Myoplasmic calcium regulation in myotubes from horses with recurrent exertional rhabdomyolysis

OBJECTIVE: To determine whether alterations in myoplasmic calcium regulation can be identified in muscle cell cultures (myotubes) and intact muscle fiber bundles derived from Thoroughbreds affected with recurrent exertional rhabdomyolysis (RER). ANIMALS: 6 related Thoroughbreds with RER and 8 clinically normal (control) Thoroughbred or crossbred horses. PROCEDURES: Myotube cell cultures were grown from satellite cells obtained from muscle biopsy specimens of RER-affected and control horses. Fura-2 fluorescence was used to measure resting myoplasmic calcium concentration as well as caffeine- and 4-chloro-m-cresol (4-CMC)-induced increases in myoplasmic calcium. In addition, intact intercostal muscle fiber bundles were prepared from both types of horses, and their sensitivities to caffeine- and 4-CMC-induced contractures were determined. RESULTS: Myotubes of RER-affected and control horses had identical resting myoplasmic calcium concentrations. Myotubes from RER-affected horses had significantly higher myoplasmic calcium concentrations than myotubes from control horses following the addition of > or = 2mM caffeine; however, there was no difference in their response to 4-CMC (> or = 1 mM). Caffeine contracture thresholds for RER and control intact muscle cell bundles (2 vs 10mM, respectively) were significantly different, but 4-CMC contracture thresholds of muscle bundles from RER-affected and control horses (500 microM) did not differ. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in caffeine sensitivity of muscle cells derived from a family of related RER-affected horses was detected in vitro by use of cell culture with calcium imaging and by use of fiber bundle contractility techniques. An alteration in muscle cell calcium regulation is a primary factor in the cause of this heritable myopathy.     

Prevalence of feline leukemia virus infection and serum antibodies against feline immunodeficiency virus in unowned free-roaming cats

OBJECTIVE: To determine prevalence of FeLV infection and serum antibodies against feline immunodeficiency virus (FIV) in unowned free-roaming cats. DESIGN: Cross-sectional serologic survey. ANIMALS: 733 unowned free-roaming cats in Raleigh, NC, and 1,143 unowned free-roaming cats in Gainesville, Fla. RESULTS: In Raleigh, overall prevalence of FeLV infection was 5.3%, and overall seroprevalence for FIV was 2.3%. In Gainesville, overall prevalence of FeLV infection was 3.7%, and overall seroprevalence for FIV was 4.3%. Overall, FeLV prevalence was 4.3%, and seroprevalence for FIV was 3.5%. Prevalence of FeLV infection was not significantly different between males (4.9%) and females (3.8%), although seroprevalence for FIV was significantly higher in male cats (6.3%) than in female cats (1.5%). CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of FeLV infection and seroprevalence for FIV in unowned free-roaming cats in Raleigh and Gainesville are similar to prevalence rates reported for owned cats in the United States. Male cats are at increased risk for exposure to FIV, compared with female cats.     

Coordinate regulation of ovine adipose tissue gene expression by propionate

The current study examined the acute effects of intravenous propionate infusion on plasma hormones and metabolites and the expression of adipose tissue lipogenic genes. Four yearling rams were assigned to one oftwo groups (saline or propionate infusion) in a crossover design. All sheep were cannulated in both jugular veins and infused with 1.2 M propionate at a rate of 64 micromol x mix(-1) x kg BW(-1) for 30 min. Blood samples were collected at -10, 0, 5, 10, 20, 30, 60, and 120 min after initiation of infusion. Subcutaneous adipose tissue biopsies were obtained from the tailhead at 0 and 2 h after propionate infusion and analyzed for gene expressions of lipoprotein lipase, acetyl CoA carboxylase, fatty acid synthase, peroxisome proliferator-activated receptor gamma, leptin, and uncoupling protein-2 using a nonisotopic ribonuclease protection assay. The partial cDNA of the enoyl reductase region of ovine fatty acid synthase was cloned and sequenced from s.c. adipose tissue of sheep. The deduced amino acid sequence (210 amino acids) was 86% identical to human, 88% identical to rat, 88% identical to mouse, and 72% identical to chicken. Plasma glucose and insulin concentrations abruptly increased 5 min after beginning propionate infusion and further increased up until 30 min but were unaffected in saline-infused sheep (P < 0.05). Plasma concentration of NEFA decreased (P < 0.05) during propionate infusion, whereas IGF-I levels were unaltered. The amounts of lipoprotein lipase, acetyl CoA carboxylase, fatty acid synthase, peroxisome proliferator-activated receptor gamma, and leptin mRNA increased (P < 0.05) in s.c. adipose tissue of propionate-infused sheep compared with those of saline-infused sheep. However, uncoupling protein-2 mRNA decreased (P < 0.05) in propionate-infused sheep. This study demonstrates that an acute nutrient challenge, in the form of i.v. propionate, can stimulate or inhibit the expression of various adipose tissue genes involved with lipogenesis and adipose tissue metabolism.     

