正常及白血病小鼠白细胞染色质和DNA的酶切分析

 本文应用的核酸酶为DNaseⅡ、微球菌核酸酶与限制性内切核酸酶BstNI、EcoRⅡ、HpaⅡ和MspⅠ,将它们作用于正常小鼠615和可移植性白血病小鼠L7712脾脏白细胞染包质及其DNA,根据酶切电泳谱及水解动力学分析表明:1.白血病小鼠染色质相对正常小鼠染色质易被DNaseⅡ微球菌核酸酶水解;2.白血病小鼠染色质比正常小鼠者易被MspⅠ水解,但其DNA的MspⅠ酶切电泳谱无明显差别;3.白血病小鼠染色质及其DNA较正常小鼠染色质及其DNA易被EcoRⅡ水解。这些观察说明,白血病小鼠脾脏白细胞染色质有较活跃的构象状态;其染色质DNA的CCATGC区段内有较低的甲基化程度。     

Studies on the biological activity of an insulin-stimulating peptide from a tryptic digest of bovine serum albumin

Summary A two-chain polypeptide, which corresponds to amino acid residues 115–143 and 144–184(185) of bovine serum albumin, connected to each other by a disulfide bridge, potentiated the effects of insulin on glucose transport and glucose metabolism in isolated rat adipocytes. Although the peptide alone had little activity, it shifted the concentration-response curves of insulin-stimulated D-[I-¹⁴C]glucose oxidation, 2-deoxyglucose transport, and lipid synthesis from D-[U-¹⁴C]glucose to lower insulin concentrations. It also increased the maximal responses of these parameters to insulin. However, it did not affect insulin binding to adipocytes. The peptide protected insulin considerably from degradation, but this effect alone cannot account for its effect in increasing the maximal responses to the hormone, and even when degradation of a submaximal concentration of insulin was suppressed by bacitracin, the peptide still had an enhancing effect. These results suggest not only that the peptide influences a step distal to receptor-mediated insulin binding but also that inhibition of insulin degradation alone cannot explain its total effect.The peptide lost its insulin-stimulating activity completely when it was further digested with V8 or lysinespecific endopeptidase, or when it was reduced and then carboxamidomethylated or oxidized with performic acid. Similar active tryptic fragments were obtained from human and rat albumins.Insulin-stimulating peptides should be useful in studies on the mechanisms of insulin action including both the sensitivities and responsiveness of target cells to the hormone.Abbreviations ISP insulin-stimulating peptide - HEPES N-(2-hydroxyethyl)piperazine-N-2-ethanesulfonic acid - HPLC high-performance liquid chromatography - SDS sodium dodecyl sulfate     

Biochemical and physiological studies of certain ticks (Ixodoidea): specific gravity of hemolymph and coxal and gut fluids of Argas (Persicargas) persicus and Argas (Persicargas) arboreus (Argasidae)

Specific gravity (sp. gr.) of cell-free hemolymph and gut and coxal fluids was determined at different states of the gonotrophic cycle (unfed + 15 days, engorgement day before and after coxal fluid emission, engorgement + 1 day, oviposition day, and oviposition completion + 1 day) of female Argas (Persicargas) persicus and A. (P.) aboreus (Argasidae). The patterns of hemolymph and gut fluid sp. gr. change differed from each other during the gonotrophic cycle, but both patterns were similar in the 2 Argas species. Hemolymph sp. gr. decreased to a minimum one day after feeding (1.0085 and 1.0081 for persicus and arboreus, respectively), and increased through oviposition to a maximum on oviposition + 1 day (1.0187 and 1.0221). Minimum gut fluid sp. gr. occurred on engorgement day before coxal fluid emission (1.0565 and 1.0697). Afterward, gut fluid sp. gr. increased to a maximum on engorgement day + 1 for persicus (1.1089) and on oviposition day for arboreus (1.0973), and then decreased during oviposition in both species. In each tested state of each species, the sp. gr. was consistently higher in gut fluid than in hemolymph. In each species, coxal fluid and hemolymph sp. gr. were the same on engorgement day.     

Distribution,properties, and functions of midgut carboxypeptidases and dipeptidases from Musca domestica larvae

Dipeptidase and carboxypeptidase A activities were determined in cells and luminal contents of the fore-, mid-, and hind-midgut of Musca domestica larvae. Dipeptidase activity was found mainly in hind-midgut cells, whereas carboxy-peptidase activity was recovered in major amounts in both cells and in luminal contents of hind-midguts. The subcellular distribution of dipeptidase and part of the carboxypeptidase A activities is similar to that of a plasma membrane enzyme marker (aminopeptidase), suggesting that these activities are bound to the microvillar membranes. Soluble carboxypeptidase A seems to occur both bound to secretory vesicles and trapped in the cell glycocalyx. Based on density-gradient ultracentrifugation and thermal inactivation, there seems to be only one molecular species of each of the following enzymes (soluble in water or solubilized in Triton X-100): membrane-bound dipeptidase (pH optimum 8.0; Km 3.7 mM GlyLeu, M_r 111,000), soluble carboxypeptidase (pH optimum 8.0; Km 1.22 mM N-carbobenzoxy-glycyl-L-phenylalanine (ZGlyPhe), M_r45,000) and membrane-bound carboxypeptidase (pH optimum 7.5, Km 2.3 mM ZGlyPhe, M_r58,000). The results suggest that protein digestion is accomplished sequentially by luminal trypsin and luminal carboxypeptidase, by membrane-bound carboxypeptidase and aminopeptidase, and finally by membrane-bound dipeptidase.     

