Association of Size‐Exclusion HPLC of Endosperm Proteins with Dough Mixing and Breadmaking Characteristics in a Recombinant Inbred Population of Hard Red Spring Wheat

Variation of polymeric proteins affects wheat end‐use quality. This research investigated associations of polymeric proteins with dough mixing strength and breadmaking characteristics in a near‐homogenous population of 139 recombinant inbred lines (RILs) derived from a cross between two hard red spring wheat breeding lines. Flours from the RILs grown at three locations were analyzed for molecular weight (MW) distribution of SDS‐extractable and unextractable proteins using size‐exclusion HPLC protocol. Correlations were calculated between mixing and breadmaking properties and HPLC absorbance data obtained a 0.01‐min retention time interval to identify protein fractions that had a significant effect on the quality traits. Very high MW polymeric proteins in the unextractable fraction had more distinct and positive associations with dough mixing strength and bread loaf volume than did other polymeric protein fractions, whereas extractable polymeric had negative influence. Consequently, the ratio of unextractable very high MW polymeric proteins to extractable polymeric proteins had greater correlations with dough mixing parameters than other HPLC absorbance area data. Covariate‐effect biplots also visually validated positive effects of unextractable very high MW polymeric proteins and negative effects of extractable polymeric proteins on mixing properties and loaf volume across three growing locations.     

Concentrations of anthocyanins in common foods in the United States and estimation of normal consumption

Anthocyanins (ACNs) are water-soluble plant pigments that have important functions in plant physiology as well as possible health effects. Over 100 common foods were screened for ACNs, and 24 of them were found to contain ACNs. Concentrations of total ACNs varied considerably from 0.7 to 1480 mg/100 g of fresh weight in gooseberry ('Careless' variety) and chokeberry, respectively. Not only does the concentration vary, but the specific anthocyanins present in foods are also quite different. Only six common aglycones, delphinidin, cyanidin, petunidin, pelargonidin, peonidin, and malvidin, were found in all of these foods. However, their sugar moieties and acylation patterns varied from food to food. Results from this study will add to the available data for the USDA Nutrient Database of flavonoids. On the basis of the concentration data and updated food intake data from NHANES 2001-2002, the daily intake of ACNs is estimated to be 12.5 mg/day/person in the United States. Of the different aglycones, cyanidin, delphinidin, and malvidin were estimated to contribute 45, 21, and 15%, respectively, of the total ACN intake. Nonacylated contributed 77% compared to 23% from acylated ACNs.     

Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis

OBJECTIVE: To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. SAMPLE POPULATION: Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. PROCEDURE: Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. RESULTS: Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% alpha1-PI, 93.6% by 0.5% alpha1-PI, and 90.0% by ES. CONCLUSIONS AND CLINICAL RELEVANCE: We documented that EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.     

Use of a three-drill-tract technique for arthrodesis of the distal tarsal joints in horses with distal tarsal osteoarthritis: 54 cases (1990-1999)

OBJECTIVE: To assess the long-term clinical outcome of horses with distal tarsal osteoarthritis (OA) in which a 3-drill-tract technique was used to induce arthrodesis of the affected joints, identify any preoperative or operative factors associated with outcome, and describe any complications associated with the technique. DESIGN: Retrospective study. ANIMALS: 54 horses. PROCEDURE: Medical records were reviewed for information on signalment, use, history, physical and lameness examination findings, surgical technique, and postoperative care. Radiographs were examined, and severity of OA was graded. Follow-up information was obtained through telephone interviews with owners at least 13 months after the procedure. RESULTS: 32 (59%) horses had a successful outcome, 6 (11%) improved but were not sound after surgery, and 16 (30%) did not improve following surgery. Outcome was negatively associated with the previous use of intra-articular injections. Few postoperative complications were evident. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that distal tarsal OA in horses can be successfully treated by means of distal tarsal arthrodesis with a 3-drill-tract technique. Horses with advanced distal tarsal OA are likely to have poorer outcomes, and the procedure will likely be of minimal benefit in horses with concomitant causes of hind limb lameness prior to surgery and in horses with preexisting proximal intertarsal joint disease.     

