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.     

Enhancement of hepatocellular proliferative activity of kojic acid in mice by a simultaneous administration of ascorbic acid

To examine the tumor modification activity of kojic acid (KA) by sodium ascorbic acid (AA), 5-week-old male ICR mice were administered intraperitoneally with N-diethylnitrosamine (DEN) as an initiation treatment. Two weeks after the initiation treatment, animals were fed basal diet containing 0 (Group 1: DEN alone) or 3% KA (Group 3: DEN+KA), drinking water containing 5,000 ppm AA (Group 2: DEN+AA) or 3% KA and 5,000 ppm AA (Group 4: DEN+KA+AA) for 6 weeks. One week after the administration of KA and/or AA, all mice were subjected to two-thirds partial hepatectomy. At the end of the experimental period, all surviving mice were sacrificed and removed the liver. The liver weights of the Groups 3 and 4 were significantly increased, and the number of proliferating cell nuclear antigen positive hepatocytes and the gene expressions of Ccnc, Ccnd1, Ercc and Cyp7a1 were significantly increased in the Group 4, as compared to the Group 1. These results of the present study suggest that AA enhances the hepatocellular proliferative activity of KA in mice.     

干旱区人工绿洲间作农田蒸散研究

在黑河流域中游的张掖绿洲区建立了大田环境下的春小麦和夏玉米间作农田能水平衡研究观测点,以气象观测资料为基础,采用波文比能量平衡法(BREB)和参考作物蒸散量—作物系数法(ET₀-K_c)对作物的蒸散进行了计算。结果表明：在一个完整的生长期内,利用波文比能量平衡法得到的间作作物蒸散量为688 mm,日均3.4 mm/d,用参考作物蒸散量—作物系数法得到的作物蒸散量为666 mm,日均3.3 mm/d,两种计算方法得到的蒸散量总值差别小。同期,水文平衡法计算结果为733 mm。利用波文比能量平衡法所得结果的分析表明,试验地在不同生长阶段,ET变化剧烈,生长初期、中期、末期分别为1.19、4.41和2.58 mm/d,其蒸散量分别占全年蒸散总量的7.79%、78.73%和13.48%。ET月变化显示,3月维持在一个较低水平;4月和5月剧烈增加;6月达到最大;此后的7月和8月降低,但仍维持在一个高水平;9月,随着作物进入生长末期,蒸散急剧减小。对ET日内变化分析可知,作物蒸散开始于早晨7∶00～8∶00,在14∶00左右达到最大,19∶00～20∶00趋于0 mm/d。不同生长阶段蒸散强度差异明显。     

The polar neutral and basic taxoids isolated from needles and twigs of Taxus cuspidata and their biological activity

Twelve basic taxoids and 22 neutral taxoids were isolated from basic and polar neutral fractions of the extracts of needles and twigs of Taxus cuspidata. Among them, taxine NA-13, 3,11-cyclotaxinine NN-1, taxinine NN-6, 11(15→1)abeo-taxinine NN-1, taxine NA-8, and taxine NA-4 were isolated first from natural sources by us. The cytotoxic activity of isolated compounds was evaluated against three human cell lines: normal human fibroblast cells (WI-38), malignant tumor cells induced from WI-38 (VA-13), and human liver tumor cells (HepG2). 7-Epitaxol, 7-epicephalomannine, taxinine NN-6, taxine NA-2, taxuspine H, and taxagifine were active toward VA-13 cells and 7-epitaxol, 7-epicephalomannine, taxinine NN-1, 9,10-deacetyltaxinine, and taxagifine were active toward HepG2 cells. The multidrug-resistant (MDR) cancer reversal activity of isolated compounds was evaluated on the basis of the amount of vincristine (VCR) accumulated in MDR human ovarian cancer 2780 AD. Taxine NA-8, taxine NA-2, 5-cinnamoyl-10-acetyltaxicin II, and taxinine NN-1 indicated stronger MDR cancer reversal activity than verapamil. The result of primary screening based on 39 human cancer cell lines suggests that taxinine NN-1 belongs to a new mechanistic class and is a new anticancer agents. 7-Epicephalomannine was found to be an effective anticancer agent with tubulin as its molecular target, which is the same as paclitaxel. Part of this work was presented at the 74th Annual Meeting of the Chemical Society of Japan, Tanabe, March 1998, and the 41st Symposium on the Chemistry of Terpenes, Essential Oils, and Aromatics, Gifu, December 1998. The results of this work were also presented as patent documents at PCT Int: Appl WO 2001007040 A1, February 2001; Appl WO 2000-Jp 5036, 27 July 2000. Priority: JP 1999-214273, 28 July 1999; JP 1999-224652, 6 August 1999; JP 2000-76404, 14 March 2000     

Natural growth of juveniles of the sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>: studying juveniles in the intertidal habitat in Hirao Bay,eastern Yamaguchi Prefecture,Japan

In order to determine the population dynamics of juvenile Apostichopus japonicus, successive surveys were conducted in the stony intertidal zone of Hirao Bay, Seto Inland Sea. The juveniles of the green and black types of this species grew well from January to June. Before this high growing season, zero year old juveniles were too small to be readily detected by visual observations during field surveys. During the high water temperature season from August to November, juveniles over 1 year of age estivated underneath rocks. They awakened from their estivation in November, and it took another month before they could eat and discharge. The juveniles mainly migrated from the intertidal zone before they reached 3 years of age. In the present work, three patterns of juvenile growth were found. It is suggested that these differences in juvenile growth are mainly caused by differences in their start times for growth initiation when they are zero years old.     

