这项工作是由宝来惠康事业奖和国家卫生部授予AI074832研究院庚寅刘的支持。

1。准备葡萄球菌感染（感染前两天） <ol><li>接种的MRSA loopful从股市文化血液琼脂（胰酪胨大豆琼脂（TSA））板。 </li><li>检查血琼脂平板上溶血表型（每个殖民地周围明确区）。 </li><li>选择一个殖民地，与其他实验是一致的，具有溶血性表型。 </li><li>接种3毫升托德休伊特肉汤（THB）的殖民地，与适当的抗生素在必要时，在单元皑皑管15毫升。 </li><li>在37 ° C晃动在220转过夜。 </li></ol> 2。准备小鼠感染（感染的前一天） <ol><li>根据老鼠的大小，削去一个3 × 4厘米的面积上的小鼠背部的皮毛。 </li><li>剃光〜5毫米3头发卸妆霜（从当地药店购买）。 </li><li>让头发卸妆霜，在皮肤表面孵育约1分钟。 </li><li> DDH 2 O湿纸巾</li><li>用湿纸巾擦去头发卸妆霜。 </li></ol> 3。准备MRSA的感染（感染当天） <ol><li>过夜的细菌培养在1:500到1:1000稀释预热泰铢10毫升，50毫升的螺丝帽管。 </li><li>在37℃孵育，用颤抖的在220转， 直到 540到达2.5℃，约2.5小时。 </li><li> 10分钟在3225 XG收集离心MRSA在4 ° C。 </li><li>上清液。 </li><li>细菌沉淀重新悬浮在10毫升贝科磷酸盐缓冲液（DPBS）。 </li><li>重复步骤3和4。 </li><li>在DPBS细菌沉淀重悬在所需浓度。 </li><li>串行稀释的菌悬液从10 1 10 8。 </li><li>板到血琼脂平板稀释的菌悬液。 </li><li>孵育37℃过夜板。 </li><li>观察和记录溶血性表型的接种。 </li><li>计数菌落形成单位（CFU）数盘上。 </li><li>板CFU数为基础计算的接种。 </li></ol> 4。皮下的MRSA感染（感染当天，感染后3天） <ol><li>皮下注入100μL，剃光面积的菌悬液。 </li><li> 3至5个小时的观察动物注射后，以确保小鼠还活着。 </li><li>每天观察病变的动物的回，并记录病变区，每日需要的时候。应包括病变的观察记录病灶的大小和形态。皮肤和肌肉病变的定量乘以病变的长度和宽度。需要被分解成更小的对称件，每件用同样的方法测量不规则形的病灶。病变的测量，也可以使用电脑辅助组织形态学评估方案（ImageJ; <a href="http://rsb.info.nih.gov/ij">开源</a> http://rsb.info.nih.gov/ij/从美国国立卫生研究院提供）2。 </li><li>异氟醚吸入颈椎脱位，感染后第3天牺牲的动物。 </li><li>观察并记录在皮肤上的病变。 </li><li>尼克的皮肤，一双无菌剪刀。 </li><li>关闭仔细剥离皮肤，观察并记录肌肉病变。 </li><li>切除病灶及周边地区约2 - 5毫米。 </li><li>收集脾和肾。 </li><li>均质使用组织匀浆的组织或反复应用到离心管中含有的组织和100μL的DPBS，从1 mL注射器柱塞。 </li><li>以每管加入900μL的DPBS。 </li><li>涡5分钟的样品。 </li><li>串行稀释与DPBS 10 1至10月6日暂停。 </li><li>在临屋板的稀释悬浮板。 </li></ol> 5。代表性的成果: <ol><li>后立即皮下注射，将观察到的气泡在皮肤表面感染时做正确（图1A）。在皮肤表面观察到，如果没有气泡注射过深，这可能会影响感染的结果（病灶的大小，CFU）的。 </li><li>需要连续稀释的菌悬液的CFU需要血琼脂平板上，每个实验研究。这一步将产生重要的信息:1）接种是否有同质化的表型; 2）感染的确切可行的含菌量。 </li><li>可能被评定在不同时间点后感染病灶和细菌生存。在这里，我们审查了感染，感染后第3天。分析病灶大小在皮肤表面（图1b:1X 10 9;图1D:1 × 10 7 CFU）和肌肉表面（图1C:1X 10月9日 </s上涨;图1E:1 × 10 7 CFU）。病灶面积等于长度（毫米）x宽（mm）可用于评估组织损伤（图2 A和B）来衡量。这一结果可能有助于确定一个给定的致病因子引起的组织损伤的程度。此外，病灶切除，并在DPBS匀浆，并镀以定量的CFU（图2C），这表明，在感染部位的细菌的可行性。 </li></ol><img src="/files/ftp_upload/2528/2528fig1.jpg" alt="图1" /> 图1。皮肤和肌肉病变。CD1小鼠皮下接种于一个MRSA（LAC）的侧翼。照片拍摄时间为0，感染后第3天。 （一）时间后感染（PI），（二） - （G）的第3天PI，B，D和F:皮损，C，E和G:肌肉病变，B和C:1X 10月9日 CFU感染; D和E:1 × 10 7 CFU感染; F和G:模拟。 <img src="/files/ftp_upload/2528/2528fig2.jpg" alt="图2" /> 图2。在感染部位，感染后第3天，皮肤和感染MRSA（LAC）的CD1小鼠肌肉病变皮肤病灶的大小和活菌计数。 （一）皮肤病灶的大小。 （二）肌肉病变的大小。 （c）总组织的CFU。 H:1 × 10 9 CFU接种，L:1 × 10 7 CFU接种。 *:P &lt;0.05，Mann - Whitney检验。

