CpG联合铝佐剂对HCV免疫原体液免疫作用的影响

目的:探讨胞苷酸鸟苷寡脱氧核苷酸(CpG ODN)联合铝佐剂对丙型肝炎病毒(HCV)重组免疫原的体液免疫作用。方法:采用高交叉HCV-HVR1和E1重组蛋白与CpG ODN、铝佐剂组合,免疫BALB/c小鼠后以ELISA、酶联免疫斑点测定、流式细胞术、免疫沉淀等方法检测相关体液免疫指标和免疫血清多抗的交叉反应性。结果:CpG联合铝佐剂激发了最高的特异性抗体滴度;佐剂通过提高抗体分泌细胞数量、增加脾脏中记忆B细胞数量、增加脾淋巴细胞IL-6、IL-10分泌浓度实现体液免疫增效;CpG则能提高免疫效率,联合铝佐剂时显著提高浆细胞数量;12份HCV阳性血清中有10份可与多抗HVR1 IgG发生免疫沉淀。结论:CpG和铝佐剂联合应用具有协同作用,多抗HVR1 IgG具有较好的交叉反应性。     

beta 2-Inhibin contains the active core of human seminal plasma beta-inhibin: synthesis and bioactivity

The complete synthesis of the C-terminal 28 residues segment 67-94 of human seminal plasma beta-Inhibin, called beta 2-Inhibin, is reported. The Inhibin-like activity of the native 94 amino acids beta-Inhibin is compared to that of the synthetic replica of beta 2-Inhibin. In all assays used both peptides effectively suppress the FSH release induced by LHRH but have little effect on the LH release. In the mouse both peptides are equipotent on a mole basis. In the rat the synthetic beta 2-Inhibin is 3-10 times more potent than beta-Inhibin. Both peptides are active in rat anterior pituitary primary culture assays where maximum suppression of FSH release induced by LHRH occurs around 300 pmol/ml of beta 2-Inhibin. In contrast, maximum suppression of FSH release in the mouse pituitary assay occurs at 10-15 pmol/ml of either Inhibin.     

The structure of type IX collagen

We present a detailed analysis both of tryptic peptides and amino-terminal sequences of the subunits of two collagenous fragments (HMW and LMW) previously isolated from pepsin extracts of chicken cartilage (Reese, C.A., and Mayne, R. (1981) Biochemistry 20, 5443-5448). This analysis and a comparison with the nucleotide sequence of the cDNApYN1738 (Ninomiya, Y., and Olsen, B.R. (1984) Proc. Natl. Acad. Sci. U. S. A. 81, 3014-3018) shows that HMW and LMW are pepsin-resistant fragments of a unique collagen composed of molecules with three different polypeptide chains (alpha-chains). This collagen has been assigned the type number IX, and the alpha-chain encoded by pYN1738 has been given the designation alpha 1 (IX). Type IX collagen contains three triple-helical domains and at least two sets of interchain disulfide bridges. At the amino and carboxyl ends are noncollagenous domains which do not appear to be homologous to amino and carboxyl propeptides of interstitial collagens.     

The Role of Anaerobic Respiration in Germinating Muskmelon Seeds: I. IN RELATION TO SEED LOT QUALITY

Five muskmelon (Cucumis melo L.) cultivar seedlots from a commercialsource and freshly produced seeds of two cultivars, when artificiallyaged, were found to differ in their viability and vigour asdetermined by germination tests. Furthermore, the various commercialseedlots without ageing also exhibit a range of deteriorationlevels. Low vigour seeds had higher respiratory quotient valuesthan the high vigour seeds as a result of a higher level ofCO₂ production. This high level of CO₂ evolution in low vigourseeds may have been due to anaerobic respiration. Levels of acetaldehyde and ethanol produced by imbibing seedswere negatively associated with seed viability and vigour. After6 h of imbibition low vigour seeds produced significantly moreethanol and acetaldehyde than high vigour seeds. After 24 hof imbibition, ethanol continued to accumulate in the commercialseedlots up to 10-fold the amount produced after 6 h of imbibition,whereas, acetaldehyde levels increased less. However, in thefreshly produced, artificially aged seeds (except the most extremeageing), ethanol levels were reduced and no acetaldehyde productioncould be detected, indicating re-utilization of ethanol. It is suggested that ethanol production in the first hours ofimbibition can be used as a reliable index to predict germinationin muskmelon seedlots. Key words: Germination, Cucumis melo L., Seeds, Anaerobic respiration     

