Bull selection and use in northern Australia. 4. Calf output and predictors of fertility of bulls in multiple-sire herds

On 10 northern Australian properties, the number of calves sired (calf output) by individual bulls in multiple-sire matings was measured by DNA typing for paternity. There were 235 bulls (92 Santa Gertrudis, 25 5/8 Brahman and 119 Brahman) from 37 multiple-sire mating groups. Number of bulls in groups ranged from 2 to 25 and ages of bulls ranged from 2 to 5 years. Mating periods were for 3-12 months and bull mating percentages were 2.5-6%. In all, there were 4251 calves tested and the resolution of paternity ranged from 92.5 to 100% and averaged 97.7% across all sites. This included 9.9% of calves with no potential sires in any of the mating groups. Of the 235 bulls mated, 58% sired 10% or less calves in each of their respective mating groups with 6% not siring any calves. In contrast, 14% sired over 30% of the calves in each of the respective mating groups. When bulls were mated in groups of 8-24, the maximum percent of calves sired by individual bulls was 26+/-7% (mean+/-S.D.) with a range 11-36%. However, when bulls were mated in groups of 2-7, the maximum percent of calves sired by individual bulls was 59+/-19% with a range 24-94%. Calf output of bulls was moderately repeatable across years at four of five sites. Multiple regression models relating pre-mating measures of physical, seminal and behavioural traits to calf output were developed for the three breed groups. In all, only 138 of the 235 bulls were included in the models (40 Santa Gertrudis, 24 5/8 Brahman and 74 Brahman). Sheath and testicular traits, such as scrotal circumference and testicular tone, were generally not related to calf output, the exceptions being sheath depth in Brahman bulls which was negatively related (P<0.05) and scrotal circumference in 5/8 Brahmans which was positively related (P=0.08) to calf output. Dominance was only included in the 5/8 Brahman model but there was no significant relationship between dominance hierarchy and calf output. Semen motility was only related (P<0.05) to calf output in 5/8 Brahmans. However, measures of semen quality based on spermatozoa morphology were important contributors to calf output in the Santa Gertrudis and Brahman models where percent normal spermatozoa was positively related (P<0.01) to calf output. In Santa Gertrudis and Brahman bulls, measures of sexual behaviour in the serving capacity test were related to calf output. In Santa Gertrudis, these were for the number of displays of sexual interest (P<0.05), and mounts (P<0.01), but not number of serves, whilst in Brahman bulls, libido score was positively related to calf output (P<0.05). The models only explained 35-57% of the variation in calf output.     

Redox State of Pentraxin 3 as a Novel Biomarker for Resolution of Inflammation and Survival in Sepsis

In an endotoxaemic mouse model of sepsis, a tissue-based proteomics approach for biomarker discovery identified long pentraxin 3 (PTX3) as the lead candidate for inflamed myocardium. When the redox-sensitive oligomerization state of PTX3 was further investigated, PTX3 accumulated as an octamer as a result of disulfide-bond formation in heart, kidney, and lung—common organ dysfunctions seen in patients with sepsis. Oligomeric moieties of PTX3 were also detectable in circulation. The oligomerization state of PTX3 was quantified over the first 11 days in critically ill adult patients with sepsis. On admission day, there was no difference in the oligomerization state of PTX3 between survivors and non-survivors. From day 2 onward, the conversion of octameric to monomeric PTX3 was consistently associated with a greater survival after 28 days of follow-up. For example, by day 2 post-admission, octameric PTX3 was barely detectable in survivors, but it still constituted more than half of the total PTX3 in non-survivors (p < 0.001). Monomeric PTX3 was inversely associated with cardiac damage markers NT-proBNP and high-sensitivity troponin I and T. Relative to the conventional measurements of total PTX3 or NT-proBNP, the oligomerization of PTX3 was a superior predictor of disease outcome.Severe sepsis is a common acute illness in intensive care units (ICUs)¹ and is associated with high mortality rates and chronic morbidity. When it is associated with hypotension (termed septic shock), the mortality rate is very high (50% to 80%). Cardiovascular dysfunction during sepsis is multifactorial and often associated with minimal loss of myocardial tissue, but with the release of myocardial-specific markers such as troponins. A key unmet clinical need is the availability of a biomarker that predicts myocardial dysfunction early, monitors response to treatment, and thus identifies a cohort of patients at higher risk of septic shock to aid in targeted interventions and improve outcome (1).In the present study, we used proteomics for biomarker discovery. Over the past decade, the field of proteomics has made impressive progress. Plasma and serum, however, are the most complex proteomes of the human body (2), and less abundant proteins tend to be missed in untargeted proteomics analyses of body fluids (3). Thus, we pursued an alternative strategy: the application of proteomics to diseased tissue (4), in which the potential biomarkers are less dilute and have a less uncertain cellular origin (5–7). We employed a solubility-based protein-subfractionation methodology to analyze inflammatory proteins that are retained with sepsis tissue. This innovative proteomics approach shall reveal inflammatory molecules that reside and persist within inflamed tissue. We hypothesized that proteins that accumulate in the susceptible tissues are more likely to be biomarker candidates for organ dysfunction than proteins that just circulate in plasma or serum. We then validated our proteomics findings in the preclinical model using samples from sepsis patients admitted to ICUs.     

