Cellular injury and lipid peroxidation induced by hexavalent chromium in isolated rat hepatocytes

In order to elucidate the relationship between cellular injury and lipid peroxidation induced by hexavalent chromium (CrVI), isolated rat hepatocytes treated with any one of scavengers of active oxygen species, antioxidants or antichromium agent were incubated with K2Cr2O7 as CrVI (1 mM Cr). After the incubation, the development of lipid peroxidation was determined as thiobarbituric acid (TBA)-reacting materials in total lipid extracts from the incubated hepatocytes. Cellular injury was observed as a leakage of lactate dehydrogenase (LDH) from hepatocytes into incubation medium. The contents of reduced glutathione (GSH) in hepatocytes were also assessed. Results obtained were as follows: (1) CrVI facilitated lipid peroxidation in isolated hepatocytes after 20 min of incubation. On the other hand, the cellular injury induced by CrVI was barely observed even after 60 min of incubation. (2) The CrVI-induced lipid peroxidation was inhibited by catalase and mannitol as scavengers of active oxygen species, or N,N'-diphenyl-p-phenylenediamine and alpha-tocopherol as antioxidants. However the cytotoxicity of CrVI could not be prevented by these chemicals. (3) CrVI depleted the contents of intracellular GSH and diminished the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) except glutathione peroxidase. (4) The scavengers of active oxygen species and the antioxidants could not prevent the depletion of intracellular GSH induced by CrVI. (6) Ascorbic acid, antichromium agent, prevented all of the lipid peroxidation, the cellular injury and intracellular GSH depletion induced by CrVI.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of Cytochrome P450-side Chain Cleavage in the Theca Interna Layers of Bovine Small Antral and Cystic Follicles

Cystic follicle is anovulatory follicular structure that is caused by an endocrine imbalance. The activity of cytochrome P450‐side chain cleavage (P450scc) is essential for the initiation of steroidogenesis in the follicle. The present study was designed to compare the frequency of cells containing P450scc between healthy and atretic small antral follicles, and among several types (I, II and III, classified based on the presence of granulosa layer) of cystic follicles. Paraffin sections of healthy (2–5 mm in diameter), atretic (2–5 mm) and cystic follicles (>25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine P450scc. The P450scc‐positive cells were counted in four different regions of the follicles from the apical to the basal side. In small antral follicles and cystic follicles, P450scc‐positive cells were localized in the theca interna layers but not granulosa layers. The P450scc‐positive cell populations decreased in the late atretic follicles compared with the early and advanced atretic follicles at all the regions of follicle. Type III cystic follicles showed significantly lower frequencies of P450scc‐positive cells than those in the types I and II cystic follicles. These results suggest that in both small and cystic follicles in cows, total loss of granulosa cells may be associated with the reduction of frequency of P450scc‐positive cells in theca interna layer.     

Intra‐uterine Infection with Babesia bovis in a 2‐day‐old Calf

Infection with Babesia bovis was diagnosed in a 2‐day‐old female calf apparently transmitted in utero. The calf was born as the second calving to a cross‐bred beef cow permanently on pasture. Diagnosis was based upon identification of B. bovis in peripheral blood smears and clinical signs which included fever, jaundice, pale mucous membranes and convulsions. Anaemia, leucocytosis, thrombocytopenia and lymphocytosis were noted at the febrile acute stage of the disease. The blood smears revealed evidence of regeneration of toxic neutrophils with a left shift, severe spherocytosis and high degree of basophilic stippling. Elevated concentration of aspartate aminotransferase, lactate dehydrogenase, and creatine kinase were also noted, and were probably the result of haemolysis, dehydration and muscle damage because of recumbancy. Elevated total bilirubin concentration following haemolysis resulted in jaundice. The neurological symptoms observed were probably caused by sludging of parasitized erythrocytes in the brain capillaries. The calf recovered following treatment with diminazene aceturate and the recovery was followed up clinically, haematologically and biochemically.     

