Antibody-inducing properties of a prototype bivalent herpes simplex virus/enterotoxigenic Escherichia coli DNA vaccine

The antibody-inducing properties of a bacterial/viral bivalent DNA vaccine (pRECFA), expressing a peptide composed of N- and C-terminal amino acid sequences of the herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) fused with an inner segment encoding the major structural subunit of enterotoxigenic Escherichia coli (ETEC) CFA/I fimbriae (CFA/I), was evaluated in BALB/c mice following intramuscular immunization. The bivalent pRECFA vaccine elicited serum antibody responses, belonging mainly to the IgG2a subclass, against both CFA/I and HSV gD proteins. pRECFA-elicited antibody responses cross-reacted with homologous and heterologous ETEC fimbrial antigens as well as with type 1 and type 2 HSV gD proteins, which could bind and inactivate intact HSV-2 particles. On the other hand, CFA/I-specific antibodies could bind but did not neutralize the adhesive functions of the bacterial CFA/I fimbriae. In spite of the functional restriction of the antibodies targeting the bacterial antigen, the present evidence suggests that fusion of heterologous peptides to the HSV gD protein represents an alternative for the design of bivalent DNA vaccines able to elicit serum antibody responses.     

Cloning of laminin gamma2 cDNA and chromosome mapping of the genes for the dog adhesion ligand laminin 5

Overexpression of the gamma2 chain of laminin-5 has been linked to tumor invasion and an unfavorable prognostic value, but the role of this adhesion molecule in cancer progression remains unclear. Because dog models of human cancers provide the opportunity of clarifying the relation between laminin-5 and tumor malignancy we have isolated and characterized the cDNA of dog gamma2 chain. Comparative analysis of the nucleotide sequence revealed high identity between the dog and the human gamma2, including the intermolecular molecule binding sites and the regulatory promoter sequences. Moreover, expression of a recombinant human gamma2 chain in dog keratinocytes results in assembly and secretion of hybrid laminin-5 molecules, which underscore the functional relevance of the gamma2 conserved domains. We have also determined the syntenic location of the dog laminin-5 loci on CFA7. Our study provides a basis for therapeutical approaches of epithelial cancers of gamma2 using dogs as large animal models.     

The analgesic effect induced by capsaicin is enhanced in inflammatory states

Agonists of the vanilloid receptor type 1 (VR1), such as capsaicin, induce an analgesic effect following an initial excitatory response. It has been demonstrated that the vanilloid system plays an important role in inflammatory hyperalgesia. In accordance, we show that the VR1 antagonist capsazepine (30 microg; i.pl.) prevented the thermal hyperalgesia induced by carrageenan or complete Freund's adjuvant (CFA) in mice. Furthermore, we studied whether this inflammation-induced activation of the vanilloid system could enhance the analgesic properties of capsaicin. A single administration of capsaicin (10 microg; i.pl.) induced in control mice an analgesic effect that lasted for 2 days. In contrast, in carrageenan-treated animals, the analgesic effect of this dose of capsaicin lasted for 6 days and in CFA-treated mice for 30 days. This prolongation of capsaicin-induced analgesia during inflammation was mediated through VR1 since it was completely blocked by coadministration of capsazepine (10 microg). Licking behavior induced by capsaicin in carrageenan- and CFA-treated mice was greater than in control animals. However, although capsaicin induced a more prolonged analgesia in CFA-treated mice, the licking behavior was greater in the carrageenan-treated group, suggesting that the prolongation of analgesia is independent of the initial nociceptive input. Overall, these results show that the analgesic effects of capsaicin are importantly enhanced during inflammation, supporting the fact that the stimulation of VR1 could perhaps constitute a suitable strategy to avoid inflammatory hyperalgesia.     

Cellular aspects of tolerance. VI. The effect of age on responsiveness and tolerance induceability of SJL mice

New born and 3-week-old SJL mice but not 8–12-week-old animals could be rendered tolerant to rabbit γ-globulin. Animals reconstituted with thymus cells from 12-week-old donors and bone marrow cells from 3-week-old donors showed resistance to tolerance induction. Animals reconstituted with bone marrow cells from 12-week-old animals and thymus cells from 3-week-old donors could be rendered tolerant. Earlier work has shown that tolerance could be induced in older animals, if they were deprived of competent accessory cells. It was suggested that a lesion in the thymus cell population is expressed through the mediation of accessory cells. The possibility of a relation between resistance to tolerance induction and lymphoid malignancies was discussed.     

