Cycloheximide resistance in Chinese hamster cells. II. Induction of Chm resistance in Chinese hamster cells by N-nitrosomethylurea.

Mutant Chinese hamster ovarian (CHO) cells with a resistance to 7-10(-7) and 8-10(-7) M cycloheximide (CHM) were induced at mutation rates of 1.9-5.2-10(-3) and 1.6-1.8-10(-3) respectively after treatment with N-nitrosomethylurea (NMU) at 100 mug/ml. The induced mutation rates differed by two orders of magnitude from the spontaneous rate of mutation to CHM resistance.     

A three-step method for isolating a few to several hundred milligrams of protein

An operationally simple general protein isolation method was devised from three previously available separation tools, and was tested by application to two demanding fractionation problems and for yield. One test system was the isolation by gel electrofocusing of two model proteins with pI values of 4.6 and 4.8, bovine serum albumin and ovalbumin, with a load of 220 mg each. The other test was the isolation of 10 mg of human growth hormone isohormone B from a mixture of closely migrating other isohormones. The three-step procedure comprises of: (1) separation into zones of homogeneous protein by gel electrofocusing; (2) excision of the zones of homogeneous protein from the gel followed by concentration of the protein to a small volume of solution by means of Steady-State Stacking; (3) purification from polyacrylamide-like contaminants and non-volatile buffers by gel filtration followed by lyophilization. The average overall recovery was 70--80%.     

Editorial: Twin peaks mission

The Y human chromosome has many ancient genes whose fidelity seems to have been preserved by tandem sequences and palindromic ‘hairpins’, compared/repaired by ‘gene conversion’. That a primary function of recombination machinery is DNA repair has been suggested, and rejected, several times; this new evidence is very persuasive. The process, better called gene conservation than gene conversion, could operate in all diploid organisms, accounting for the retention of long gene sequences without ‘informational meltdown’ ('concerted evolution'). It resembles rocket‐science computer‐redundancy error‐checking, comparison of three or four sequences, not just two. If recognition of errors in ‘converted’ sequences can be followed by either repair or rejection, the rejection option can account for the vast wastage of meiotic products. The repair option might be used in Drosophila oocytes and even zygotic nuclei, possibly other oocytes, ancient asexual lineages such as mycorrhizal fungi, perhaps the Y itself. Both evolutionary stasis (conservatism) and development and deployment of complex developmental modules can be understood in these terms so both the evolution of biodiversity and the practice of systematics may have these mechanisms as their bases. The main individual‐fitness and evolutionary advantages of diploidy were not primarily cloaking of recessive al‐leles, or allelic recombination and Mendelism, but conserving long DNA sequences.     

Molecular Characteristics of Bovine Serum Albumin-Dextran Conjugates

Bovine serum albumin (BSA)-dextran conjugates were prepared by using the Maillard reaction; depending on the ratio of dextran to BSA used, about 0.5–1 mol of dextran could be bound to 1 mol of native BSA. SDS–PAGE patterns revealed that BSA and dextran had been covalently bonded. Structural analyses by fluorescence spectroscopy and circular dichroism indicated that the BSA surface in each conjugate was covered with dextran without any great disruption of the native conformation. The conjugates could be grouped into two fractions on the basis of the weight-average molecular mass measured: the main fraction at 1.95–2.35×10⁵ g/mol and a less-abundant fraction with aggregates greater than 1.50×10⁶ g/mol. High-performance size-exclusion chromatography in conjunction with multi-angle laser light scattering detection revealed that the BSA-dextran conjugates prepared by using the Maillard reaction had various molar masses and radii.     

Comparison of effects of four weaning methods on health and performance of beef calves

Weaning of beef calves is a stressful event that negatively impacts health and performance. A variety of interventions have been proposed to reduce stress and improve gains following weaning. This study used 288 7- to 8-month-old calves from two separate locations, to examine four different weaning strategies, as well as the impact of shipment. Calves were blocked by weight and sex, and then randomly assigned to one of four treatments: abrupt weaning (AW), where calves were separated from the dam on day 0 (D0) and allowed no further contact with the dam; fence line (FL), where calves were weaned on D0 but had fence line contact with dams for 7 days; nose flap (NF), where on day -6 calves received a nose flap that interferes with suckling, then had the flap removed and were weaned from the dam on D0; and intermittent separation (SEP), where calves were removed from dams for 24-h intervals on day -13 and day -6, then weaned on D0, but allowed fence line contact with the dam for 7 days. Each treatment group was further divided into two subgroups, one of which was shipped early (D0 for AW, day 7 for others) or shipped later (day 28). Body weight and sickness were recorded for all groups. Results showed a negative impact on gain for early shipping compared to later shipping, and poorer gain in AW calves than most other treatments. Results of the analyses of morbidity were inconclusive. This study found that delayed shipment following FL weaning improves performance under common management conditions for the US cow–calf industry.     

