Influence of death struggle on the structural changes in chub mackerel muscle during chilled storage

ABSTRACT: Chub mackerel (34–35 cm, approximately 500 g), which were caught by fishing with a rod and line at the Bungo Channel, Oita prefecture, were rested overnight in a fish preserve and either killed by decapitation (control group) or allowed to struggle in air for 30 min (struggled group). Muscle samples were excised every 4 h, and measurements on breaking strength and histological observations were done for both groups. The breaking strength of muscle in the control group was significantly higher than that in the struggled group, whereby a decrease in breaking strength was delayed for 12 h compared to the struggled group. Light microscopy showed space extension among muscle cells in association with a decrease in breaking strength. Especially in the struggled group, the extended area was larger and the difference in area was significant at the time when breaking strength showed a significant difference. Using electron microscopy, the extended area showed cut and/or disappeared collagen fibrils. From these results, it was demonstrated that struggling to death promoted the degradation of collagen fibrils and the weakening of connective tissue and, resultantly, led to the faster softening of muscle of chub mackerel.     

A direct enzyme immunoassay for the measurement of furosemide in horse plasma

A new enzyme immunoassay (EIA) for the measurement of furosemide in horse plasma is described. The lower limit of detection of this EIA method was 7.8 ng/ml. The intra-and inter-assay coefficients of variation ranged from 2.5% to 4.9% and 7.5% to 9.8%, respectively. Cross-reactivity with other compounds was not observed. There was a high correlation (r2=0.987) between the high-performance liquid chromatography and EIA results obtained for furosemide concentrations in horse plasma. These results indicate that the newly developed EIA method is useful for the quantitative analysis of furosemide in horse plasma.     

Studies on the properties of organic matfer in buried humic horizon derived from volcanic ash

To investigate the relationship between the change of humus property and time factor, the humus composition in the buried humic horizons with the age from 28,000 years B.P. to the present was studied, according to the method presented by Kumada et al. (7).

The fH decreased with the elapse of time and in case of most of humus in the buried humic horizons with the age over 10,000 years, the fH became lower than 40 and were in the form of so-called ‘combined’ humus.

When 3,000-4,000 years have passed since the surface soil buried, the humic acid first moved . to ‘combined form.’ Second, after over 10,000 years, the fulvic acid moved to ‘combined form.’

The humification of ‘free’ humic acid proceeded with the elapse of time till around 5,000 years but of 'combined' humic acid till around 7,000 years. Both the humification-degree of ‘free’ humic acid after around 5,000 years and of ‘combined’ humic acid after around 7,000 years decreased. The humification-degree of ‘free’ and ‘combined’ humic acid from the samples with the age about 15,000 years B.P. to 28,000 years B.P. may be the same.     

Isolation and measurement of carbonic anhydrase isoenzyme III in plasma, sera, and tissues of dogs

OBJECTIVE: To purify canine carbonic anhydrase isoenzyme III (CA-III) and determine plasma, serum, and tissue concentrations of CA-III in healthy dogs and dogs with experimentally induced muscle damage. ANIMALS: 121 healthy Beagles. PROCEDURE: Muscle was obtained from 2 Beagles after euthanasia, and CA-III was purified and characterized by use of column chromatography and electrophoresis, respectively. A CA-III-specific ELISA was developed to determine concentrations of CA-III in plasma of 116 dogs and tissues of 1 dog. Serum creatine kinase (CK) activity and CA-III concentration were also determined before and after induction of muscle damage by IM injection of 2 ml of 10% lidocaine to 2 dogs. RESULTS: Canine CA-III had a molecular weight of 28 kd and an isoelectric point of 8.2. Mean (+/- SD) concentration of CA-III in plasma of healthy dogs was 16.91 +/- 9.55 ng/ml. The highest tissue concentration of CA-III was detected in skeletal muscle. Serum concentration of CA-III increased and peaked within the first 2 to 3 hours after induction of muscle damage. The increase in CA-III concentration was more rapid than that of CK activity, and concentration reached its maximum and returned to baseline sooner than did CK activity. CONCLUSIONS AND CLINICAL RELEVANCE: The CA-III ELISA we developed was a sensitive method for determining CA-III concentrations in plasma, serum samples, and tissue specimens of dogs. Use of this ELISA requires only a small volume of serum and may enable the study of changes in CA isoenzyme concentrations associated with muscle disorders in dogs.     

