不同方法生产的短棒状杆菌生物学活性比较

提高短棒状杆菌疫苗临床疗效,解决接种后局部硬结,发烧等不良副反应。在菌体菌苗的基础上,采用超声、破碎、离心、胰酶消化脱脂的方法提取细胞壁做脾激活抑瘤试验生物学检定。结果显示,纯化的短棒状杆菌细胞壁可使小鼠脾激活指数达到4.06。能抑制艾氏腹水瘤的生长,而菌体脾指数3.18。实验证实,纯化的短棒杆菌细胞壁菌苗的活性好于菌体菌苗。     

短棒状杆菌外用制剂安全性的评价

研究短棒状杆菌外用剂型,为体外用药开辟新途径。将短棒状杆菌接种在适宜培养基上,连续扩增传代,培养终液稀释成不同浓度。依照新药管理办法,在动物体内进行各种安全性试验。结果显示:经急性毒性试验、长期毒性试验、生殖毒性试验,皮肤光敏试验、局部刺激试验等测定,在动物体内未见到毒性反应。该试验为短棒状杆菌外用制剂的开发提供了安全性依据。     

厌氧棒状杆菌群抗血清的血清学分析

厌氧棒状杆菌具有刺激网状内皮系统、抗感染、调节免疫反应、抑制肿瘤等一系列生物学活性。近年来制备的厌氧棒状杆菌菌苗作为肿瘤免疫治疗剂受到了重视。这类细菌的分类较混乱,1966年Prevot将其分为12个属,但其中不少为同义名。1972年Johnson和Cummins用细胞壁分析、DNA同源性以及血清学研究分为3大群4个血清学群;[即疮疱丙酸杆菌     

细胞壁缺陷对白喉棒状杆菌致病性影响的研究

检测白喉棒状杆菌稳定L型对动物的致病性,探讨细胞壁缺陷对白喉棒状杆菌致病性的影响及其可能的分子机制。采用氨苄青霉素在非高渗培养基内诱导并获得产毒性白喉棒状杆菌稳定L型纯培养物。收集白喉棒状杆菌稳定L型纯培养物及其代谢产物,将收集的高于细菌型10 000倍浓度的白喉棒状杆菌稳定L型纯培养物及其代谢产物皮内注射家兔,观察局部注射部位皮肤或全身的病理改变。分别采用对流免疫电泳(CIEP)和SDS-不连续聚丙烯酰胺凝胶电泳(SDS-PAGE)检测白喉棒状杆菌稳定L型可溶性代谢产物中的白喉毒素蛋白质。结果显示,白喉棒状杆菌稳定L型不能引起动物局部或全身发生异常表现,在其可溶性代谢产物中并未检测到白喉毒素蛋白质。提示细胞壁缺陷变异可影响白喉棒状杆菌产生白喉毒素蛋白质,从而使其丧失了产生外毒素致病的作用。     

细胞壁缺陷白喉棒状杆菌致病作用的观察及其毒力基因检测

目的比较白喉棒状杆菌亲代细菌型及其稳定L型动物致病性的差异,了解白喉棒状杆菌稳定L型变异的特点,探讨其变异的性质及其与细胞壁缺陷突变的关系。方法用氨苄青霉素在非高渗培养基内人工诱导产毒性白喉棒状杆菌为稳定L型。采用白喉棒状杆菌稳定L型纯培养物及其代谢产物皮内感染家兔,观察局部感染部位皮肤或全身的病理改变。采用微量法提取白喉棒状杆菌稳定L型的染色体DNA,用Tox基因特异性引物进行PCR扩增以检测毒素蛋白结构基因,并进行序列测定和分析。结果白喉棒状杆菌在氨苄青霉素作用下可发生细胞壁缺陷而成为L型,白喉棒状杆菌稳定L型不能引起动物局部或全身发生异常表现,该稳定L型的传代培养物可仍然保留同其亲代细菌型一致的Tox基因及其核苷酸序列。结论提示细胞壁缺失将导致白喉棒状杆菌与产生毒素蛋白有关结构基因在宿主菌细胞内的表达受到抑制,以致使白喉棒状杆菌稳定L型丧失了产生外毒素致病的作用。     

