In vivo apoprotein catabolism of high density lipoproteins in copper-deficient, hypercholesterolemic rats

High density lipoprotein (HDL) apoprotein catabolism was examined in male Sprague-Dawley rats deficient in dietary copper. Twenty-four rats were randomly divided into two groups: copper-adequate (control, 5 mg of copper/kg diet) and copper-deficient (0.6 mg of copper/kg diet). After 5 weeks, animals were administered a tracer dose of iodinated HDL protein previously isolated from donor rats that were subjected to the same dietary treatments as the test animals. Copper-deficient rats exhibited a 54% increase in plasma volume and a 26% increase in HDL protein concentration above controls. Consequently, the intravascular pool of total HDL protein was increased 2-fold. The fractional catabolic rate of total HDL protein was similar between groups. However, because of the increased intravascular HDL pool in copper-deficient animals, the absolute catabolic rate was greater (640 +/- 49 micrograms/hr vs 316 +/- 12 micrograms/hr in controls). Tissue uptake of total HDL protein in copper-deficient rats tended to be greater in the kidneys, spleen, and testes compared with controls; the heart exhibited a significant 2.3-fold increase. In contrast, the catabolic rate of HDL protein in the liver and adrenal gland were not different between treatment groups. That an obligatory increase in HDL protein uptake was not observed in the liver and adrenal gland (organs which are sensitive to and can further metabolize cholesterol) suggests that these organs may be regulated, possibly contributing to the observed hypercholesterolemia in this model. These data imply that total HDL apoprotein catabolism is increased in response to the increased intravascular pool of HDL in copper-deficient rats.     

辣椒素引起脊髓P物质释放及其对血压的影响

为进一步研究脊髓 P 物质(SP)在调节心血管活动中的作用,在大鼠脊髓蛛网膜下腔注射(ith)辣淑素(cap),以刺激脊髓 SP 能神经末梢释放 SP,结果引起血浆去甲肾上腺素(NA)和肾上腺素(AD)含量增高,及具有剂量依赖性的动脉血压上升,心率升高。ith 具有高度特异性的 SP 受体拮抗剂或 SP 抗血清均可阻断 cap 引起的升压效应,免疫组化测定也观察到注入的cap 剂量越大,脊髓胸段 SP 样免疫阳性反应物的致密度越低,这些观察结果支持 cap 可以引起脊髓内 SP 的释放的说法。在第一颈段(C_1)横断脊髓后 ith cap 所引起的升压效应与完整动物 ith cap 的升压效应无显著差异。以上结果提示脊髓 SP 能神经末梢释放的 SP 可以通过交感肾上腺髓质系统引起心血管兴奋效应,SP 可能是引起交感节前神经元兴奋的神经递质。     

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.     

白魔芋和花魔芋葡甘露聚糖研究

从白魔芋和花魔芋的块茎中分离纯化出两种魔芋葡甘露聚糖(Konjac Glucomannan,简称KGM),分别名为白魔芋葡甘露聚糖(aKGM)和花魔芋葡甘露聚糖(rKGM)。通过超离心、玻璃纤维纸电泳和凝胶过沪证明aKGM和rKGM都是均一的多糖。这两种多糖都由甘露糖(M)和葡萄糖(G)组成,其克分子比G/M:aKGM为1:1.69,rKGM为1:1.60,分子量前者为8.09×10~5,后者为7.37×10~5。酶水解实验和红外光谱分析说明aKGM和rKGM都是由甘露糖和葡萄糖以β-1,4-糖苷键连接的杂多糖;有O-乙酰基的特征吸收峰,说明在某些糖残基上可能有乙酰基团。     

韦氏鳞(虫八)的精子结构(双尾目)

韦氏鳞(虫八)(Lepidocampa weberi)精子为细长的鞭毛精子,成束排列成精索。顶体由三层结构组成;核圆柱形,接于顶体后;鞭毛轴丝微管式为“9+9+2”,9条副微管排列不整齐,集中于轴丝的一侧;两条细长的线粒体衍生物位于顶体、核与轴丝之间,贯穿于精子中部;精子尾部围有大量片层结构。结果表明韦氏鳞(虫八)精子与康(虫八)精子较为接近,但可能比康(虫八)精子更为特化。     

高稳定人胰岛素的晶体学研究:Ⅲ,A21-Asp突变体的结晶和初步晶体学分析

研究一系列有明确意义改性胰岛素的结构将有益于新型胰岛素分子的设计。经蛋白质工程制备的AspA21—人胰岛素(ADHI)基本保持了生物活力,而在酸性溶夜(pH3—4)中的稳定性比天然胰岛素在中性溶液中高5—10倍。本文报道ADHI的结晶、初步晶体学研究及中等分辨率衍射数据的收集。ADHI晶体属R3空间群:a_H=b_H=82.72埃,c_H=34.02埃。     

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.