. Male Wistar rats were fed with or without OA supplementation for 30, 60, and 90 days. The NAFLD group showed increased liver lipid at 30, 60, and 90 days; glucose intolerance was noted at 60 and 90 days. Furthermore, partial liver proteins and gene expressions related to upstream signaling of insulin were decreased. However, the liver glycogen content was elevated, and gluconeogenesis genes expressions were obviously decreased at 90 days. The occurrence of fatty liver preceded insulin resistance in OA-induced NAFLD without the interference of obesity and hyperlipidemia, and hepatic insulin resistance may not play a conclusive role in insulin resistance in this model.     

Comparative characteristics of 2 morphometric methods of assessing the degree of liver fatty distrophy

The fat content in the liver of white male Wistar rats was measured over time (1, 2, 4, 8 and 14 days) after a single exposure to CCl4 within a wide concentration range (from 30 to 25700 mg/m3) by the two morphometric techniques: subjective-graduation and semi-automatic evaluation of the volumetric fat fraction, using a TAS unit (Leitz-Wetzlar). The results obtained with both the methods indicate that the CCl4 concentration equal to 260 mg/m3 is the threshold one as shown by the morphological signs of liver fatty dystrophy. The semi-automatic method offered enables an objective evaluation of the findings, making the research time shorter. To obtain adequate results with the method, it is necessary to observe the rules of the morphological material treatment (contrast selective color, uniform and sufficient thickness of the sections).     

The dynamics of the morphological changes in the tissues after the subcutaneous administration of vaccines against Q fever]

The comparative study of the dynamics of morphological changes in tissues of guinea pigs after the subcutaneous injection of chemical, live and combined vaccines against Q fever during the period from 12 hours to 90 days was made. All vaccines under study were shown to produce a pronounced local damaging effect. Two periods were tentatively discriminated in the dynamics of changes: the early phase (till 48 hours) and the late phase (days 2-90). At the early stage the most pronounced changes were registered after the injection of the combined vaccine. At the late phase the use of the chemical and combined vaccines was accompanied by the appearance of secondary hemorrhages into newly formed connective tissue. Starting from day 30, practically no deviation from the normal state of tissues were registered at the site of injection.     

Analysis of morphometric indices in mice for registration of induced mutations in germ cells

The new-born mice of the first generation have been examined for changes in morphometric indices after the influence of fopurine, cyclophosphamide and cadmium chloride on their fathers. It is shown that fopurine and cyclophosphamide, the mutagens for mammals, increase the variability of morphometric indices. The method to calculate the morphometric indices of mice may be used for evaluating genetic risk of the compounds for man.     

The peculiarities of separate and combined exposure to low dose-rate gamma-radiation and 232Th nitrate on thyroid gland

We studied the effects of both: of the separate and of the combined exposure to gamma-radiation (absorbed doses for 30 days were 1.4-1.8 cGy, for 90 days were 4.3-5.4 cGy) and 232Th nitrate (in concentrations 6.0; 20.0 and 60.0 mg/l) on the CBA line mice thyroid gland. Hat is shown the thyroid gland response to the combined exposure is significant in an appearance of non-linear effects of interaction. The intensiveness of the radiation influence (time and doses) is revealed to affect on the biological response direction in thyroid gland. The intensity of this effect was depended on the correlation between the radiation activity and the chemical components concentration. The dispersive analyses showed that all the studied factors caused structural changes in thyroid gland in this experiment, but the gamma-radiation influence causes the primary contribution.     

Structural and circulatory responses to hypoxia in the secondary lamellae ofSalmo gairdneri gills at two temperatures

Summary The first gill arch ofSalmo gairdneri was fixed from normoxic (O₂ saturation of the water >90%) and hypoxic (O₂ saturation 25–30% for 5 days) fish at 10 °C and 18 °C, and from fish after one-day recovery from hypoxia at 18 °C. The secondary lamellae of the gills were analysed with morphometric methods for structural, haemotological and circulatory changes. During hypoxia a marked vascular distension took place at both temperatures. At both temperatures the vascular distension coincided with a shortening of the diffusion distance (36% at 10 °C and 21% at 18 °C) and a swelling of the erythrocytes (60% at 10 °C and 42% at 18 °C). The effects of these changes on the oxygen uptake of the gills are discussed.     

