Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.     

Peripartal Endocrinology in the Mare and Foetus

The endocrine profiles in the periparturient mares are dominated by increasing concentrations of progestagens and decreasing oestrogens. These hormones are produced by precursors from the foetus, metabolized by the placenta and act primarily on the maternal uterus. The circulating concentrations of hormones in maternal plasma, generally, represent a small proportion of those metabolized by the foetus and utero-placental tissues. There is clear evidence that the foetal hypothalamo-pituitary-adrenal (HPA) axis initiates the process of foetal maturation and the hormonal cascade which culminates in parturition at term. The endocrine changes associated with abnormal pregnancy and abortion in late pregnancy are less well understood, as are the hormonal treatments needed to avert these problems. Further work is needed to establish the biological role of the various hormones present in pregnant mares and, in particular, those hormones which control myometrial quiescence.     

Effectiveness of passive gas-scavenging canisters attached to isoflurane anesthesia systems under standard-use conditions in a laboratory animal facility

Chronic exposure to trace levels of waste anesthetic gases has been linked to higher incidences of neurologic and reproductive dysfunction, hepatic and renal toxicity, and neoplasia in health care professionals. We have shown that low levels of isoflurane emissions are likely in conventional laboratory animal treatment rooms during the use of standard anesthesia delivery systems equipped with activated charcoal canisters for passive gas scavenging. In the present study, we surveyed the effectiveness of canisters (attached to well‐maintained precision isoflurane vaporizers) in current use throughout our AAALAC‐accredited laboratory animal facility. Canisters (Omnicon f/air) had been weighed prior to use and then attached to dual‐loop systems (face mask and induction box circuits) from 1 week to 6 months of service. Isoflurane emissions were measured using a pre‐calibrated, portable infrared spectrophotometer by attaching each canister to the face‐mask circuit, occluding the face mask and closing the stopcock to the induction circuit, and running the system at uniform isoflurane concentration (2%) and oxygen flow rate (1 L minute^?1). Samples were taken in animal procedure rooms (size range, 45–80 m³) in which the air turnover rate ranged between 20 and 30 nonrecirculating changes per hour. Nine of the 60 canisters (15%) in current use were found to have exceeded the manufacturer's recommended use‐life (defined as a weight increase of 50 g). Of these nine, seven canisters did not scavenge isoflurane at all (indicated by emissions greatly exceeding 100 ppm). Isoflurane was not detected in the operator's breathing zone under normal use conditions (i.e. gas directed to both circuits at once). Of the 51 canisters that had not exceeded their rated use‐life, 12 (23.5%) exhausted isoflurane at >2 ppm. Our data show that (i) the potential for exposure to waste isoflurane emissions will depend on the configuration of the delivery system and that (ii) enhanced attention to canister surveillance may be warranted even in well‐run facilities.     

The isolation of salmonella from kangaroos and feral pigs processed for human consumption

A total of 154 feral pig carcases and 81 kangaroo carcases were examined for the presence of Salmonella, coliforms and total aerobic counts. Approximately 34% of pig carcases yielded one or more serotypes of Salmonella, while about 11% of kangaroo carcases were contaminated with salmonella. The results differed widely between sampling occasions. A total of 13 serotypes were isolated from feral pigs with S. anatum (31 isolates) and S. typhimurium (9 isolates) being the predominant serotypes. Coliforms were isolated from approximately 90% of carcases. The mean log10 coliform count on feral pigs was 4.39 +/- 1.45/g and the mean log10 total count was 6.15 +/- 1.15/g. About 21% of carcases were contaminated with more than 100,000 coliforms/g. A total 3 serotypes were isolated from kangaroos (S. bahrenfeld, S. binza, and S. onderstepoort). The mean log10 coliform count on kangaroos was 3.54 +/- 1.04. More than 50% of kangaroo carcases were contaminated with less than 100 coliforms/g. About 15% of carcases were contaminated with more than 10,000 coliforms/g. The mean log10 total count was 5.2 +/- 1.01/g.     

Profile of Steroid Receptors and Increased Aromatase Immunoexpression in Canine Inflammatory Mammary Cancer as a Potential Therapeutic Target

Canine inflammatory mammary cancer (IMC) has been proposed as a model for the study of human inflammatory breast cancer (IBC). The aims of this study were to compare the immunohistochemical expression of aromatase (Arom) and several hormone receptors [estrogen receptor α (ERα), estrogen receptor β (ERβ), progesterone receptor (PR) and androgen receptor (AR)], in 21 IMC cases vs 19 non‐IMC; and to study the possible effect of letrozole on canine IMC and human inflammatory breast cancer (IBC) in vitro using IPC‐366 and SUM‐149 cell lines. Significant elevations of the means of Arom Total Score (TS), ERβ TS and PR TS were found in the IMC group (p = 0.025, p = 0.038 and p = 0.037, respectively). Secondary IMC tumours expressed higher levels of Arom than primary IMC (p = 0.029). Non‐IMC PR‐ tumours contained higher levels of Arom than non‐IMC PR+ tumours (p = 0.007). After the addition of letrozole, the number of IMC and IBC cells dropped drastically. The overexpression of Arom found and the results obtained in vitro further support canine IMC as a model for the study of IBC and future approaches to the treatment of dogs with mammary cancer, and especially IMC, using Arom inhibitors.     

Reticular contraction attributes and passage of digesta from the ruminoreticulum in cattle fed roughage diets.

Four nonlactating Holstein dairy cows (BW = 692 +/- 49 kg) were used in a 4 x 4 Latin square design experiment to determine how changes in attributes of reticular contractions were related to changes in passage of digesta from the ruminoreticulum in cattle fed a forage-based diet. Frequencies of reticular contraction during eating, ruminating, and resting phases of the feeding cycle were not affected (P = .09; P = .07; P = .09, respectively) by feeding level. Duration and amplitude of reticular contractions both increased linearly during eating (P = .03; P = .01), ruminating (P = .01; P = .02), and resting (P = .04; P = .02), respectively, as feed intake level increased. After removing the effects of intake, squared partial correlations computed within analyses of covariance indicated that NDF flow was not related to frequency (r2 = .15, P = .20) but was positively correlated with duration (r2 = .76, P = .0001) and amplitude (r2 = .56, P = .003) of contraction, indicating that duration and amplitude of reticular contraction were more associated with duodenal NDF flow than frequency. However, duration of reticular contraction (r2 = .47, P = .014) explained a greater variation in duodenal NDF flow than amplitude (r2 = .011, P = .74). Stepwise multiple regression also indicated that duration of reticular contraction was the attribute most associated (r2 = .76, P = .0001) with duodenal NDF flow. It was concluded that the changes in digesta passage from the ruminoreticulum were associated primarily with changes in the duration of reticular contraction.