新生隐球菌生物膜动物模型构建及PMT4对生物膜形成的影响

目的构建新生隐球菌的PMT4基因缺陷株及新生隐球菌生物膜的体内、外模型;研究PMT4基因对生物膜形成的影响。方法采用PCR介导的长侧翼同源重组的方法敲除新生隐球菌H99的PMT4基因;采用基础培养基96孔板培养的方法建立生物膜体外模型;兔中心静脉插管、管内放置聚苯乙烯薄膜的方法建立生物膜动物模型;用倒置显微镜、共聚集激光扫描显微镜、MTT、CFU计数等方法研究PMT4缺陷株与野生株生物膜的异同。结果新生隐球菌在体内、外模型中均能形成生物膜;PMT4缺陷株与野生株生物膜在生物量和结构方面存在明显差异。结论本实验的生物膜动物模型可行;PMT4基因缺陷可造成隐球菌生物膜代谢活性下降,并形成假菌丝样结构。     

植入物动物感染模型在葡萄球菌生物膜研究中的应用与进展

葡萄球菌生物膜引起的持续性感染及耐药性问题一直是临床治疗的难题,围绕生物膜形成分子机制的研究成为防治葡萄球菌生物膜相关感染的关键。建立葡萄球菌感染动物模型有利于研究体内生物膜形成、扩散、致病机制及药物的体内抗生物膜效果评估等。然而,动物体内生物膜形成的影响因素多,如动物种类、植入材料、接种部位、感染剂量、观察时间及评估方法等均会影响体内生物膜形成。结合本课题研究,系统地总结了近40年来葡萄球菌生物膜感染动物模型,重点综述动物模型的建立方法、适用范围及优缺点,为葡萄球菌生物膜感染的防治提供理论依据。     

生态安全评价研究述评

生态安全评价是可持续发展研究的一个新领域,也是建立生态安全预警系统及进行环境管理的基础。首先介绍了与生态安全评价相关的几个概念:生态风险评价、生态系统健康评价、生态系统服务功能评价及生态承载力分析,进而对当前生态安全评价的指标体系进行了评述,并在此基础上总结了国内外生态安全评价的方法,包括数学模型法、生态模型法、景观生态学法及数字地面模型法;最后,提出在评价实践中存在的问题及今后发展的3个方向,静态的现状评价转向动态的预测和预警、与现代科学技术相结合、与基础理论相结合。     

实验性自身免疫性心肌炎动物模型的建立与评价

实验性自身免疫性心肌炎动物模型（ experimental autoimmune myocarditis, EAM）作为自身免疫性心肌炎机制、诊断、药物治疗等实验研究的重要载体,在心肌炎科学研究中发挥着重要的作用。本文针对EAM模型的建立及评价予以综述,分别对心肌自身抗原、心肌自身抗原表位、活化自身免疫细胞、干酪乳杆菌细胞壁成分等介质诱导的EAM模型进行整理总结,分析比较各造模方法的特点并阐述EAM模型的评价方法,为建立客观的EAM模型提供有效依据。     

口腔种植材料表面特性与生物膜形成研究进展

人类口腔环境为微生物提供了适宜生存的条件,多种微生物在牙齿表面形成了由基质包裹相互粘附的口腔生物膜,口腔生物膜是口腔微生物生存、代谢和致病的基础。随着1965年Brnemark种植体在临床上的成功应用,种植相关材料周围致病菌导致的种植体周围炎成为种植修复最常见的并发症之一,影响种植修复的远期效果。种植体周围炎引起了许多关注,并且口腔种植材料表面的特性和口腔生物膜的形成密切相关。本文就种植材料及天然牙齿表面的生物膜形成、种植材料表面特性对口腔生物膜及细菌粘附的影响因素、增强种植材料抗菌性能的方法以及未来的研究方向等作一综述。     

白念珠菌生物膜体内外模型研究进展

白念珠菌引起的慢性难治性感染与其形成的生物膜密切相关,生物膜的诸多生物学特征明显不同于游离菌.合适的生物膜模型的构建对于研究白念珠菌生物膜的形成机制以及药物干预至关重要.该文结合作者自身的研究,对目前国内外关于白念珠菌生物膜体外、体内多种模型的构建方法作一综述.     

