Effects of heat stress on markers of skeletal muscle proteolysis in dairy cattle.

Heat stress (HS) markedly affects postabsorptive energetics and protein metabolism. Circulating urea nitrogen increases in multiple species during HS and it has been traditionally presumed to stem from increased skeletal muscle proteolysis; however, this has not been empirically established. We hypothesized HS would increase activation of the calpain and proteasome systems as well as increase degradation of autophagosomes in skeletal muscle. To test this hypothesis, lactating dairy cows (~139 d in milk; parity ~2.4) were exposed to thermal neutral (TN) or HS conditions for 7 d (8 cows/environment). To induce HS, cattle were fitted with electric blankets for the duration of the heating period and the semitendinosus was biopsied on d 7. Heat stress increased rectal temperature (1.3°C) and respiratory rate (38 breaths per minute) while it decreased dry matter intake (34%) and milk yield (32%). Plasma urea nitrogen (PUN) peaked following 3 d (46%) and milk urea nitrogen (MUN) peaked following 4 d of environmental treatment and while both decreased thereafter, PUN and MUN remained elevated compared with TN (PUN: 20%; MUN: 27%) on d 7 of HS. Contrary to expectations, calpain I and II abundance and activation and calpain activity were similar between groups. Likewise, relative protein abundance of E3 ligases, muscle atrophy F-box protein/atrogin-1 and muscle ring-finger protein-1, total ubiquitinated proteins, and proteasome activity were similar between environmental treatments. Finally, autophagosome degradation was also unaltered by HS. Counter to our hypothesis, these results suggest skeletal muscle proteolysis is not increased following 7 d of HS and call into question the presumed dogma that elevated skeletal muscle proteolysis, per se, drives increased AA mobilization.

Autophagy in the heart is enhanced and independent of disease progression in mus musculus dystrophinopathy models.

BACKGROUND: Duchenne muscular dystrophy is a muscle wasting disease caused by dystrophin gene mutations resulting in dysfunctional dystrophin protein. Autophagy, a proteolytic process, is impaired in dystrophic skeletal muscle though little is known about the effect of dystrophin deficiency on autophagy in cardiac muscle. We hypothesized that with disease progression autophagy would become increasingly dysfunctional based upon indirect autophagic markers. METHODS: Markers of autophagy were measured by western blot in 7-week-old and 17-month-old control (C57) and dystrophic (mdx) hearts. RESULTS: Counter to our hypothesis, markers of autophagy were similar between groups. Given these surprising results, two independent experiments were conducted using 14-month-old mdx mice or 10-month-old mdx/Utrn± mice, a more severe model of Duchenne muscular dystrophy. Data from these animals suggest increased autophagosome degradation. CONCLUSION: Together these data suggest that autophagy is not impaired in the dystrophic myocardium as it is in dystrophic skeletal muscle and that disease progression and related injury is independent of autophagic dysfunction.

Lichens or endophytes? The enigmatic genus Leptosillia in the Leptosilliaceae fam. nov. (Xylariales), and Furfurella gen. nov. (Delonicicolaceae).

Based on DNA sequence data, the genus Leptosillia is shown to belong to the Xylariales. Molecular phylogenetic analyses of ITS-LSU rDNA sequence data and of a combined matrix of SSU-ITS-LSU rDNA, rpb1, rpb2, tef1 and tub2 reveal that the genera Cresporhaphis and Liberomyces are congeneric with Leptosillia. Coelosphaeria fusariospora, Leptorhaphis acerina, Leptorhaphis quercus f. macrospora, Leptorhaphis pinicola, Leptorhaphis wienkampii, Liberomyces pistaciae, Sphaeria muelleri and Zignoëlla slaptonensis are combined in Leptosillia, and all of these taxa except for C. fusariospora, L. pinicola and L. pistaciae are epitypified. Coelosphaeria fusariospora and Cresporhaphis rhoina are lectotypified. Liberomyces macrosporus and L. saliciphilus, which were isolated as phloem and sapwood endophytes, are shown to be synonyms of Leptosillia macrospora and L. wienkampii, respectively. All species formerly placed in Cresporhaphis that are now transferred to Leptosillia are revealed to be non-lichenized. Based on morphology and ecology, Cresporhaphis chibaensis is synonymised with Rhaphidicyrtis trichosporella, and C. rhoina is considered to be unrelated to the genus Leptosillia, but its generic affinities cannot be resolved in lack of DNA sequence data. Phylogenetic analyses place Leptosillia as sister taxon to Delonicicolaceae, and based on morphological and ecological differences, the new family Leptosilliaceae is established. Furfurella, a new genus with the three new species, F. luteostiolata, F. nigrescens and F. stromatica, growing on dead branches of mediterranean fabaceous shrubs from tribe Genisteae, is revealed to be the closest relative of Delonicicola in the family Delonicicolaceae, which is emended. ITS rDNA sequence data retrieved from GenBank demonstrate that the Leptosilliaceae were frequently isolated or sequenced as endophytes from temperate to tropical regions, and show that the genus Leptosillia represents a widely distributed component of endophyte communities of woody plants.