Bull selection and use in northern Australia. 3. Serving capacity tests

Serving capacity studies were conducted on Santa Gertrudis, 5/8 Brahman, Brahman and Belmont Red bulls at 10 sites. Modifications, such as providing prior sexual experience, using females in oestrus and comparing restrained with unrestrained females, were made to the standard Bos taurus serving capacity test in a series of experiments with Santa Gertrudis bulls. Providing sexual experience to virgin bulls prior to testing improved the number of serves and libido scores in a serving capacity test. In serving capacity tests that used restrained females, expressions of sexual behaviour of bulls were not improved by having females in oestrus compared with non-oestrus females. There were more sexual behaviours, particularly mounts, in serving capacity tests involving restrained rather than unrestrained females. Summary statistics were collated from over 1100 serving capacity tests, in the 2 months prior to mating, across the 10 sites. In serving capacity tests using restrained females, in bulls of the same age, there were more expressions of sexual behaviours (except interest) in Belmont Red than in Brahman or 5/8 Brahman bulls with Santa Gertrudis bulls intermediate. The percentage of bulls displaying serves tended to increase with age; 82, 83 and 86% of 2-, 3- and > or =4-year-old Belmont Red bulls, respectively, completed 1 or more serves. This compared with 50, 50 and 66% of 2-, 3- and > or =4-year-old Santa Gertrudis bulls and 31% of 2-year-old Brahman bulls. Between 2 and 9% of bulls did not display any sexual behaviour in serving capacity tests with restrained females. Across genotypes, there were few and inconsistent relationships between either mounts or serves and liveweight, condition score, scrotal circumference, testicular tone and sheath measurements. The exceptions were in 2-year-old Brahmans where mounts were negatively correlated (r=-0.61; P<0.05) with umbilicus thickness and in 3-year-old Santa Gertrudis bulls, where serves (r=-0.53; P<0.01) and mounts+serves (r=-0.49; P<0.01) were negatively correlated with navel score. Measures of sexual behaviour were unable to consistently predict calf output of bulls in multiple-sire mating groups in extensive pastoral conditions. The main value of the serving capacity test in these genotypes is to identify whether a bull is capable of natural service.     

Host and parasite genetics shape a link between Trypanosoma cruzi infection dynamics and chronic cardiomyopathy

Host and parasite diversity are suspected to be key factors in Chagas disease pathogenesis. Experimental investigation of underlying mechanisms is hampered by a lack of tools to detect scarce, pleiotropic infection foci. We developed sensitive imaging models to track Trypanosoma cruzi infection dynamics and quantify tissue‐specific parasite loads, with minimal sampling bias. We used this technology to investigate cardiomyopathy caused by highly divergent parasite strains in BALB/c, C3H/HeN and C57BL/6 mice. The gastrointestinal tract was unexpectedly found to be the primary site of chronic infection in all models. Immunosuppression induced expansion of parasite loads in the gut and was followed by widespread dissemination. These data indicate that differential immune control of T. cruzi occurs between tissues and shows that the large intestine and stomach provide permissive niches for active infection. The end‐point frequency of heart‐specific infections ranged from 0% in TcVI‐CLBR‐infected C57BL/6 to 88% in TcI‐JR‐infected C3H/HeN mice. Nevertheless, infection led to fibrotic cardiac pathology in all models. Heart disease severity was associated with the model‐dependent frequency of dissemination outside the gut and inferred cumulative heart‐specific parasite loads. We propose a model of cardiac pathogenesis driven by periodic trafficking of parasites into the heart, occurring at a frequency determined by host and parasite genetics.     

Master Settlement Agreement (MSA) Spending and Tobacco Control Efforts

We investigate whether the distributions to the states from the Tobacco Master Settlement Agreement (MSA) in 1998 is associated with stronger tobacco control efforts. We use state level data from 50 states and the District of Columbia from four time periods post MSA (1999, 2002, 2004, and 2006) for the analysis. Using fixed effect regression models, we estimate the relationship between MSA disbursements and a new aggregate measure of strength of state tobacco control known as the Strength of Tobacco Control (SoTC) Index. Results show an increase of $1 in the annual per capita MSA disbursement to a state is associated with a decrease of −0.316 in the SoTC mean value, indicating higher MSA payments were associated with weaker tobacco control measures within states. In order to achieve the initial objectives of the MSA payments, policy makers should focus on utilizing MSA payments strictly on tobacco control activities across states.     

