Response of Biochemical Markers of Bone Metabolism to Exercise Intensity in Thoroughbred Horses

We studied the response of biochemical markers of bone metabolism to exercise intensity in horses. Four horses were walked on a mechanical walker for one week (pre-exercise). Then they performed low-speed exercise on a high-speed treadmill in the first week and medium-speed exercise in the second week and high-speed exercise in the third week of training. We measured two indices of bone resorption, serum hydroxyproline concentration and the urinary deoxypyridinoline/creatinine ratio, and serum osteocalcin (OC) concentration as an index of bone formation. Both indices of bone resorption gradually decreased during the experiment. Serum OC concentration did not change in the first week but was significantly lower in the second and the third weeks compared to in the pre-exercise period and in the first week. These results suggest that the low-speed exercise decreased bone resorption but did not affect bone formation, which possibly results in increasing bone mineral content and strengthening of bones. The high-speed exercise decreased bone formation and bone resorption, i.e., bone turnover was suppressed. The low-speed exercise may be preferable for increasing bone mineral content.     

Cell wall degradation of tropical and temperate forage grasses measured by nylon bag and in vitro digestion techniques

Four tropical and four temperate grasses were subjected to nylon bag fiber degradation study with fistulated Holstein cattle as well as in vitro digestion with broad spectrum cellulase, and the degradability of cell wall carbohydrate and lignin for the respective forage grasses were determined. The degradability of neutral detergent fiber and acid detergent fiber for the same plant materials appeared to be similar, but that of acid detergent lignin was constantly lower. As for the plant materials used in the present study, Italian ryegrass, orchardgrass and guineagrass showed high degradability of fibers in nylon bag incubation, while setaria, cogongrass and wheat showed smaller degradability. In both nylon bag incubation and in vitro digestion experiments, the forage samples showing relatively lower degradability exhibited notably lower disappearance of xylose than that of glucose or arabinose. It was also observed that dry matter degradability correlated negatively with the initial content of esterified and/or etherified p‐coumaric acid. These results were assumed to be caused by the difference in resistance towards rumen microbial degradation due to the concentration of lignin‐carbohydrate complex.     

Involvement of cytochrome P‐450 enzyme activity in the selectivity and safening action of pyrazosulfuron‐ethyl

To investigate the selectivity and safening action of the sulfonylurea herbicide pyrazosulfuron‐ethyl (PSE), pyrazosulfuron‐ethyl O‐demethylase (PSEOD) activity involving oxidative metabolism by cytochrome P‐450 was studied in rice (Oryza sativa L cv Nipponbare) and Cyperus serotinus Rottb. Cytochrome P‐450‐dependent activity was demonstrated by the use of the inducers 1,8‐naphthalic anhydride and ethanol, the herbicides PSE, bensulfuron‐methyl, dimepiperate and dymron, or the inhibitor piperonyl butoxide (PBO). Growth inhibition in C serotinus seedlings was more severe than that in rice seedlings. O‐Dealkylation activities of PSE were induced differently in rice and in C serotinus, with distinctly higher activity in rice seedlings. The induced PSEOD activities were slightly inhibited by PBO in rice seedlings, whereas they were strongly inhibited in C serotinus seedlings. Dimepiperate and dymron were effective safeners of rice against PSE treatment. Treatments with herbicide alone resulted in less induction of PSEOD activity compared with combined treatments of the herbicide and safener. PSEOD activity in rice seedlings induced with herbicide alone was strongly inhibited by PBO, whereas it was weakly inhibited in rice seedlings induced with combinations of PSE and two safeners. These results suggest that O‐demethylation by cytochrome P‐450 enzymes may be involved in the metabolism of PSE and may contribute to its selectivity and safening action. Furthermore, these results suggest the existence of a multiple form of cytochrome P‐450 in plants. © 2001 Society of Chemical Industry     

Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.     

The cumulative methane production from dairy cattle slurry can be explained by its volatile solid,temperature and length of storage

In order to refine the national estimate of methane emission from stored cattle slurry, it is important to comprehend the basic characteristics of methane production. Two dairy cattle slurries were obtained from livestock farms located in Hokkaido (a northern island) and Kyushu (a southern island). The slurries were diluted with water into three levels: undiluted, three times diluted, and 10 times diluted. Three hundred mL of the slurries were put into a bottle with a headspace volume of 2.0 L, which was filled with nitrogen gas and then sealed by butyl rubber. Four levels of temperature were used for incubation: 35, 25, 15 and 5 °C. The time course of the cumulative methane production per volatile solid (VS) was satisfactorily expressed by an asymptotic regression model. The effect of dilution on the methane production per VS was not distinctive, but that of temperature was of primary importance. In particular, higher temperature yields a higher potential production and a shorter time when the cumulative production reaches half of the potential production. The inclusive and simple models obtained in this study indicate that the cumulative methane production from stored cattle slurry can be explained by VS, temperature and length of storage.     

