Quantification of plasma reduced glutathione, oxidized glutathione and plasma total glutathione in healthy cats

Glutathione is an important intracellular tripeptide with multiple functions. Abnormal glutathione metabolism is thought to play an important role in various diseases of cats. However, no data regarding concentration of plasma glutathione are available for domestic cats. This study discusses the development of a rapid, simple high pressure liquid chromatography method for measurement of reduced glutathione (GSH), oxidized glutathione (GSSH) and total glutathione (GSHt) in plasma, for the purpose of establishing baseline data for future studies. The mean concentrations of GSH, GSSH and GSHt were 4.51+/-1 microM; 19.44+/-3.79 microM (expressed as GSH equivalent) and 23.59+/-3.89 microM, respectively. This is the first report of plasma glutathione concentrations in this species.     

Effect of non-linear plasma protein binding on unbound and total plasma phenylbutazone concentrations in cows

Boudinot, F.D., Williams, R.J. & Smith, J. A. Effect of non-linear plasma protein binding on unbound and total plasma phenylbutazone concentrations in cows. J. vet. Pharmacol. Therap. 13, 132–136.
The influence of plasma protein binding on unbound and total phenylbutazone concentrations in cows was examined employing data from the literature. Protein binding parameters (number of binding sites and affinity constants) were generated by computer analysis to characterize the concentration-dependent plasma protein binding of phenylbutazone. Unbound plasma phenylbutazone concentrations were calculated from total plasma drug concentrations observed after administration of a single dose of phenylbutazone to cows. Pharmacokinetic parameters for unbound phenylbutazone were obtained. Parameters characterizing die plasma protein binding and pharmacokinetics of unbound phenylbutazone derived from single-dose administration were then used to predict unbound and total drug concentrations after multiple-dose administration of phenylbutazone. Total plasma phenylbutazone concentrations predicted from single-dose pharmacokinetic parameters agreed well with observed values following multiple-dose administration of the drug. Thus, the results of this analysis demonstrate that the non-linear pharmacokinetics of phenylbutazone in the cow can be attributed to the concentration-dependent plasma protein binding of the drug.
F. Douglas Boudinot, Department of Pharmaceutics, College of Pharmacy, University of Georgia, Athens, GA 30602, USA.     

Evaluation of total plasma bile acid concentrations for the diagnosis of hepatobiliary disease in horses

Plasma bile acid concentrations were measured in normal horses. There was no diurnal variation in values, and age and sex had no effect. There was no significant difference between serum and plasma bile acid concentrations in clinically normal horses. Plasma bile acids were stable on storage for one month at -20 degrees C. The total plasma bile acid concentrations together with total and direct bilirubin concentrations and plasma activities of aspartate aminotransferase, glutamate and iditol dehydrogenase were evaluated in horses with various types of hepatobiliary disease (hepatic necrosis, lipidosis, neoplasia and cirrhosis), gastrointestinal disease, cardiovascular, orthopaedic and various other conditions not affecting the liver. Total plasma bile acids together with plasma glutamate and iditol dehydrogenase activities were the best indicators of liver disease. Total plasma bile acid concentrations were the most sensitive indicator of a wide variety of hepatic diseases but alone were unhelpful in differential diagnosis and were of more value when combined with the other tests of hepatic disease.     

Fluctuations in total calcium and magnesium in the plasma and uterine fluid of the domestic fowl

Fluctuations in calcium and magnesium were measured in the shell gland fluid and blood from the inferior oviducal vein of the domestic fowl throughout the laying cycle.

It was concluded that:

1. The variations in calcium concentration reflected both the known uptake of this ion by the albumen and medullary bone formation and destruction.

2. The variations in magnesium concentration did not parallel the changes in calcium concentration.

3. Magnesium was released into the blood during medullary bone formation.

     

Increase in plasma total antioxidant capacity of grazing Japanese Black heifers and cows in forestland in Japan

Blood total antioxidant capacity (TAC) has become a key bio‐marker for animal health. Forest‐grazing cattle are known to forage various native plants that have high TAC. This study evaluated differences of plasma TAC between forest‐grazing (FG) and pasture‐grazing cattle (PG). Experiment 1 monitored the plasma TAC levels of 32 Japanese Black cattle. The level in PG did not change throughout the grazing period. However, that in FG, which increased from summer, was significantly higher than that in PG through fall (P < 0.05). In experiment 2, we used nine Japanese Black heifers and investigated their blood antioxidant parameters and the TAC in plants that the cattle consumed in late June and September. The plasma TAC levels in FG were significantly higher than those in PG in both periods (P < 0.05). Plasma levels of lipid peroxidation in FG tended to be lower than that in PG (P = 0.098). Furthermore, the TAC levels in various species of shrubs and trees consumed by FG were higher than those in pasture grasses. Results of this study show that plasma TAC of grazing Japanese Black cattle in forestland increase from summer through fall.     

