Structural differences between myofibrillar protein,paratropomyosin, and tropomyosin as revealed by high‐performance liquid chromatography

Paratropomyosin (PTM) composes myofibril functions to weaken the rigor linkages formed between actin and myosin during postmortem aging of muscles. PTM has the similar physico‐chemical properties as tropomyosin (TM) that is a regulatory protein of myofibrils. So far, it is unclear whether PTM is definitely different from TM, because the primary structure of PTM has not been determined yet. The aim of this study was to clarify structural difference of PTM from TM. PTM was prepared by column chromatography immediately after slaughter from broiler breast muscle, and purified by high‐performance liquid chromatography (HPLC). Purified PTM was successfully separated from TM, and the recovered PTM molecule was reduced with dithiothreitol to separate again by HPLC. Two subunits were obtained and peptides from each digested subunit by V8 protease were recovered by HPLC, and then amino acid sequences of the peptides were analyzed by protein sequencing. As a result, some amino acid residues were replaced from that of TMα1 isoform which is the major isoform of TM, and also was different between the two subunits. Therefore, it is concluded that PTM clearly differs from TM and it is suggested that functional difference in PTM from TM is attributed to amino acid replacements in subunits composing PTM.     

Predation on glass eels of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> in the Tone River Estuary,Japan

Aquaculture of Japanese eel Anguilla japonica relies upon the natural recruitment of their glass eels (juveniles); however, predation that could influence glass eel recruitment remains unknown. In the present study, we aimed to elucidate the proportion of predation on A. japonica glass eels through stomach content analysis of predatory fishes collected in the estuary region of the Tone River system and its vicinity in Japan. Species of the predated glass eels were identified by DNA barcoding. A total of 270 predatory fishes of 15 taxa was collected over 2 years. The overall proportion of predation on glass eels, genetically identified as Japanese eel, was 0.7%, but this rose to 2.0% when data were limited to fishes caught during the peak months of glass eel recruitment. A glass eel was found in the stomach contents of a channel catfish Ictalurus punctatus, an invasive species in this river system, and a blackfin sea bass Lateolabrax latus. These fishes are therefore considered potential predators of A. japonica glass eels. However, as the proportion of predation was low, and the glass eels represented only small proportions of predator stomach contents, further investigation is needed for a better understanding of predation on A. japonica glass eels, and its effects on the early life stages of this endangered species.     

In vitro measurement of post‐natal changes in proliferating satellite cell frequency during rat muscle growth

Satellite cells, resident myogenic stem cells found in postnatal skeletal muscle, are most abundant during early postnatal development and sharply decline in frequency thereafter to adult levels in mice and rats. Therefore, postnatal changes in satellite cell mitotic activities are important aspects for further understanding a muscle growth strategy. In large meat‐production animals, however, the traditional in vivo proliferation assay may be less realistic because it requires intra‐peritoneal (ip) injection of huge dosage of mutagenic nucleosides, ³H‐labeled thymidine or bromodeoxyuridine (BrdU), at each age‐time of sacrifice. We report in the present pilot study using rats that in vivo proliferation activity of satellite cells can be evaluated by an in vitro BrdU‐incorporation assay in early cultures. Briefly, satellite cells were prepared from upper hind‐limb and back muscles and maintained for 24 h with imposing by BrdU addition for the last 2 h, followed by the regular immunocytochemistry for determining BrdU‐incorporated cell percentage. This in vitro assay demonstrated a rapid decrease in proliferating satellite cell frequency to the adult level during about 3‐month period after birth, and yielded a high correlation to the measurements by the in vivo BrdU ip‐injection method during the postnatal period examined from day‐2 to month‐11. The in vitro proliferation assay may be further adaptable for large domestic animals by the combination with a muscle biopsy technique that enables age‐interval sampling from the same growing animals.     

Spontaneous vacuolar degeneration of the thyroid follicular epithelium in cynomolgus monkeys

Vacuolar degeneration of the thyroid follicular epithelium was observed in two untreated female cynomolgus monkeys assigned to control groups. In light microscopy, large vacuoles containing a homogenous substance occupied the basal region of the epithelium, and the nuclei had shifted toward the apical region. The vacuoles showed negative reactions to PAS and thyroglobulin. Electron microscopic observation revealed dilatation of the rough endoplasmic reticulum corresponding to the vacuoles. The plasma TSH, T3 and T4 levels determined for the samples kept frozen were within the normal ranges, suggesting that the thyroid function was kept intact.     

Long-term respiration in relation to growth and maintenance processes of the aboveground parts of a hinoki forest tree

Over a three-year period, CO(2) exchange rates were measured continuously on the aboveground parts of a 13-year-old hinoki (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) tree in the field, using an open gas-exchange system tracking ambient temperature. The relationship between daily aerial respiratory consumption and daily gross production, which was analyzed monthly, closely resembled McCree's equation. The value of the coefficient of growth respiration ranged between 0.0 in winter and 0.16 in summer and increased with increasing monthly mean temperature. A clockwise loop was observed for monthly change in the relationship between the coefficient of growth respiration and temperature. Maintenance respiration could be formulated as a power function of aboveground dry weight. The exponent of the equation ranged from 0.3 to 1.1. A value of 1.1 in May and June, when trees were growing most actively, indicated that maintenance respiration was directly proportional to aboveground weight. In March, April, July, and August, maintenance respiration was not proportional to aboveground weight, but it was closely proportional to surface area. The exponent value exhibited seasonal change with a clockwise loop in response to monthly average temperature. During the dormant season, respiration was used only for maintenance purposes, whereas during the growing season both growth and maintenance respiration occurred. Annual growth and maintenance respiration increased with increasing tree age. The average annual contribution to total respiratory consumption was 21% for growth respiration and 79% for maintenance respiration.     