<ol>
	<li>Ahn, J. Y., Song, J. Y., Yun, Y. S., Jeong, G., Choi, I. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+of+Staphylococcus+aureus-infected+septic+mice+by+suppression+of+early+acute+inflammation+and+enhanced+antimicrobial+activity+by+ginsan.">Protection of Staphylococcus aureus-infected septic mice by suppression of early acute inflammation and enhanced antimicrobial activity by ginsan.</a> FEMS Immunol Med Microbiol. 46, 187-197 (2006).</li><li>Girish, V., Vijayalakshmi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Affordable+image+analysis+using+NIH+Image/ImageJ.">Affordable image analysis using NIH Image/ImageJ.</a> Indian J Cancer. 41, 47-47 (2004).</li><li>Hahn, B. L., Onunkwo, C. C., Watts, C. J., Sohnle, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+dissemination+and+cutaneous+damage+in+a+mouse+model+of+staphylococcal+skin+infections.">Systemic dissemination and cutaneous damage in a mouse model of staphylococcal skin infections.</a> Microb Pathog. 47, 16-23 (2009).</li><li>Hruz, P., Zinkernagel, A. S., Jenikova, G., Botwin, G. J., Hugot, J. P., Karin, M., Nizet, V., Eckmann, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NOD2+contributes+to+cutaneous+defense+against+Staphylococcus+aureus+through+alpha-toxin-dependent+innate+immune+activation.">NOD2 contributes to cutaneous defense against Staphylococcus aureus through alpha-toxin-dependent innate immune activation.</a> Proc Natl Acad Sci U S A. 106, 12873-12878 (2009).</li><li>Ji, Y., Zhang, B., Van, S. F., Horn, P. W. a. r. r. e. n., Woodnutt, G., Burnham, M. K., Rosenberg, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+critical+staphylococcal+genes+using+conditional+phenotypes+generated+by+antisense+RNA.">Identification of critical staphylococcal genes using conditional phenotypes generated by antisense RNA.</a> Science. 293, 2266-2269 (2001).</li><li>Somerville, G. A., Beres, S. B., Fitzgerald, J. R., DeLeo, F. R., Cole, R. L., Hoff, J. S., Musser, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+serial+passage+of+Staphylococcus+aureus:+changes+in+physiology,+virulence+factor+production,+and+agr+nucleotide+sequence.">In vitro serial passage of Staphylococcus aureus: changes in physiology, virulence factor production, and agr nucleotide sequence.</a> J Bacteriol. 184, 1430-1437 (2002).</li><li>Tseng, C. W., Kyme, P., Low, J., Rocha, M. A., Alsabeh, R., Miller, L. G., Otto, M., Arditi, M., Diep, B. A., Nizet, V., Doherty, T. M., Beenhouwer, D. O., Liu, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staphylococcus+aureus+Panton-Valentine+leukocidin+contributes+to+inflammation+and+muscle+tissue+injury.">Staphylococcus aureus Panton-Valentine leukocidin contributes to inflammation and muscle tissue injury.</a> PLoS One. 4, e6387-e6387 (2009).</li><li>von Kockritz-Blickwede, M., Rohde, M., Oehmcke, S., Miller, L. S., Cheung, A. L., Herwald, H., Foster, S., Medina, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunological+mechanisms+underlying+the+genetic+predisposition+to+severe+Staphylococcus+aureus+infection+in+the+mouse+model.">Immunological mechanisms underlying the genetic predisposition to severe Staphylococcus aureus infection in the mouse model.</a> Am J Pathol. 173, 1657-1668 (2008).</li></ol>

<ol><li>小鼠的皮肤及软组织感染模型是一个强大的工具，在体内的病原体毒力评估。 s 的致病性皮肤及软组织感染的金黄色葡萄球菌可能会有所不同，取决于一些参数。这些措施包括:接种量，细菌的生长阶段，接种的深入，对小鼠的年龄，和小鼠的遗传背景 7 ，8。研究毒力的功能，使用这种模式时，关键是这些参数控制，以确保结果将是一致的。我们已经研究了几株小鼠，包括SKH1（无...

