Bam HI DNA甲基化酶酶学性质的研究

以Lineweave-Burk plot双倒数作图法测得该酶对底物S-腺苷酰甲硫氨酸(SAM)的K_m=7.69×10~(-6)mol/L,在1mmol/LS-腺苷酰高半胱氨酸(SAH)存在下,Ki=7.33×10~(-4)mol/L,两条直线相交于纵轴,证明SAH是该酶的竞争性抑制剂。该酶最适pH为7.8,对热不稳定。同时还测定了该酶对不同DNA底物的专一性及盐浓度、代谢相关物’两价阳离子、某些酸根等对该酶调节性质的影响。以碘代乙酰胺修饰该酶的SH基’及用二硫苏糖醇(DTT)和巯基乙醇(MSH)保护该酶SH基所作的实验表明SH基是该酶活性中心所必需的,用高效液相色谱(HPLC)法证明该酶所甲基化的碱基为刘氏小球菌(M·L、DNA)分子中的胞嘧啶,且求得甲基化30min后所得甲基化水平为2.39％。同时也证明当用该酶将λDNA甲基化后,可使BamHI限制性核酸内切酶对甲基化后的λDNA丧失切割作用。     

Participation of breast and leg muscles in shivering thermogenesis in young turkeys and guinea fowl

Turkey (Meleagris gallopavo) and guinea fowl (Numida meleagris) chicks (0–27 days posthatch) were exposed to decreasing or increasing ambient temperatures. Root mean square electromyographic activity of musculus pectoralis (m. pect.) and musculus iliotibialis (m. iliot.) was recorded simultaneously with O₂ consumption and CO₂ production. From both muscles, relative mass, water fraction and fibre type were determined. M. iliot. participated in shivering from hatching onwards. The relationship between its root mean square electromyographic activity and ambient temperature resembled that of metabolic rate and ambient temperature, and the shivering threshold temperature was indistinguishable from the lower critical temperature. This suggests that the leg muscles are major contributors to shivering thermogenesis. M. pect. participated in shivering only at days 6–20 in turkeys and at days 6–10 in guinea fowl. Both water fraction and histological analysis indicated that m. pect. was less developed than m. iliot. at hatching. We hypothesize that a minimal level of maturity is required before a muscle can participate in shivering, which is probably represented by a water fraction of about 0.85. Both species recruited the aerobic leg muscles first; the anaerobic breast muscle was recruited only when the rate of mass-specific heat loss was high. Accepted: 20 March 1997     

昆明山海棠根部水抽提物对体外微管蛋白聚合的影响

本研究利用猪脑中分离纯化的微管蛋白聚合和解聚反应,分析了非整倍体诱发剂昆明山海棠根部水抽提物(THH)对微管蛋白聚合状态的影响,从该角度探讨了THH诱发哺乳动物非整倍体的机制。秋水仙素(COL)为本研究的阳性对照物。结果发现THH能显著抑制体外微管蛋白的聚合,该抑制效应呈明显的剂量——效应关系。研究结果与我们以往关于THH为非整倍体诱发剂的实验证据相吻合并进一步提示THH可以抑制微管蛋白聚合作为诱发非整倍体的途径之一。     

Alternative risk-taking styles: The case of time-budgeting strategies of terrestrial gastropods

The terrestrial slugs Arion ater, Limax maximus and Ariolimax columbianus have similar morphological designs but differ remarkably in their life history tactics and behavioural time budgets. The adaptive value of particular risk-taking styles was investigated using a comprehensive computer simulation model. The model allowed each species' success (growth rates, food acquisition) and costs (distance travelled, hydration deficits, injury from aggressive encounters) to be evaluated in various types of weather (benign, harsh and surprise). This was interpreted in terms of the species' life history design. In addition, each species was simulated with the behavioural strategy of the other two species substituted for its natural programming. The simulation experiments demonstrated how variations in a few simple rules could lead to divergent cost-benefit consequences. The model also illustrated that coarsely-tuned sensitivity to the environment may be better than finely-tuned responses, depending on the animal's ability to respond quickly and the degree of risk. The simulations suggested that competition has shaped the time-budgeting tactics of A. ater and L. maximus. Each species actually performed better in harsh weather, using the behavioural program of the other. L. maximus is an aggressive species with a narrow, nocturnal activity period. The concentrated activity of L. maximus may allow it to displace competitors more effectively, whereas the broader time span of A. ater's activity may be necessary to avoid L. maximus.     

