Purification to homogeneity of human placental acid sphingomyelinase

Acid sphingomyelinase was purified to homogeneity from human placenta in the presence of a dialyzable detergent, n-octyl-beta-D-glucopyranoside. The major steps in the procedure included column chromatographies with Con A-Sepharose, sphingosylphosphorylcholine-Sepharose 4B, hexyl-agarose, and Mono P. The purified enzyme with pI 7.4 had a specific activity of approx 170,000 units/mg protein with a yield of 3.6%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single protein band of Mr 62,000. Gel filtration with a Superose 12 column gave a single peak, and the enzyme in the presence 50 mM n-octyl-beta-D-glucopyranoside was of Mr 123,000, indicating that the native enzyme occurs in a dimeric form. The optimal pH was 5.5 with both sphingomyelin and an artificial substrate, 2-N-hexadecanoylamino-4-nitrophenylphosphorylcholine. The Km values were 55 microM with sphingomyelin and 340 microM with the artificial substrate. The enzyme activity was not affected by Mg2+ (1-5 mM), confirming that the enzyme is acid sphingomyelinase. The enzyme was stable at -80 degrees C for more than 4 months. In addition to the enzyme with pI 7.4, the Mono P chromatofocusing gave two peaks (pI 7.0 and 6.7) possessing the enzymatic activity.     

中国盲蝽科两新种六新纪录(半翅目:盲蝽科)

本文共记述盲蝽科叶盲蝽亚科(Phylinae)新种两个,计为:宽束盲蝽 Pilophorus latus sp.nov.和黄平盲蝽 Zanchius vitellinus sp.nov.。两新种的模式产地均为云南。并记录了6个中国新纪录种,计为:棕二带束盲蝽 Pilophorus alstoni Schuh、长黑束盲蝽 Pilophorus dailahn Schuh、细毛束盲蝽 Pilophorus setulosus Horvath、朝束盲蝽 Pilophorus koreanus Josifov、褐束盲蝽 Pilophorusgallicus Remane、亮束盲椿 Pilophorus lucidus Linnavuori。     

蚕豆根端细胞核中微核仁的研究

以蚕豆(Vicia faba)根端分生组织细胞为材料研究了微核仁的超微结构和细胞化学特点。结果表明;微核仁是直径0.3—0.5μm 的卵圆形或球形结构。常规染色时,微核仁与集缩染色质的电子密度相仿,但两者之间在结构上没有任何联系。细胞化学研究指出,微核仁含有 RNA 和蛋白质,其结构成分主要是与核仁颗粒组分十分相似的 RNP 颗粒。报道了植物细胞核中微核仁发生于核仁的过程并对微核仁的本质和功能进行了讨论。     

南京地区繁缕和小繁缕群体的研究

繁缕(Stellaria media)和小繁缕(S.apetala)是两个形态相似的近缘种,有人把后者作为前者的亚种或变种来处理。本文通过对南京地区不同生境的三个自然群体和三个人工控制栽培群体的取样,以群体为单位,分别测算了叶、萼片、雄蕊、花瓣、果实和种子的8个数量性状的变异,绘制了多角形图;对花粉粒和种子进行了扫描;还通过花蕾套袋试验对种子活力作了检查。结果发现繁缕和小繁缕都是近亲繁殖植物,在形态上区别明显,对生态环境的要求基本相同,但小繁缕似更能耐受人为的践踏和刈割。     

Reactivity of anti-human milk fat globule antibodies with synthetic peptides

The nucleotide sequence of partial cDNA clones coding for the core protein of a human polymorphic epithelial mucin has recently been obtained, this mucin consists of a highly conserved 60 bp tandem repeat and the amino acids commonly found are PDTRPAPGSTAPPAHGVTSA. We synthesized three peptides, 1) P1.24 containing the 20 amino acids and four amino acids (PDTR) of the adjoining repeat; 2) P1.15 consisting of the first fifteen (PDTRPAPGSTAPPAH) and P1.09 the second nine amino acids (GVTSAPDTR) of peptide P1.24. The reactivities of the synthetic peptides with mAb known to react with breast cancer (BC1, BC2, BC3, HMFG-1, 3E1.2, and RCC-1) were studied. The synthetic peptide, P1.24, corresponding to the antigenic sequence predicted from the tandem repeat reacted with antibodies BC1, BC2, and BC3 (known to react with human milk mucin and mucin expressed in breast cancer) and the antibody HMFG-1 which was used to select the cDNA clones. In addition, the epitopes recognized by BC1, BC2, and BC3 appear to be in the same region of the molecule represented by their reactions with the nine amino acids in peptide P1.09 (GVTSAPDTR). By contrast, other antibodies such as 3E1.2 which reacts only weakly with components of human milk, and RCC-1 that detects a low Mr component (95 kDa) in breast cancer, had no specific reaction with the synthetic peptides, indicating that their epitopes are distinct from those of BC1, BC2, BC3, and HMFG-1. Inasmuch as the antibodies HMFG-1, BC1, BC2, and BC3 react with the fully processed milk mucin, it is likely that some of the peptide is exposed, even in the fully glycosylated molecule. Identification of the different epitopes could lead to the development of "second generation" mAb with enhanced specificity for breast carcinoma using the appropriate synthetic peptides as immunogens.     