<table><tbody><tr><td>THB</td><td>VWR international</td><td>95025-314</td><td></td></tr><tr><td>DPBS</td><td>Invitrogen</td><td>21-031-CV</td><td></td></tr><tr><td>1 ml syringe</td><td>BD Biosciences</td><td>309602</td><td></td></tr><tr><td>27G1/2 needle</td><td>BD Biosciences</td><td>305109</td><td></td></tr><tr><td>Sheep Blood Agar (TSA)</td><td>VWR international</td><td>90004-328</td><td></td></tr></tbody></table>

subcutaneous infection of methicillin resistant staphylococcus aureus (mrsa)

MRSA是一种全球范围内对公众健康构成威胁，现在占一半以上在美国的所有软组织感染MRSA的皮肤及软组织感染。在软组织感染，肌炎，pyomyositis，坏死性筋膜炎已被越来越多的报道在协会与MRSA从社会中产生的。要了解MRSA和宿主免疫导致更严重的感染之间的相互作用，动物模型的可用性是至关重要的，允许主机和细菌因素的研究。几个感染模型已被引入到评估的S.发病在浅表皮肤感染的金黄色葡萄球菌。在这里，我们描述，检查皮肤，皮下和肌肉病理的皮下感染模型。

Watch this Scientific Journal Video about Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus MRSA at JoVE.com

Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus MRSA

利用小鼠的皮肤及软组织感染模型，以评估耐甲氧西林金黄色葡萄球菌（MRSA）和宿主免疫反应的毒性作用。在这里，我们提出了一个皮肤及软组织感染，皮下感染模型。

皮下感染耐甲氧西林金黄色葡萄球菌（MRSA）的

MRSA is a worldwide threat to public health, and MRSA skin and soft-tissue infections now account for more than half of all soft-tissue infections in the ...

subcutaneous-infection-methicillin-resistant-staphylococcus-aureus

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus (MRSA)

22.9K 次观看.  Cedars-Sinai Medical Center. 利用小鼠的皮肤及软组织感染模型，以评估耐甲氧西林金黄色葡萄球菌（MRSA）和宿主免疫反应的毒性作用。在这里，我们提出了一个皮肤及软组织感染，皮下感染模型。

视频: Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus MRSA

Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria

A structurally transformed lytic bacteriophage having a broad host range of Staphylococcus aureus strains and a penicillin-binding protein (PBP 2a) antibody conjugated latex beads have been utilized to create a biosensor designed for discrimination of methicillin resistant (MRSA) and sensitive (MSSA) S. aureus species 1,2. The lytic phages have been converted into phage spheroids by contact with water-chloroform interface. Phage spheroid monolayers have been moved onto a biosensor surface by Langmuir-Blodgett (LB) technique 3. The created biosensors have been examined by a quartz crystal microbalance with dissipation tracking (QCM-D) to evaluate bacteria-phage interactions. Bacteria-spheroid interactions led to reduced resonance frequency and a rise in dissipation energy for both MRSA and MSSA strains. After the bacterial binding, these sensors have been further exposed to the penicillin-binding protein antibody latex beads. Sensors analyzed with MRSA responded to PBP 2a antibody beads; although sensors inspected with MSSA gave no response. This experimental distinction determines an unambiguous discrimination between methicillin resistant and sensitive S. aureus strains. Equally bound and unbound bacteriophages suppress bacterial growth on surfaces and in water suspensions. Once lytic phages are changed into spheroids, they retain their strong lytic activity and show high bacterial capture capability. The phage and phage spheroids can be utilized for testing and sterilization of antibiotic resistant microorganisms. Other applications may include use in bacteriophage therapy and antimicrobial surfaces.

Lytic phage biosensors and antibody beads are able to discriminate between methicillin resistant (MRSA) and sensitive staphylococcus bacteria. The phages were immobilized by a Langmuir-Blodgett method onto a surface of a quartz crystal microbalance sensor and worked as broad range staphylococcus probes. Antibody beads recognize MRSA.

