Evaluation of jitter by stimulated single-fiber electromyography in normal dogs

Single-fiber electromyography (SFEMG), a technique used to investigate neuromuscular transmission, has been described previously in the pelvic limb of dogs. Because preferential involvement of isolated muscle groups can occur in disorders of neuromuscular transmission, SFEMG was done in the peroneus longus (PL), extensor carpi radialis (ECR), and orbicularis oculi (OO) muscles of 10 adult, clinically normal dogs. Jitter was calculated as the mean absolute value of the consecutive differences in latency of 50 single muscle fiber action potentials after stimulation of intramuscular nerve bundles at the level of the motor point in at least 20 muscle fibers per muscle. Bilateral recordings were performed in 3 dogs. Mean jitter values were determined for each muscle, and differences among muscle groups and among dogs were compared. The upper limits of mean consecutive difference (mean plus 3 standard deviations) for the PL, ECR, and OO muscles were 21.94, 22.53, and 23.39 micros, respectively, and the upper limit of mean consecutive difference for individual muscle fibers in the respective fiber pools was 28.62, 36.39, and 35.68 micros. Jitter values for the ECR and OO were significantly higher than the jitter value for the PL muscle (P < .05). Significant differences among muscles or dogs or between sides were not observed for the ECR. Significant differences among dogs were observed for OO jitter values and were attributed to extremely low jitter values in 1 dog. Significant differences were demonstrated between sides for the PL and were attributed to small sample size. Results of this study provide normative data that can be used in the application of the stimulated SFEMG technique to dogs with suspected disorders of neuromuscular transmission.     

Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

Effect of fast protein liquid chromatography fractionated salivary gland extracts from different ixodid tick species on interleukin-8 binding to its cell receptors

Interleukin-8 plays a critical role in inflammatory processes. Hence generation of molecules with anti-IL-8 activity is likely to be important for successful feeding and for survival of the ticks. Anti-IL-8 activity was studied in saliva of three ixodid tick species--Dermacentor reticulatus (Fabricius, 1794), Rhipicephalus appendiculatus Neumann, 1901, and Amblyomma variegatum (Fabricius, 1794). The greatest activity was shown in saliva prepared from D. reticulatus. The activity was attributed to tick salivary gland molecules that bind to IL-8, preventing binding of the chemokine to its specific receptor, rather than to occupation of the IL-8 cell receptor by the tick molecules. The distribution of anti-IL-8 activity in fast protein liquid chromatography (FPLC) fractions of salivary gland extracts (SGE) derived from adult female D. reticulatus, R. appendiculatus and A. variegatum was compared directly by both ELISA and receptor-binding inhibition assays. The correspondence in results with fractions of SGE from ELISA is consistent with detection of tick molecules that inhibit IL-8 binding to its receptor. As IL-8 is an important chemoattractant and activator of neutrophils, the presence of an anti-IL-8 activity in tick saliva indicates that neutrophils play an important role in the host response to parasitism by ticks.     

Liquid Formulation of the Postharvest Biocontrol Agent Candida sake CPA-1 in Isotonic Solutions

ABSTRACT Viability of the postharvest biocontrol agent Candida sake CPA-1 stored as liquid formulation was evaluated by studying the effect of growth, preservation medium, and temperature. C. sake was grown in molasses medium with unmodified water activity (a(w)) and in the same with a(w) modified to 0.98 with the addition of several solutes. Cells were preserved with isotonic solutions of different substances. Efficacy of liquid formulations stored for different periods was tested against infection by Penicillium expansum on apples. The best growth media were the (unmodified one and those modified to 0.98 a(w) with the addition of glycerol or sorbitol. For all growth media, the best preservation medium was the isotonic solution prepared with trehalose. When the effect of trehalose concentration in the preservation medium was studied, generally, at trehalose concentrations below the isotonic one, C. sake viabilities increased with increased trehalose. However, the best results were obtained when cells were preserved with the trehalose solution which was isotonic with cells. After 7 months of storage at 4 degrees C, cells that were grown in the sorbitol-modified medium and preserved with the isotonic solution of trehalose (0.96 M) maintained their viability and efficacy against P. expansum infection of apples.     

Evaluation of the role of lower urinary tract disease in cats with urine-marking behavior