棒状乳杆菌FZU63发酵对红茶汤风味品质改善的作用

乳酸菌发酵可赋予茶饮料独特的香气与滋味,且可改变其物质组成,产生益生因子等。目前,针对乳酸菌在不同发酵阶段对茶汤中风味物质形成影响的研究较少。本研究以从中国传统泡菜中筛选获得的棒状乳杆菌FZU63为发酵菌株,对不同发酵阶段红茶汤中的挥发性香气成分、还原糖、游离氨基酸、有机酸等含量的变化过程进行分析,并对发酵红茶汤的感官品质进行评价。结果表明,棒状乳杆菌FZU63以红茶汤中的葡萄糖、果糖、甘露糖和木糖作为发酵过程中的主要碳源物质。红茶汤经棒状乳杆菌FZU63发酵作用后,香气成分丰度显著增加,且主要香气组分结构发生改变,发酵红茶汤在花香、坚果香的基础上增添了水果香;此外,部分苦味氨基酸含量下降,甜味和鲜味氨基酸含量增加;并且,乳酸、苹果酸、柠檬酸等有机酸含量在发酵过程中呈现积累。同时,感官评定结果表明棒状乳杆菌FZU63发酵可改善红茶汤的感官品质,且在发酵48h后达到较优。本文系统分析了经棒状乳杆菌发酵不同阶段对红茶汤风味的影响,可为乳酸菌发酵茶饮料的品质控制与产业化应用提供理论参考。     

N—CWS对纯系小鼠Lewis肺癌移植瘤的抗癌效应——不同菌种及不同剂型的筛选研究

诺卡氏放线菌细胞壁骨架(Nocardia-Cell wall skeleton),以下简称N-CWS)作为恶性肿瘤的免疫治疗剂,具有疗效较好而副作用少的特点,国内外已有报导,并认为优于卡介苗及厌氧棒状杆菌细胞壁骨架制剂。诺卡     

植物病原真菌广谱拮抗菌的筛选鉴定及发酵条件初步研究

通过对峙培养法筛选到对8种植物病原真菌具有拮抗作用的芽孢杆菌菌株,通过测量10-30℃下的抑菌圈直径发现AFB037菌株在不同温度下抑菌效果最好。通过表型和16S rDNA序列分析将AFB037菌株鉴定为多粘类芽孢杆菌。AFB037菌株抗菌成分的产生需要真菌菌体成分的诱导,提示其有效成分可能为细胞壁降解酶类。     

百日咳杆菌生物学活性成分研究与应用

本文介绍了百日咳杆菌产生的各种生物学活性成分的研究及应用进展，主要涉及这些成分的理化性质，生物学活性，对有效成分的应用情况也作了介绍。     

酚处理珊瑚色奴卡菌对小鼠S—180实体瘤的抗瘤效应

奴卡放线菌是介于细菌和真菌之间的革兰氏阳性微生物。据山村雄一、东市郎等报导,奴卡放线菌的细胞壁骨架(CWS)具有较强的抗癌作用[2.3.4.5]。其抗癌的有效成分与BCC—CWS相同。主要有分枝酸、阿拉怕-半乳聚糖、粘多肽。其抗癌机制尚不十分清楚。目前认为主要在于诱生干扰素,调动T淋巴细胞及巨噬细胞的活性。其抗癌作用比BCG、CP(短小棒状杆菌)为强,副作用比     

Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography

The bacterial cell wall is critical for the determination of cell shape during growth and division, and maintains the mechanical integrity of cells in the face of turgor pressures several atmospheres in magnitude. Across the diverse shapes and sizes of the bacterial kingdom, the cell wall is composed of peptidoglycan, a macromolecular network of sugar strands crosslinked by short peptides. Peptidoglycan’s central importance to bacterial physiology underlies its use as an antibiotic target and has motivated genetic, structural, and cell biological studies of how it is robustly assembled during growth and division. Nonetheless, extensive investigations are still required to fully characterize the key enzymatic activities in peptidoglycan synthesis and the chemical composition of bacterial cell walls. High Performance Liquid Chromatography (HPLC) is a powerful analytical method for quantifying differences in the chemical composition of the walls of bacteria grown under a variety of environmental and genetic conditions, but its throughput is often limited. Here, we present a straightforward procedure for the isolation and preparation of bacterial cell walls for biological analyses of peptidoglycan via HPLC and Ultra Performance Liquid Chromatography (UPLC), an extension of HPLC that utilizes pumps to deliver ultra-high pressures of up to 15,000 psi, compared with 6,000 psi for HPLC. In combination with the preparation of bacterial cell walls presented here, the low-volume sample injectors, detectors with high sampling rates, smaller sample volumes, and shorter run times of UPLC will enable high resolution and throughput for novel discoveries of peptidoglycan composition and fundamental bacterial cell biology in most biological laboratories with access to an ultracentrifuge and UPLC.     

The safety of the preparation viroden for vertebrate animals]

Toxicity-pathogenicity test of viroden, a new preparation, and its acting agent--a mosquito densonucleosis virus (MDV) has been carried out on warm-blooded animals. It is shown that the preparation is not toxic for laboratory animals (white common mice, rats, guinea pigs, rabbits), chicken embryos and cell cultures of warm-blooded animals. The MDV is not adapted to a warm-blooded organism with different ways of introduction and in passages. Using electron and luminescent microscopy, serological reactions, specific test systems and a biological test for sensitive insects no explicit or latent infection was found in animals, chicken embryos and cell cultures of vertebrates with primary infection and in passages. Sensibilized animals shown an immunological rearrangement of the organism proceeding by the retarded hypersensitivity type.     

Whole plant cell wall characterization using solution-state 2D NMR

Recent advances in nuclear magnetic resonance (NMR) technology have made it possible to rapidly screen plant material and discern whole cell wall information without the need to deconstruct and fractionate the plant cell wall. This approach can be used to improve our understanding of the biology of cell wall structure and biosynthesis, and as a tool to select plant material for the most appropriate industrial applications. This is particularly true in an era when renewable materials are vital to the emerging bio-based economies. This protocol describes procedures for (i) the preparation and extraction of a biological plant tissue, (ii) solubilization strategies for plant material of varying composition and (iii) 2D NMR acquisition (for typically 15 min-5 h) and integration methods used to elucidate lignin subunit composition and lignin interunit linkage distribution, as well as cell wall polysaccharide profiling. Furthermore, we present data that demonstrate the utility of this new NMR whole cell wall characterization procedure with a variety of degradative methods traditionally used for cell wall compositional analysis.     

Mutations of the secondary cell wall

It has not been possible to isolate a number of crucial enzymes involved in plant cell wall synthesis. Recent progress in identifying some of these steps has been overcome by the isolation of mutants defective in various aspects of cell wall synthesis and the use of these mutants to identify the corresponding genes. Secondary cell walls offer numerous advantages for genetic analysis of plant cell walls. It is possible to recover very severe mutants since the plants remain viable. In addition, although variation in secondary cell wall composition occurs between different species and between different cell types, the composition of the walls is relatively simple compared to primary cell walls. Despite these advantages, relatively few secondary cell wall mutations have been described to date. The only secondary cell wall mutations characterised to date, in which the basis of the abnormality is known, have defects in either the control of secondary cell wall deposition or secondary cell wall cellulose or lignin biosynthesis. These mutants have, however, provided essential information on secondary cell wall biosynthesis.     