Determination of free and total cyst(e)ine in plasma of dogs and cats

BACKGROUND: In human blood, the amino acid cysteine forms disulfide bonds with itself and with other sulfhydryl compounds in their free form and with sulfhydryls in protein. Protein-bound cysteine is lost when plasma proteins are removed before amino acid analysis. OBJECTIVE: The purpose of this study was to assess the time course and extent of cyst(e)ine (cysteine + half-cystine) loss in dog and cat plasma. METHODS: An equal volume of 6% sulfosalicylic acid was added to plasma aliquots at 0, 2, 4, 10, 16, 24, 36, 48, 60, and 72 hours after separation of blood cells. Tris-2-carboxyethyl-phosphine hydrochloride (TCEP - HCl), a reducing agent, was used to regenerate total plasma cyst(e)ine after 3 months of sample storage (-20 degrees C). RESULTS: Initial free cyst(e)ine concentrations (mean +/- SEM) were higher in canine plasma (77 +/- 4 micromol/L) than in feline plasma (37 +/- 3 micromol/L). Free plasma cyst(e)ine concentrations in dogs and cats decreased after first-order kinetics, with a half-life of 23 and 69 hours, respectively. Total plasma cysteine after TCEP - HCl treatment was similar for dogs (290 micromol/L) and cats (296 micromol/L), but the percentage of free cysteine was higher (P = .02) in dogs (27%) than in cats (13%). Over half of the cyst(e)ine, homocysteine, cysteinylglycine, and glutathione were bound in vivo to plasma proteins. CONCLUSION: These results emphasize the importance of removing plasma proteins within 1 hour after blood collection for reliable assay of free plasma cyst(e)ine.     

Changes in the level of plasma cortisol, progesterone and total testosterone in developing hairless dogs.

Changes in the levels of plasma cortisol, progesterone and total testosterone were examined in developing hairless and haired dogs. Cortisol levels in the hairless dogs seemed to be higher than those in haired dogs within the age of 4-5 weeks. No apparent changes were seen in the level of plasma progesterone between the groups of hairless females and haired females. Total testosterone levels in hairless males showed to be significantly lower than those in haired males at the age of 13-21 weeks.     

Interdependence of the level of total proteins, magnesium and beta-carotene in the plasma of dairy cows

The degree of dependence of the concentration of total protein, magnesium and beta-carotene in the blood plasma of over 3,400 dairy cows in the first and second phases of lactation and in months 8 and 9-9.5 of pregnancy was attested statistically. It was shown that in late pregnancy the concentration of all three indices drops both with winter and summer feed rations and that the connection between all three pairs of indices is positive. The highest values of the linear correlation coefficient were ascertained between the total protein and magnesium of blood plasma (+0.87 in the winter period, +0.63 in the summer period and +0.75 in both periods), the correlation was somewhat lower between magnesium and beta-carotene (+0.56, +0.75 and +0.36) and it was lowest between total protein and beta-carotene (+0.45, +0.41 and +0.42). The discussion deals with the causes of the stochastic correlation of these components of blood plasma.     

Seminal plasma insulin-like growth factor I and total protein concentration in peripubertal period of the Gyr bulls

This study aimed to evaluate the correlation of seminal plasma IGF-I and total proteins from peripubertal bulls with semen characteristics and onset of puberty. Sixteen dairy Gyr bulls were separated into early (n = 8) and regular (n = 8) groups, according to the onset of puberty. Semen was monthly collected by electroejaculation from 14 to 26 months of age, and the onset of puberty was retrospectively determined (17.0 ± 1.6 and 19.2 ± 1.5 months, to early and regular group, respectively). Five time points were evaluated (day −60, day −30, day 0, day 30 and day 60 days of puberty), being day 0 considered as beginning of puberty. Scrotal circumference and body weight were also assessed. Semen characteristics were evaluated, the seminal plasma was separated by centrifugation and total protein and IGF-I concentrations were determined. There was no difference between groups to concentration of the seminal plasma total proteins, but we found an interaction between group and age. Seminal plasma IGF-I concentrations were not different between early and regular groups; thus, the data were combined for analysis. Combined IGF-I concentrations were positively correlated with sperm motility and concentration, age, body weight and scrotal circumference. Negative correlation was found between IGF-I concentration and total sperm defects. Increased IGF-I was observed in day −30 and day 0 of puberty in early and regular groups, respectively. Seminal plasma total proteins are involved in peripubertal modifications and IGF-I from Zebus dairy bulls can influence the seminal characteristics and the growth factor increase occurs concomitantly with body growth but cannot be used to define puberty bulls earlier.     

