Characterization of seminal plasma proteins and sperm proteins in ejaculates from normospermic bulls and bulls with thermally-induced testicular degeneration

Semen was collected from 5 mature beef bulls by electroejaculation before, during, and after 20 days of scrotal insulation. Thermally-induced testicular degeneration was irreversible in 3 of the bulls. Analysis of sperm and seminal plasma polypeptides revealed that 15 to 30 sperm polypeptides and 25 to 30 seminal plasma polypeptides were indistinguishable between bulls prior to the insulation treatment. Changes in the sperm polypeptides pattern appeared as early as 2 days after the insulation treatment and persisted for at least 11 months in 2 of the bulls. In the spermatozoa, there was a detectable loss of 31, 34, 49 and 58 kDa polypeptides and an appearance of 6 to 8 new major polypeptides, ranging from 32 to 83 kDa. The 83 kDa polypeptide was most prominent in the 2 bulls that regained normal sperm motility and morphology following the insulation period. The post-insulation appearance of a seminal plasma polypeptide (circa 60 kDa) was also identified in these 2 bulls. Seminal plasma polypeptides remained qualitatively unaltered by the insulation treatment in the 3 bulls with irreversible testicular degeneration.     

Bacteriological evaluation of groups of beef carcasses before the wash at six Alberta abattoirs

K.W.F. JERICHO, J.A. BRADLEY AND G.C. KOZUB. 1994. A method has been developed for the bacteriological evaluation of groups of beef carcasses which can be used to measure the degree of control over hygiene during hide removal and carcass dressing in abattoirs. This method, which enumerates aerobic mesophilic bacteria automatically using a hydrophobic grid membrane filter, was applied at six abattoirs. Two hundred excision samples (5 × 5 × 0.5 cm) were taken at 10 sites on the external surface of a group of 20 carcasses (five carcasses were sampled on each of four consecutive daily visits) for group-carcass evaluation at each abattoir. For each abattoir, the mean log₁₀ Most Probable Number of Growth Units (MPNGU) and between-carcass variance component were obtained for each site and the average over sites. Using the average within-abattoir variance of this study and previously published studies involving 76 additional carcasses (Jericho et al. 1993), it was determined that 20 carcasses are more than adequate to estimate the mean log₁₀ MPNGU per cm² within 0.5 units at a site. The distribution of the log₁₀ MPNGU per cm² over the 10 sites was compared for the abattoirs, and sites were found to cluster into 2–4 homogenous groups. The means over sites of log₁₀ MPNGU per cm² for the abattoirs ranged from 1.52 to 2.64 and were unrelated to line speed     

The influence of sampling method on the classification of wetland macroinvertebrate communities

Macroinvertebrate communities sampled by a corer, plankton net and sweep net from five wetlands on the Swan Coastal Plain were compared. The composition of the fauna collected in sweeps and tows was generally similar and differed from that collected in the cores. Cores caught fewer species than tows and sweeps at all wetlands and did not capture fast swimming hemipterans or less abundant taxa. The highest species richness was recorded in sweep samples in four out of the five wetlands. Classification (TWIN-SPAN) and ordination (SSH) of the samples collected in sweeps and tows gave good separation of the wetlands, whereas classification of core samples did not. Coring appeared to be the least suitable sampling method for describing the major components of the macroinvertebrate communities of these wetlands. Plankton tows were useful if the time available for sorting was limited as these samples were free of sediments and generally gave similar results to those obtained with sweeps. Sweeps appeared to be the most useful method for a large classification study as they collected more species and resulted in the best discrimination amongst wetlands.     