The detection of the meq gene in chicken infected with Marek's disease virus serotype 1

In the genome of strains of very virulent Marek's disease virus serotype 1(vvMDV1), such as Md5 and RB1B, the meq open reading frame (ORF) encoding a 339-amino-acid bZIP protein, is present, while a slightly longer meq ORF, termed as L-meq, in which a 180-bp sequence is inserted into the meq ORF is found in other strains of MDV1, such as CV1988/R6 and attenuated JM. When chickens were infected with vvMDV1 strains and the meq gene was amplified by nested polymerase chain reaction (PCR), the meq gene was detected throughout the experimental period for 7 weeks post inoculation (pi). However, the L-meq gene was also detected at 3 to 5 weeks and 3 to 4 weeks pi. in Md5-infected and RB1B-infected chickens, respectively. In the case of chickens infected with an attenuated MDV1, the JM strain, the L-meq gene was detected at 2 to 7 weeks pi., and the meq gene was also detected at 2 to 6 weeks pi. Both L-meq and meq genes were detected in chickens infected with an attenuated nononcogenic vaccine strain of MDV1 (CVI988/R6), throughout the experimental period. Though quantitative PCR was not performed, a larger amount of the PCR products corresponding to the L-meq than the meq gene was amplified from chickens infected with JM or CVI988/R6. These results suggest that a dynamic population shift between the MDV subpopulations displaying meq and L-meq genes occurs in chickens during the course of MDV infection. Since the MDV subpopulation that displays the L-meq gene only displays it during the latent phase, the L-meq and its gene product, if any, might contribute to the maintenance of the MDV latency.     

Low expression of cyclin D2 in G2/M-arrested and transformed proliferating Balb/3T3 cells

At present, the effect of cyclin D2 implicated in cell cycle regulation, differentiation, and oncogenic transformation is not fully confirmed. To better elucidate the role of cyclin D2 in controling the cell proliferation, cyclin D2 expression level was determined at the early initiation and promotion stages during the in vitro two-stage transformation process of Balb/3T3 A31 cells. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced G2/M-arrested cells expressed low level of cyclin D2 mRNA, while the contact-inhibited nonproliferating cells expressed high level of cyclin D2 mRNA. In the transformed proliferating cells at the promotion stage, cyclin D2 mRNA was not expressed. These data suggest that cyclin D2 expression may be associated with the type of growth arrest and nonproliferating state, but not with the cell proliferation and transformation.     

Evaluation of canine gastric motility with ultrasonography

For the evaluation of canine gastric motility with ultrasonography, contraction number of pyloric antrum and gastric emptying time (GET) by area and volume method developed by Bolondi et al.'s method were studied in 14 dogs. All experimental dogs were administered with saline and soup solution (10 ml/kg, B.W.). The mean values of contraction number of pyloric antrum in saline and soup group were 4.19 +/- 1.30/min and 4.82 +/- 0.65/min before feeding, and overall mean values were 4.66 +/- 1.37/min and 5.13 +/- 1.71/min, respectively. The mean values of the GET by area and volume method were 36.73 +/- 11.27, 40.00 +/- 8.87 min in saline group and 61.35 +/- 17.58, 59.11 +/- 14.46 min in soup group. In the GET in saline and soup groups, there was no significant difference between the area and volume method (p>0.05). Therefore, Bolondi et al.'s method by ultrasound can be used to evaluate the antropyloric motility and gastric emptying time with area and volume methods. The area method is easier to determine the GET than the volume method, but the latter is more accurate.     