Approximate digestibility of fibre for a graminivorous caterpillar

Final instar Persectania ewingii (Westwood) (Lepidoptera: Noctuidae) were fed seedling Triticum aestivum L. for 2 days to determine the approximate digestibility of the cell wall and cell content fractions. Cell wall content was estimated using a micro-analytical neutral detergent fibre technique. Approximate digestibilites of neutral detergent fibre, neutral detergent solubles and dry matter were calculated for individuals and pooled samples. P. ewingii larvae digested a small but significant proportion of the fibre ingested (13–21%), higher than that previously reported for herbivorous insects. The micro-analytical and previously used macro-analytical techniques produced similar estimates of digestibility although both techniques have inherent shortcomings, the latter requiring the pooling of samplex and the former limiting the number of replicates during chemical analysis. Differences in the amount of larval frass collected during the feeding trial (corrected for consumption) explained much of the variation in digestibility values, while there were no effects of larval mass, overall consumption and total frass produced on digestibility estimates. These results confirm that plant cell contents are the major source of nutrients to larval Lepidoptera although there is some chemical disruption of the plant cell wall.     

部分酶解酵母高效电击转化研究

以酵母质粒YCp50为外源DNA,电击转化部分酶解酵母宿主菌AB1380,转化效率稳定在10~6转化子/μg质粒DNA左右,比不酶解酵母或酵母原生质球作受体的电击转化效率高一个数量级以上,也比PEG介导的酵母原生质球转化高3～5倍,而且适合于大片段DNA如水稻YAC分子的转化。达最佳转化时的有关技术参数为:新接菌种通气培养至细胞密度1×10~8～1.5×10~8个/ml;转化时细胞密度控制在1×10~9～1.5×10~9个/ml;每毫升酶解缓冲液加15u溶菌酶(lyticase),30℃下处理酵母5min进行部分酶解;电击时,电场设置在6.25kV/cm、电容25μF,电击后直接铺板。     

Anaerobic acidogenesis of primary sludge: the role of solids retention time

This research investigates the effect of solids retention time (SRT) on the acid-phase anaerobic digestion of primary sludge. A series of experiments were conducted using two continuous-flow 3-L units with the following configuration: a completely mixed reactor (CMR) with clarifier and solids recycle and an upflow anaerobic sludge blanket (UASB) reactor. Results show that C(2) to C(5) volatile fatty acids (VFA) were the predominant compounds formed. At a constant hydraulic retention time (HRT) of 12 h, variation in SRT from 10 to 20 days resulted in a slight increase in VFA production in both systems, but at a shorter SRT (5 days) a drastic drop in acid production was observed. In addition, the percent distribution of VFA was to some extent affected by the change in SRT. On the other hand, organic matter degradation [measured by the chemical oxygen demand (COD) specific solubilization rate or the percent volatile suspended solids (VSS) reduction] appeared to be independent of SRT, at least in the range investigated. The percent soluble COD in the form of VFA, however, increased steadily with increasing SRT, approaching the 90% level at 20 days. The remaining soluble COD in the effluent from these systems may be mainly attributed to metabolic intermediates and unused soluble substrate. (c) 1994 John Wiley & Sons, Inc.     

Avian daily foraging patterns: Effects of digestive constraints and variability

Summary Birds show a typical daily pattern of heavy morning and secondary afternoon feeding. We investigate the pattern of foraging by a bird that results in the lowest long-term rate of mortality. We assume the following: mortality is the sum of starvation and predation. The bird is characterized by two state variables, its energy reserves and the amount of food in its stomach. Starvation occurs during the day if the bird's reserves fall to zero. The bird starves during the night if the total energy stored in reserves and the stomach is less than a critical amount. The probability that the bird is killed by a predator is higher if the bird is foraging than if it is resting. Furthermore, the predation risk while foraging increases with the bird's mass. From these assumptions, we use dynamic programming techniques to find the daily foraging routine that minimizes mortality. The principal results are (1) Variability in food finding leads to routines with feeding concentrated early in the day, (2) digestive constraints cause feeding to be spread more evenly through the day, (3) even under fairly severe digestive constraints, the stomach is generally not full and (4) optimal fat reserve levels are higher in more variable environments and under digestive constraints. This model suggests that the characteristic daily feeding pattern of small birds is not due to digestive constraints but is greatly influenced by environmental variability.