Nonenzymatic Glycosylation of Lepidopteran-Active Bacillus thuringiensis Protein Crystals

We used high-pH anion-exchange chromatography with pulsed amperometric detection to quantify the monosaccharides covalently attached to Bacillus thuringiensis HD-1 (Dipel) crystals. The crystals contained 0.54% sugars, including, in decreasing order of prevalence, glucose, fucose, arabinose/rhamnose, galactose, galactosamine, glucosamine, xylose, and mannose. Three lines of evidence indicated that these sugars arose from nonenzymatic glycosylation: (i) the sugars could not be removed by N- or O-glycanases; (ii) the sugars attached were influenced both by the medium in which the bacteria had been grown and by the time at which the crystals were harvested; and (iii) the chemical identity and stoichiometry of the sugars detected did not fit any known glycoprotein models. Thus, the sugars detected were the product of fermentation conditions rather than bacterial genetics. The implications of these findings are discussed in terms of crystal chemistry, fermentation technology, and the efficacy of B. thuringiensis as a microbial insecticide.     

Neutron structure factors of in-vivo deuterated amorphous protein C-phycocyanin

Neutron powder diffraction measurements of fully deuterated protein C-phycocyanin have been made at three temperatures, 295, 200, and 77 K, using dry and partially hydrated samples. The average coherent structure factors and the corresponding radial distribution functions d(r) are determined. The changes in d(r) functions observed in hydrated samples depend strongly on the level of hydration and most of these changes are due to water-protein interactions. At 0.365 gram D₂O per gram of protein, the water crystallized into hexagonal ice at 200 K and below, but at 0.175 gram D₂O per gram of protein, no crystallization of water was observed. At the higher hydration a peak appears in the radial distribution function which indicates that the average distance of the water molecule in the first hydration shell from the amino acid residues is 3.5 Å.     

A population model for the alkali fly at Mono Lake: depth distribution and changing habitat availability

The densities of alkali fly larvae and pupae were measured in relation to depth and substrate type at six locations around Mono Lake. Samples representing a mixture of different bottom features were taken to a depth of 10 m (33 ft) using SCUBA. This is at or near the depth limit of fly larvae and pupae. The biomass of larvae and pupae on hard substrate were maximum and approximately equal at depths of 0.5 m and 1 m, substantially lower at intermediate depths of 3 m and 5 m, and over an order of magnitude further reduced at 10 m. Densities of flies on hard or rocky substrates (mainly calcareous tufa deposits), were significantly greater than those found on soft substrates such as mud or sand, at all but the greatest depth surveyed.Bathymetric maps of the areas of hard and soft substrate occurring at different lake depths were used to estimate the fly population size over the whole lake, based on the density distribution of larvae and pupae with depth on different substrates. The mapped areas of soft and hard substrates were also calculated for different lake levels, and applying the same procedure, a population model comparing the abundance of flies at different lake levels was developed. This habitat-based population model predicts that the abundance of the alkali fly is maximized at 6380 ft (1945 m) lake surface elevation. Most of the tufa substrate submerged at this lake level will become exposed and unavailable as habitat as the lake declines to 6370 ft (1942 m). In late 1991, the lake level was just over 6374 ft (1943 + m).     

Understanding the links between snoring,OSA and aortic root pathologies in Marfan syndrome

Sleep and Biological Rhythms -     

Enrichment of adeno-associated virus serotype 5 full capsids by anion exchange chromatography with dual salt elution gradients

Adeno-associated virus-based gene therapies have demonstrated substantial therapeutic benefit for the treatment of genetic disorders. In manufacturing processes, viral capsids are produced with and without the encapsidated gene of interest. Capsids devoid of the gene of interest, or “empty” capsids, represent a product-related impurity. As a result, a robust and scalable method to enrich full capsids is crucial to provide patients with as much potentially active product as possible. Anion exchange chromatography has emerged as a highly utilized method for full capsid enrichment across many serotypes due to its ease of use, robustness, and scalability. However, achieving sufficient resolution between the full and empty capsids is not trivial. In this work, anion exchange chromatography was used to achieve empty and full capsid resolution for adeno-associated virus serotype 5. A salt gradient screen of multiple salts with varied valency and Hofmeister series properties was performed to determine optimal peak resolution and aggregate reduction. Dual salt effects were evaluated on the same product and process attributes to identify any synergies with the use of mixed ion gradients. The modified process provided as high as ≥75% AAV5 full capsids (≥3-fold enrichment based on the percent full in the feed stream) with near baseline separation of empty capsids and achieved an overall vector genome step yield of >65%.