钐在小鼠肝脏细胞中的动态观察

It is generally considered that the rare earth compounds are plasma membrane-impermeable, thus affecting the cells only on their surface. Recently, we found that after repeated injections to mice of large dose of samarium trichloride, a soluble compound of rare earth, samarium aggregates appeared in Kupffer cells and hepatocytes of liver. In this study, we aimed at observing the route by which samarium enters the liver cells and the process of the formation of samarium aggregates. Samarium trichloride was given to Swiss mice at one dose of 70 mg/kg intravenously. Thereafter, at different intervals from 15 min to 48 h after the injection, the samarium in liver was traced dynamically by electron microscopy and X ray microanalysis. From 15 min to 2 h both Kupffer cells and hepatocytes endocytosed samarium-containing particles and formed phagosomes, in which the ingested particles were progressively concentrated. Besides, the small phagosomes fused with each other. Phagocytosis was especially active in Kupffer cells. During the 4 h to 24 h many Kupffer cells were degenerated and broken. In hepatocytes the phagosomes gathered mostly around the bile canaliculi. Groups of highly electron-dense particles were found in the lumen of bile canaliculi, implying the excretion of samarium by bile. At the 48 h, the samarium-containing phagosomies were found still in both kinds of cells in the liver.     

铃兰族rbcL基因的PCR—RFLP分析

本文用7属9种铃兰族Tribe convallarieae植物的叶绿体DNA中rbcL基因片段的PCR产物的RFLP结果进行聚类分析。结果表明:开口箭属与蜘蛛抱蛋属关系密切,夏须草属与族内其余各属亲缘关系稍远,与外部器官形态、核型和孢粉学资料所得出的结论基本一致。此外,本文对铃兰属的系统位置也进行了讨论。     

Gene conversion plays the major role in controlling the stability of large tandem repeats in yeast.

The genomic stability of the rDNA tandem array in yeast is tightly controlled to allow sequence homogenization and at the same time prevent deleterious rearrangements. In our study, we show that gene conversion, and not unequal sister chromatid exchange, is the predominant recombination mechanism regulating the expansion and contraction of the rDNA array. Furthermore, we found that RAD52, which is essential for gene conversion, is required for marker duplication stimulated in the absence of the two yeast type I topoisomerases. Our results have implications for the mechanisms regulating genomic stability of repetitive sequence families found in all eukaryotes.     

Auxin levels and auxin binding protein availability inrolB transformedBeta vulgaris cells

The final biological effect of auxin depends both on free auxin levels and on auxin perception capacity.RolB transformedBeta vulgaris L. hairy roots provide a system for studying both factors. Highly purified plasma membrane fractions were prepared with aqueous two-phase partitioning. Individual hairy root clones were assessed for the binding activities of plasma membrane-bound auxin binding proteins and for their free intracellular indole-3-acetic acid levels. The presence of a high affinity auxin binding protein with a dissociation constant of 9.07 x 10^?7 M was detected in the plasma membrane fractions isolated from non-transformed seedling roots and the six clones ofrolB transformed hairy roots. However, the levels of specific IAA binding considerably varied among different hairy root clones and between transformed and non-transformed roots. The levels of the detectable polypeptide in immunoblotting with an antibody against maize 22-kD auxin binding protein subunit were in good agreement to the levels that were detected in auxin binding assays. Differences in the indole-3-acetic acid levels were found between transformed and non-transformed roots and also between different transformed hairy root clones. A negative correlation was observed between free intracellular IAA levels and its specific binding to the plasma membrane-bound auxin binding proteins. A latency study indicated that the binding site for auxin may be located on the exterior face of the plasma membrane