Sequential changes in plasma progesterone, total oestrogens and corticosteroids in cow throughout pregnancy and around parturition

Blood samples were taken thrice a week for 2 weeks from 33 pregnant cows in 9 groups, 1 month apart. In addition, blood samples were collected twice daily 1 week from 8 cows before parturition. Plasma progesterone and total oestrogens were quantified by validated radioimmunoassays and total corticosteroids by a competitive protein binding technique. During the 1st 3 month of pregnancy, no remarkable changes were observed from the 3 hormones studied. From the 4th month on, concentrations of progesterone and total corticosteroids decreased significantly (P less than 0.05) and remained fairly constant until the 9th month. Values of total oestrogens increased significantly (P less than 0.05) from the 4th to 6th months and stayed unchanged afterwards. Progesterone levels in periparturient cows decreased linearly (P less than 0.01) during 3 days preceding delivery. Concentrations of total oestrogens rose sharply from day -5 and peaked at calving. Values of total corticosteroids increased dramatically on the 6th day and 24 hours before parturition. The results obtained suggest that in contrast to studies cited earlier plasma corticosteroids could play a key role for triggering parturition in cows.     

The value of plasma fibrinogen estimations in dogs. A comparison with total leucocyte and neutrophil counts

Plasma fibrinogen levels were measured by a simple refractometry method on 552 dog blood samples submitted from clinical cases for routine haematological examination. Sixty samples with total leucocyte and neutrophil counts within the defined normal range had elevated fibrinogen levels. The majority of these samples were from animals with a disease condition. It was concluded, because of this, and because of the simplicity of the method, that fibrinogen estimations are a worthwhile addition to the routine haematological examination of dog blood.     

The sensitivity, specificity and predictive value of total plasma protein in the diagnosis of traumatic reticuloperitonitis.

We analyzed the value of total plasma protein in the diagnosis of traumatic reticuloperitonitis in dairy cattle using the concept of sensitivity, specificity and predictive value of a test. Cows with traumatic reticuloperitonitis surgically confirmed at the New York State College of Veterinary Medicine served as cases, while cows surgically explored for other abdominal diseases that might be confused with traumatic reticuloperitonitis during differential diagnosis were controls. The sensitivity of the total plasma protein in diagnosis of traumatic reticuloperitonitis decreased from 97% to 6.3% as the cutoff value for a positive test rose from 65 g/L to 110 g/L, while specificity increased from 11.3% to 99.1% over the same range of cutoff values. Application of these figures to predictive values positive and negative at various prevalence levels of disease is discussed. For example at the measured prevalence level of 37%, 76% of cattle with plasma protein concentrations in excess of 100 g/L had traumatic reticuloperitonitis.     

Sequential changes in plasma progesterone, total oestrogens and corticosteroids in the cow throughout pregnancy and around parturition

Blood samples were taken thrice weekly for 2 weeks from 33 pregnant cows in nine groups, 1 month apart, and twice daily for 8 days commencing 1 week before parturition from eight cows. Plasma progesterone and total oestrogens were quantified by validated radioimmunoassays and total corticosteroids by a competitive protein binding technique. During the first 3 months of pregnancy, no remarkable changes were observed for the three hormones studied. From the fourth month on, concentrations of progesterone and total corticosteroids decreased significantly (P less than 0.05) and remained fairly constant until the ninth month. Values of total oestrogens increased significantly (P less than 0.05) from the fourth to the sixth month and thereafter remained unchanged. In the periparturient cows, progesterone levels decreased linearly (P less than 0.01) during the 3 days preceding delivery. Concentrations of total oestrogens rose sharply from day -5 and peaked at calving. Values of total corticosteroids increased dramatically on the sixth day and also at 24 hours before parturition. These results suggest that, contrary to earlier studies, plasma corticosteroids could have a primary key role for triggering parturition in cows.