Purification and characterisation of relevant natural and recombinant apple allergens

Apple (Malus domestica) is the most widely cultivated fruit crop in Europe and frequently causes allergic reactions with a variable degree of severity. So far, four apple allergens Mal d 1, Mal d 2, Mal d 3 and Mal d 4 have been identified. Mal d 1, a Bet v 1 related allergen, and Mal d 4, apple profilin, are sensitive to proteolytic degradation, whereas Mal d 2, a thaumatin-like protein and Mal d 3, a nonspecific lipid transfer protein, are rather stable to proteolytic processes. Mal d 1 and Mal d 4 were purified after expression in Escherichia coli expression system, while Mal d 2 and Mal d 3 were purified from apple fruit tissue. All purified proteins were subjected to detailed physicochemical characterisation to confirm their structural integrity and maintained IgE binding capacity. Detailed investigations of carbohydrate moieties of Mal d 2 demonstrated their involvement in the overall IgE binding capacity of this allergen. It was concluded that the folded structure and IgE binding capacity of all four allergens were preserved during purification.     

Proteome profiling of exosomes derived from plasma of heifers with divergent genetic merit for fertility

The current study evaluated exosomes isolated from plasma of heifers bred to have high or low fertility through developing extreme diversity in fertility breeding values, however, key animal traits (e.g., body weight, milk production, and percentage of North American genetics) remained similar between the 2 groups. The exosomes were isolated by a combined ultracentrifugation and size exclusion chromatography approach and characterized by their size distribution (nanoparticle tracking analysis), morphology (transmission electron microscopy), and presence of exosomal markers (immunoblotting). In addition, a targeted mass spectrometry approach was used to confirm the presence of 2 exosomal markers, tumor susceptibility gene 101 and flotillin 1. The number of exosomes from plasma of high fertility heifers was greater compared with low fertility heifers. Interestingly, the exosomal proteomic profile, evaluated using mass spectrometry, identified 89 and 116 proteins in the high and low fertility heifers respectively, of which 4 and 31 were unique, respectively. These include proteins associated with specific biological processes and molecular functions of fertility. Most notably, the tetratricopeptide repeat protein 41-related, glycodelin, and kelch-like protein 8 were identified in plasma exosomes unique to the low fertility heifers. These proteins are suggested to play a role in reproduction; however, the role of these proteins in dairy cow reproduction remains to be elucidated. Their identification underscores the potential for proteins within exosomes to provide information on the fertility status and physiological condition of the cow. This may potentially lead to the development of prognostic tools and interventions to improving dairy cow fertility.     

Occurrence of wax esters in the tissues of the organge roughy (Hoplostethus atlanticus)

The skin, skeleton and a fat-filled swim bladder of the orange roughy (Hoplostethus atlanticus) each contained greater than 20% lipid by wet weight which was almost entirely wax esters. These had carbon numbers of 34–40 consistent with the major fatty acid being 18∶1 and the major fatty alcohols being 16∶0, 18∶1, 20∶1 and 22∶1. In contrast, the liver and the roe contained appreciable quantities of glycerolipids with 18∶1 and 22∶6 as the major fatty acids.     

Dynamics of ion beam modification of polymer films

Thin polymer films were irradiated in a high vacuum environment with energetic (～2 MeV) ions. The emitted molecular species were studied with a quadrupole mass spectrometer during bombardment. The emitted species are predominantly small molecules. The efficiency of emission depends strongly upon the electronic energy loss of the incident ions in the film and, hence, upon the velocity and atomic number of the incident ion. The emission efficiency of all species decreases with increasing damage in the film. By pulsing the ion beam, the time dependence of irradiation induced emission of molecular species from the films has been studied. Emission delays of hundreds of milliseconds are observed in some cases. These delays are found to depend upon the emitted species as well as the host film and seem to be associated with diffusion of the species in the films.     

The effects of various post-mortem treatments on certain physical and sensory properties of three different bovine muscles

Sixty crossbred steers with chronological ages approximating 15 months, carcass weights approximating 270 kg and subcutaneous fat thickness measurements approximating 1·2 cm, were maintained for approximately 210 days on a constantly increasing concentrate ration (13 to 75%, according to NAS nutrient requirements for beef cattle; NAS, 1976). These animals were utilized to evaluate the effects of delayed chilling, hot-boning and electrical stimulation on certain physical, histological, biochemical and sensory properties of three different muscles (Semimembranosus, SM; Longissimus dorsi, LD; and Triceps brachii, TB). Results indicated that neither delayed chilling nor electrical stimulation produced consistent meaningful alterations in any of the physical, cooking or palatability attributes evaluated. However, early post-mortem excision of muscle (hot-boning) consistently produced toughening in all of the muscles evaluated. This was observed to a much greater degree in the TB than in either the SM or LD. Based upon sarcomere length determinations, such toughening could be attributed to cold shortening only in the TB. Electrical stimulation of excised muscle was ineffective in preventing or offsetting this toughening. The delayed chilling treatment imposed (2 h at 12·5°C) was ineffective in enhancing potential effects of electrical stimulation on tenderness.     