Down regulation of a novel protein,PSP, in rat hepatoma cdRLh 84-beared tumor

Perchloric acid-soluble protein (PSP) is a members of a new hypothetical family (YER057c/YJGF family) of small proteins with presently unknown function. The high degree of evolutionary conservation of these proteins reflect an involvement in basic cellular regulation. The expression of PSP was examined in rat hepatoma cell dRLh 84-beared rats. The tumor weight increased to 4.24 g at 3 weeks after the transplantation of hepatoma cells and hepatoma which has less differentiated characteristics were observed in rat liver. The expression of PSP in rat hepatoma was down regulated as compared with intact tissue. Thus the expression of PSP seems to be associated with the differentiation process in these transformed cells. On the other hand, some positive cells against the PSP-antibody were observed in the central region of tumor tissue by immunohistochemistry. These cells were shown to be the lymphocytes and the macrophages. The involvement of PSP to cellular growth and differentiation is discussed.     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

The effect of pretreatment with different doses of GnRH to synchronize follicular wave on superstimulation of follicular growth in dairy cattle

The objective of this study was to evaluate the efficiency of gonadotropin releasing hormone (GnRH) and GnRH doses in synchronizing follicular wave emergence as a pretreatment for superovulation in cattle. Fourteen Holstein-Friesian cows 6 days from estrus were randomly assigned to receive 100 microg (n=4), 50 microg (n=5), or 25 microg (n=5) of GnRH. Superovulation was induced with injections of porcine FSH (pFSH) twice daily, decreasing the dose (total 42 AU) over 5 days beginning 2.5 days after receiving GnRH. On the 7th and 8th injections of pFSH, 750 microg of PGF(2alpha) was also given. With the exception of one cow that was given 50 microg of GnRH, ovulation was induced in all cows from the three groups and the new follicular wave emergence was observed. The total number of follicles for the 25 microg GnRH group was less than that observed for the 100 microg GnRH group (P<0.05), although there were no differences between the 100 microg, 50 microg and 25 microg GnRH groups with respect to the number of preovulatory follicles (>or=10 mm) and CL. The numbers of normal embryos were greater for the 25 microg GnRH group than the 100 or 50 microg GnRH groups (P<0.01); however, the numbers of ova/embryos did not differ significantly between the three groups. These results suggest that 25 microg of GnRH was sufficient to induce ovulation and follicular wave emergence. On day 6 of the estrous cycle, a reduction of the dose of GnRH to synchronize follicular wave emergence as a pretreatment for superstimulation promotes transferable embryos.     

Quantitative trait loci associated with lodging tolerance in soybean cultivar ‘Toyoharuka’

Lodging tolerance (LT) is an important trait for high yield and combine-harvesting efficiency in soybean [Glycine max (L.) Merr.]. Many previous studies have investigated quantitative trait loci (QTLs) for lodging score (LS) in soybean. Most of the investigated QTLs were located in the proximal region of maturity or growth habit loci. The aim of this study was to identify genetic factors for LT not associated with maturity or growth habit. QTL analysis was performed using a recombinant inbred line (RIL) population derived from a cross between ‘Toyoharuka’ (TH), a lodging-tolerant cultivar, and ‘Toyomusume’ (TM). The genotypes of TH and TM were estimated as both e1e2E3E4 and dt1. The average LS over 4 years was used for QTL analysis, identifying a major and stable QTL, qLS19-1, on chromosome 19. The LS of the near-isogenic line (NIL) with the TH allele at Sat_099, the nearest marker to qLS19-1, was significantly lower than the NIL with the TM allele at that position. The TH allele at Sat_099 rarely had a negative influence on seed yield or other agronomic traits in both NILs and the TM-backcrossed lines. Our results suggest that marker-assisted selection for qLS19-1 is effective for improving LT in breeding programs.     

