Effect of Protease Inhibitors on Early Events of Apoptosis

Proteolysis is an early event of apoptosis which appears to be associated with activation of the endonuclease which is responsible for internucleosomal DNA cleavage. The present study was designed to reveal the possible role of proteolysis in other early events, such as chromatin condensation, nuclear breakdown, and destabilization ofin situDNA double-stranded structure. Apoptosis of human leukemic HL-60 cells and rat thymocytes was induced by different agents, including DNA topoisomerase inhibitors, an RNA antimetabolite, and the glucocorticosteroid, prednisolone. DNA degradation was evaluated by pulsed field and conventional gel electrophoresis and by the presence ofin situDNA strand breaks. DNA stability was estimated by the measure of its sensitivityin situto denaturation. Chromatin condensation, nuclear breakdown, and other morphological changes were monitored by interference contrast and UV microscopy following cell staining with the DNA-specific fluorochrome 4′,6-diamidino-2-phenylindole. Several irreversible or reversible serine protease inhibitors prevented internucleosomal DNA degradation, nuclear breakdown, and destabilization of DNA double-stranded structure. The effective inhibitors, however, did not prevent the onset of chromatin condensation, nor the loss of the fine structural framework, nor the initial step of DNA cleavage generating DNA fragments of ≥50 kb in size. The data indicate that in both cell systems the activity of proteases sensitive to the inhibitors tested is needed for internucleosomal DNA cleavage to occur. The data also suggest that these proteases may be involved in dissolution of the nuclear envelope. Because nuclear matrix proteins and histones stabilize DNAin situ,and the decrease in DNA stability which occurs during apoptosis is precluded by the inhibitors, it is likely that serine proteases may degrade DNA stabilizing proteins. The activity of these proteases, however, appears needed neither for DNA cleavage to ≥50-kb fragments nor for the onset of chromatin condensation which is associated with dissolution of the structural framework of the nucleus.     

A Bacillus subtilis malate dehydrogenase gene.

A Bacillus subtilis gene for malate dehydrogenase (citH) was found downstream of genes for citrate synthase and isocitrate dehydrogenase. Disruption of citH caused partial auxotrophy for aspartate and a requirement for aspartate during sporulation. In the absence of aspartate, citH mutant cells were blocked at a late stage of spore formation.     

Comparison of fish assemblages among an artificial reef, a natural reef and a sandy-mud bottom site on the shelf off Iwate, northern Japan

Synopsis Fish assemblages at an artificial reef site, a natural reef site and a sandy-mud bottom site, on the shelf (depth 130 m) off Iwate Prefecture, northern Japan, were surveyed by using a bottom trammel net from May 1987 to March 1993. A total of 12 173 fishes of 48 species were recorded. Physiculus maximowiczi was dominant and comprised 69% of the total numerical abundance. Total fish number was lowest in March at all the 3 sites when P. maximowiczi migrated to deeper and warmer waters. Assemblage equitability and species diversity also varied seasonally in accordance with the abundance fluctuation of P. maximowiczi. P. maximowiczi, Alcichthys alcicornis and Hexagrammos otakii were more abundant at the artificial reef and natural reef sites, while Dexistes rikuzenius and Hemitripterus villosus were more abundant at the sandy-mud bottom site; total fish abundance was largest at the artificial reef site mainly due to the large number of P. maximowiczi. Species richness was similar among sites, but equitability, and consequently species diversity, was lowest at the artificial reef site. The main effect of the artificial reef seemed the attraction of P. maximowiczi from nearby bottoms, especially from natural rocky reefs; its large abundance determined the structure of the artificial reef fish community.     

Charactererization of novel non-toxic Bacillus thuringiensis isoloates from Korea

J.Y. ROH, H.W. PARK, B.R. JIN, H.S. KIM, Y.M. YU AND S.K. KANG. 1996. Four Bacillus thuringiensis isolates from soil samples produced parasporal inclusions which were non-toxic to insects. The isolates were named B. thuringiensis NTB-1, NTB-2, NTB-3 and NTB-4. The parasporal inclusions were shown to be ovoid by phase contrast and scanning electron microscopy. The serotypes of the four isolates were determined by agglutination using 33 antisera; NTB-1 and NTB-4 seemed to be subsp. isruelensis ,and NTB-2 seemed to be subsp. pondzcheriensis . NTB-3 did not react with the 33 antisera. However, comparison of parasporal protein and plasmid DNA patterns of the four isolates with those of 15 known non-toxic B. thuringiensis strains demonstrated that the four isolates are novel.     