Persistence of Oral Structures in the Sexual Process of Amicronucleate Paramecium tetraurelia

ABSTRACT In the sexual process, amicronucleate Paramecium tetraurelia , unlike micronucleates, fail to produce an oral apparatus, but resorb the pre-existing one. Exceptions were found in some amicronucleate cell lines in which about 1% of the cells possessed oral structures, including pieces of oral membranelles, sometimes complete with buccal cavity, after autogamy or conjugation. By following oral development in the sexual process in some detail, the present study supports the view that these oral structures are derived from the pre-existing oral apparatus and not newly developed from the oral primordium. The possible involvement of the micronucleus and the pre-existing oral apparatus in oral resorption is discussed. The possession of a functional oral apparatus after the sexual process may open up a new evolutionary avenue to the amicronucleates.     

Amino acid sequence of rabbit kidney neutral endopeptidase 24.11 (enkephalinase) deduced from a complementary DNA.

Neutral endopeptidase (EC 3.4.24.11) is a major constituent of kidney brush border membranes. It is also present in the brain where it has been shown to be involved in the inactivation of opioid peptides, methionine- and leucine-enkephalins. For this reason this enzyme is often called 'enkephalinase'. In order to characterize the primary structure of the enzyme, oligonucleotide probes were designed from partial amino acid sequences and used to isolate clones from kidney cDNA libraries. Sequencing of the cDNA inserts revealed the complete primary structure of the enzyme. Neutral endopeptidase consists of 750 amino acids. It contains a short N-terminal cytoplasmic domain (27 amino acids), a single membrane-spanning segment (23 amino acids) and an extracellular domain that comprises most of the protein mass. The comparison of the primary structure of neutral endopeptidase with that of thermolysin, a bacterial Zn-metallopeptidase, indicates that most of the amino acid residues involved in Zn coordination and catalytic activity in thermolysin are found within highly honmologous sequences in neutral endopeptidase.     

The complete amino acid sequence of rat submaxillary gland tonin does contain the aspartic acid at the active site: confirmation by protein sequence analysis

The revised amino acid sequence of rat submaxillary gland tonin, a serine protease, does contain the active site Asp residue. The active site of this kallikrein-related enzyme is thus made up of the same catalytic triad (Asp, Ser, and His) found in all known serine proteases. The important Asp residue has now been localized in a 16 amino acid peptide previously reported as missing in the tonin sequence. The complete amino acid sequence thus contains 235 residues corresponding to a molecular weight of 25,658, more in agreement with previously reported molecular weights. Moreover, the revised structure led (a) to the assignment of Arg, Asn, and Val residues instead of His, Asp, and Gly at positions 63, 165, and 169, respectively; (b) to the assignment of residues occupying an overlapping sequence at positions 165-171, and finally (c) to the localization of two N-glycosylation sites at positions 82 and 165. These results further document the close relationship of tonin to the ever expanding kallikrein family.     

Expression of XET-related genes and its relation to elongation in leaves of barley (Hordeum vulgare L.)

Five cDNA clones were isolated from barley (Hordeum vulgare L.) that encoded mRNAs related to xyloglucan endotransglycosylase (XET). One of the clones encoded a protein with XET activity in vitro. Sequence comparisons revealed five families of XET-related sequences, one of which (containing two of the barley genes) was novel. Hybridization studies using clone-specific probes indicated that the corresponding genes were represented once, or possibly twice, in the barley genome. Treatment of dwarf mutants with gibberellic acid (GA₃), or homozygosity at the ‘slender’ (sln1) locus, resulted in a 2.5-fold (approximately) stimulation of blade elongation rate. Three of the five clones detected mRNAs that were maximally expressed towards the base of the blade, and present in greater quantities in GA₃-treated or slender seedlings. The remaining two clones detected mRNAs that were maximally expressed in the middle of the blade. Relative elemental growth rate (REGR) profiles of leaves growing with or without GA₃ treatment revealed similar maximal REGR values despite a 2.5-fold difference in leaf elongation rate. Segments of GA₃-treated leaves attained their maximal REGR values more rapidly, this being associated with enhanced expression of the three ‘basal’ XET-related mRNAs. Highest XET activities were detected in the base of the elongation zone, and in GA₃-treated seedlings a second activity peak was observed near the distal end of the elongation zone. We conclude that there are likely to be several XET isoenzymes with different expression patterns, and identify those XET-related proteins potentially involved in leaf elongation.