Synchronization of 9L rat brain tumor cells by centrifugal elutriation

Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic data indicated that fractions containing greater than or equal to 97% G1 cells, greater than or equal to 80% S cells, and 70-75% G2 cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle was a constant 60-70%, which was comparable to the 60-80% usually found for asynchronous 9L cells. The percentage of cells in the G1, S, and G2 phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of the DNA histogram obtained from flow cytometry. In genereal, the degree of synchrony was related to the coefficient of variation (CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all elutriated fractions. When the greater than or equal to 97% G1 population was allowed to progress to S and G2, the CVs were about 17 and 20.2%, respectively. Thus, the best nonperturbing method for obtaining synchronous 9L cells in the S or G2 phases was direct elutriation with the long collection method.     

固氮蓝藻“红圈病”的研究——Ⅰ.“红圈病”在大面积培养蓝藻中的危害以及防范措施

对蓝藻“红圈病”的病症、发病条件、传播程度方式、危害进行了研究。结果表明,发病条件与蓝藻最佳生长条件基本一致;病原通过水体和带病藻体传播,对藻的产量和质量均造成严重危害。同时探讨了综合防治措施。     

显微高速摄影技术及其在血液微流变学研究中的应用

本文选用金黄地鼠的微血管,利用显微高速摄影技术,放大微观流场和血细胞,连续地“冻结”短瞬间的变化状态,把物理图象呈现在胶片上,经图像分析和数据处理,从定性及定量方面研究血液微流变学,为生命科学和医学研究提供了又一种新的方法。     

基因定点诱变删除及天然α型心钠素的分泌型表达

用基因定点诱变技术,删除了pO_1α ANF表达质粒中的33对碱基,使人α型心钠素结构基因直接与大肠杆菌分泌型表达质粒pIN-Ⅲ-OmPA中的信号肽酶切位点编码区相连,构成天然人α型心钠素的表达质粒pANF,在IPTG诱导下表达28肽的天然人α型心钠素。纯化后的表达产物具有天然心钠素的放免活性和很强的舒张血管的生物活性。     

Description of the neotype ofRepomucenus sagitta (Callionymidae) with comments on the species

The neotype ofRepomucenus sagitta is described. Specimens conspecific with the neotype from India, the Gulf of Thailand, the South China Sea, the Yellow Sea and Australia are described. Sexual characters, geographic variation, habitat and geographic distribution were noted.Repomucenus macdonaldi is a junior synonym ofR. sagitta as defined by the neotype.     

Mathematical modelling of the effect of sole elasticity distribution on pronation.

A coronal plane model of a distributed elastic sole has been proposed and analyzed with respect to the effects of different medial-lateral elasticity distribution on pronation under quasi-static conditions. The distributed model consists of an array of linear vertical line springs. Under minimum energy assumption, the behavior of the top surface of the interface under resultant force and moment loading was shown to be equivalent to that of a rigid-body mechanism under the same loading. The model was then combined with a rigid-link model of the lower limb. Expressions that describe the relationship of the interface aggregate parameters with pronation and the center of pressure were obtained. These expressions were confirmed by an experiment in which the elastic distribution in the interface was systematically varied and the pronation angle and the center of pressure measured. The model has the potential of being a useful analytical tool in the design of elastic soles in running shoes.     

The genes encoding the two carboxyltransferase subunits of Escherichia coli acetyl-CoA carboxylase.

We report characterization of the component proteins and molecular cloning of the genes encoding the two subunits of the carboxyltransferase component of the Escherichia coli acetyl-CoA carboxylase. Peptide mapping of the purified enzyme component indicates that the carboxyltransferase component is a complex of two nonidentical subunits, a 35-kDa alpha subunit and a 33-kDa beta subunit. The alpha subunit gene encodes a protein of 319 residues and is located immediately downstream of the polC gene (min 4.3 of the E. coli genetic map). The deduced amino acid composition, molecular mass, and amino acid sequence match those determined for the purified alpha subunit. Six sequenced internal peptides also match the deduced sequence. The amino-terminal sequence of the beta subunit was found within a previously identified open reading frame of unknown function called dedB and usg (min 50 of the E. coli genetic map) which encodes a protein of 304 residues. Comparative peptide mapping also indicates that the dedB/usg gene encodes the beta subunit. Moreover, the deduced molecular mass and amino acid composition of the dedB/usg-encoded protein closely match those determined for the beta subunit. The deduced amino acid sequences of alpha and beta subunits show marked sequence similarities to the COOH-terminal half and the NH2-terminal halves, respectively, of the rat propionyl-CoA carboxylase, a biotin-dependent carboxylase that catalyzes a similar carboxyltransferase reaction reaction. Several conserved regions which may function as CoA-binding sites are noted.     

Heterogeneity of high affinity uptake of L-glutamate and L-aspartate in the mammalian central nervous system.

Characteristics of high affinity uptake of L-glutamate are examined in order to evaluate the possible use of the uptake of [3H]L-glutamate, [3H]L-aspartate or any other suitable [3H]-labelled substrate as a marker for glutamatergic and aspartergic synapses in autoradiographic studies in the mammalian brain. Review of data on substrate specificity indicates the presence of at least two high affinity uptake systems specific for acidic amino acids in the central nervous tissue; one which takes up L-glutamate and L-aspartate and the other which is selective for L-glutamate only. Studies on ionic requirements, too, point to the existence of at least two distinct uptake systems with high affinity for L-glutamate. The Na(+)-dependent uptake system(s) handle(s) both L-glutamate and L-aspartate whereas the Na(+)-independent uptake system(s) show(s) selectivity for L-glutamate only. Available data do not favour the Na(+)-dependent binding of [3H]D-aspartate to thaw-mounted sections of frozen brain tissue as a suitable marker for glutamatergic/aspartergic synaptic nerve endings. However, there are reasons--such as the results of lesion studies and the existence of uptake sites which have a higher affinity for L-aspartate than for D-aspartate--to suggest that Na(+)-dependent binding of [3H]L-aspartate, rather than that of [3H]D-aspartate, should be further investigated as a possible marker for the glutamatergic/aspartergic synapses in the autoradiographic studies using sections of frozen brain.