Role of acetylation on metal induced precipitation of alginates

Acetylation dramatically effects both the solution properties and the metal induced precipitation of alginates. The presence of acetyl groups on both bacterial and seaweed alginate polymers marginally increased the weight average molecular weight (M_w) of each polymer by 7% and 11%, respectively. Acetylated bacterial alginate showed a significant increase in solution viscosity compared to its deacetylated counterpart. However microbial acetylation of seaweed alginate did not change its solution viscosity. Acetylation altered the calcium induced precipitation of both alginates. The presence of acetyl groups decreased the ability of each polymer to bind with calcium but increased their ability to bind with ferric Ion (Fe³⁺). By controlling the degree of acetylation on the alginate chains, it was possible to modify solution viscosity and cation induced precipitation of these polymers.     

Cellobiose-oxidizing enzyme from a newly isolated cellulolytic bacterium Cytophaga sp. LX-7

Summary The cellobiose oxidizing enzyme of the newly isolated cellulolytic bacterium Cytophaga sp. LX-7 was produced extracellularly when grown on cellulose or other saccharides, which was previously noted only in fungi. The enzyme could use not only cellobiose, maltose, glucose and other saccharides but also cellulose as substrates, and use dichlorophenol indophenol and oxygen as electron acceptors.     

Comparative study of nickel toxicity to growth and photosynthesis in nickel-resistant and -sensitive strains of Scenedesmus acutus f. alternans (Chlorophyceae)

Nickel (Ni) toxicity to growth and photosynthesis was studied in four strains of Scenedesmus acutus f. alternans. Effects of Ni dosage and duration of exposure on growth and photosynthesis were strain specific. Large differences in responses of both growth and photosynthesis to Ni were detected between three resistant strains (B4, Cu-Tol, and Ni-Tol) and one sensitive strain (UTEX 72). Growth of UTEX 72 was 18 times more sensitive to Ni than those of the three resistant strains. The order of Ni dosages (fmol Ni/pg cell dry weight) causing 50% inhibition (D₁₅₀) of growth rates in the four strains was Ni-Tol (10.5) > B4 (8.19) > Cu-Tol (4.60) > UTEX 72 (0.25). The effect of Ni dosage on photosynthetic rate as percentage of control corresponded to a saturation curve and was a strong function of duration of exposure. The DI₅₀s of photosynthetic rates were 3.5 times lower in UTEX 72 than in the three resistant strains, and in all four strains they decreased sharply with the increase in duration of exposure. The order of the four strains in DI₅₀s of photosynthetic rate was B4 (58.2) > Cu-Tol (38.0) > Ni-Tol (28.9) > UTEX 72 (8.24) for 6-h exposure and Ni-Tol (2.88) > Cu-Tol (1.30) > B4 (1.01) > UTEX 72 (0.15) for 24-h exposure. The DI₅₀s of photosynthetic rate for 6-h exposure were higher than those of growth rate in all four strains, and for 24-h exposure they were lower, except in UTEX 72. Thus, the relative Ni sensitivity of growth and photosynthesis of the four strains depends on the duration of exposure. The results of factorial analysis of variance suggested that Ni toxicity to photosynthesis is a consequence of a strong interaction among strain, Ni dosage, and duration of exposure.     

生物复苏——大绝灭后生物演化历史的第一幕

生命史是一部生物界短期,快速剧变与长期,慢速稳定相互交替的历史。大绝灭（即集群绝灭）事件反映了全球环境的大突变,点断了地质历史中的生命记录及其发展历程,预示着生物界的演化出现了最有意义的飞跃,近年来尝试研究大绝灭后全球生物界的残存－复苏及其基本型式,并探索复苏的控制因素,标志着地质科学中一个重心的转移（即从大绝灭转向其后的生物残存与复苏的研究）。生物复苏揭示了大绝灭后生物演化历史的第一幕,其研究的