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Detection and antigenicity of chlamydial proteins that blind eukaryotic cell membrane proteins
1992
Baghian, A. | Schnorr, K.L.
Chlamydia psittaci proteins capable of binding eukaryotic cell membranes were identified and antigenically characterized. Cell membrane proteins (CMP) of noninfected cells were labeled with biotin (B-CMP), then were extracted with 1% Triton X-100. Nitrocellulose membrane strips containing sodium dodecyl sulfate-polyacrylamide gel electrophoresis-resolved proteins of chlamydial elementary bodies (EB) were reacted with the B-CMP extract, followed by addition of streptavidin-conjugated horse radish peroxidase. Among the various strains of chlamydiae examined, a protein of approximately 16 to 18 kDa consistently bound B-CMP. A second larger protein, ranging in molecular mass from 24 to 32 kDa, also bound B-CMP. Immunoblotting techniques were used to analyze the reactions of antisera from immunized and experimentally infected animals to these proteins. A rabbit polyclonal antiserum produced against the 18-kDa adhesin of a serovar-1 strain of C psittaci (B577) reacted strongly with 18-kDa proteins of aH C psittaci strains, but weakly with that of C trachomatis. Mouse antisera raised against the serovar-2 (FCc-Stra) 28-kDa protein reacted only with proteins of the homologous serovar. Sera from experimentally infected animals did not react with the C trachomatis 18-kDa adhesion protein, but did react in 2 patterns with related and nonrelated C psittaci isolates. Two rabbits inoculated with infective serovar-1 EB and 1 rabbit inoculated with a serovar-2 strain reacted specifically with the 18-kDa proteins of their homologous serovars. In contrast, 2 other rabbits inoculated with the same serovar-2 strain produced antisera that reacted with all C psittaci 18-kDa proteins, as did serum from a similarly inoculated buff. The reason for the 2 types of responses to infection remains to be determined.
Afficher plus [+] Moins [-]Measurement of serum myoglobin concentrations in horses by immunodiffusion
1992
Holmgren, N. | Valberg, S.
Quantitative immunodiffusion in one dimension was performed in 6-mm Duran tubes containing a 1% Nobel agar solution and various dilutions of antisera. A series of dilutions of pure myoglobin in equine sera as well as plasma from horses with rhabdomyolysis were tested. Standard curves were prepared of the migration distance of the formed precipitate from the meniscus of the gel after 3, 6, 12, and 24 hours. The clearest line of precipitate was formed with a 1:20 dilution of antisera in agar. Standard curves were nonlinear and plasma myoglobin could be detected at 2 micrograms of myoglobin/ml or greater. The test was optimal, with an error of 5.6%, when read at 24 hours at approximately 25 C. Tubes with agar could be stored for 6 months at 4 C without affecting the accuracy of the test. The specificity of myoglobin for skeletal or cardiac muscle, and its rapid clearance from serum after muscle necrosis, make it ideally suited for evaluating acute muscle damage and for testing the susceptibility of horses for rhabdomyolysis following an exercise test.
Afficher plus [+] Moins [-]Potential attachment area of the first mandibular molar in dogs
1992
Smith, M.M. | Massoudi, L.M.
Tooth root and root trunk surface area measurements were taken from 20 first mandibular molar (M1) teeth obtained randomly from canine cadaver specimens. The mean root surface area for all teeth was 497.1 +/- 116.2 mm2. The cumulative surface areas of the mesial (MR) and distal (DR) roots were 251.4 +/- 70.2 mm2 and 211.7 +/- 51.9 mm2, respectively. The root trunk surface area was 34.0 +/- 21.4 mm2. Surface area measurements for individual roots and the root trunk were significantly different. Cumulative individual root surface area (MR + DR) was 93.2% of total root surface area. The MR surface area contributed most to total root surface area (50.6%) and cumulative individual root surface area (54.3%). The mean distance from the beginning of the root trunk to the furcation was 2.1 mm. On the basis of the linear variation of the percentage of root surface area of the M1, 44.6% of the total root surface area was 3 to 7 mm apical to the most coronal aspect of the cementoenamel junction. The MR was heart-shaped, compared with the more cylindrical shape of the DR.
Afficher plus [+] Moins [-]Impact on bovine somatotropin administration beginning at day 70 of lactation on serum metabolites, milk constituents, and production in cows previously exposed to exogenous somatotropin
1992
Lean, I.J. | Baldwin, R.L. | Troutt, H.F. | Bruss, M.L. | Galland, J.C. | Farver, T.B. | Rostami, J. | Weaver, L.D. | Holmeberg, C.A.