Production and Deformation of Clonorchis sinensis Eggs during In Vitro Maintenance

Clonorchis sinensis is a carcinogenic human liver fluke. The present study monitored eggs produced by long-term maintained adult worms of C. sinensis to confirm their egg productivity in vitro. The worms from infected rabbits were incubated in vitro in 1× Locke’s solution and broth media (RPMI-1640, DMEM and IMDM). Numbers of expelled eggs were counted sequentially and their morphological changes were monitored by microscopy after 1, 30, 60, and 90 days of cultivation. On the 1–3 days of cultivation, the eggs counted maximum 4,756±202 eggs/worm/day in IMDM medium. The number of eggs gradually decreased less than 1,000 at 7–14 days and below 100 at 21days but continued to pass eggs after 56 days in all media. Length of the eggs were reduced about 1 µm at 30 days, and the length/width ratio was maintained around 1.8 at 30 days but decreased to 1.7 at 60 days and 1.5 at 90 days. Faust-Meleney index (FMI) decreased as the cultivation duration increased and lowest FMI (5662.9±974.7) observed in IMDM media at day 90 (P = 0.001). Microscopic findings of the eggs recognized the miracidium in most of eggs at 60 days but not in those at 90 days. Instead, the eggs contained dark granules or vacuoles in the deformed shell at 90 days. Scanning electron microscopy revealed partial loss of wrinkles on the deformed egg surface and prominent abopercular knob. Eggs viability decreased as the cultivation progressed and showed significant positive correlation with FMI and length/width ratio. In conclusion, the cultivated worms pass only the eggs which are preformed in their uterus before cultivation. One gravid C. sinensis contains about 37,000 eggs in its uterus and produces about 4,000 eggs every day. The deformed eggs with FMI less than 7,000 and length/width ratio lower than 1.7 are non-viable.     

Testicular morphometry as a tool to evaluate the efficiency of immunocastration in lambs

This study aimed to evaluate the efficiency of immunocastration in lambs using testicular morphometry. Thirty lambs were randomly divided into two treatments (subcutaneous administration of 1.0 mL and 0.5 mL of an anti-GnRH vaccine) and a control group (1.0 mL saline solution). The animals were vaccinated at four months of age, received a second dose 30 days later, and were slaughtered 90 days after the first vaccine dose. After slaughter, testicles were collected, and samples were removed for histological processing and evaluation of testicular morphometric parameters. Analysis of variance, Tukey’s test, and Kruskal–Wallis test were performed, with a 5% level of significance. There was a reduction in testicular weight, gonadosomatic index, seminiferous tubule diameter, germinal epithelium height, leydigosomatic index, and total tubule length. The total length per testicular gram increased in the immunocastrated group. Intrinsic spermatogenesis yield, Sertoli cell indices, and estimates of sperm and Sertoli cell production were reduced in the immunized groups (P < 0.05). The anti-GnRH vaccine in lambs at doses of 1.0 mL and 0.5 mL is sufficient to promote immunocastration, verified through severe changes in testicular morphometry from animals.     

Effect of short photoperiod on organ growth kinetics and serum hormone profile in pullets of domestic fowl, Gallus gallus domesticus

Effect of short photoperiod (SP; LD 6:18) treatment on serum hormone profile and growth rate of adrenal, thyroid, ovary, oviduct, liver and lymphoid organ was studied in rearing pullets (RIR breed) of 1 to day 90 old. Body weight and growth index of SP pullets were lesser as compared to pullets reared under LD 12:12. Except for ovary (recorded marginal increment), weights and growth indices of thyroid, adrenal and oviduct decreased under SP. Weight of liver and lymphoid organs was higher at 30 and 90 days, in SP pullets as compared to LD 12:12. Histometric data suggested that the transition from small to big follicles was slow in ovary of SP pullets, and also reduced follicular atresia was noted in SP pullets. Except for higher corticosterone level at 30 days and higher progesterone level at 30 and 60 days, relative levels of all the hormones at all other ages were lower in SP pullets. In general, the present observations suggested intraovarian changes in pullets exposed to SP.     