Short communication: administering an appeasing substance to Bos indicus-influenced beef cattle at weaning and feedlot entry

The bovine appeasing substance (BAS) is expected to have calming effects in cattle experiencing stressful situations. Therefore, this study investigated the impacts of BAS administration during two of the most stressful events within beef production systems: weaning and feedlot entry. In experiment 1, 186 Bos indicus-influenced calves (73 heifers, 113 bulls) were weaned at 211 ± 1 days of age (day 0). At weaning, calves were ranked by sex and BW, and assigned to receive BAS (Nutricorp, Araras, SP, Brazil; n = 94) or water (CON; n = 92). Treatments (5 ml) were topically applied to the nuchal skin area of each animal. Calf BW was recorded and samples of blood and tail-switch hair were collected on days 0, 15 and 45. Calves that received BAS had greater (P < 0.01) BW gain from day 0 to 15 compared with CON. Overall BW gain (days 0 to 45) and BW on days 15 and 45 were also greater (P ≤ 0.03) in BAS v. CON. Plasma haptoglobin concentration was less (P < 0.01) in BAS v. CON on day 15, whereas cortisol concentrations in plasma and tail-switch hair did not differ between treatments (P ≥ 0.13). In experiment 2, 140 B. indicus-influenced bulls (∼27 months of age) from 2 different pasture-based systems (70 bulls/origin) were transported to a commercial feedlot (≤ 200-km transport; day -1). On day 0, bulls were ranked by source and BW, and assigned to receive BAS (n = 70) or CON (n = 70) and the same sampling procedures as in experiment 1. Bulls receiving BAS had greater (P = 0.04) BW gain from day 0 to 15, but less (P < 0.01) BW gain from day 15 to 45 compared to CON. No other treatment effects were detected (P > 0.14). Therefore, BAS administration to beef calves alleviated the haptoglobin response associated with weaning, and improved calf growth during the subsequent 45 days. Administration of BAS to beef bulls at feedlot entry improved BW gain during the initial 15 days, but these benefits were not sustained throughout the 45-day experiment.     

Genotype by environment interaction in response to cold stress in a composite beef cattle breed

Extreme weather conditions such as cold stress influence the productivity and survivability of beef cattle raised on pasture. The objective of this study was to identify and evaluate the extent of the impact of genotype by environment interaction due to cold stress on birth weight (BW) and weaning weight (WW) in a composite beef cattle population. The effect of cold stress was modelled as the accumulation of total cold load (TCL) calculated using the Comprehensive Climate Index units, considering three TCL classes defined based on temperature: less than −5°C (TCL5), −15°C (TCL15) and −25°C (TCL25). A total of 4221 and 4217 records for BW and WW, respectively, were used from a composite beef cattle population (50% Red Angus, 25% Charolais and 25% Tarentaise) between 2002 and 2015. For both BW and WW, a univariate model (ignoring cold stress) and a reaction norm model were implemented. As cold load increased, the direct heritability slightly increased in both BW and WW for TCL5 class; however, this heritability remained consistent across the cold load of TCL25 class. In contrast, the maternal heritability of BW was constant with cold load increase in all TCL classes, although a slight increase of maternal heritability was observed for TCL5 and TCL15. The direct and maternal genetic correlation for BW and maternal genetic correlation for WW across different cold loads between all TCL classes were high (r > 0.99), whereas the lowest direct genetic correlations observed for WW were 0.88 for TCL5 and 0.85 for TCL15. The Spearman rank correlation between the estimated breeding value of top bulls (n = 79) using univariate and reaction norm models across TCL classes showed some re-ranking in direct and maternal effects for both BW and WW particularly for TCL5 and TCL15. In general, cold stress did not have a big impact on direct and maternal genetic effects of BW and WW.     

Staphylococcus aureus persistence properties associated with bovine mastitis and alternative therapeutic modalities

Staphylococcus aureus is an important agent of contagious bovine intramammary infections in dairy cattle. Its ability to persist inside the udder is based on the presence of important mechanisms such as its ability to form biofilms, polysaccharide capsules small colony variants, and their ability to invade professional and nonprofessional cells, which will protect S. aureus from the innate and adaptive immune response of the cow, and from antibiotics that are no longer considered to be sufficient against S. aureus bovine mastitis. In this review, we present the recent research outlining S. aureus persistence properties inside the mammary gland, including its regulation mechanisms, and we highlight alternative therapeutic strategies that were tested against S. aureus isolated from bovine mastitis such as the use of probiotic bacteria, bacteriocins and bacteriophages. Overall, the persistence of S. aureus inside the mammary gland remains a pressing veterinary problem. A thorough understanding of staphylococcal persistence mechanisms will elucidate novel ways that can help in the identification of novel treatments.