Benzodiazepines inhibit the acetylcholine receptor-operated potassium current (IK.ACh) by different mechanisms in guinea-pig atrial myocytes

The anticholinergic effects of 7 benzodiazepines, bromazepam, camazepam, chlordiazepoxide, diazepam, lorazepam, medazepam and triazolam, were compared by examining their inhibitory effects on the acetylcholine receptor-operated potassium current (I(K).(ACh)) in guinea-pig atrial myocytes. All of these benzodiazepines (0.3-300 μM) inhibited carbachol (1 μM)-induced I(K).(ACh) in a concentration-dependent manner. The ascending order of IC(50) values for carbachol-induced I(K).(ACh) was as follows; medazepam, diazepam, camazepam, triazolam, bromazepam, lorazepam and chlordiazepoxide (>300 μM). The compounds, except for bromazepam, also inhibited I(K).(ACh) activated by an intracellular loading of 100 μM guanosine 5'-[γ-thio]triphosphate (GTPγS) in a concentration-dependent manner. The ascending order of IC(50) values for GTPγS-activated I(K).(ACh) was as follows; medazepam, diazepam, camazepam, lorazepam, triazolam chlordiazepoxide (>300 μM) and bromazepam (>300 μM). To clarify the molecular mechanism of the inhibition, IC(50) ratio, the ratio of IC(50) for GTPγS-activated I(K).(ACh) to carbachol-induced I(K).(ACh), was calculated. The IC(50) ratio for camazepam, diazepam, lorazepam, medazepam and triazolam was close to unity, while it for chlordiazepoxide could not be calculated. These compounds would act on the GTP binding protein and/or potassium channel to achieve the anticholinergic effects in atrial myocytes. In contrast, since the IC(50) ratio for bromazepam is presumably much higher than unity judging from the IC(50) values (104.0 ± 30.0 μM for carbachol-induced I(K).(ACh) and >300 μM for GTPγS-activated I(K).(ACh), it would act on the muscarinic receptor. In summary, benzodiazepines had the anticholinergic effects on atrial myocytes through inhibiting I(K).(ACh) by different molecular mechanisms.     

Relationship between pH and Temperature in the Ruminal Fluid of Cows, Based on a Radio-Transmission pH-Measurement System

To assess the relationship between pH and temperature in the ruminal bottom fluid, circadian changes were monitored using cows fed a control diet (C diet) or a rumen acidosis-inducing diet (RAI diet) by using a wireless radio-transmission pH- measurement system. These two parameters were measured simultaneously at 10-min intervals on day 14 after commencement of feeding. Compared to the mean ruminal pH for 60 min immediately after the morning feeding (0 hr), significantly lower pH was noted 3-13 hr later (P<0.05) and 4-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively, although the reduction in the latter was much higher than that in the former. In contrast, significantly higher ruminal temperature was found at 8 and 12-14 hr later (P<0.05) and 6, 8, and 10-19 hr later (P<0.01) in cows fed the C and RAI diets, respectively. A significant negative correlation was observed between the lowest ruminal pH and its corresponding ruminal temperature in cows fed the C and RAI diets (r=-0.722 and -0.650, P<0.01, respectively), suggesting active fermentation and volatile fatty acid production in the rumen. However, ruminal pH profiles may not be predictable by measuring only ruminal temperature because decreases in ruminal pH were preceded by increases in ruminal temperature, and circadian changes in pH and temperature were associated with ruminal fermentation.     

Superoxide dismutase activity as a measure of hepatic oxidative stress in cattle following ethionine administration

The goal of this study was to assess if oxidative stress, as measured by alterations in the concentrations of antioxidant enzymes in the liver and erythrocytes of cattle, could be induced following dl-ethionine administration. Whole blood, serum and liver biopsy samples were collected 0, 4, 7 and 10 days after intra-peritoneal ethionine administration to five cows. The activities of the antioxidant enzymes copper zinc superoxide dismutase (Cu, Zn SOD) and catalase were assessed in the liver biopsies which were also examined histopathologically.Significant increases in hepatic Cu, Zn SOD concentrations (P < 0.01) were noted on days 7 and 10 post-treatment. Hepatic catalase activity decreased significantly (P < 0.01) on days 4, 7 and 10 post-treatment and erythrocyte Cu, Zn SOD activity was significantly increased on day 10. Serum biochemical analysis revealed a significant increase (P < 0.01) in non-esterified fatty acid concentrations on day 4 and significant decreases in total cholesterol and phospholipid levels on days 4 (P < 0.05), 7 (P < 0.01) and 10 (P < 0.01). In this model system, dl-ethionine administration was effective in inducing oxidative stress particularly reflected in the liver.     

Lipid peroxides and antioxidants in serum of neonatal calves

OBJECTIVE: To examine the association between oxidative stress and antioxidants in neonatal calves after birth. SAMPLE POPULATION: Sera from 6 healthy Holstein-Friesian cows and 7 of their newborn calves were obtained at various intervals after birth. PROCEDURE: Lipid peroxides in serum of cows and their newborn calves were estimated by measuring concentrations of thiobarbituric acid-reactive substances (TBARS). The antioxidative activities of neonatal sera were evaluated by measuring their superoxide-scavenging activities, ferroxidase activities, and the concentration of bilirubin-associated albumin. RESULTS: Concentration of TBARS in neonatal sera within 1 day after birth was significantly higher than concentrations > or = 2 days after birth and concentrations in serum of the dams. In contrast, antioxidative activities of neonatal sera, evaluated on the basis of superoxide-scavenging activities, ferroxidase activities, and concentration of bilirubin-associated albumin 3 hours after birth, were significantly lower than antioxidative activities in sera of the dams. CONCLUSIONS: Susceptibility of calves to oxidative stress during the neonatal period may be explained by the immature defense system against superoxide radicals.     