Total RNA content and blood flow in rat brain after RNA administration

The changes in blood flow through selected brain structures and the changes in the total RNA content of cells of these structures were examined after a single administration of yeast RNA to 6-month-old male rats. The total content of ribosomal RNA in cells of the limbic system (septum, hippocampus, hypothalamus) increased 48 hrs after the administration of 100 mg i.p. yeast RNA , dropped after 7 days (in hypothalamus), 21 and 30 days (in hippocampus), 30 days (in septum). In cells of the limbic system as a whole there is a higher total RNA content in experimental rats. No changes were observed in the cells of parietal brain cortex. Blood flow increased in limbic structures 21 and 30 days after RNA administration and in septum and in hippocampus also 90 days after application. No changes were observed in parietal brain cortex, bulbi olfactorii, cerebellum and brain stem. Histochemical changes correlated positively with blood flow changes in the limbic system 14, 21, 30 and 90 days after RNA application. The body weight of experimental rats did not differ from that of control animals. The changes in haemodynamic parameters were transient and were demonstrated as fluctuations in heart rate, cardiac output, and peripheral resistance. Blood pressure experienced no changes.     

The effects of long-term food deprivation on respiration and haematology of the neotropical fish Hoplias malabaricus

Growth of adult traíras Hoplias malabaricus ceased and body mass ( M ) decreased during starvation periods of 30, 60, 90, 150, 180 and 240 days. Hepatic reserves were mobilized in fish starved for 30 days, but liver mass of fish starved for longer periods was not significantly different from those starved for 30 days. Perivisceral fat bodies were consumed gradually, being completely exhausted after 240 days of food deprivation. Length of starvation was associated with a significant decrease in the oxygen uptake ( V o₂). In spite of this reduction, the respiratory frequency ( f _R) was kept nearly constant during the starvation periods. The haematocrit and the number of red blood cells decreased after 150 and 240 days of starvation, respectively. These parameters did not recover after refeeding (after 90 and 240 days of starvation). This hypometabolic state in response to food deprivation contributed to energy conservation during these periods. Traíras can survive food deprivation for periods of up to 180 days without reductions in metabolism and when they do become hypometabolic, normal metabolic rates are rapidly restored upon refeeding.     

Effects of clofibrate (alpha-p-chlorophenoxyisobutyryl-ethyl-ester) on male rat liver

In male rats, fed 0.5% clofibrate in their diet for 8 days and 21 days, the ultrastructural morphometric alterations of the hepatocytes were evaluated and compared with the biochemical data. The morphologic alterations of the microbodies were particularly related to the changes of the catalase activity of the liver homogenates. The results showed a marked hypertrophy of the liver and an increase in the volume of the individual hepatocyte. The numerical density and, even more pronounced, the volume density of the microbodies increased excessively during the treatment. The numerical density of the mitochondria decreased markedly after 21 days of administration. The surface of the rough endoplasmic reticulum showed a significant decrease, whereas the surface of the smooth endoplasmic reticulum showed a hypertrophy. The catalase activity of the liver homogenates increased 2-fold after 8 days and remained at this new steady-state after 21 days of treatment. The results suggest that the enzyme content of the microbodies changed after treatment with clofibrate, and support the suggestion that clofibrate may induce the synthesis of a yet unidentified peroxisomal protein.     

Influences of crude extract of tea leaves,Camellia sinensis,on streptozotocin diabetic male albino mice

Natural remedies from medicinal plants are considered to be effective and safe alternative treatment for diabetes mellitus. The aim of the present study was to investigate the hypoglycemic activity of the crude tea leaves extract on streptozotocin (STZ)-induced diabetic mice. The average body weight of animals with diabetes and their percentage changes of body weight gain after 15 and 30 days were significantly lower than that of the normal control mice. In diabetic mice, supplementation with tea leaves extract decreased the loss of body weight. After 15 and 30 days, significant increases in the levels of serum glucose, triglycerides, cholesterol, creatinine, urea, uric acid, glutamic pyruvic acid transaminase (GPT) and glutamic oxaloacetic acid transaminase (GOT) were noted in STZ-diabetic mice fed with normal diet. Also, the values of total protein in this group were statistically declined after 15 and 30 days. The levels of serum glucose and GPT were significantly elevated after 15 and 30 days in diabetic mice supplemented with tea leaves extract. Moreover, the level of serum GOT was notably increased after 30 days. Insignificant alterations were observed in the levels of serum triglycerides, cholesterol, total protein, creatinine, urea and uric acid in diabetic mice supplemented with tea leaves extract. Thus, the present results have shown that tea leaves extract has the antihyperglycemic, antihyperlipidemic, and antihyperproteinemic effects and consequently may alleviate liver and kidney damage associated with STZ-induced diabetes in mice.     

