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.     

Patterns of antimicrobial susceptibility in Michigan wildlife and bovine isolates of Mycobacterium bovis.

The state of Michigan has recognized the presence of Mycobacterium bovis in its free-ranging white-tailed deer population since 1994. This endemic infection is primarily located in a 12-county area in the northeastern lower peninsula of Michigan. A statewide surveillance and eradication program of the disease has been in effect since 1994. Worldwide, Mycobacterium tuberculosis complex organisms have a known predilection toward development of antimicrobial resistance. The objective of this study was to investigate the antimicrobial susceptibility of M. bovis isolates from white-tailed deer in Michigan and detect any changes in susceptibility over time. M. bovis isolates from 2 fall hunting seasons (1999 and 2004) were used in this study. The fall season of 2004 marked the first documented case of direct transmission of M. bovis from a wild deer to a human in Michigan. Since M. bovis is a zoonotic disease, knowledge of susceptibility can expedite treatment options in humans. M. bovis isolates were obtained from 58 deer, 4 coyotes, 3 cattle, 2 raccoons, and 1 human case from the 2 years combined. Methods of susceptibility testing included 1% proportion agar plates and Bactec radiometric broth testing. M. bovis was found to be uniformly resistant to the antibiotic pyrazinamide; this resistance is common to all M. bovis isolates. No other antimicrobial resistance was found in any of the tested M. bovis isolates, which may be, in part, attributed to the lack of any significant treatment pressure in wildlife.     

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.     

ATM engages autodegradation of the E3 ubiquitin ligase COP1 after DNA damage

The ataxia telangiectasia mutated (ATM) protein kinase is a critical component of a DNA-damage response network configured to maintain genomic integrity. The abundance of an essential downstream effecter of this pathway, the tumor suppressor protein p53, is tightly regulated by controlled degradation through COP1 and other E3 ubiquitin ligases, such as MDM2 and Pirh2; however, the signal transduction pathway that regulates the COP1-p53 axis following DNA damage remains enigmatic. We observed that in response to DNA damage, ATM phosphorylated COP1 on Ser(387) and stimulated a rapid autodegradation mechanism. Ionizing radiation triggered an ATM-dependent movement of COP1 from the nucleus to the cytoplasm, and ATM-dependent phosphorylation of COP1 on Ser(387) was both necessary and sufficient to disrupt the COP1-p53 complex and subsequently to abrogate the ubiquitination and degradation of p53. Furthermore, phosphorylation of COP1 on Ser(387) was required to permit p53 to become stabilized and to exert its tumor suppressor properties in response to DNA damage.     

Feline thyroid carcinoma: diagnosis and response to high-dose radioactive iodine treatment

This study reports the scintigraphy, histopathology, sole treatment with high-dose radioactive iodine and outcome of eight cases of feline thyroid carcinoma. Scintigraphic findings were variable and in 7/8 cases scintigraphic features could not reliably distinguish whether the thyroid tissue was malignant. Histopathology revealed typical criteria of malignancy in all cases, with mitotic activity described most frequently (7/8 cases), followed by infiltration of local tissues (4/8 cases). Cellular pleomorphism was infrequently observed. Single high-dose (1100MBq I(131)) radioiodine therapy was successful in 6/8 cases, with complete resolution of hyperthyroidism, and was associated with prolonged survival times (181-2381 days). Sole treatment with high-dose radioiodine is a safe and effective treatment for functional thyroid carcinoma. The prognosis for feline thyroid carcinoma successfully treated with radioiodine is good, with extended survival times commonly achieved.