Changes of serum alkaline phosphatase activity in dry and lactational cows

Serum alkaline phosphatase (ALP) activities were measured during dry and lactational periods to investigate the influence of lactation on serum ALP activity in cows. Higher levels of serum ALP activity were seen in lactational periods than in dry periods. The serum activities of bone-specific ALP (BALP), liver ALP (LALP), tartrate resistant acid phosphatase (TRAP) and aspartate aminotransferase also increased in lactational periods. ALP activities in the bone extract and in whey were decreased at similar rates by the addition of lectin. Moreover, since the ALP band in whey was observed to have the same migration in polyacrylamide gel (PAG) disk electrophoresis as that of the bone extract, analysis of ALP isoenzymes by lectin affinity or PAG disk electrophoresis could not distinguish ALP originating from the mammary gland from that of bone. In this study, it was clear that the increased level of serum ALP activity was due to increases of BALP and LALP in lactational periods. However, the extent of the influence of ALP originating from the mammary glands on serum ALP activity was unknown. Judging from changes of BALP and TRAP activities in the serum and the correlation between the both, it was guessed that ALP originating from the mammary glands influenced serum ALP activity.     

Traumatic intracerebral hematoma in a dog: MR images and clinical findings

A nine-year-old, male, mongrel dog was unsteady after falling down a set of stairs. The dog exhibited a mildly abnormal gait 2 days after injury, and was not able to stand, with spasticity of the right limbs, 4 days after injury. MR imaging revealed a clearly demarcated mass on the top of the left lateral ventricle that showed mild hyperintensity on T1-weighted images and hyperintensity on T2-weighted images. The authors diagnosed the dog as having a traumatic intracerebral hematoma. This type of case, in which the clinical signs deteriorated due to edema associated with hematoma, is extremely rare.     

Bovine leukemia virus high tax molecular clone experimentally induces leukemia/lymphoma in sheep

Sheep were inoculated with high tax coded pBLV-IF (H group, Nos.1-5) of bovine leukemia virus (BLV), wild tax coded pBLV-IF (W group, Nos. 6-11), or control plasmid (C group, Nos. 12-14). During the observation period (4 to 46 months), 5 of 5 cases in H group and 3 of 6 cases (Nos. 6, 7, 9) in W group became positive for gp 51. Only 1 case in H group became leukemic, and one case each of H and W groups developed lymphoma. In No. 3, lesions were found in multiple organs including the lymph nodes, gastrointestinal tract following abomasum, and heart. In No. 6, lesions of lymphoma were found only in the jejunum and heart. Morphologically, small to middle-sized lymphocytic neoplastic (NP) cells were found in both cases, but lymphoblastic NP cells were found only in No. 3. By immunohistochemical examination, the phenotypes of NP cells were determined as CD1-, CD4-, CD5- -, CD8alpha-, sIgM+, lambda light chain+, B-B4+, MHC class II+ in both case. The results of this study indicate that inoculation of pBLV-IF can induce lymphocytic and lymphoblastic leukemia/lymphoma in sheep. Additionally, it is suggested that the expression rate of tax gene is not associated with the development of leukemia/lymphoma in sheep experimentally inoculated with pBLV-IF.     

Adenoviral gizzard erosion in commercial broiler chickens

Pathologic and immunohistochemical changes caused by group I of the fowl adenovirus (FAV) serotype-1 99ZH strain, isolated from broiler chickens exhibiting gizzard erosion, were investigated in commercial broiler chickens. One hundred twenty-two chickens were inoculated with the strain by both oral and ocular routes at 1, 3, or 5 weeks of age and euthanatized for necropsy within 4-18 days of inoculation. Focal gizzard erosions were observed in the inoculated chickens of each age group. A histologically degenerative koilin layer, necrotic mucosa, intranuclear inclusion bodies in the glandular epithelial cells, inflammatory cell infiltrations in the lamina propria, submucosa, and a muscle layer were seen in the gizzards. Immunohistochemical staining showed evidence of FAV antigens in the intranuclear inclusion bodies. These findings were recognized regardless of their maternal antibody levels for FAV serotype-1. Gizzard lesions appeared later in the lower-dose-inoculated chickens than in the higher-dose-inoculated chickens. Numerous CD3-positive cells and IgY-positive plasma cells were seen in the gizzard lesions. In 5-week-old chickens the heterophil infiltrations in the lesions were milder than in younger chickens. Intranuclear inclusion bodies also were observed in the epithelial cells of the ileum or cecal tonsils of some chickens. Thus, this study shows that FAV-99ZH causes adenoviral gizzard erosion in broiler chickens without hepatic or pancreatic lesions and that cell infiltration is more severe than in dietary gizzard erosions.     

Association between changes in plasma calcium concentration and plasma tartrate-resistant acid phosphatase activity in periparturient cows

Eight periparturient Holstein Friesian cows were examined for plasma tartrate-resistant acid phosphatase (TRAP) activity to assess the degree of bone metabolic activity and to evaluate the association between the change in calcium (Ca) concentration and bone metabolism during the periparturient period. Milk fever occurred in 1 of 8 cows just after parturition. Plasma TRAP activities did not markedly change in 5 of 8 cows during the experimental period. The changing rate of Ca between preparturition and just after parturition was under -20% in 3 of 8 cows, and low TRAP activities were observed in 2 of these 3 cows. This study suggests that cows with a low TRAP activity are at risk of developing milk fever in comparison to cows with high TRAP activity. Temporary increases of parathyroid hormone were observed in 7 cows, but not in the cow with milk fever.