Metabolic and production responses are reported for 72 cows treated with bovine somatotropin (BST) for 30 days starting at day 70 of lactation. Of these 72 cows, 48 had been exposed in the preceding lactation to long-term treatment with BST at 3 dosages and 24 (controls) had not been given BST. Approximately half of the cows in each group were parity-2 cows, the rest were older. Comparisons between groups were made separately for parity-2, and older cows. Analyses, using pretreatment values of each variable as a covariate, indicated that older cows, but not parity-2 cows, significantly (P < 0.05) increased milk production during treatment. Parity-2 cows, however, had a significantly higher milk fat percentage than controls following treatment. Cows treated with 51.6 or 86 mg BST/d in both parity groups has significantly higher serum-free fatty acids than controls. Estimated net energy balances were significantly lower for older treated cows, but did not significantly differ from controls from for parity-2 treated cows. Older cows in the 86 mg of BST/d group tended to have higher concentrations of blood glucose than did older control-group cows. Treatment with BST did not significantly increase serum ketone concentrations in any group of animals, and none of the cows developed clinical ketosis during this period. Estimated net energy balance (ENEB) during treatment was a significant (P < 0.05) covariate for free fatty acid concentrations in older cows and for milk fat percentage in parity-2 cows. Covariate adjusted analyses, using ENEB during treatment as a covariate, indicated that lipolytic stimuli already acting may be enhanced by treatment with BST, but a negative energy balance was not a necessary precondition for free fatty acid concentrations to increase following somatotropin treatment. Similarly, milk fat percentages for parity-2 treated cows were significantly (P < 0.05) higher during treatment than controls when ENEB during treatment was used as a covariate. Increased milk fat concentrations in parity-2 treated cows were not associated with significant increases in the ratio of C(18):C(4-10) milk fatty acids, indicating that increased milk fat resulted from either an increase in incorporation of C(18) fatty acids into milk fat coupled with an increase in de novo mammary synthesis of C(4-10) milk fatty acids or an increase in C(12-16) fatty acids that may arise either from increased tissue mobilization, from diet, or from de novo mammary synthesis.
Afficher plus [+] Moins [-]Determination of sensitivity to metocurine in exercised horses
1992
White, D.A. | Hildebrand, S.V. | Jones, J.H. | Fung, D.L. | Gronert, G.A.
On the basis of results in dogs, conditioning exercise may increase sensitivity to nondepolarizing muscle relaxants. Five Thoroughbreds were exercised/conditioned 3 times weekly on a treadmill for 8 months. Increasing maximal rate of O2 consumption verified that the horses were responding to exercise conditioning. Six nonexercised Thoroughbreds served as the control group. Studies were done with horses under general anesthesia by use of halothane during partial paralysis by a brief constant-rate infusion with the muscle relaxant, metocurine iodide. Quantification of degree of paralysis of the hoof twitch (eg, digital extensor) occurred with simultaneous quantification of blood values of metocurine. Pharmacokinetic and pharmacodynamic analyses of the data were done by a nonlinear regression program, using the Hill equation. There were no differences in findings between exercised and nonexercised horses. The mean blood concentration for the 50% paralyzing dose of metocurine was 0.44 +/- 0.11 (SD) micrograms/ml in exercised horses, and 0.58 +/- 0.22 micrograms/ml in nonexercised horses. Despite evidence for a response to conditioning, a significant change in the sensitivity of the neuromuscular junction to metocurine was not found.
Afficher plus [+] Moins [-]Effect of metoclopramide on luteinizing hormone secretion in postpartum anestrous cows
1992
Thompson, F.N. | Jones, R.D. | Stuedemann, J.A. | Mizinga, K.M. | Smith, C.K.
The effect of metoclopramide (MC), a dopamine antagonist on luteinizing hormone (LH), was examined in anestrous primaparous cows. Metoclopramide has been found to be beneficial in overcoming fescue toxicosis; increasing LH secretion stimulates return to ovulatory function after parturition. Consequently, if MC had negative effect on LH secretion, it would indicate that administration of MC to reproducing animals might be limited. Of 14 postpartum (47 to 66 days) cows, 7 were given MC (4 mg/kg of body weight, IV), and 7 served as controls. Blood was obtained via jugular cannulas at 15-minute intervals for 8 hours; MC was given at the end of the first hour, and gonadotropin-releasing hormone (GnRH, 7 mg/kg), was given IV at the end of hour 7 as a challenge stimulus for LH secretion. Prior to GnRH administration, MC did not have significant effect on LH secretion, as judged by mean serum LH concentration, LH pulse frequency, and LH pulse amplitude. Administration of MC resulted in greater (P < 0.05) LH response to GnRH, indicating enhanced secretory ability when the pituitary gland was challenged. Serum prolactin concentration was increased (P < 0.01) by MC administration. Therefore, MC did not have adverse effect on LH secretion in postpartum cows.
Afficher plus [+] Moins [-]Protection of neonatal calves against fatal enteric colibacillosis by administration of egg yolk powder from hens immunized with K99-piliated enterotoxigenic Escherichia coli
1992
Ikemori, Y. | Kuroki, M. | Peralta, R.C. | Yokoyama, H. | Kodama, Y.