Ultrastructure of thymus and liver following adrenalectomy and replacement adrenal hormone therapy in rats

Prior research in this laboratory has shown that dexamethasone, aldosterone, and epinephrine interact in regulating the activity of ornithine decarboxylase (EC 4.1.1.17, ODC) in rat thymus and liver. The three primary adrenal hormones were administered alone and in various combinations to adrenalectomized rats. Liver and thymus samples were removed, prepared for electron microscopy, and morphometric analysis of thymic micrographs was performed. It was found that both corticosteroids induced thymic lympholysis and that concurrent administration of epinephrine 'rescued' the lymphocytes. Observations of liver micrographs indicated that changes in liver glycogen deposition vary in response to the hormone treatment regimen. The liver response to a combination of the glucocorticoid and catecholamine was different from the response to the mineralocorticoid and catecholamine, which indicated that the liver response to the two steroids may be mediated via different mechanisms. Evidence is provided to support the conclusion that the influence of the adrenal gland on rat thymus and liver is not restricted to glucocorticoids but may also involve mineralocorticoids and catecholamines.     

Multiple dexamethasone treatment affects morphometric parameters of gonadotrophic cells in adult female rats

Exposure to glucocorticoids leads to numerous changes in various biological systems including the reproductive system. The aim of the present work was to find out whether dexamethasone (Dx) treatment of adult female rats would influence the histological and morphometric characteristics of the pituitary gonadotrophic cells (luteinizing--LH cells and follicle stimulating--FSH cells). One group of female Wistar rats received Dx injections on three consecutive days in doses 1.0, 0.5 and 0.5 mg/kg b.w. respectively, while the control rats were treated with equivalent volumes of saline. Experimental and control animals were sacrificed 24 h and 72 h after the last injection. The peroxidase-antiperoxidase (PAP) immunocytochemical procedure was used to study the LH and FSH cells. The stereological and morphometric analyses showed that multiple Dx treatments of female rats significantly decreased the volume of LH cells and the volume of their nuclei 24 h and 72 h after the last Dx injection in comparison with control values. At 24 h after Dx treatment, the volume density of LH cells was significantly increased, but at 72 h differences between the experimental and control groups were insignificant. The increase in number of LH cells per unit area (mm2) was significant at both timepoints (24 h and 72 h). Stereologic and morphometric characteristics of FSH cells was changed after Dx treatment in the same manner as that of LH cells, except for the volume density, where a significant increase was established 24 h and 72 h after the last Dx application. These results clearly demonstrate that 24 h and 72 h after the last of three Dx injections there were changes in the immunocytochemical and morphometric features of gonadotrophic cells.     

Ascorbic acid supplementation causes faster restoration of reduced glutathione content in the regression of alcohol-induced hepatotoxicity in male guinea pigs

Abstract

Alcoholic liver disease is caused mainly by free radicals. Ascorbic acid (AA) and glutathione (GSH) are the major water-soluble antioxidants in the liver. The impact of AA supplementation on GSH, AA and activities of GSH-dependent enzymes in alcoholic guinea pigs was studied and was compared with alcohol abstention. Guinea pigs were administered ethanol at a dose of 4 g/kg body weight (b.wt)/day for 90 days. After 90 days, alcohol administration was stopped and one-half of the ethanol-treated animals were supplemented with AA (25 mg/100 g b.wt) for 30 days and the other half was maintained as the abstention group. There was a significant increase in the activities of alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transpeptidase in the serum of the ethanol group. In addition, a significant decrease in the GSH content, activities of GSH peroxidase, GSH reductase, and increased activity of GSH-S-transferase were observed in the liver of the ethanol group. Histopathological analysis and triglycerides content in the liver of the ethanol group showed induction of steatosis. But AA supplementation and abstention altered the changes caused by ethanol. However, maximum protective effect was observed in the AA-supplemented group indicating the ameliorative effect of AA in the liver.