Epidemiological studies on the expectation of life for cats computed from animal cemetery records

Based on the Chiang's method, the life table for cats was constructed from the death data of 3936 cats. They died in the Kanto area and were buried in an animal cemetery in Tokyo from June 1981 through May 1982. This life table seems to be the first one for domestic pet cats. The expectation of life for cats was 4.2 years at age 0, 5.0 years at age 1, 5.4 years at age 4, 5.3 years at age 5, 3.5 years at age 10, and 2.2 years at age 15. The maximum age at death was 22 years. From age 0 to age 5, the probability of dying for cats was higher than that for dogs, but over 6 years of age it seemed that Gompertz's equation was applicable to this life table for cats. From these results, if the probability of dying for cats at early ages decreases, the fundamental pattern of dying curve for cats seems to be a similar figure of dogs. The life table was constructed for different breeds and localities. Comparing the expectation of life at age 1 (e1) of the two populations divided by breeds or localities, there was significant difference in the e1 among different localities but not among different breeds. These facts suggest the existence of some factors which may influence the life span of cats among different localities.     

Detection of early pregnancy factor in superovulated mice

Rosette inhibition tests for the detection of early pregnancy factor (EPF) were performed on naturally ovulated and superovulated mice from day 2 of pregnancy up to 4 days after parturition. In both groups of mice, the rosette inhibition titre (RIT) increased on day 2 of pregnancy, and persisted at high levels until day 15. Thereafter, the RITs of both groups of mice decreased to the non-pregnancy range. No significant differences of the mean RITs between these two groups were observed during the high RIT period. These results showed that the superovulatory treatment did not cause any changes or interference in the detection of EPF. In order to investigate the initial time of appearance of EPF in the maternal circulation in relation to the stage of fertilization, measurement of RIT and examination of the fertilization stage were carried out on superovulated mice 1 day after mating. The mean RIT of mice with pronucleus stage ova was significantly (p less than 0.01) higher than that of mice with sperm-penetrated ova. EPF was considered to appear in the maternal peripheral blood at the pronucleus stage.     

In vitro evaluation of protein degradability in the rumen and digestibility of undegraded protein

A three-phase laboratory procedure suitable for predicting protein degradability in the rumen and digestibility of undegraded protein is reported. In the first phase the feed was incubated with starch and buffered rumen fluid. In the incubation mixture the viability of protease-active bacteria was checked by anaerobic culturing, whereas changes in protease activity were monitored by azocasein degradation. In the second and third phase rumen undegradable protein (UDP) was digested with pepsin and pancreatin, respectively. The measurements showed that 63.2, 5.2 and 4.7% of the crude protein of green lucerne was decomposed by rumen fluid, pepsin and pancreatin, respectively. Degradability of the crude protein of extracted sunflower meal was 68.3, 17.7 and 5.5% in the three phases, respectively. Repeated determination yielded crude protein degradabilities of 66.7, 27.1 and 5.1% for the three phases, respectively.     

Dynamics of changes in the cytological picture of vaginal smears and circulating ovarian hormones during the puerperal period in ewes]