Swimming angle and target strength of larval Japanese anchovy (<Emphasis Type="Italic">Engraulis japonicus</Emphasis>)

The swimming angle of larval Japanese anchovy (Engraulis japonicus) was measured in a tank, and target strength (TS) was calculated using a theoretical scattering model. The mean swimming angle was 12.8° (SD ±22.1). Increased speeds of flow led to increased mean swimming angles. The mean swimming angle at flow of 5 cm s⁻¹ was higher than at other speeds. TS values were estimated using a distorted-wave Born approximation model for two cases. Average values were 1–3 cm s⁻¹ (11.5° ± 22.1) and 5 cm s⁻¹ (16.6° ± 21.7) for cases 1 and 2, respectively. For case 1, TS ranged from −92.0 to −74.7 dB with a mean of −79.4 dB at 120 kHz. For case 2, TS ranged from −92.2 to −75.2 dB with a mean of −79.9 dB. The mean TS in case 2 was lower than that in case 1, with the maximum difference being 1.0 dB at 120 kHz (standard length 22.0 mm). However, there were no significant differences between the regression lines of cases 1 and 2. Thus, changes in flow speed altered the swimming angle of larval Japanese anchovy, but had little influence on TS.     

A novel mutant with triple the dietary fiber content in white rice

Dietary fiber has high functional value in relation to gut flora. We searched for a high-lysine mutant of the most popular rice cultivar in Japan, ‘Koshihikari’, as a route to a higher dietary fiber content like a success case in new barley cultivar, ‘Beau-fiber’. We found several promising high-lysine mutants with high dietary fiber content. One of these, ‘WFE5’, has three times the dietary fiber content in white rice. Two rounds of backcrossing to Koshihikari produced a near-isogenic line with a high fiber content. The line’s agronomic traits were close to those of Koshihikari except for yield and eating quality. As these two traits are critical, we discuss how to improve them.     

Pathology of female mice experimentally infected with an in vitro cultured strain of Trypanosoma equiperdum

Dourine, caused by infection with Trypanosoma equiperdum, is one of the trypanosomiasis in equids. The clinical course of dourine is long-term, ranging from 1–2 months to several years. Since the pathogenesis of dourine has not yet been elucidated, experimental studies using mouse infection models are needed. Although mice are not susceptible to most T. equiperdum strains, some strains can infect mice. Even in such strains, infected mice develop rapidly transient parasitemia and die within 2–8 days. Therefore, mice experimentally infected with these T. equiperdum strains are not suitable for mouse infection models to analysis the pathogenesis of dourine. A sequential method of isolating parasites from dourine-affected horses and adapting them to in vitro cultures using soft agarose media was recently developed. Various T. equiperdum strains adapted to in vitro conditions have been established using this technique. We used one of these strains, the T. equiperdum IVM-t2 strain. In the present study, T. equiperdum IVM-t2 strain inoculated mice developed periodic parasitemia during the experimental period of 60 days. Histopathologically, vaginitis and dermatitis were observed. These findings were comparable to those of dourine-affected horses. Therefore, mice infected with T. equiperdum IVM-t2 strain may be a valuable tool for pathological, immunological, and parasitological in vivo research, and will contribute to investigations on the mechanisms underlying the disease process and the host-parasite relationship.     

Time dependence of Poisson’s effect in wood I: the lateral strain behavior

To understand the viscoelasticity of wood three dimensionally, a longitudinal tensile creep test for 12 species was conducted to examine the changes with time in the lateral strain and the viscoelastic, i.e., apparent Poisson’s ratio. The changes in the lateral strain (ɛ _T and ɛ _R) were similar to those in the longitudinal strain (ɛ _L). That is, during creep, the absolute value of lateral strain continued to increase with the gradual reduction in the increase rate; immediately after the removal of the load, it recovered abruptly; then, it recovered slowly and finally reached a certain value. The rate of increase in the longitudinal strain during creep was smaller than that in the absolute value of lateral strains. The apparent Poisson’s ratio became large during creep because the lateral strain increased more than the longitudinal strain. The analysis of lateral strain by decomposition into three components, that is, instantaneous strain, delayed elastic strain, and permanent strain, has revealed that the lateral permanent strain in the transverse direction contributes most to the increase in the apparent Poisson’s ratio during creep.