Autophagic dysfunction and autophagosome escape in the mdx mus musculus model of Duchenne muscular dystrophy.

AIM: Duchenne muscular dystrophy is caused by the absence of functional dystrophin protein and results in a host of secondary effects. Emerging evidence suggests that dystrophic pathology includes decreased pro-autophagic signalling and suppressed autophagic flux in skeletal muscle, but the relationship between autophagy and disease progression is unknown. The purpose of this investigation was to determine the extent to which basal autophagy changes with disease progression. We hypothesized that autophagy impairment would increase with advanced disease. METHODS: To test this hypothesis, 7-week-old and 17-month-old dystrophic diaphragms were compared to each other and age-matched controls. RESULTS: Changes in protein markers of autophagy indicate impaired autophagic stimulation through AMPK, however, robust pathway activation in dystrophic muscle, independent of disease severity. Relative protein abundance of p62, an inverse correlate of autophagic degradation, was dramatically elevated with disease regardless of age. Likewise, relative protein abundance of Lamp2, a lysosome marker, was decreased twofold at 17 months of age in dystrophic muscle and was confirmed, along with mislocalization, in histological samples, implicating lysosomal dysregulation in this process. In dystrophic muscle, autophagosome-sized p62-positive foci were observed in the extracellular space. Moreover, we found that autophagosomes were released from both healthy and dystrophic diaphragms into the extracellular environment, and the occurrence of autophagosome escape was more frequent in dystrophic muscle. CONCLUSION: These findings suggest autophagic dysfunction proceeds independent of disease progression and blunted degradation of autophagosomes is due in part to decreased lysosome abundance, and contributes to autophagosomal escape to the extracellular space.

Effect of pharmacological lowering of plasma urate on exercise-induced oxidative stress.

Urate is a metabolic end product of purine metabolism that contributes about 66% of the antioxidant capacity of plasma. The objective of this study was to evaluate the importance of plasma urate as an antioxidant using pharmacological lowering and examining the impact on plasma antioxidant capacity and oxidative stress after intense exercise. Fifteen subjects ran for 45 min at approximately 80% VO2 max under the influence of probenecid (1 g/d) (PRO) or placebo (PLA) in a double-blind, crossover design. Blood samples obtained at baseline, pre-exercise, and immediately post-exercise were analyzed for F2-isoprostanes, lipid hydroperoxides (LHs), ferric-reducing ability of plasma (FRAP), urate, ascorbate (AA), and nitrite. A 2 (group)x2 (time) repeated-measures analysis of variance (ANOVA), one-way ANOVA, Tukey-Kramer multiple comparison tests, and Student's t tests were used for statistical analysis. PRO exhibited lowered urate and FRAP compared with baseline (p

A prospective clinical study of Mineral Trioxide Aggregate and IRM when used as root-end filling materials in endodontic surgery.

