Time‐coordinated prevalence of extracellular HGF,FGF2 and TGF‐β3 in crush‐injured skeletal muscle

Successful regeneration and remodeling of neuromuscular junctions are critical for restoring functional capacities and properties of skeletal muscle after damage, and axon‐guidance molecules may be involved in the signaling that regulates such restoration. Recently, we found that early‐differentiated satellite cells up‐regulate a secreted neural chemorepellent Sema3A upon in vivo muscle‐crush injury. The study also revealed that Sema3A expression is up‐regulated in primary satellite‐cell cultures in response to hepatocyte growth factor (HGF) and basic fibroblast growth factor (FGF2) and is prevented by transforming growth factor (TGF)‐β2, 3. In order to verify the physiological significance of this regulation in vitro, the present study was designed to estimate the time‐course of extracellular HGF, FGF2 and TGF‐β3 concentrations after crush‐injury of Gastrocnemius muscle in the rat lower hind‐limb, using a combination of a non‐homogenization/non‐spin extraction of extracellular wound fluids and enhanced chemiluminescence–Western blotting analyses. Results clearly demonstrated that active HGF and FGF2 are prevalent in 2–8 days post‐crush, whereas active TGF‐β3 increases after 12 days, providing a better understanding of the time‐coordinated levels of HGF, FGF2 and TGF‐β3 that drive regulation of Sema3A expression during regenerative intramuscular moto‐neuritogenesis.     

Gas chromatography–mass spectrometry associated global analysis of rice root exudates under aseptical conditions

Components of rice root exudates were surveyed by gas chromatography–mass spectrometry (GC-MS) and the effect of phosphate (P) starvation was investigated. Rice seedlings were aseptically grown in controlled environments under P-supplied (+P) or P-deficient (–P) conditions. Root exudates were collected from the culture solution 4 and 8 days after treatment (D4 and D8, respectively), which contain the first and second 4-day intervals. The collected solution was lyophilized and then eluted with methanol. In the present study, primary metabolites, such as sugars, amino acids, organic acids and fatty acids, were mainly targeted. After derivatization with methoxyamine hydrochloride and N-methyl-N-trifluoroacetamide, the samples were analyzed by GC-MS. Sugars and various amino acids (such as aspartic acid and l -isoleucine) were mainly detected and the components changed with the growth stage rather than the P content in the culture solution. The relative amount of sugars (such as glucose and fructose) and amino acids (such as aspartic acid and l -isoleucine) decreased in the D8 samples. Of the 78 detected compounds, orthogonal partial least-squares discrimination analysis (O-PLS-DA) showed that four compounds were related to the P content of the nutrient solution and 58 were related to the sampling date (days after treatment). The results suggest that the physiological change during seedling development was unexpectedly large and that the response to the environment was rather small.     

Characterization of lignin-derived products from various lignocellulosics as treated by semi-flow hot-compressed water

To elucidate the decomposition behaviors of lignin from different taxonomic groups, five different lignocellulosics were treated with hot-compressed water (230 °C/10 MPa/15 min) to fractionate lignins into water-soluble portions, precipitates, and insoluble residues. The lignin-derived products in each fraction were characterized and compared. The delignification of monocotyledons [nipa palm (Nypa fruticans) frond, rice (Oryza sativa) straw, and corn (Zea mays) cob] was more extensive than that achieved for Japanese cedar (Cryptomeria japonica, gymnosperm) and Japanese beech (Fagus crenata, dicotyledon angiosperm). The water-soluble portions contained lignin monomers like coniferyl alcohol and phenolic acids, while the precipitates contained higher molecular weight lignin with high content of ether-type linkages. Lignin in the insoluble residues was rich in condensed-type structures. In all five lignocellulosics, ether-type linkages were preferentially cleaved, while condensed-type lignin showed resistance to hot-compressed water. In the monocotyledons, lignin–carbohydrate complexes were cleaved and gave lignins that had higher molecular weights than those eluted from the woods. These differences would facilitate the delignification in monocotyledons. Such information provides useful information for efficient utilization of various lignocellulosics.     

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for mtDNA typing in hokkaido brown bear (Ursus arctos yesoensis)

To develop an easy method of typing of mitochondrial DNA (mtDNA) in Hokkaido brown bears (Ursus arctos yesoensis), the PCR-RFLP technique was improved using four restriction enzymes: Mbo 1, Cfr 13 I, TspE 1, and Fok 1. This approach identified seven groups of mtDNA haplotypes, HB1/2/5-7, HB 3, HB4, HB8/9, HB10/11, HB12 and HB13 from 102 brown bears of northern, central and eastern Hokkaido.     

Annual changes in fecal estradiol-17beta concentrations of the sun bear (Helarctos malayanus) in Sarawak,Malaysia

Fecal estradiol concentrations were measured in three captive unmated female sun bears (Helarctos malayanus) from August 1998 to July 1999 in Sarawak, Malaysia and vaginal smears from one of the females was observed in August 1998 and March 1999. A single peak in fecal estradiol concentration was obvious for each bear in August or September 1998, and there was a much higher percentage of superficial vaginal anuclear cells in August 1998 than in March 1999. These results suggest that sun bears in Sarawak are likely to be a seasonal breeder associated with a peak of estrogen production in August or September.     