固体载体支承的双层膜系统

固态载体支承的双层膜的各种制备过程都简便易行,较脂质体等系统有重复性好、其物化性质可严格控制等优越性,并可将膜蛋白质分子镶嵌其中,是研究生物膜的良好模型．由于其研究方法日益成熟,固态载体支承的双层膜系统越来越成为研究生物膜与膜蛋白的有利工具之一．对固态载体支承的双层膜的制备技术和研究方法进行了系统的综述,并列举了一些在膜生物物理化学领域的应用．     

两种检测方法评价厚朴对白念珠菌黏附的影响

正白念珠菌是人体常见的机会致病性真菌,可存在于正常人口腔、上呼吸道、肠道及阴道,易在义齿、人工插管如尿道插管、气管插管等植入性设备表面定植并进一步形成生物膜。生物膜的形成可保护菌体免受药物的杀伤和免疫系统的攻击。黏附是生物膜成膜的关键,也是白念珠菌致病的关键步骤[1]。本实验拟通过结晶紫法与XTT法观察中药厚朴对白念珠菌黏附的影响,并对两种检测方法进行评价。1材料与方法     

生物燃料系统分析模型

定量化系统分析模型是生物燃料潜力预测、影响分析及制定技术路线图和政策目标的重要工具。生物燃料供应链涉及很多行业,需要进行基于农 (林) 业、能源、经济和环境等多学科领域的综合分析。以下从生物燃料系统分析所需解决的问题出发,梳理了国内外生物燃料系统分析的一般方法,重点对农 (林) 业系统模型、能源系统模型、综合评价模型、微观成本、能耗和排放分析模型以及生物燃料专项宏观分析模型的主要优缺点和适用性进行了分析,给出了相应的应用实例,并强调根据特定的研究问题,选择具有不同适用性的模型和研究路线。有助于研究人员和政策制定者更好地了解生物燃料系统分析方法,也为我国学术研究机构建立生物燃料分析模型提供了参考。     

土壤重金属污染评价方法

土壤中的重金属长期停留和积累在环境中,对生态环境和人体健康存在诸多现实和潜在风险,受到越来越多的关注.因此,评价土壤中的重金属污染程度对于环境和健康问题有着重要意义.评价土壤中重金属的污染程度需选用一种或几种正确的评价方式.本文综述了目前国内外常用于土壤中重金属评价的指数法、模型指数法和基于重金属形态分析、有效态含量和总量、人体健康风险以及GIS和地统计学的评价方法.其中,指数法主要包括内梅罗指数法、富集因子法、地累积指数法、潜在生态危害指数法;模型指数法则主要包括模糊数学模型、灰色聚类模型及层次分析法等.文章对各种评价方法进行了综述,并指出使用各种方法的优越性、局限性.     

动物模型在细菌生物被膜研究中的应用与展望

细菌生物被膜的形成与其致病性、耐药性密切相关,在许多由细菌导致的慢性、亚慢性感染中发挥着重要作用。动物模型广泛应用于细菌生物被膜相关感染的研究中,为其致病机理和控制策略的探究提供了强有力的科学工具。因此,本文系统阐述了哺乳类(鼠、兔、猪等)和非哺乳类(黑腹果蝇、斑马鱼、秀丽隐杆线虫等)动物模型在细菌生物被膜相关研究中的应用,并对动物模型在细菌生物被膜研究中的应用前景进行了展望,以期为研究由生物被膜导致的相关感染而选择理想动物模型提供理论支撑,从而对生物被膜感染导致的潜在危害进行防控。     

Predicting biofilm deformation with a viscoelastic phase-field model: Modeling and experimental studies

Biofilms commonly develop in flowing aqueous environments, where the flow causes the biofilm to deform. Because biofilm deformation affects the flow regime, and because biofilms behave as complex heterogeneous viscoelastic materials, few models are able to predict biofilm deformation. In this study, a phase-field (PF) continuum model coupled with the Oldroyd-B constitutive equation was developed and used to simulate biofilm deformation. The accuracy of the model was evaluated using two types of biofilms: a synthetic biofilm, made from alginate mixed with bacterial cells, and a Pseudomonas aeruginosa biofilm. Shear rheometry was used to experimentally determine the mechanical parameters for each biofilm, used as inputs for the model. Biofilm deformation under fluid flow was monitored experimentally using optical coherence tomography. The comparison between the experimental and modeling geometries, for selected horizontal cross sections, after fluid-driven deformation was good. The relative errors ranged from 3.2 to 21.1% for the synthetic biofilm and from 9.1 to 11.1% for the P. aeruginosa biofilm. This is the first demonstration of the effectiveness of a viscoelastic PF biofilm model. This model provides an important tool for predicting biofilm viscoelastic deformation. It also can benefit the design and control of biofilms in engineering systems.     

