Transgenic expression of bean alpha-amylase inhibitor in peas results in altered structure and immunogenicity

The development of modern gene technologies allows for the expression of recombinant proteins in non-native hosts. Diversity in translational and post-translational modification pathways between species could potentially lead to discrete changes in the molecular architecture of the expressed protein and subsequent cellular function and antigenicity. Here, we show that transgenic expression of a plant protein (alpha-amylase inhibitor-1 from the common bean (Phaseolus vulgaris L. cv. Tendergreen)) in a non-native host (transgenic pea (Pisum sativum L.)) led to the synthesis of a structurally modified form of this inhibitor. Employing models of inflammation, we demonstrated in mice that consumption of the modified alphaAI and not the native form predisposed to antigen-specific CD4+ Th2-type inflammation. Furthermore, consumption of the modified alphaAI concurrently with other heterogeneous proteins promoted immunological cross priming, which then elicited specific immunoreactivity of these proteins. Thus, transgenic expression of non-native proteins in plants may lead to the synthesis of structural variants possessing altered immunogenicity.     

Dietary supplementation of grape skin extract improves glycemia and inflammation in diet-induced obese mice fed a Western high fat diet

Dietary antioxidants may provide a cost-effective strategy to promote health in obesity by targeting oxidative stress and inflammation. We recently found that the antioxidant-rich grape skin extract (GSE) also exerts a novel anti-hyperglycemic activity. This study investigated whether 3-month GSE supplementation can improve oxidative stress, inflammation, and hyperglycemia associated with a Western diet-induced obesity. Young diet-induced obese (DIO) mice were randomly divided to three treatment groups (n = 12): a standard diet (S group), a Western high fat diet (W group), and the Western diet plus GSE (2.4 g GSE/kg diet, WGSE group). By week 12, DIO mice in the WGSE group gained significantly more weight (24.6 g) than the W (20.2 g) and S groups (11.2 g); the high fat diet groups gained 80% more weight than the standard diet group. Eight of 12 mice in the W group, compared to only 1 of 12 mice in the WGSE group, had fasting blood glucose levels above 140 mg/dL. Mice in the WGSE group also had 21% lower fasting blood glucose and 17.1% lower C-reactive protein levels than mice in the W group (P < 0.05). However, the GSE supplementation did not affect oxidative stress in diet-induced obesity as determined by plasma oxygen radical absorbance capacity, glutathione peroxidase, and liver lipid peroxidation. Collectively, the results indicated a beneficial role of GSE supplementation for improving glycemic control and inflammation in diet-induced obesity.     

Comparison of four test procedures to identify Staphylococcus aureus isolated from bovine intramammary infections

A comparison was made between conventional tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test systems for identification of Staphylococcus aureus of bovine origin. A total of 303 gram-positive, catalase-positive cocci of bovine origin were tested. Agreement between each pair of 4-hour tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test results within isolates was greater than 95.0. Seventeen (5.6%) isolates were test negative for 4-hour tube coagulase, but test positive for 24-hour tube coagulase. Thirteen (76.5%) of these isolates were identified as S hyicus, 3 were S aureus, and 1 was not identified.     

Costs associated with selected preventive practices and with episodes of clinical mastitis in nine herds with low somatic cell counts

Nine dairy herds (mean size, 149 cows) with bulk-tank milk somatic cell counts of less than 300,000 cells/ml and greater than 80% of cows with Dairy Herd Improvement Association linear somatic cell counts less than or equal to 4 were selected for study. Each herd was monitored for 12 consecutive months. Duplicate quarter-milk specimens were collected from each cow for bacteriologic culturing at beginning of lactation, cessation of lactation, and at the time of each clinical episode of mastitis. Streptococcus agalactiae was never isolated and Staphylococcus aureus was isolated from less than 1% of all quarters. There were 554 episodes of clinical mastitis. During the year of study, the incidence rate of clinical mastitis varied from 15.6 to 63.7% of cows among the 9 herds. Mean costs per cow per year in herd for mastitis prevention were: $10 for paper towels, $3 for nonlactating cow treatment, and $10 for teat disinfectants. Mean cost associated with clinical mastitis was $107/episode. Approximately 84% ($90) of the costs attributed to a clinical episode were associated with decreased milk production and nonsalable milk. Costs of medication and professional veterinary fees per clinical episode varied significantly among the 9 herds. Three of the herds did not have a veterinarian treat a clinical episode of mastitis during the year of study even though 2 of these herds had the first and third highest incidence rates of clinical mastitis. When calculated on a per cow in herd basis, mean costs of $40/cow/year were attributed to clinical mastitis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Salmonella Brandenburg in non-pregnant 8-month-old sheep