生物传感器检测抗生素耐药金黄色葡萄球菌细菌

A structurally transformed lytic bacteriophage having a broad host range of Staphylococcus aureus strains and a penicillin-binding protein (PBP 2a) ...

科研

生物工程

Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus (MRSA) in Rat

Endovascular infections, including endocarditis, are life-threatening infectious syndromes1-3. Staphylococcus aureus is the most common world-wide cause of such syndromes with unacceptably high morbidity and mortality even with appropriate antimicrobial agent treatments4-6. The increase in infections due to methicillin-resistant S. aureus (MRSA), the high rates of vancomycin clinical treatment failures and growing problems of linezolid and daptomycin resistance have all further complicated the management of patients with such infections, and led to high healthcare costs7, 8. In addition, it should be emphasized that most recent studies with antibiotic treatment outcomes have been based in clinical settings, and thus might well be influenced by host factors varying from patient-to-patient. Therefore, a relevant animal model of endovascular infection in which host factors are similar from animal-to-animal is more crucial to investigate microbial pathogenesis, as well as the efficacy of novel antimicrobial agents. Endocarditis in rat is a well-established experimental animal model that closely approximates human native valve endocarditis. This model has been used to examine the role of particular staphylococcal virulence factors and the efficacy of antibiotic treatment regimens for staphylococcal endocarditis. In this report, we describe the experimental endocarditis model due to MRSA that could be used to investigate bacterial pathogenesis and response to antibiotic treatment.

Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus MRSA in Rat

Experimental rat endocarditis model due to methicillin-resistant S. aureus.

耐甲氧西林实验性心内膜炎模型<em&gt;金黄色葡萄球菌</em&gt;（MRSA）的大鼠

Endovascular infections, including endocarditis, are life-threatening infectious syndromes1-3. Staphylococcus aureus is the most common world-wide cause ...

免疫与感染

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

Staphylococcal Cassette Chromosome mec (SCCmec) typing is a very important molecular tool for understanding the epidemiology and clonal strain relatedness of methicillin-resistant Staphylococcus aureus (MRSA), particularly with the emerging outbreaks of community-associated MRSA (CA-MRSA) occurring on a worldwide basis. Traditional PCR typing schemes classify SCCmec by targeting and identifying the individual mec and ccr gene complex types, but require the use of many primer sets and multiple individual PCR experiments. We designed and published a simple multiplex PCR assay for quick-screening of major SCCmec types and subtypes I to V, and later updated it as new sequence information became available. This simple assay targets individual SCCmec types in a single reaction, is easy to interpret and has been extensively used worldwide. However, due to the sophisticated nature of the assay and the large number of primers present in the reaction, there is the potential for difficulties while adapting this assay to individual laboratories. To facilitate the process of establishing a MRSA SCCmec assay, here we demonstrate how to set up our multiplex PCR assay, and discuss some of the vital steps and procedural nuances that make it successful.

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

We demonstrate a simple multiplex PCR assay for quick-screening and typing of Staphylococcal Cassette Chromosome mec (SCCmec) types I-V for methicillin-resistant Staphylococcus aureus, and provide some of the vital steps and procedural nuances that make it successful for adapting this assay to individual laboratories.

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Staphylococcal Cassette Chromosome mec (SCCmec) typing  is a very important molecular tool for understanding the epidemiology and  clonal strain ...

Deep Dermal Injection As a Model of Candida albicans Skin Infection for Histological Analyses

The skin is an extremely extended organ of the body and, due to this large surface, it is continuously exposed to microorganisms. Skin damage can easily lead to infections in the dermis which can, in turn, result in the dissemination of pathogens into the bloodstream. Understanding how the immune system fights infections at the very early stage and how the host can eliminate the pathogens is an important step to set the base for future therapeutic interventions. Here we describe a model of Candida albicans infection that can visualize the processes that occur early during an infection, including when the pathogen has passed the epithelial barrier, as well as the immune response elicited by the C. albicans invasion. We used this infection model to perform histological analyses that show the immune cells that infiltrate the skin as well as the presence and localization of the pathogen. Samples collected after the infection can be processed for RNA extraction.

Deep Dermal Injection As a Model of Candida albicans Skin Infection for Histological Analyses

Here we describe a protocol that allows histological and molecular analysis of skin samples after Candida albicans intradermal injection. This protocol maintains the structural integrity of the skin and allows for the localization of tissue-resident or newly recruited immune cells as well as the pathogen distribution.

深部真皮注射液作为白色念珠菌皮肤感染模型的组织学分析

The skin is an extremely extended organ of the body and, due to this large surface, it is continuously exposed to microorganisms. Skin damage can easily ...