A postparturient period is characterized by low basal secretion of adenohypophysis gonadotropins with the following appropriate changes in ovarian hormones and their response to the morphology of vaginal epithelium. In this study the dynamics of the cytological picture of vaginal swabs and ovarian hormones 17 beta-oestradiol (E2) and progesterone was investigated in the puerpery of ewes. The objective was to obtain and extend the knowledge of cytological changes in vaginal epithelium and levels of ovarian hormones of ewes after parturition and of their relationships from the first several days after lambing until the 51st day of the period of observation. Vaginal swabs for vaginal cytology were taken from nine ewes on days 1, 4, 7, 14, 17, 21, 25, 34, 42 and 51 after parturition. These swabs were fixed in ether-alcohol 1:1, stained according to the Faltínová-Zidovsky method, embedded in Canada balsam and evaluated by differentiation of cells according to Luksh (1953). Blood samples for E2 and P4 determinations were taken from the jugular vein in the same intervals as vaginal swabs. The serum was centrifuged and stored at -18 degrees C until use. E2 and P4 concentrations were determined radioimmunologically, using kits RIA-test ESTRA and RIA-test PROG from URVJT Kosice. A statistically significant decline (P < 0.05) of percentual representation of basal and parabasal cells (Fig. 1, Tab. I) on day 7 after lambing was replaced by their multiplication from day 14 reaching the values of 66.07 +/- 3.95 on day 42. A statistically significant decrease in intermediary flat cells (Fig. 2, Tab. II) was observed on days 14 (P < 0.001), 34 and 42 (P < 0.01; P < 0.001), in comparison with the first day after lambing. An evaluation of intermediary convoluted cells revealed their highest percentage on days 1 and 17 after parturition (34.65 +/- 4.77-20.62 +/- 12.57) and their decline to values in the range of 6.77 +/- 1.46-7.66 +/- 2.25 on the remaining days of the period of observation. Percent occurrence of superficial flat cells (Fig. 3, Tab. I) ranged from 3.9 +/- 1.10 to 10.63 +/- 7.23 from day 1 to day 51 after lambing. The lowest percentual representation (1.32 +/- 0.79-4.10 +/- 1.89) was recorded for superficial convoluted cells. Multiplication of the evaluated cells was observed, reaching the highest but insignificant representation (P > 0.05) on day 25 of postparturient investigation: 4.10 +/- 1.89 (Fig. 3, Tab. I). 17 beta-oestradiol (E2) concentrations were compared to the -1st day before parturition, when its values varied at the level of 2.45 +/- 0.64 nmol/l serum.(ABSTRACT TRUNCATED AT 400 WORDS)     

Type-I hypersensitivity as a component of eosinophilic myositis (muscular sarcocystosis) in cattle

Eight bovine hearts with lesions of eosinophilic myositis (EM) and 2 bovine hearts without EM lesions were collected at slaughter. Blood samples from these 10 hearts, and the heart of a newborn calf also were collected. Histologically, Sarcocystis cruzi was identified in the 8 hearts with EM lesions and the 2 hearts without EM lesions, but not in the heart of the newborn calf. Serum was harvested from the 10 blood samples and was used in homologous, modified, passive cutaneous anaphylaxis tests. Antigen was prepared from S cruzi bradyzoites isolated from the 2 hearts without EM lesions. Serum samples from the 8 cattle with EM lesions reacted positively to S cruzi antigen. When heat-inactivated IgE in serum (56 C for 4 hours) was used, all passive cutaneous anaphylaxis responses were considered negative. Using ELISA, serum IgE concentrations from the 10 cattle with and without EM lesions were 2.2 to 9 U/ml. As determined by radial immunodiffusion, IgM concentrations were 80 to 215 mg/dl. Immunoglobulin G concentrations were 420 to 2,050 mg/dl, but most were less than or equal to 1,700 mg/dl. Immunoglobulin A concentrations were 0 to 62 mg/dl; 1 steer with EM lesions had 0 mg/dl. Double-gel immunodiffusion confirmed the presence of Sarcocystis-specific precipitating antibodies. Sera from the 10 cattle with and without EM lesions formed at least 1 precipitin band.     

Effects of abiotic variables on the response of carnation cultivars to Fusarium oxysporum f.sp. dianthi

The effects of abiotic variables on the response of carnation cultivars to Fusarium oxysporum f.sp. dianthi ( F.o. dianthi  ) were examined in experiments conducted under semi-controlled environments. The abiotic variables examined were solar radiation intensity, temperature and growth substrate. Temperature was not controlled, but differed markedly among experiments, thus, its effect was not determined quantitatively. Disease incidence and disease severity varied significantly among the experiments (due mainly to differences in temperature), among the solar radiation treatments and among the cultivars tested. The three-way interaction term (i.e. cultivar × shade treatment × experiment) was highly significant ( P  < 0.001) when both disease incidence and disease severity were considered, indicating that no single variable was predominant in determining disease intensity. The effects of the growth substrate on disease progress was examined in plants grown in tuff or in tuff mixed with peat (1 : 1 and 1 : 3) substrates. The growth substrate had a potent effect on disease development in the less susceptible cultivars. Severe epidemics developed in all cultivars when they were grown in the tuff/peat mixture, although some were resistant when grown in tuff alone. These results led to the conclusion that the carnation response to F.o. dianthi is substantially influenced by the environmental conditions of the test.