The protective effects of egg yolk powder prepared from hens vaccinated with heat-extracted antigens from K99-piliated enterotoxigenic Escherichia Coli (ETEC) strain 431 were evaluated in a colostrum-fed calf model of ETEC-induced diarrhea caused by a heterologous strain (B44). The antibody powder was obtained by spray-drying the water-soluble protein fraction of egg yolks after removing the lipid and fatty components by precipitation with hydroxypropylmethylcellulose phthalate. A total of 16 colostrum-fed calves were studied to determine whether the orally administered antibody powder would prevent fatal bovine colibacillosis caused by a virulent ETEC strain. Clinical response of individual calves was monitored and evaluated in the context of these variables: fecal consistency score, intestinal colonization, weight loss, and mortality. Control calves that were treated with vehicle (milk with egg yolk powder from nonimmunized hens) had severe diarrhea and dehydration and died within 72 hours after infection was manifested. In contrast, calves fed milk containing egg yolk powder with antipili agglutinin titers of 1:800 and 1:1,600 had transient diarrhea, 100% survival, and good body weight gain during the course of the study. Results indicate that the orally administered egg yolk powder protected against ETEC-induced diarrhea in neonatal calves and that the protective components may have been the antibodies raised by vaccination of chickens against ETEC.
Afficher plus [+] Moins [-]Use of computertized interactive morphometry in the diagnosis of mamary adenoma and adenocarcinoma in dogs
1992
Ciurea, D. | Wilkins, R.J. | Shalev, M. | Liu, Z. | Barba, J. | Gil, J.
We attempted to define quantitative objective criteria for diagnosis of mammary adenoma and adenocarcinoma in dogs. To that end, correlation between diagnosis made by conventional histologic examination and morphometric descriptors, obtained by computerized histologic analysis, was assessed in biopsy specimens of mammary tissue from 63 dogs. We used 2 interactive computer procedures: one assessed mean nuclear perimeter, mean nuclear area, and circularity factor; and the other evaluated nuclear stratification and cell crowding. A data base was generated with 11 specimens from normal mammary tissue, 17 specimens from mammary adenoma, 18 specimens from low-grade mammary adenocarcinoma, and 15 specimens from mammary adenocarcinoma. The mean values of nuclear perimeter, nuclear area, nuclei per millimeter of basement membrane, and minimal distances from cells to basement membrane gradually increased from normal to high-grade malignancy. Distributions of nuclear areas and of minimal distances from cells to basement membrane were shifted in specimens from malignant tumors. Multivariate analysis confirmed the homogeneity of the diagnostic groups.
Afficher plus [+] Moins [-]Effects of lactated Ringer solution and prednisolone sodium succinate on dogs with induced hemorrhagic shock
1992
Hankes, G.H. | Dillon, A.R. | Ravis, W.R.
Hemorrhagic shock was induced in nonsplenectomized dogs by removing 41% of their blood volume over a 15-minute period. Hemodynamic and metabolic variables were determined prior to and for 3 hours after completion of hemorrhage. One group of 5 dogs was not treated. After the 30-minute sample was collected, a second group of 5 dogs was given lactated Ringer solution (LRS) at 88 ml/kg of body weight, IV. A third group of 5 dogs was given LRS (88 ml/kg, IV) and prednisolone sodium succinate (11 mg/kg, IV) 30 minutes after hemorrhage. The IV administration of LRS was completed within 15 minutes. The glucocorticoid was administered as an IV bolus after 500 ml of LRS had been given. The large volume and administration of LRS significantly (P = 0.05) improved many of the hemodynamic and metabolic effects of acute hemorrhage and hemorrhagic shock. At one time or another during the 2.5-hour observation period after the initiation of treatment, mean arterial pressure, cardiac index, systemic vascular resistance, heart rate, respiratory rate, lactate, glucose, and arterial and venous blood gas values were significantly (P = 0.05) improved, compared with baseline values. The addition of prednisolone sodium succinate to the treatment regimen improved the effectiveness of LRS alone only in some dogs at random sampling times. Significant trends were not observed except, possibly, the improvement of venous pH and A-V pH and P(CO)2 differences.
Afficher plus [+] Moins [-]Effects of clinically apparent and subclinical Ostertagia ostertagi infections on nitrogen and water metabolism in calves
1992
Xiao, L. | Gibbs, H.C.
Four balance trials were conducted in 3 groups of 5 calves each at 0, 4, 8, and 14 weeks after initial inoculation with Ostertagia ostertagi. Group-1 calves were inoculated with 100,000 third-stage larvae (L3) of O. ostertagi/wk for 14 weeks. Group-2 calves were inoculated with 10,000 L3/wk for 14 weeks, and group-3 calves were not inoculated. Effects of infection on apparent digestibilities of dry matter and nitrogen, and balances of nitrogen and water were evaluated. Neither clinically apparent (group 1) nor subclinical (group 2) O. ostertagi infections had observable effects on the apparent digestibility of dry matter. Subclinical infection also had no significant effects on nitrogen digestibility or nitrogen and water balance. Clinically apparent infection, however, decreased the apparent digestibilities of nitrogen significantly (P < 0.05) at 4, 8, and 14 weeks after inoculation, and decreased nitrogen balance at 4 and 8 weeks after inoculation. Group-1 calves also had lower water intake, fecal-water excretion, and apparent water balance, but higher urinary water output at 4, 8, and 14 weeks after inoculation.
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