Cervical transposition test using Hegar dilator at oestrus as a tool to select ewes for transcervical embryo collection

This study evaluated the cervical transposition method as a tool to select ewes for embryo collection by transcervical route. Adult Santa Inês ewes (n = 50) received Day 0 protocol for superovulation treatments. The cervix transposition test was performed both at oestrus and at the embryo collection time. The latter was preceded by hormonal cervical dilation. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 85.7%, 66.6%, 85.7%, 66.6% and 80.0%, respectively. The kappa index yielded a moderate score (κ = 0.52). In conclusion, the high sensitivity and accuracy indicate that the cervical transposition test is a screening option to select ewes for embryo collection by transcervical route.     

Partitioning of C-photosynthates from different current shoots neighboring with fruiting spur in later-maturing Japanese pear during the period of rapid fruit growth

To elucidate the fate of photosynthates from different current shoots and their influence on fruit growth and bud differentiation in neighboring spur complex during the period of rapid fruit growth in two late-maturing Japanese pear cultivars: ‘Atago’ and ‘Shinkou’ with contrasting fruit size, ¹³C labeling of single shoot was done to investigate of C-relations in fruit branches of eight shoot-combinations. The results showed that all of the current shoots investigated (bourse shoots of nonfruiting spur, bourse shoots of fruiting spur, extension shoot, nonfruiting spur, vegetative shoot, and water sprout) could export photosynthates to the neighboring fruit and buds. Water sprouts together with vegetative shoots, bourse shoots, and extension shoots are important source for fruit growth after shoot growth termination during the period of rapid fruit growth in production of late-maturing pears. The carbon transfer rate from the neighboring to the fruit bearing spur is depent (i) on the types of shoot which acts as C source, (ii) on the position of the fruiting spur and (iii) on the source-sink distance. Furthermore, the cultivar difference in carbon partitioning from different current shoot-combinations confirmed that the movement of photosynthates into the fruit was determined by sink strength of the fruit, and ‘Atago’ exhibited a greater relative sink strength of fruit than ‘Shinkou’. In addition, vegetative shoots are very important C sources for fruit growth in ‘Atago’ and the growth pattern of bourse shoot seriously affects C allocated to fruit in ‘Shinkou’.     

Characterization of myofibrillar adenosine triphosphatase activity and liberation of actin from myofibrils upon heating chicken breast meat

Denaturation of actin and myosin in myofibrils induced by heating at 50°C was investigated to reveal the mechanism of irreversible liberation of actin from myofibrils on heating at lower temperatures than conventional cooking. Denaturation of these proteins was determined by Mg²⁺‐ATPase (adenosine triphosphatase) and Ca²⁺‐ATPase activities. When minced meat was heated for 20 min, actin was liberated accompanying denaturation of 80% of actin and 50% of myosin. Heating of the myofibrillar fraction (MFF) isolated from meat homogenate induced much slower denaturation of actin than myosin. When MFF was heated with sarcoplasmic fractions, denaturation of actin was facilitated, suggesting that sarcoplasmic fractions contain factors to facilitate actin denaturation. Inosine‐5′‐monophosphate, a component of sarcoplasmic fractions, was shown to have no effect on actin and myosin denaturation. These results suggest that heating meat at 50°C dissociates binding (‘Bond A’) between actin and myosin participating in ATPase activities, resulting in denaturation of both proteins under influence of sarcoplasmic components. Although denaturation of actin and myosin disrupted Bond A, actin was not liberated simultaneously, suggesting the presence of another bond (‘Bond B’, more heat‐stable than Bond A) between both proteins and necessity of disruption of Bond B for actin release from myofibrils.     

Pool‐based genome‐wide association study identified novel candidate regions on BTA9 and 14 for oleic acid percentage in Japanese Black cattle

Fatty acid composition is an important indicator of beef quality. The objective of this study was to search the potential candidate region for fatty acid composition. We performed pool‐based genome‐wide association studies (GWAS) for oleic acid percentage (C18:1) in a Japanese Black cattle population from the Hyogo prefecture. GWAS analysis revealed two novel candidate regions on BTA9 and BTA14. The most significant single nucleotide polymorphisms (SNPs) in each region were genotyped in a population (n = 899) to verify their effect on C18:1. Statistical analysis revealed that both SNPs were significantly associated with C18:1 (p = .0080 and .0003), validating the quantitative trait loci (QTLs) detected in GWAS. We subsequently selected VNN1 and LYPLA1 genes as candidate genes from each region on BTA9 and BTA14, respectively. We sequenced full‐length coding sequence (CDS) of these genes in eight individuals and identified a nonsynonymous SNP T66M on VNN1 gene as a putative candidate polymorphism. The polymorphism was also significantly associated with C18:1, but the p value (p = .0162) was higher than the most significant SNP on BTA9, suggesting that it would not be responsible for the QTL. Although further investigation will be needed to determine the responsible gene and polymorphism, our findings would contribute to development of selective markers for fatty acid composition in the Japanese Black cattle of Hyogo.