Growth performance and health status in weanling piglets fed spray-dried porcine plasma under typical Northern European conditions

The effect of inclusion of spray-dried porcine plasma (SDPP) in diets for weanling piglets was studied. The objectives were to determine whether SDPP would have positive effects on post-weaning piglet performance and health under typical Northern European conditions. In experiment 1, 160 weanling piglets were assigned randomly to a control diet or a diet containing 3% SDPP, which was added at the expense of both fishmeal and dried skim milk. In experiment 2, 264 weanling piglets were assigned to a control diet containing whey protein, a diet without whey protein but with SDPP or a diet containing both whey protein and SDPP. In essence, SDPP was added to the test diets at the expense of either whey protein or fishmeal. Piglets were fed the diets for 3 weeks. In experiment 1, the piglets fed the SDPP diet had a 7% higher average daily gain (ADG) and a 4% lower feed conversion ratio (FCR) (p < 0.05) during the first 3 weeks after weaning than did those fed the control diet. There were no differences in leucocyte counts or γ-globulin. In experiment 2 there were no significant differences in ADG and FCR among the dietary treatments. It is concluded that low amounts of SDPP in weanling diets can have positive effects on growth performance under Northern European conditions.     

Improved monospermic fertilization and subsequent blastocyst formation of bovine oocytes fertilized in vitro in a medium containing NaCl of decreased concentration

The objective of this study was to evaluate whether changes in NaCl concentration in a fertilization medium could improve normal fertilization and preimplantation development of bovine oocytes. In vitro matured bovine oocytes were inseminated with frozen-thawed semen for 18 hr in a Tyrode's medium with albumin, lactate and pyruvate (TALP), to which 114 (TALP-114), 96 (TALP-96) or 78 (TALP-78) mM NaCl was added. Presumptive zygotes were cultured for 192 hr in a modified TALP containing 90 mM NaCl, 1.5 mM glucose, 0.3% (w/v) BSA, minimal essential medium (MEM) essential and nonessential amino acids, and insulin-transferrin-selenium complex. Lower polyspermy rate was obtained by the insemination in TALP-96 (7.8 +/- 2.3%) than by the insemination in TALP-114 (25.6 +/- 1.4%), without decrease in male pronucleus (MPN) formation. Fertilization in TALP-78 also yielded decreased polyspermic fertilization (3.8 +/- 1.5%), but significant decrease in MPN formation was found (63.1 +/- 3.1%). In preimplantation development, more blastocysts developed from oocytes inseminated in TALP-96 (24.1 +/- 1.7%) than from oocytes inseminated in TALP-114 (16.8 +/- 1.4%). TALP-78, however, did not improve preimplantation development beyond the 8-cell stage compared with TALP-114. Mean cell number of blastocyst was higher when oocytes were fertilized in TALP-96 (137.0 +/- 4.5) than in TALP-114 (123.1 +/- 5.1) and in TALP-78 (102.3 +/- 4.5). These results demonstrate that insemination of bovine oocytes in a TALP with decreased NaCl concentration (96 mM) improves blastocyst formation and embryo viability. Decrease in NaCl concentration below 96 mM, however, may delay or inhibit MPN formation, and inhibits subsequent development in vitro.     

Positive intraoperative cultures and canine total hip replacement: risk factors,periprosthetic infection,and surgical success

The results of closing intraoperative cultures from 27 canine total hip replacements (THR) were reviewed. The relationship between these culture results and presurgical and surgical factors, and the short- and long-term success of THR were assessed. Eleven out of 27 cases had a positive culture, but none of these 11 cases were diagnosed with periprosthetic infection at follow-up examination. The duration of the clinical signs of hip disease prior to THR was significantly greater for those cases with a positive culture (P=0.034). The incidence of positive cultures was not related to surgical success.