OBJECTIVE: To determine whether findings of urinalyses could be used to reliably distinguish gonadectomized cats with urine-marking behavior from those with no problem urination. DESIGN: Case control study. ANIMALS: 58 gonadectomized cats (47 males and 11 females) with urine-marking behavior (ie, marking of vertical surfaces) and 39 (26 males and 13 females) without problem urination or urinary tract-associated conditions. PROCEDURE: Urine was collected by cystocentesis from all cats. Findings of urinalyses of cats with urine-marking behavior were analyzed statistically for sex-related differences and differences between cats that marked vertical surfaces only and those that marked both vertical and horizontal surfaces; findings of urinalyses of control cats were compared between sexes. Subsequently, results of urinalyses of cats with urine-marking behavior were compared with those of control cats. RESULTS: With regard to variables measured via urinalysis, there were no differences between male and female cats within either group. Among cats with urine-marking behavior, there were no differences between those that only marked vertically and those that marked vertically and horizontally. Analyses of data from all cats with urine-marking behavior and control cats revealed no differences that could be associated with urine marking. CONCLUSIONS AND CLINICAL RELEVANCE: These data suggest that urine-marking behavior by gonadectomized cats is an aspect of normal behavior. Clinicians are advised to focus on behavioral history of house-soiling cats to differentiate between urine-marking behavior and inappropriate urination; for the latter, urinalysis is appropriate to rule out lower urinary tract disorders.     

Babesia odocoilei infection in elk

Two male North American elk from a commercial herd were evaluated because of a sudden onset of lethargy, anorexia, and voiding of red urine. These 2 elk were kept in the same pen as 4 other male elk that had died during the preceding 2 months. Laboratory analyses revealed anemia and intraerythrocytic parasites, later confirmed as Babesia odocoilei (a protozoal hemoparasite of cervids). Of the 240 elk remaining in the herd, 59 were screened for B odocoilei by microscopic evaluation of blood smears, protozoal culture of blood, and immunofluorescent antibody testing of serum. Of those 59 elk, 34 (58%) were infected with B odocoilei. Babesia odocoilei infection in elk can be fatal and should be considered in cases of sudden death or acute hemolytic anemia. Familiarity with the disease in elk is essential for practitioners because of the increasing popularity of commercial elk farming.     

Estimates of Breed Average Direct,Maternal and Heterosis Effects for Some Pre-weaning Traits of Zebu Breeds and Their Crosses in Southeastern Mexico

Information on 936 birth weights (BW), adjusted weaning weights (AWW) and average daily gains (ADG) from Brahman (B), Indubrazil (I), Gyr (G) and commercial Zebu (C) cattle and their crosses were obtained from the records of a farm in the State of Yucatan, Mexico. Two statistical models were used to analyse the data: the first included the effects of year and season of birth, sex of the calf, cow's parity number and breed group. The other model included the same environmental effects as the previous model and the genetic components of breed group (breed additive, breed maternal and heterosis effects). Additive and maternal effects were expressed as deviations from those for the B breed. The means of BW, AWW and ADG for the B breed were 33.3 ± 0.64 kg, 204.0 ± 4.9 kg and 760.06 ± 19.24 g/day, respectively. Both models gave similar results in terms of the variance explained. Additive effects were significant (p < 0.05) for AWW and ADG but not for BW. The minor additive effect was for the C breed (−10.75 kg and −40.77 g for WW and ADG, respectively). Maternal and heterosis effects were not significant (p > 0.05) for any of the traits. The correlation between the breed group means and the means estimated by the prediction equation was r = 0.88. Finally, there was no significant genetic effect on pre-weaning calf performance among the genotypes evaluated. The genetic model can be used to predict the genotype means with a high degree of accuracy. This revised version was published online in July 2006 with corrections to the Cover Date.     

Quantitative changes in the normal and apoptotic thymocytes of pigs treated with anabolic doses of the beta2 adrenergic agonist clenbuterol

Although the beta2 adrenergic agonists have been seen to have important effects on the mechanisms regulating the development and death of T-cells in the thymus, the side-effects on the immune system of anabolic treatments of these substances have hardly been considered. In order to evaluate the effects exerted by the beta2 adrenergic agonist clenbuterol on the thymocyte population, the thymus of eight pigs treated with anabolic doses of this substance was studied by morphometric methods, regarding apoptotic (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive) and normal (TUNEL-negative) cells. The thymus of another eight pigs fed without clenbuterol served as a control. The clenbuterol treatment had a clear effect on the thymocyte size, decreasing their mean nuclear area. The T-cell apoptosis index was also affected by the clenbuterol, significantly increasing the apoptosis percentage in the treated group with respect to the control. In the light of our results, the clenbuterol induced thymocyte apoptosis throughout the thymus and caused morphometric changes in the thymocyte population, which was in line with the immunosuppressive role attributed to other beta2 adrenergic agonists.     

Jojoba seed meal proteins associated with proteolytic and protease inhibitory activities

The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. The jojoba meal after oil extraction is rich in protein. The major jojoba proteins were albumins (79%) and globulins (21%), which have similar amino acid compositions and also showed a labile thrombin-inhibitory activity. SDS-PAGE showed two major proteins at 50 kDa and 25 kDa both in the albumins and in the globulins. The 25 kDa protein has trypsin- and chymotrypsin-inhibitory activities. In vitro digestibility of the globulins and albumins resembled that of casein and soybean protein concentrates and was increased after heat treatment. The increased digestibility achieved by boiling may be attributed to inactivation of the protease inhibitors and denaturation of proteins.