Abstract

Extract

S. Brandenburg has been reported as a cause of abortion and often subsequent death of ewes from areas of the South Island of New Zealand (Bailey, 1998 Bailey, K.M. 1997. Sheep abortions associated with Salmonella Brandenburg. Surveillance, 24(4): 10–11.  [Google Scholar] and Clark et. al., 1999 Clark, G, Fenwick, S, Boxall, N, Swanney, S and Nicol, C. 1999. “Salmonella Brandenburg abortions in sheep, pathogenesis and pathology”. In Proceedings of the 29^pth Seminar of the Society of Sheep and Beef Cattle Veterinarians NZVA 13–22. Palmerston North, , New Zealand: Veterinary Continuing Education, Massey University,.  [Google Scholar]). This bacterium has also been isolated from a bitch with metritis, a dog with diarrhoea, cattle with diarrhoea and black-backed gulls (Laurus dominicanus) (Clark et. al., 1999 Clark, G, Fenwick, S, Boxall, N, Swanney, S and Nicol, C. 1999. “Salmonella Brandenburg abortions in sheep, pathogenesis and pathology”. In Proceedings of the 29^pth Seminar of the Society of Sheep and Beef Cattle Veterinarians NZVA 13–22. Palmerston North, , New Zealand: Veterinary Continuing Education, Massey University,.  [Google Scholar]). It has also been isolated from pigs, goats, a takahe (Notornis mantelli) and a deer (Bailey, 1998 Bailey, K.M. 1997. Sheep abortions associated with Salmonella Brandenburg. Surveillance, 24(4): 10–11.  [Google Scholar]). Until recently S. Brandenburg was thought only to affect sheep in the later stages of pregnancy.     

Phylogenetic analysis and molecular methods for the detection of lymphocystis disease virus from yellow perch, Perca flavescens (Mitchell)

Lymphocystis disease is a prevalent, non-fatal disease that affects many teleost fish and is caused by the DNA virus lymphocystis disease virus (LCDV). Lymphocystis-like lesions have been observed in yellow perch, Perca flavescens (Mitchell), in lakes in northern Alberta, Canada. In an effort to confirm the identity of the virus causing these lesions, DNA was extracted from these lesions and PCR with genotype generic LCDV primers specific to the major capsid protein (MCP) gene was performed. A 1357-base pair nucleotide sequence corresponding to a peptide length of 452 amino acids of the MCP gene was sequenced, confirming the lesions as being lymphocystis disease lesions. Phylogenetic analysis of the generated amino acid sequence revealed the perch LCDV isolate to be a distinct and novel genotype. From the obtained sequence, a real-time PCR identification method was developed using fluorgenic LUX primers. The identification method was used to detect the presence/absence of LCDV in yellow perch from two lakes, one where lymphocystis disease was observed to occur and the other where the disease had not been observed. All samples of fin, spleen and liver tested negative for LCDV in the lake where lymphocystis disease had not been observed. The second lake had a 2.6% incidence of LCD, and virus was detected in tissue samples from all individuals tested regardless of whether they were expressing the disease or not. However, estimated viral copy number in spleen and liver of symptomatic perch was four orders of magnitude higher than that in asymptomatic perch.     

System behavior of commonly used cranial cruciate ligament reconstruction autografts

Biomechanical analysis was performed on the cranial cruciate ligament (CCL) and three autogenous tissues used for CCL reconstruction in the canine stifle. The autogenous tissues were patellar ligament-based autografts described for over-the-top CCL replacement and included the central one third of the patellar ligament, the medial one third of the patellar ligament, and the lateral one third of the patellar ligament with fascia lata. Tension testing produced abrupt failure of the central and medial autografts but sequential failure of the lateral autograft. Structural properties were determined for the overload condition and within the load range of normal activity for the CCL (physiologic range). None of the autograft systems approached the stiffness, maximum load, and energy absorbed to maximum load of the CCL. The central and lateral autografts were stiffer, had greater maximum loads, and absorbed more energy to maximum load than the medial autograft. The central and lateral autografts had an elastic range, as defined by proportional limit, which corresponded to the physiologic range of loading for the CCL. Loads that corresponded to physiologic displacement of the lateral and central autografts were near the maximum load of the fixation site, which underscored the need for postoperative support of the repaired stifle.     

EFFECT OF SEDATION ON TRANSIT TIME OF FELINE GASTROINTESTINAL CONTRAST STUDIES

The effect of commonly used sedatives on gastrointestinal motility and transit time in cats was evaluated using barium sulfate in gastrointestinal contrast studies. Control studies were performed in nonsedated animals, and the results were compared with those obtained from each of five sedation studies (ANOVA; p < 0.05). The ketamine/acepromazine transit time (18 minutes) was shortened significantly compared with the control group (42 minutes), and both ketamine/acepromazine and ketamine alone resulted in significant increase in the number of gastric contractions. The level of sedation was evaluated subjectively and compared with the transit times to determine a chemical restraint method for potential clinical use that would have the least effect on transit time and motility yet provide adequate sedation. When sedation is necessary and motility is not a primary concern, the ketamine/acepromazine combination if recommended. If a gastrointestinal motility problem is suspected, the ketamine/valium combination should be used.     

Radiation treatment for thymoma in a dog

A 10-year-old Toy Poodle was admitted for evaluation of a chronic cough caused by a cranial mediastinal mass. The mass was diagnosed, by needle biopsy, as a thymoma. Detection of serum acetylcholine receptor antibodies suggested that the dog had subclinical myasthenia gravis. The dog underwent orthovoltage radiation treatment, which resulted in an approximate 60% reduction in tumor mass and freedom from clinical signs for 6 months. Concurrent use of prednisolone may have been associated with disappearance of the acetylcholine receptor antibodies.