Evaluation of an alkaline phosphatase-labeled oligonucleotide probe for the detection and enumeration of the thermostable-related hemolysin (trh) gene of Vibrio parahaemolyticus

Reliable methods are needed to detect total and pathogenic Vibrio parahaemolyticus. One marker of V. parahaemolyticus virulence is the thermostable-related hemolysin. We developed an alkaline phosphatase-labeled DNA probe method for the specific detection and enumeration of trh-positive V. parahaemolyticus by colony hybridization. The probe was tested against a panel of 200 bacterial strains and determined to be specific for trh-positive V. parahaemolyticus. Additionally, the trh alkaline phosphatase probe colony hybridization was successfully used to detect and enumerate trh-positive V. parahaemolyticus in seafood and water samples collected from the United States and the United Kingdom.     

Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland

The potential for applying biotechnology to benefit animal agriculture and food production has long been speculated. The addition of human milk components with intrinsic antimicrobial activity and positive charge to livestock milk by genetic engineering has the potential to benefit animal health, as well as food safety and production. We generated one line of transgenic goats as a model for the dairy cow designed to express human lysozyme in the mammary gland. Here we report the characterization of the milk from 5 transgenic females of this line expressing human lysozyme in their milk at 270 μg/mL or 68% of the level found in human milk. Milk from transgenic animals had a lower somatic cell count, but the overall component composition of the milk and milk production were not different from controls. Milk from transgenic animals had a shorter rennet clotting time and increased curd strength. Milk of such nature may be of benefit to the producer by influencing udder health and milk processing.     

Heat and anaerobic treatments affected physiological and biochemical parameters in tomato fruits

The aim of the present work was to evaluate the effect of thermal and anaerobic treatments on physiological and biochemical parameters in a variety of tomatoes (Lycopersicon esculentum Mill. cv. Colt 45). Treatments applied to mature green tomatoes were: (A) heat treatments by water immersion at 42 °C for 30 min (HS30′) or for 60 min (HS60′); or by air at 38 °C for 72 h (HS72h); and (B) anaerobic treatments carried out at 20 °C under humidified nitrogen atmosphere for 3 days (ANA3d) or 6 days (ANA6d). After treatments, fruits were stored at 2 or 14 °C. Parameters evaluated were: colour, total acidity, major organic acids, firmness, and ethanol and acetaldehyde concentration. Anaerobic and long-term heat shock treatments inhibited colour development irrespective of storage temperature. Air heat treatment reduced tritratable acidity by increasing malic acid metabolism. Anaerobic treatments induced ethanol accumulation, which could be reversed during storage for the short treatment (3 days), but not for the longer treatment (6 days). Acetaldehyde concentration was increased by anaerobic treatments, but also by immersion in hot water for 60 min, which would produce a “low-aerobic” environment.     

Evaluation of gaseous chlorine dioxide as a sanitizer for killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and yeasts and molds on fresh and fresh-cut produce

Gaseous chlorine dioxide (ClO2) was evaluated for effectiveness in killing Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on fresh-cut lettuce, cabbage, and carrot and Salmonella, yeasts, and molds on apples, peaches. tomatoes, and onions. Inoculum (100 microl, ca. 6.8 log CFU) containing five serotypes of Salmonella enterica, five strains of E. coli O157:H7, or five strains of L. monocytogenes was deposited on the skin and cut surfaces of fresh-cut vegetables, dried for 30 min at 22 degrees C, held for 20 h at 4 degrees C, and then incubated for 30 min at 22 degrees C before treatment. The skin surfaces of apples, peaches, tomatoes, and onions were inoculated with 100 microl of a cell suspension (ca. 8.0 log CFU) containing five serotypes of Salmonella, and inoculated produce was allowed to dry for 20 to 22 h at 22 degrees C before treatment. Treatment with ClO2 at 4.1 mg/liter significantly (alpha = 0.05) reduced the population of foodborne pathogens on all produce. Reductions resulting from this treatment were 3.13 to 4.42 log CFU/g for fresh-cut cabbage, 5.15 to 5.88 log CFU/g for fresh-cut carrots, 1.53 to 1.58 log CFU/g for fresh-cut lettuce, 4.21 log CFU per apple, 4.33 log CFU per tomato, 1.94 log CFU per onion, and 3.23 log CFU per peach. The highest reductions in yeast and mold populations resulting from the same treatment were 1.68 log CFU per apple and 2.65 log CFU per peach. Populations of yeasts and molds on tomatoes and onions were not significantly reduced by treatment with 4.1 mg/liter ClO2. Substantial reductions in populations of pathogens on apples, tomatoes, and onions but not peaches or fresh-cut cabbage, carrot, and lettuce were achieved by treatment with gaseous ClO2 without markedly adverse effects on sensory qualities.