Benefits of prophylactic gastropexy for dogs at risk of gastric dilatation-volvulus

The lifetime probability of death from gastric dilation–volvulus (GDV) for five dog breeds was estimated based on published breed-specific longevity and GDV incidence. These breeds were Great Dane, Irish Setter, Rottweiler, Standard Poodle and Weimaraner. Lifetime risk (95% CI) of GDV in these breeds ranged from 3.9% (0–11.2%) for Rottweiler to 36.7% (25.2–44.6%) for Great Dane.

A decision-tree analysis for prophylactic gastropexy—using lifetime probability of death from GDV and expected cost savings for veterinary services as outcome measures—was undertaken to determine the preferred course of action in several dog breeds. Prophylactic gastropexy was the preferred choice of action for all breeds examined, with the reduction in mortality (versus no gastropexy) ranging from 2.2-fold (Rottweiler) to 29.6-fold (Great Dane). Assuming a prophylactic gastropexy costs US$ 400, the procedure was cost-effective when the lifetime risk of GDV was ≥34%. The maximum and minimum estimated breakeven costs for the gastopexy procedure ranged from US$ 20 (Rottweiler) to US$ 435 (Great Dane). The cost-effectiveness of prophylactic gastropexy was most sensitive to the cost of treating GDV (US$ 1500). Prophylactic gastropexy raises ethical issues that need to be considered by veterinarians and dog breeders.     

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.     

New technologies for studying immune regulation in ruminants

There remains a great need for the development of reagents and techniques to study immune regulation in ruminants. These tools are fundamental to our understanding of the mechanisms of immune modulation and underpin the rational design of disease control strategies in livestock. Technological advances in molecular biology have resulted in the development of a new range of techniques to measure and quantify gene expression in normal and disease states. However, the ability to measure protein expression, produce proteins (usually in recombinant form) and to block receptor-ligand interactions remains essential for elucidating the function of the molecules of interest. The aims of this workshop were to look at the current status of reagents for studying ruminant immunology, the prospects for future developments, and also to discuss reagents availability between different groups worldwide.     

Plasma pharmacokinetics of selamectin after a single topical administration in the American bullfrog (Rana catesbeiana).

Parasitism is common in wild and captive amphibians; however, pharmacologic data are lacking for anthelmintic drugs. This study was developed to determine the plasma pharmacokinetics of selamectin after topical administration in bullfrogs. Thirty-two adult American bullfrogs (Rana catesbeiana) were randomly assigned into eight groups of four with each group representing a different collection time point. Seven groups received selamectin (6 mg/ kg) topically and the remaining group served as the untreated control group. One group of frogs was euthanized and blood samples immediately collected on days 0 (control), 1, 5, 10, 15, 20, 25, and 30. Plasma was analyzed for selamectin using high performance liquid chromatography with fluorescence detection. Individual samples were analyzed, then data were reported as the mean of the four frogs at each time point. A histologic evaluation of the lung, liver, kidney, and skin tissues was performed and none of the frogs showed histologic evidence of toxicity due to selamectin administration. The mean peak plasma concentration was 162.5 +/- 42.3 ng/ml, area under the curve was 2,856 ng day/ml, mean residence time was 12.2 days, and disappearance half-life was 1.87 days. Based on the plasma pharmacokinetics, bullfrogs appear to absorb selamectin very efficiently, concentrations reach high levels in the plasma, and there were no apparent histologic effects from single dose administration.     

Performing the field necropsy examination.

This article is designed to aid the practitioner by maximizing the effectiveness of field postmortem diagnostic investigations. Contents include an outline of the procedure for field necropsy of ruminants, recommended tools and supplies, and guidelines for sample collection and submission.