AIM: To assess the success rate of the root-end filling material, Mineral Trioxide Aggregate (MTA). METHODOLOGY: Referred adult patients were recruited using strict entry criteria and randomly allocated to receive MTA or IRM. A standardized surgical technique was employed: the root end was resected perpendicularly and a root-end cavity was prepared ultrasonically and filled. A radiograph taken immediately after surgery was compared with those taken at 12 and 24 months. Customised film holders and the paralleling technique were used; radiographs were assessed by two trained observers using agreed criteria. The results from 122 patients (58 in IRM group, 64 in MTA group) after 12 months and 108 patients (47 in IRM group, 61 in MTA group) for the 24-month review period were analysed using the chi2 test. RESULTS: The highest number of teeth with complete healing at both times was observed when MTA was used. When the numbers of teeth with complete and incomplete (scar) healing, and those with uncertain and unsatisfactory healing were combined, the success rate for MTA was higher (84% after 12 months, 92% after 24 months) compared with IRM (76% after 12 months, 87% after 24 months). However, statistical analysis showed no significant difference in success between materials (P > 0.05) at both 12 and 24 months. CONCLUSIONS: In this study, the use of MTA as a root-end filling material resulted in a high success rate that was not significantly better than that obtained using IRM.

Predictors of hospital postdischarge infant mortality: implications for high-risk infant follow-up efforts.

The association between maternal/infant characteristics and postdischarge mortality was determined for 19,573 infants from a single regional perinatal center. Postdischarge mortality was not associated with infant sex or ethnicity, mother's marital status, education, method of delivery, or income status. Mortality risk was statistically increased for infants who were neonatal intensive care unit graduates, of low birth weight, preterm, or small for gestational age. Infants who were either of low birth weight, neonatal intensive care unit graduates, or preterm represented 38% of hospital discharges and 64% of postdischarge deaths. Mortality odds for low-birth-weight infants exceeded that of neonatal intensive care unit graduates or preterm infants. Within these high-risk groups, mortality was further increased for infants with pulmonary interstitial emphysema, patent ductus arteriosus, bronchopulmonary dysplasia, hyaline membrane disease, apnea, or intraventricular hemorrhage III/IV. There were no postdischarge deaths among infants with meconium aspiration, persistent fetal circulation, or necrotizing enterocolitis. Partitioning infant mortality attributable to a single tertiary center suggested that 73.5% of the deaths occurred in infants who were continuously hospitalized from birth; 1.7% of the deaths occurred after discharge in infants who had lethal anomalies and were not expected to survive; 9.0% of the deaths occurred after discharge in infants without a selected risk factor; and 15.8% of the deaths occurred after discharge in infants with at least one high-risk characteristic.

Changing patterns of acute myocardial infarction: decline in period prevalence and delay in onset.

The rural Pee Dee area of South Carolina has had the highest mortality rate in the nation for coronary heart disease. Community surveillance shows a 22.7% (p = 0.0008) decline in fatal and nonfatal acute myocardial infarction (AMI) rates during the period 1978 to 1985 in the Pee Dee area. Rates for white men decreased 32% (p = 0.001), whereas in other race-sex groups changes were not significant, although there was a downward trend (p = 0.18) among black men. Among white men decreases in the AMI rates occurred in each of the decades of age 35 to 64 years. In all race-sex groups there was a sharp increase in rates between 1978 and 1985 for those 65 to 74 years of age, suggesting that the incidence of AMI was delayed to a later age. Out-of-hospital AMI death rates declined markedly in all race-sex groups: 63% for white men, 62% for white women, 49% for black men, and 39% for black women. Overall case fatality rates declined from 14% (27 of 193) to 10% (23 of 232), but the decrease was not statistically significant.

Acute myocardial infarction: period prevalence, case fatality, and comparison of black and white cases in urban and rural areas of South Carolina.

Community surveillance revealed 1085 prevalent cases of acute myocardial infarction (AMI) during 1978 in urban metropolitan Columbia and rural Pee Dee areas of South Carolina. Six hundred fifty-eight hospitalized cases met our criteria and were classified as definite or probable. Death certificates identified 427 who died before admission to the hospital and who were classified as unvalidated. However, there is need to verify death certificate diagnosis in out-of-hospital deaths which account for approximately two thirds of total cases in blacks and about one third of white cases. Other findings were: White males had higher AMI rates in the rural Pee Dee area than in urban Columbia, while black males and black females had higher rates in Columbia than in the Pee Dee area and white females had similar rates in both areas. Rates for out-of-hospital AMI mortality were higher in blacks than in whites. Out-of-hospital AMI mortality rates in Columbia and the Pee Dee area were four times higher than in Minneapolis-St. Paul in 1978. For definite and all criteria AMI, white males had the highest rates, double the black male rate except for all criteria AMI in Columbia, where white male and black male rates were similar. Urban cases of both races experienced more anterior infarctions than rural cases.