Effects of ammonium hydroxide treatment on the in vitro dry matter digestibility and gas production of wheat straw, sugarcane bagasse medium and konara oak rotted by edible basidiomycetes

Wheat straw, sugarcane bagasse medium and konara oak logs used to cultivate edible basidiomycetes were treated with ammonium hydroxide to investigate the relationship between the degree of rotting and the effect of ammonium hydroxide treatment on digestibility. Wheat straw was sampled before inoculation and on days 67 and 97 after inoculation with Pleurotus salmoneostramineus. Similarly, bagasse media were sampled before inoculation and on days 47 and 96 after inoculation with Pleurotus abalonus. Konara oak logs were sampled before inoculation and at 2 and 4 years after inoculation with Lentinula edodes. Aliquots of the samples were treated with ammonium hydroxide (NH₄‐N, 3% dry matter) at 45°C for 10 days. Ammonium hydroxide treatment increased the in vitro dry matter digestibility (IVDMD) and gas production (IVGP) of all the samples, except the konara oak samples collected before inoculation. The degree of increased digestibility was largest for the samples collected after short cultivation periods, followed by those collected after long cultivation periods and those collected before inoculation. However, the IVDMD and IVGP values after short cultivation periods were lower than those after long cultivation periods. Ammonium hydroxide treatment is effective at improving the digestibility of white‐rotted materials, although the effect is lessened as rotting proceeds.     

Estimation of direct and maternal genetic parameters for growth and carcass traits in a herd of Japanese Black cattle in Miyagi prefecture, using a multitrait animal model

Data from 1170 records of fattening calves were collected on growth and carcass traits from a Japanese Black cattle herd located in Miyagi prefecture, Japan. The objective was to determine direct and maternal heritabilities, direct and maternal genetic correlations and phenotypic correlations between bodyweight at the beginning of the fattening period (BWS), bodyweight at the end of the fattening period (BWF), carcass weight (CW), average daily gain during the fattening period (ADG), rib eye area (REA), rib thickness, subcutaneous backfat thickness (SFT), yield estimate (YE) and beef marbling score (BMS). Direct heritability estimates of 0.16 (SFT) and 0.07 (BMS) were low, whereas estimates of the other traits were medium to high and ranged between 0.44 (REA) and 0.78 (CW). Direct genetic correlations were all positive, except those that were between BWS and SFT, and between BWS and YE (?0.49 and ?0.14, respectively). The lowest positive genetic correlation was between BWS and BMS (0.04) and the highest was between BWF and CW (0.99). The phenotypic correlation coefficients ranged between ?0.41 (between SFT and YE) and 0.96 (between BWF and CW). Maternal heritability estimates were generally low and ranged between 0.00 for BMS and 0.08 for BWS, CW and ADG. Selection programs comprising information on growth and carcass traits of calves and maternal traits of dams were suggested.     

Effects of grazing on deposition of chemical body components, energy retention and plasma hormones in steers

The objective of this study was to evaluate the effects of grazing on the accretion of energy and chemical components in steers, using the comparative slaughter technique and taking into consideration the growth rate. Twelve Holstein steers aged 6 months and weighing 160 kg were divided into three treatment groups – an initial, grazing and control group. The initial group was slaughtered at the beginning of the experiment. The grazing group was grazed on timothy pastures for 18 weeks. The control group was kept in a stable and offered concentrate and timothy hay in a proportion of 2:1 throughout the 18 weeks. The amount of ration offered to the control group was adjusted in order to obtain a growth rate similar to that of the grazing group. The grazing and control groups were slaughtered at the end of the experiment. The grazing and the control groups had similar growth rates of approximately 0.8 kg/day throughout the experimental period and similar final live weights of approximately 260 kg. The grazing group had lower fat and energy deposition rates than the control group (P < 0.05). Deposition rates of protein, fat, ash, moisture and energy were 130, 32, 30, 374 g/day and 4.4 MJ/day, respectively, in the grazing group and 117, 85, 32, 381 g/day and 6.2 MJ/day, respectively, in the control group. The net energy for growth was lower in the grazing group (7.7 MJ/kg) than in the control group (9.9 MJ/kg, P < 0.05). The grazing group had higher plasma concentrations in growth hormone and lower plasma concentrations in insulin and insulin‐like growth factor 1 than those in the control group (P < 0.05).     

Effects of fat coated rumen bypass lysine and methionine on performance of dairy cows fed a diet deficient in lysine and methionine

Two experiments were conducted to evaluate the effect of fat coated rumen bypass lysine (RPLys) and methionine (RPMet) on the lactation performance of dairy cows. In Experiment 1, three lactating cows were supplied with RPLys and fat coated DL‐Met which was highly protected (H‐RPMet) as an indigestible marker, and total fecal emission was collected for 72 h following administration. Measuring the proportional difference in fecal excretion of lysine derived from RPLys relative to methionine derived from H‐RPMet, the intestinal availability of RPLys was estimated to be 66.2%. In Experiment 2, 20 multiparous Holstein cows producing approximately 40 kg/day of milk were assigned to two treatments; fed RPLys (16 g/day as lysine) and RPMet (6.5 g/day as methionine) or none (control) from 5 to 21 weeks postpartum. The consumption of dry matter, organic matter, crude protein, neutral detergent fiber and acid detergent fiber were significantly more in the control cows throughout the experimental period. Their milk protein yield, the contents of their milk protein and milk fat were higher by 0.03 kg (P = 0.03), 0.06% (P < 0.001) and 0.11% (P = 0.07), respectively, in the treatment group compared to the control. These results suggest that the RPLys and RPMet used in this study improved the lactation performance of dairy cows.