Linoleic acid inhibits Pseudomonas aeruginosa biofilm formation by activating diffusible signal factor‐mediated quorum sensing

Bacterial biofilm formation causes serious problems in various fields of medical, clinical, and industrial settings. Antibiotics and biocide treatments are typical methods used to remove bacterial biofilms, but biofilms are difficult to remove effectively from surfaces due to their increased resistance. An alternative approach to treatment with antimicrobial agents is using biofilm inhibitors that regulate biofilm development without inhibiting bacterial growth. In the present study, we found that linoleic acid (LA), a plant unsaturated fatty acid, inhibits biofilm formation under static and continuous conditions without inhibiting the growth of Pseudomonas aeruginosa. LA also influenced the bacterial motility, extracellular polymeric substance production, and biofilm dispersion by decreasing the intracellular cyclic diguanylate concentration through increased phosphodiesterase activity. Furthermore, quantitative gene expression analysis demonstrated that LA induced the expression of genes associated with diffusible signaling factor‐mediated quorum sensing that can inhibit or induce the dispersion of P. aeruginosa biofilms. These results suggest that LA is functionally and structurally similar to a P. aeruginosa diffusible signaling factor (cis‐2‐decenoic acid) and, in turn, act as an agonist molecule in biofilm dispersion.     

In vivo biofilm formation on a soft denture liner in elderly patients with controlled diabetes

doi: 10.1111/j.1741‐2358.2010.00428.x In vivo biofilm formation on a soft denture liner in elderly patients with controlled diabetes Objectives: This in vivo study evaluated the influence of controlled diabetes on biofilm formation on a soft denture liner in elderly patients. Background: Soft denture lining materials are more susceptible to microbial colonisation than denture base acrylic resins. Especially in the elderly, several predisposing factors may accumulate leading to an increased probability of biofilm development that may result in candidiasis, a significant clinical oral disease. Materials and methods: Volunteers wearing complete dentures were divided into two groups (n = 20): diabetic patients with controlled glycaemia, and healthy patients. In both groups, a silicone‐based soft liner was placed in a recess created at the base of the maxillary dentures. Subjects cleaned the prosthesis three times a day. Biofilm formed on the liner was quantified at various time points (baseline, two, four and six weeks). Data were analysed by two‐way repeated measures anova and Tukey’s test (α = 0.05). Results: There was no statistical difference in biofilm formation for any of the time points between controlled diabetes patients and healthy patients. Conclusion: The results suggest that the control of diabetes in elderly patients provides the same levels of biofilm formation when compared to healthy individuals.     

The needs of denture-brushing in geriatrics: clinical aspects and perspectives

doi: 10.1111/j.1741‐2358.2011.00557.x
The needs of denture‐brushing in geriatrics: clinical aspects and perspectives Introduction: Oral and denture hygiene are often defective in particular with dependent persons such as geriatric subjects. The reasons are the lack of hygiene education of the subjects or those caring for them. Consequently, oral hygiene is often neglected, resulting in poor oral health and an increase in the presence of local or general infections. Objective: This paper is a report of brushing effectiveness on microbial biofilm deposits on dentures of subjects participating in a specific oral hygiene programme. Materials and methods: Thirty‐nine dentures of 30 subjects were assessed for 2 weeks following an educational brushing programme. Microbial biofilm was recovered using fluoresceine and then scanned and quantified by ‘Mesurim’ software three times: before study, after 1 and 2 weeks. Results: The repeated measurement procedures showed a decrease in the percentage of biofilm present (F = 15, p < 0.001) whatever the type of denture (partial or complete) and for all biomaterials. Conclusion: Regular denture‐brushing can improve local hygiene. Consequently, decreasing the biofilm surface can reduce the prevalence of oral pathogens, thereby contributing to the general prevention of the risks of infections such as pneumotisis.     