Growth analysis on the competition–density effect in Chinese fir (Cunninghamia lanceolata) and Masson pine (Pinus massoniana) stands

The competition–density (C–D) effect for Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) and Masson pine (Pinus massoniana Lamb.) stands was analyzed using the reciprocal equation of the C–D effect, i.e. 1/w=Aρ+B, where w and ρ, respectively, represent mean stem volume and stand density. The C–D curve, which is given by the reciprocal equation on logarithmic coordinates, shifted upward with the progress of time. The coefficients A and B included in the reciprocal equation were calculated at each growth stage. The biological time τ defined as the integral of λ(t), the coefficient of growth in the general logistic growth curve, with respect to physical time t, tended to increase to an asymptote with increasing t. With increasing τ, the coefficient A, the reciprocal of which means the asymptote of yield (=wρ), increased abruptly up to a maximum value and then tended to decrease gradually to a constant level. On the other hand, the coefficient B, the reciprocal of which means the asymptote of mean stem volume, decreased exponentially and tended to close to zero with increasing τ. The λ(t) decreased with increasing stand age, whereas the final yield Y(t) defined as W(t) ρ, where W(t) is the asymptote of w in the general logistic growth curve, tended to increase gradually with increasing stand age.     

Self-thinning in prince Rupprecht's larch (Larix principis-rupprechtii Mayr) stands

Prince Rupprecht's larch (Larix principis-rupprechtii Mayr) stands growing at three different densities were investigated to determine characteristics of self-thinning. Tree density decreased with increasing stand age, and the higher density stand had higher mortality than the lower one. Mean stem volume increased with increasing stand age, and the higher density stand had higher relative growth rate of mean stem volume than the lower one. Mean stem volume (ν) increased with decreasing tree density (ρ), resulting in self-thinning line being expressed as ln ν=lnK-α ln ρ, whereK and α are coefficients. The slope of self-thinning line, —α, over the whole study period for all sites was similar with a mean value of —2.13. The ν-ρ trajectories before reaching the self-thinning slope of—3/2 could be described by Tadaki's model. The phase self-thinning line tended to decrease toward a slope of—3/2 with increasing stand age, which trends agreed with those of the published data of aPinus strobus stand andP. densiflora stands.     

Stand structure,phenology and litterfall dynamics of a subtropical mangrove <Emphasis Type="Italic">Bruguiera gymnorrhiza</Emphasis>

We evaluated the phenology and litterfall dynamics of the mangrove Bruguiera gymnorrhiza (L.) Lamk along the Okukubi River, Okinawa Island, Japan. Over 3 years, this species showed the highest litterfall of leaves and stipules in summer and the lowest litterfall in winter. From Kendall’s coefficient of concordance, the monthly changes in leaf, stipule, and branch were strongly and significantly concordant among years. Leaf and stipule litterfall could be governed by monthly maximum wind speed, monthly day length, and monthly mean air temperature. Branch litterfall depended on monthly maximum wind speed and monthly rainfall, and increased exponentially with increasing monthly maximum wind speed. Mean total litterfall was 11.8 Mg ha^?1 yr^?1, with the largest component being leaf litterfall (65.8 %). Annual leaf litterfall per plot was almost constant regardless of the tree density of the plot. Mean leaf longevity was 18 months. Flower and mature propagule litterfall might be influenced by monthly mean air temperature, monthly day length and monthly mean air temperature. The average development periods from flower buds to flowers and flower buds to mature propagules were 1 and 8 months, respectively. Except for leaf and branch, all vegetative and reproductive organ litterfall had clear annual cycles. B. gymnorrhiza showed a positive correlation between leaf production and reproductive organ production.     

Discovering the dominance of the non-native European eel in the upper reaches of the Tone River system,Japan

To investigate the presence of non-native anguillid eels in Japanese waters, 141 eels were collected from seven sampling sites throughout the Tone River system. Genetic species identification showed an extraordinary dominance of the non-native European eel Anguilla anguilla in the uppermost site of the study area. Estimation of age from otoliths suggested that the European eels were introduced into the river in the 1990s, corresponding to previous reports from other Japanese water systems. Comparison of the von Bertalanffy growth curve parameters indicated that the European eels caught in the Tone River system appeared to have a similar or even higher growth rate than the same species in the original habitats in Europe. The long-term inhabitation and the normal development of European eels in Japanese waters suggest that regulations prohibiting the release of non-native eels and safeguards against accidental escape from culture ponds must be strictly maintained in order to ensure the conservation of the native Japanese eel.     

Spontaneous polyploidization in critically endangered <Emphasis Type="Italic">Acipenser mikadoi</Emphasis>

We investigated the source of spontaneous polyploidization in the critically endangered Acipenser mikadoi. Fourteen sib progeny of A. mikadoi and 11 hybrids between an A. mikadoi female and a Huso dauricus male, all showing atypically high ploidy, were analysed. Parent assignment based on five highly polymorphic microsatellite markers confirmed spontaneous duplication of maternal chromosome sets via retention of the second polar body to be the source of spontaneous polyploidization. To our knowledge, this provides the first evidence of the maternal origin of spontaneous polyploidization in A. mikadoi. Factorial correspondence analysis of the multilocus microsatellite genotypes placed the parent fish of spontaneous polyploids in clearly delineated clusters of A. mikadoi and H. dauricus, and parent fish had mitochondrial control region haplotypes corresponding to their presumed species. Thus, parent fish were confirmed to be of pure genetic origin, and hybridization did not promote the observed spontaneous polyploidization.