Specific binding and uptake of apolipoprotein E-free high density lipoproteins by cultured liver parenchymal cells of copper-deficient rats

The influence of copper deficiency on the binding and uptake of apolipoprotein E-free high density lipoprotein (apo E-free HDL) in cultured rat hepatic parenchymal cells was examined in this study. Male weanling Sprague-Dawley rats were randomly divided into two treatments, a Cu-adequate (7.33 mg Cu/kg diet) or a Cu-deficient (1.04 mg Cu/kg diet) group. After 7 weeks, plasma apo E-free HDL were isolated by a combination of ultracentrifugation, gel filtration, and heparin-Sepharose affinity chromatography. Parenchymal cells were isolated from collagenase perfused liver of Cu-deficient and adequate rats and cultured for 16 hours at 37 degrees C prior to incubation with iodinated apo E-free HDL from the same treatment group. Cells were incubated with 5 microg/ml(125) I-apo E-free HDL for 2, 6, or 12 hours in the presence or absence of 200 microg/ml (40-fold) excess unlabeled apo E-free HDL. Increases in specific binding at 4 degrees C and specific cell-associated uptake at 37 degrees C as a function of time were observed with cells and HDL from Cu-deficient rats. Cells were also incubated for 6 hours with 8 concentrations of (125)I-apo E-free HDL in the presence or absence of excess unlabeled HDL. Although no significant increase in specific binding was detected at 4 degrees C as a function of ligand concentration, the response tended to be higher at 5 to 15 microg HDL/ml for the Cu-deficient treatment. However, at 37 degrees C the specific cell-associated uptake was increased markedly with cells and HDL from Cu-deficient rats. The observed increases in HDL binding and uptake indicate that these processes may be enhanced in Cu-deficient rats. These data are also consistent with recent in vivo results which indicate that plasma clearance and tissue uptake of HDL are increased in Cu-deficient rats.     

电刺激兔延髓腹侧化学敏感区和压力敏感区对呼吸、血压,的影响及其中枢递质机制

电刺激麻醉兔延髓腹侧化学敏感区头端区引起潮气量(V_T)增加,呼吸频率(f)增快;电刺激压力敏感区(中间区)则使V_T减小,f亦增快。弱刺激时,两者均产生降压反应;刺激增强可诱发双相或升压反应。在出现周期性呼吸时,电刺激化学敏感区可使呼吸节律正常化、V_T增大,而电刺激压力敏感区则导致呼吸暂停。电刺激压力敏感区时,吸气时间(TI)和呼气时间(T_E)均缩短,以T_E变化更明显;由于V_T减小和T_I缩短,V_T/T_I保持相对不变,提示吸气终止的中枢阈值降低。在准备刺激的相应局部预先应用阿托品,可使电刺激化学敏感区产生的通气增强效应翻转,而对电刺激压力敏感区引起的通气抑制无明显影响;用印防己毒素则可选择性消除电刺激压力敏感区的通气抑制和降压效应。本工作表明延髓腹侧存在两个不同的中枢机制,其中化学敏感区产生的通气增强与胆碱能系统有关;压力敏感区产生的通气减弱效应与GABA系统有关。     

Genomic DNA sequence for human C-reactive protein

The gene for the prototype acute phase reactant, C-reactive protein, has been isolated from two lambda phage libraries containing inserted human DNA fragments using synthetic oligonucleotide probes. Nucleotide sequence analysis indicates that after coding for a signal peptide of 18 amino acids and the first two amino acids of the mature protein, there is an intron of 278 base pairs followed by the nucleotide sequence for the remaining 204 amino acids. The intron is unusual in that it contains on the positive strand a poly(A) stretch 16 nucleotides long and a poly(GT) region 30 nucleotides long which could adopt the Z-form of DNA. The nucleotide sequence reported here confirms the amino acid sequence of mature C-reactive protein as originally reported except that it codes for an additional 19 amino acids beginning at position 62. Thus DNA sequence analysis predicts that the mature protein consists of 206 amino acids rather than 187 as originally reported. The mRNA cap site is located 104 nucleotides from the start of the signal peptide and there is a 3' noncoding region 1.2 kilobase pairs in length. The gene has a typical promoter containing the sequences TATAAAT and CAAT 29 and 81 base pairs upstream, respectively, of the cap site.     

An improved DNA sequencing strategy

A modification of Hong's systematic DNA sequencing strategy is described. The original procedure has been simplified and transfectant yield increased. After DNase I limited cleavage in the presence of Mn2+, the single-cut linear DNA does not have to be separated from supercoiled or open circular DNA on an agarose gel. After ligation, the DNA is digested with a second restriction endonuclease for which a unique cleavage site resides between the insert and the first restriction endonuclease cutting site. The original intact DNA is linearized whereas the deleted subclone is not. The background is decreased to an undetectable level. This DNA sequencing strategy was tested on a 1.4-kb DNA fragment containing the araC regulatory gene from Erwinia carotovora. A set of subclones sufficient to sequence the fragment on both strands was produced in 2 days and the yield was at least 60-fold higher than in the original protocol.     