Statistical optimization of enzymatic degradation process for oil palm empty fruit bunch (OPEFB) in rotary drum bioreactor using crude cellulase produced from Aspergillus niger EFB1

Oil palm empty fruit bunch (OPEFB) was pretreated with 2% (v/v) HNO₃ and degraded by Aspergillus niger EFB1 crude cellulase. Through 2 Level Factorial Design (2LFD), it was found that OPEFB concentration, temperature, incubation time, concentration of Tween 80 and agitation speed have significant effect in reducing sugar production. A standard Response Surface Methodology (RSM) design known as Central Composite Design (CCD) was used to optimize the enzymatic degradation condition of OPEFB in rotary drum bioreactor. Reducing sugar level of 1.183 g/L was obtained with the following optimized degradation conditions: 1.95% (w/v) OPEFB, 0.5% (v/v) Tween 80, 55 °C, 87.5 rpm in the incubation period of 3 days and 16 h. The optimal degradation condition improved reducing sugar production by 1.07 fold compared to that before optimization in shake flasks culture. The optimization strategy of enzymatic degradation of OPEFB inside rotary drum bioreactor led to increase in glucose, xylose, arabinose, galactose and mannose production by 3, 2.5, 1.64, 19.37 and 22.52 fold, respectively. The improvement in reducing sugar and polyoses production were comparable with the reduction in OPEFB cellulose and hemicellulose content by 89.32% and 48.17% respectively after enzymatic degradation in optimized condition.     

多色荧光原位杂交对昆明山海棠和丙烯酰胺诱发微核染色体组成的研究

采用着丝粒和端粒DNA探针多色荧光原位杂交,分析了昆明山海棠和丙烯酰胺诱导的 NIH3T3细胞微核的染色体组成。结果表明,昆明山海棠和丙烯酰胺诱导的由整条染色体组成的微核分别可达70.7％和65.9％,提示昆明山海棠和丙烯酰胺均有较强的非整倍体毒性。     

Natural mortality at different population densities of a pine shoot moth,Evetria cristata

A field population of Evetria cristata was studied in 10 plots in 1962 and in 6 plots in 1963. These plots were divided into 2 or 3 groups of different population levels of the shoot moth in respective years. The survival of the insect was then analysed in these different groups of plots. The survival rate of E. cristata from eggs to adults in the first generation was found always higher in the group with low population density, which indicates the existence of some factors that affect the population more severely when the insect is more abundant. Lissonota evetriae and Pediobius sp. seemed to have killed more proportion of the hosts where the shoot moth density was high. However, the total effect of the all natural enemies was not always great in the plots with high density of the moth. The survival of the second generation of the moth in 1963 was observed to be much higher at any population level than in the other generations.     

昆明山海棠对中国仓鼠V79细胞二酰基甘油含量的影响

本研究以昆明山海棠根部水抽提物(Tripterygium Hypoglaucum(Level)Hutch,THH)处理中国仓鼠V79细胞,通过检测V79细胞C-M细胞频率以及二酰基甘油(1,2-diacylgcerol,DAG)的含量测定,分析了THH诱发非整倍体与细胞醇磷酯信号通路的关系.结果指出THH能在1mg/ml、2mg/ml两个剂量上使V79细胞的DAG含量显著升高(P＜0.001),并明显的提高C-M细胞频率(P＜0.05),提示肌醇酯信号通路是介导THH诱发非整倍体的途径之一.     

Spinal loads and trunk muscles forces during level walking – A combined in vivo and in silico study on six subjects

During level walking, lumbar spine is subjected to cyclic movements and intricate loading of the spinal discs and trunk musculature. This study aimed to estimate the spinal loads (T12–S1) and trunk muscles forces during a complete gait cycle.Six men, 24–33 years walk barefoot at self-selected speed (4–5 km/h). 3D kinematics and ground reaction forces were recorded using a motion capturing system and two force plates, implemented in an inverse dynamic musculoskeletal model to predict the spinal loads and trunk muscles forces. Additionally, the sensitivity of the intra-abdominal pressure and lumbar segment rotational stiffness was investigated.Peak spinal loads and trunk muscle forces were between the gait instances of heel strike and toe off. In L4–L5 segment, sensitivity analysis showed that average peak compressive, antero-posterior and medio-lateral shear forces were 130–179%, 2–15% and 1–6%, with max standard deviation (±STD) of 40%, 6% and 3% of the body weight. Average peak global muscles forces were 24–55% (longissimus thoracis), 11–23% (iliocostalis thoracis), 12–16% (external oblique), 17–25% (internal oblique) and 0–8% (rectus abdominus) of body weight whereas, the average peak local muscles forces were 11–19% (longissimus lumborum), 14–31% (iliocostalis lumborum) and 12–17% (multifidus). Maximum ± STD of the global and local muscles forces were 13% and 8% of the body weight.Large inter-individual differences were found in peak compressive and trunk muscles forces whereas the sensitivity analysis also showed a substantial variation.