Settling and raising postlarval abalone Haliotis diversicolor supertexta (Lischke) on microparticulate diets embedded in a layer of alginate

Two 21 day experiments were conducted to determine whether microparticulate diets could be used to substitute for diatoms for raising (Experiment 1) as well as settling and on‐growing (Experiment 2) Haliotis diversicolor supertexta postlarvae. The experiments were conducted in 10 L flow‐through tanks and each diet had three replicate tanks. In experiment 1, three experimental microparticulate diets supplied by a commercial feed company, labelled SF‐15, SF‐15 (T) and JFY‐13–1, were used, and in experiment 2, SF‐15 and SF‐15 (T). Artificial diets were compared to diatoms in terms of supporting growth and survival. The survival rate for experiment 1 was lowest for JFY‐13–1 (0%) and highest for diatoms (12.9±0.8%). The final size of postlarvae fed SF‐15 and SF‐15 (T) was significantly greater than that of postlarvae fed diatoms, 973±193 μm and 996±220 μm compared to 786±141 μm. In experiment 2, a total number of 501 larvae settled onto plates covered with SF‐15, 254 on SF‐15 (T) and 1085 on diatoms. The highest survival rate was 33.1±6.9% for SF‐15 and the lowest was 17.3±5.0% for diatoms. However, the final length of postlarvae fed diatoms was significantly greater than SF‐15 and SF‐15 (T); 1018±379 μm compared to 728±89 μm and 717±90 μm. The artificial micro diets used in this study were comparable to diatoms in terms of grow and survival of postlarval H. diversicolor supertexta.     

Seasonal and yearly variations in light use and nitrogen use by seedlings of four deciduous broad-leaved tree species invading larch plantations

Several deciduous broad-leaved tree species, differing in leaf phenology, invade larch (Larix kaempferii (Lamb.) Carrière) plantations in Japan. The understory light environment of larch forests changes drastically between the leafy and leafless periods. To determine how the invading seedlings exploit the changing light environment, and if phenological differences reflect the light- and nitrogen-use traits of the seedlings, we measured leaf phenology, seasonal changes in light-saturated photosynthetic rate (P(sat)), leaf nitrogen (N) content (N(area)), chlorophyll/nitrogen ratio (Chl/N), specific leaf area (SLA) and N remobilization rate (NRMR) over 3 years. The mid-successional or gap-phase species, Magnolia hypoleuca Siebold & Zucc., had a short leafy period and high P(sat) and NRMR. In contrast, two late-successional tree species, Prunus ssiori Friedr. Schmidt, which undergoes leaf flush before larch, and Carpinus cordata Blume, which maintains green leaves until frost, both had low P(sat) and NRMR but exploited the opportunity for growth during the period when the larch canopy trees were leafless. Quercus mongolica Fisch. ex Ledeb. var. crispula (Blume) Ohashi, a mid-late-successional species that underwent leaf flush at the same time as the overstory larch, had values of photosynthetic parameters between those of the gap-phase and late-successional species. Among species, M. hypoleuca and Q. mongolica had higher photosynthetic rates and photosynthetic N-use efficiencies. In all species, the relationship between N(area) and P(sat) showed species-specific yearly fluctuations; however, there was no yearly fluctuation in the relationship between N(area) and P(sat) at CO2 saturation. Yearly fluctuations in the N(area)-P(sat) relationship appeared to be induced by changes in SLA and N-use characteristics, which in turn are affected by climatic variations.