Regeneration of the arterial wall in microporous,compliant, biodegradable vascular grafts after implantation into the rat abdominal aorta

Summary The ultrastructure of a new type of vascular graft, prepared from a mixture of polyurethane (95 weight %) and poly-L-lactic acid (5 weight %), was examined six weeks after implantation into the abdominal aorta of rats. These microporous, compliant, biodegradable, vascular grafts function as temporary scaffolds for the regeneration of the arterial wall.Smooth muscle cells, covering the grafts, regenerated a neo-media underneath an almost completely regenerated endothelial layer (neo-intima). These smooth muscle cells varied in morphology from normal smooth muscle cells to myofibroblasts. They were surrounded by elastic laminae and collagen fibers.Macrophages, epithelioid cells, multinucleated giant cells, fibroblasts and capillaries were present in the disintegrating graft lattices. The epithelioid cells and multinucleated giant cells engulfed polymer particles of the disintegrating grafts.The regeneration of the endothelial and smooth muscle cells is similar to the natural response of arterial tissue upon injury. The presence of macrophages, epithelioid cells, multinucleated giant cells, fibroblasts and capillaries in the graft lattices resembles the natural response of tissue against foreign body implants. Both of these responses result in the formation of a neo-artery that possesses sufficient strength, compliance and thromboresistance to function as a small caliber arterial substitute.Supported by Grant nr. 82.042 from the Dutch Heart Foundation     

Cloning of the pectate lyase genes from Erwinia carotovora and their expression in Escherichia coli

A hybrid cosmid coding for pectate lyase (PL) activity was identified from an Erwinia carotovora genomic library by an immunological screening method. A 7-kb DNA fragment was identified which codes for three proteins identical in size to proteins with PL activity purified from E. carotovora culture supernatants. The three proteins had apparent Mrs of 41, 44 and 44 X 10(3) as estimated by SDS-PAGE. None of the PLs were exported from Escherichia coli strain HB101 but all were found in the periplasmic space. Plant tissue was macerated by the PLs made in E. coli.     

The araBAD operon of Salmonella typhimurium LT2. I. Nucleotide sequence of araB and primary structure of its product, ribulokinase

Hybrid plasmids containing the araBAD operon of Salmonella typhimurium LT2 were characterized by Southern blot and genetic analyses. The nucleotide sequence of araB was determined. The araB gene product, ribulokinase (EC 2.7.1.16), was purified and the results of amino acid composition analysis and partial amino acid sequence are in agreement with predictions from the DNA sequence. Ribulokinase is 569 amino acid residues long and has a calculated Mr of 61 793. Ribulokinase shares significant homology with xylulose kinase from Escherichia coli. Codon usage in the araB gene does not favor those codons which have intermediate codon-anticodon binding energy.     

细菌(Pseudomonas maltophilia)之绒毛膜促性腺激素(hCG)结合物质之研究

 细菌(Pseudomonas moltophilia)与hCG及LH有特异的亲和力,实验发现,细菌之生长曲线与hCG结合活性成平行关系,96小时达高峰,细菌之培养液中含有可溶性结合蛋白,该蛋白经硫酸铵沉淀(80％饱和度)、Sephadex G-100柱层析、DEAE-纤维素柱0.5mol/L NaCl梯度洗脱,再过Sepharose CL-AB柱,收集之活性部分经SDS电泳测得其分子量为70,000,凝胶层析测Stokes radius为41A,Schiff氏染色未见着色带。     

东亚飞蝗生殖的研究:雌蝗成虫卵巢发育过程中核酸和蛋白质的代谢与激素调节

本文报道东亚飞蝗交配后雌蝗卵巢中核酸和蛋白质的含量变化,以及孤雌生殖和雄蝗促性腺因子对卵巢中核酸和蛋白质代谢的影响。雌蝗羽化后7—10天进行交配,卵巢开始迅速发育,卵巢鲜重、总磷量以及蛋白质含量皆迅速增长,同时末端卵母细胞长度亦不断增加。到羽化第15天时卵巢已接近发育完成。末端卵母细胞长达6.2毫米。在各种磷化合物中,酸溶性部分在羽化15天时占总磷量的70%,磷脂磷可达20%。这表明酸溶性磷化合物和磷脂在卵巢发育过程中有较高的代谢和积累。卵巢中RNA-P增长28倍,DNA-P,增长6倍。RNA/DNA比值随着卵巢的发育而增加,这标志着蛋白质在卵巢中合成;对蛋白质含量的测定也证实了这一点。如果以RNA-P和DNA-P占总磷量的百分含量或以每百毫克卵巢鲜重计算其含量,则在卵巢发育过程中反而皆相对降低,表明卵巢中其他含磷化合物的积累优于核酸磷的增长。当雌蝗第一次产卵后,卵巢的各组成成份迅即减少,此后四天内卵巢中核酸和蛋白质的含量复可再度迅速增长,末端卵母细胞(即原未产卵前之末端第二卵母细胞)亦进一步长大,从而表现了卵巢发育的周期性变化。 人为隔离的孤雌生殖的雌蝗在羽化后40天内,卵巢发育缓慢,其末端卵母细胞长度增长缓慢,卵蜒中核酸和蛋白质的含量皆较低,相当于正常发育卵巢5—10天的水平。如将雄蝗脂肪体脂类提取物涂抹于羽化后5—7天雌蝗体表,则表现出促进雌蝗卵巢发育的作用,卵巢鲜重、卵巢中核酸等磷化合物和蛋白质含量,以及卵管中末端卵母细胞长度,在短时期内,基本上相同于正常交配对照的卵巢发育水平。因此证实:雄媓促健腺因子对卵巢发育过程中核酸和蛋白质代谢和合成起调节作用。这种调节因子可能是与保幼激素共同起作用的。     

蝗卵的研究Ⅳ.浸水对于蝗卵胚胎发育和死亡的影响

一、引言 东亚飞蝗常生长在湖滨、已干涸的河道、沿海边等长着禾本科杂草的低洼荒地;这些地方也就成为它们的产卵场所。蝗卵在产入土中之后须经一定时间方始孵化;夏蝗所产卵在自然情况下须经12天以上的孵育时间,秋蝗卵在越冬时在土中常长达7—8个月。蝗区每当雨季或湖、河的水位上涨时其有卵的地区常被淹没,使蝗卵在水中经过长短不一的