Efficient generation of antigen-specific cellular immunity by vaccination with poly(gamma-glutamic acid) nanoparticles entrapping endoplasmic reticulum-targeted peptides

Because antigen-specific cytotoxic T-lymphocytes (CTLs) are major effector cells in tumor immunity, more efficient delivery of tumor-associated antigens to the major histocompatibility complex class I-presentation pathway in antigen-presenting cells (APCs) will substantially contribute to establish more effective cancer immunotherapy. Herein, we demonstrated that a combinational approach based on the antigen-delivery system using poly(gamma-glutamic acid) nanoparticles (gamma-PGA NPs) and an endoplasmic reticulum (ER)-transport system containing an ER-insertion signal sequence (Eriss) significantly enhanced the ability of a peptide vaccine to induce cellular immune responses, including CTL activity. Immunization with gamma-PGA NPs entrapping Eriss-conjugated antigenic peptides markedly amplified and activated CTLs and interferon-gamma-secreting cells specific for the antigen, whereas no cellular immune responses were detected following vaccination with only one of the systems alone. Our data provide evidence that efficient delivery of antigenic peptides into APCs, as well as active ER-translocation of antigenic peptides in APCs should be considered in the development of peptide-based cancer immunotherapy.     

Enrichment of bitumen-degrading microorganisms

Enrichment cultures in a percolation apparatus with two different soil types supplemented with bitumen, were used to follow growth on this complex substrate. Microscopic techniques allowed visualisation of bacteria on solid surfaces of bitumen and soil particles; quantification was only possible in the percolation fluid. In the latter, changes in pH and amount of organic material provided evidence for metabolic activities and degradation of the substrate bitumen.     

鸡羽毛制备复合氨基酸铁铵的研究

本文介绍了以鸡羽毛为原料,采用盐酸水解,经与铁络合、调ＰＨ值制备复合氨基酸铁铵的方法。收率为４４．２％,总铁含量９．６％,游离铁含量０．４２％。废物充分利用,工艺简单易行。     

肠毒素大肠杆菌定居因子抗原基因CFA-I在痢疾杆菌中的表达

通过体外重组的方法 ,构建了包含asd基因的重组表达质粒 pZHY2 1,与福氏志贺氏菌Fwl0 1构成了宿主 载体平衡致死系统 ,Western印迹结果表明 ,在没有抗生素条件选择的情况下 ,稳定表达肠毒素大肠杆菌定居因子抗原CFA I。电镜结果显示 ,在重组菌株的菌体表面 ,表达产物能够装配成菌毛。重组菌通过口服和鼻饲免疫小鼠后 ,可以诱生CFA I的抗体 ;同时可以检测到分泌型IgA产生 ,表明重组菌可以诱导相应的粘膜免疫反应     

Tissue distribution of cytochrome c oxidase isoforms in mammals. Characterization with monoclonal and polyclonal antibodies

Monoclonal and polyclonal antibodies specific to the two isoforms of subunit VIa of bovine cytochrome c oxidase were generated and used to study the tissue distribution of this subunit pair in beef, human and rat. The so-called H-(heart) form was found exclusively in heart and skeletal muscle, whereas the so-called L-(liver) form was the only isoform present in brain, kidney, liver and smooth muscle. Little or no L-form was detected in skeletal muscle. In bovine heart no subunit VIa-L was detected, while in human heart the subunit VIa-H and VIa-L isoforms were present in roughly equal proportions. These results imply that, in humans, the deficiency of a subunit VIa isoform may have a different effect on the physiology of heart then on the physiology of skeletal muscle.     

Nonparametric Comparison of Longitudinal Response Patterns from Paired Samples Using Configural Frequency Analysis

Nonparametric methods of Configural Frequency Analysis (CFA) for comparing longitudinal response patterns are discussed. Characteristics of diagonally symmetric frequency distributions are mapped with hypotheses on the locus of treatment effects. Test statistics for the analysis of local dependencies are discussed. In particular, applications to paired samples are considered. A biomedical example is given from research on the effects of spasmolytic drugs. Local CFA-testing is compared with global testing. Alternative strategies of testing and the respective underlying assumptions are discussed.