Isolation and biochemical characterization of a basic myrosinase from ripe Crambe abyssinica seeds,highly specific for epi-progoitrin

On the basis of previous studies on the mechanism-based inhibition, activation, and active site structure of myrosinase(s) isolated from Sinapis alba and other cruciferous seeds, crambe myrosinase shows uncommon properties and behavior. For this reason homogeneous crambe myrosinase was isolated and investigated to establish the most important physicochemical features, including kinetic properties determined with the epimers progoitrin (R) and epi-progoitrin (S) as substrates, with and without ascorbate as an activator. The results of this study demonstrate that crambe myrosinase is highly specific for epi-progoitrin due to a better stabilization of the enzyme-substrate complex. This stabilization is caused by additional hydrogen bonding that only epi-progoitrin can set up between its hydroxyl group and a suitable residue in the hydrophobic pocket where the "docking" of the glucosinolates side chain takes place.     

Kiwellin, a modular protein from green and gold kiwi fruits: evidence of in vivo and in vitro processing and IgE binding

Kiwellin, an allergenic protein formerly isolated from green kiwi fruit, has been identified as the most abundant component of the gold kiwi species. A protein named KiTH, showing a 20 kDa band on reducing SDS-PAGE and 100% identity with the C-terminal region of kiwellin, has been identified in the extract of the ripe green species. In vitro treatment of purified kiwellin with the protease actinidin from green kiwi fruit originated KiTH and kissper, a recently described pore-forming peptide. Primary structure analysis and experimental evidence suggest that kiwellin is a modular protein with two domains. It may undergo in vivo proteolytic processing by actinidin, thus producing KiTH and kissper. When probed with sera recognizing kiwellin from green kiwi fruit, KiTH showed IgE binding, with reactivity levels sometimes different from those of kiwellin. The IgE-binding capacity of kiwellin from gold kiwi fruit appears to be similar to that of the green species.     

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.     

Ovarian Structure in Mice Lines Selected for Weight

Selection for body weight at 49 day of age ( s and h , downward selected lines; s' and h' , upward selected lines) affected reproductive traits in CF1 mice lines. The objective of this study was to compare ovarian structures in females of these lines, as well as in unselected controls (Line t ). The number of ovarian follicles ( N ), follicle diameter (FD), number of corpora lutea (CL), litter size (LS), and body weight ( W ), were recorded. There were significant differences among lines for N , FD, CL, LS and W ; means values for the lines with the greatest difference for post-pubertal females were: N _s  = 19.3 and N _s'  = 32.7; FD _h'   = 161.7 and FD _s'  = 178.2; CL _h  = 10.3 and CL _s'  = 21.9; LS _s  = 6.0 and LS _h'  = 11.1; W _h  = 18.9 and W _s'  = 32.4. There were also differences between positive lines; Line s' had a higher proportion of large follicles in pre-pubertal females, a greater capacity to convert these follicles into CL, but a lower capacity to maintain embryos until term than Line h' . For negative lines, Line h apparently had a reduced incidence of embryonic loss when compared with Line s . In conclusion, selection for body weight modified ovarian structure, as well as reproductive efficiency.     

Mycoparasitic Trichoderma isolates as a biocontrol agent against Valsa ceratosperma,the causal agent of apple valsa canker

European Journal of Plant Pathology - Apple valsa canker, caused by the fungus Valsa ceratosperma, is one of the most destructive diseases of this crop. Conventional fungicide treatments are not...