Preparative laser capture microdissection and single-pot cell wall material preparation: a novel method for tissue-specific analysis

In adaptation to their function the walls of plant cell display tissue-specific variations of composition according to their developmental stage, cell type and stress of various origin. It is therefore important to obtain a precise analytical data describing the cell wall composition with respect to these different factors. In the present work, laser capture microdissection (LCM) was used for isolating different tissues from the stem of Urtica dioica L. at a semi-preparative scale. The technique was associated for the first time to a one-pot sequential cell wall preparation and hydrolysis for the carbohydrate analysis of each cell type. The results demonstrate that the combination of LCM and micro-analytical methods can provide individual cell type composition and should improve our knowledge of the biochemical diversity of cell walls in plants. This approach will be of potential interest for the understanding of the effects of stress or genetic engineering on the composition of the cell walls.     

透射电子显微镜(TEM)在孢粉学研究中的应用

透射电子显微镜作为常规的显微镜工具,已被广泛运用于观察研究生物组织超微构造,20世纪60年代开始在生物化石的研究中得到应用,特别是孢粉学,如大孢子、花粉、疑源类等的研究。通过对处理后的化石样品进行超薄切片,可观察到生物组织中保存下来的有机质壁及内含物的显微构造,对孢粉系统分类学及个体发育学的研究有重要的作用。即使是古生代的或更早期的生物样品也有一些保存下来的有价值的组织可供于超微构造的研究,只是样品在进行前期处理及超薄切片过程中会遇到一些技术问题。文章简要阐述这些具体技术问题。     

Protein-lipid-lipopolysaccharide association in the superficial layer of Spirillum serpens cell walls.

The backing layer of the Spirillum serpens VHA cell wall, which supports and is bonded to the outer, structured protein layer, was isolated and shown to be similar in composition to the same elements of the outer membrane. It contained a lipopolysaccharide that was similar, but not identical, to that of the intact wall and the same phospholipids. The interaction of the isolated wall lipopolysaccharide with the loosely bound wall lipids provided lamellae, whose surfaces were an effective template for a lifelike reassembly of the isolated outer-layer hexagonal protein in the presence of Ca2+. Assembly did not take place on pure lipopolysaccharide, which dispersed in differing forms. A lipid-lipopolysaccharide-water interface appeared to be required as a template surface for the assembly. Lipopolysaccharide from Pseudomonas aeruginosa was able to replace that of S. serpens in the template. These observations suggest that lipid-lipopolysaccharide complexes are highly ordered, and this order is important to the nucleation and assembly of the protein array.     

Mutanase induction in Trichoderma harzianum by cell wall of Laetiporus sulphureus and its application for mutan removal from oral biofilms

The cell wall material from fruiting bodies of Laetiporus sulphureus has been suggested as a new alternative to mutan for the mutanase induction in Trichoderma harzianum. Structural analyses revealed that the alkali-soluble wall fraction from this polypore fungus contained 56.3% of (1-->3)-linked alpha-glucans. When the strain T. harzianum F-340 was grown on a cell wall preparation from L. sulphureus, the maximal enzyme productivity obtained after 3 days of cultivation was 0.71 U/ml. This yield was about 1.8-fold higher than that achieved on mutan, known so far as the best, but expensive and inaccessible, inducer of mutanase production. Cell-wall-induced mutanase showed a high hydrolytic potential in reaction with a dextranasepretreated mutan, where maximal degrees of saccharification and solubilization of this biopolymer (80% and 100%, respectively) were reached in 3 h at 45oC. The mutanase preparation was also effective in degradation of streptococcal mutan and its removal from oral biofilms, especially in a mixture with dextranase.     

The extensins

The plant cell was includes a matrix which is species specific and which chages in composition during growth and development. Characterization of the protein component of the wall matrix has resulted in the purification of extensin and the genes which encode it. Analysis of the protein sequences for the extensins has provided clues about the types of interactions which may occur as the chemistry and architecture of the cell wall accommodate growth and development.