Biofilm formation of Candida albicans on the surface of a soft denture‐lining material

Background: Soft denture lining‐materials are more susceptible to microbial adhesion than hard denture base acrylic resin. Poor oral hygiene and Candida albicans infection are common among elderly denture wearers as these patients usually have difficulty in keeping them clean. Purpose: To evaluate the influence of the oral hygiene methods on the formation of a biofilm over a soft denture‐lining material. Material and methods: Twenty volunteers were randomly separated into two groups: G1 and G2. Ten volunteers performed daily hygiene of the prostheses with a soft toothbrush and toothpaste. The G2 performed a treatment identical to G1 but also immersed the prostheses in sodium hypochlorite 0.5% for 20 min, once a week. Quantification of the mean score values of biofilm formation at different times were statistically analysed using analysis of variance and Tukey’s test (α = 0.05). Results: G1 (0.65 ± 0.52) showed the lowest mean score values of biofilm formation. There was statistical difference between G1 and G2. The highest mean score values were found at 6 weeks (1.3 ± 1.08) and were statistically different from other times. Conclusion: The oral hygiene methods had a significant effect in the formation of the biofilm over a soft denture‐lining material.     

Rapid depletion of dissolved oxygen in 96‐well microtiter plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration

Biofilm‐related research using 96‐well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96‐well plates irrespective of the oxygen concentration in the gaseous environment in which the plates are incubated. These data indicate that depletion of dissolved oxygen during growth of bacterial biofilm cultures in 96‐well plates may significantly influence biofilm production. Furthermore higher inoculum cell concentrations are associated with more rapid consumption of dissolved oxygen and higher levels of S. epidermidis biofilm production. Our data reveal that oxygen depletion during bacterial growth in 96‐well plates may significantly influence biofilm production and should be considered in the interpretation of experimental data using this biofilm model. Biotechnol. Bioeng. 2009;103: 1042–1047. © 2009 Wiley Periodicals, Inc.     

Experimental and statistical methods to evaluate antibacterial activity of a quaternary pyridinium salt on planktonic,biofilm-forming,and biofilm states

Robust evaluation and comparison of antimicrobial technologies are critical to improving biofilm prevention and treatment. Herein, a multi-pronged experimental framework and statistical models were applied to determine the effects of quaternary pyridinium salt, 4-acetyl-1-hexadecylpyridin-1-ium iodide (QPS-1), on Streptococcus mutans in the planktonic, biofilm-forming and biofilm cell states. Minimum inhibitory and bactericidal concentrations (MIC and MBC, respectively) were determined via common methods with novel application of statistical approaches combining random effects models and interval censored data to estimate uncertainties. The MICs and MBCs for planktonic and biofilm-forming states ranged from 3.12 to 12.5 μg ml^?1, with biofilm values only ≈ 8 times higher. Potent anti-biofilm activity and reactive structural features make QPS-1 a promising antibacterial additive for dental and potentially other biomedical devices. Together, the experimental framework and statistical models provide estimates and uncertainties for effective antimicrobial concentrations in multiple cell states, enabling statistical comparisons and improved characterization of antibacterial agents.     

Comparative analysis of biofilm levels in complete upper and lower dentures after brushing associated with specific denture paste and neutral soap

Objectives: The objective of this study was to compare and correlate biofilm levels in complete upper and lower prosthesis after brushing, associated with specific paste and soap, by means of computerised methodology. Materials and methods: Forty‐five complete denture wearers were selected and instructed to brush their prostheses (Soft Oral B 40) three times a day for 3 weeks with water (Control), specific paste for complete dentures (Corega Brite) (Experiment 1) and neutral soap (Experiment 2). The study was based on a cross‐over model and a wash‐out period was not included. For biofilm quantification, the internal surfaces were dyed (neutral red 1%), photographed (Canon EOS Digital) and the disclosed biofilm was measured with the Image Tool 2.0 software. The products were assessed by means of a questionnaire regarding their hygiene properties and acceptance. Results: The variance analysis indicated that the lower prostheses exhibited a mean biofilm percentage, significantly higher than the upper prostheses and that brushing with paste (Experiment 1) was more effective than soap (Experiment 2) and, in turn, this was more effective than water (Control). There was a high biofilm correlation (Pearson correlation) between both prostheses. Both products were well accepted by the patients, but the most favoured one was the paste. Conclusions: This was effective in controlling the biofilm and can be used preventatively in the maintenance of oral health by wearers of complete dentures. This is important where the lower prosthesis can harbour microorganisms which may act as a reservoir for other areas of the mouth and thus enhance the importance of proper hygiene.