Stochastic study of the effect of ionic strength on noncovalent interactions in protein pores

Salt plays a critical role in the physiological activities of cells. We show that ionic strength significantly affects the kinetics of noncovalent interactions in protein channels, as observed in stochastic studies of the transfer of various analytes through pores of wild-type and mutant α-hemolysin proteins. As the ionic strength increased, the association rate constant of electrostatic interactions was accelerated, whereas those of both hydrophobic and aromatic interactions were retarded. Dramatic decreases in the dissociation rate constants, and thus increases in the overall reaction formation constants, were observed for all noncovalent interactions studied. The results suggest that with the increase of salt concentration, the streaming potentials for all the protein pores decrease, whereas the preferential selectivities of the pores for either cations or anions drop. Furthermore, results also show that the salt effect on the rate of association of analytes to a pore differs significantly depending on the nature of the noncovalent interactions occurring in the protein channel. In addition to providing new insights into the nature of analyte-protein pore interactions, the salt-dependence of noncovalent interactions in protein pores observed provides a useful means to greatly enhance the sensitivity of the nanopore, which may find useful application in stochastic sensing.     

Gel-based methods in redox proteomics

Background

The key to understanding the full significance of oxidants in health and disease is the development of tools and methods that allow the study of proteins that sense and transduce changes in cellular redox. Oxidant-reactive deprotonated thiols commonly operate as redox sensors in proteins and a variety of methods have been developed that allow us to monitor their oxidative modification.

Scope of the review

This outline review specifically focuses on gel-based methods used to detect, quantify and identify protein thiol oxidative modifications. The techniques we discuss fall into one of two broad categories. Firstly, methods that allow oxidation of thiols in specific proteins or the global cellular pool to be monitored are discussed. These typically utilise thiol-labelling reagents that add a reporter moiety (e.g. affinity tag, fluorophore, chromophore), in which loss of labelling signifies oxidation. Secondly, we outline methods that allow specific thiol oxidation states of proteins (e.g. S-sulfenylation, S-nitrosylation, S-thionylation and interprotein disulfide bond formation) to be investigated.

Major conclusions

A variety of different gel-based methods for identifying thiol proteins that are sensitive to oxidative modifications have been developed. These methods can aid the detection and quantification of thiol redox state, as well as identifying the sensor protein.

General significance

By understanding how cellular redox is sensed and transduced to a functional effect by protein thiol redox sensors, this will help us better appreciate the role of oxidants in health and disease. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.     

Bull selection and use in northern Australia. 1. Physical traits

A systematic breeding soundness examination, including assessment of serving capacity was conducted on 5/8 Brahman, Brahman and Belmont Red bulls, most aged 2-4 years, at 12 different cattle properties across northern Australia. A subset of bulls (n=235) were subsequently mated in various groups, as multiple-sires, to cows and heifers at bull:female ratios of 2.5-6%. The number of calves sired by individual bulls (calf output) was determined by DNA typing for paternity. Overall, the incidence of physical abnormalities that were judged likely to result in reduced reproductive performance was low (5.6-12% of bulls). Measures of key physical traits scrotal circumference, sheath depth (vertical distance from ventral abdominal wall to preputial orifice), and sheath score were moderately to highly repeatable. Except in Belmont Red bulls, liveweight was positively correlated with scrotal circumference (r=0.36-0.78; P<0.01), and both traits increased with age. In 2-year-old Brahman bulls only, percent normal spermatozoa was correlated with scrotal circumference (r=0.34; P<0.05). However, 12 and 15% of Santa Gertrudis and Brahman bulls, respectively, with greater than or equal to the recommended threshold values for scrotal circumferences, had less than 50% morphologically normal spermatozoa. Size and conformation of the umbilicus was associated with conformation of the sheath, and influenced mating ability in 2-year-old Brahman and 3-year-old Santa Gertrudis bulls. In 2-year-old Brahman bulls umbilical cord thickness was positively related (r=0.36; P<0.05) to sheath depth, and negatively related (r=-0.65; P<0.05) to number of mounts and serves in a serving capacity test. In 3-year-old Santa Gertrudis bulls navel (external umbilical scar) score was negatively related to the number of serves (r=-0.53; P<0.01) in a serving capacity test. None of the physical traits measured were consistently included in the final regression model for calf output for each breed.