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

Staphylococcus aureus (S. aureus)&#160;infections, including methicillin resistant stains, are an enormous burden on the healthcare system. With incidence rates of S. aureus infection climbing annually, there is a demand for additional research in its&#160;pathogenicity. Animal models of infectious disease advance our understanding of the host-pathogen response and lead to the development of effective therapeutics. Neutrophils play a primary role in the innate immune response that controls S. aureus infections by forming an abscess to wall off the infection and facilitate bacterial clearance; the number of neutrophils that infiltrate an S. aureus skin infection often correlates with disease outcome. LysM-EGFP mice, which possess the enhanced green fluorescent protein (EGFP) inserted in the Lysozyme M (LysM) promoter region (expressed primarily by neutrophils), when used in conjunction with in vivo whole animal fluorescence imaging (FLI) provide&#160;a means of quantifying neutrophil emigration noninvasively and longitudinally into wounded skin. When combined with a bioluminescent S. aureus strain and sequential in vivo whole animal bioluminescent imaging (BLI), it is possible to longitudinally monitor both the neutrophil recruitment dynamics and in vivo bacterial burden at the site of infection in anesthetized mice from onset of infection to resolution or death. Mice are more resistant to a number of virulence factors produced by S. aureus that facilitate effective colonization and infection in humans. Immunodeficient mice provide a more sensitive animal model to examine persistent S. aureus infections and the ability of therapeutics to boost innate immune responses. Herein, we characterize responses in LysM-EGFP mice that have been bred to MyD88-deficient mice (LysM-EGFP&#215;MyD88-/- mice) along with wild-type LysM-EGFP mice to investigate S. aureus skin wound infection. Multispectral simultaneous detection enabled study of neutrophil recruitment dynamics by using in vivo FLI, bacterial burden by using in vivo BLI, and wound healing longitudinally and noninvasively over time.

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

An approach is described for real-time detection of the innate immune response to cutaneous wounding and Staphylococcus aureus infection of mice. By comparing LysM-EGFP mice (which possess fluorescent neutrophils) with a LysM-EGFP crossbred immunodeficient mouse strain, we advance our understanding of infection and the development of approaches to combat infection.

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

Staphylococcus aureus (S. aureus)&#160;infections, including methicillin resistant stains, are an enormous burden on the healthcare system. With incidence ...

Standardized In vitro Assays to Visualize and Quantify Interactions between Human Neutrophils and Staphylococcus aureus Biofilms

Neutrophils are the first line of defense deployed by the immune system during microbial infection. In vivo, neutrophils are recruited to the site of infection where they use processes such as phagocytosis, production of reactive oxygen and nitrogen species (ROS, RNS, respectively), NETosis (neutrophil extracellular trap), and degranulation to kill microbes and resolve the infection. Interactions between neutrophils and planktonic microbes have been extensively studied. There have been emerging interests in studying infections caused by biofilms in recent years. Biofilms exhibit properties, including tolerance to killing by neutrophils, distinct from their planktonic-grown counterparts. With the successful establishment of both in vitro and in vivo biofilm models, interactions between these microbial communities with different immune cells can now be investigated. Here, techniques that use a combination of traditional biofilm models and well-established neutrophil activity assays are tailored specifically to study neutrophil and biofilm interactions. Wide-field fluorescence microscopy is used to monitor the localization of neutrophils in biofilms. These biofilms are grown in static conditions, followed by the addition of neutrophils derived from human peripheral blood. The samples are stained with appropriate dyes prior to visualization under the microscope. Additionally, the production of ROS, which is one of the many neutrophil responses against pathogens, is quantified in the presence of a biofilm. The addition of immune cells to this established system will expand the understanding of host-pathogen interactions while ensuring the use of standardized and optimized conditions to measure these processes accurately.

Standardized In vitro Assays to Visualize and Quantify Interactions between Human Neutrophils and Staphylococcus aureus Biofilms

The present protocol describes the study of neutrophil-biofilm interactions. Staphylococcus aureus biofilms are established in vitro and incubated with peripheral blood-derived human neutrophils. The oxidative burst response from neutrophils is quantified, and the neutrophil localization within the biofilm is determined by microscopy.

标准化 体外 测定，以可视化和量化人中性粒细胞与 金黄色葡萄球菌 生物膜之间的相互作用

Neutrophils are the first line of defense deployed by the immune system during microbial infection. In vivo, neutrophils are recruited to the site of ...

Evaluating the Immune Response of a Nanoemulsion Adjuvant Vaccine Against Methicillin-Resistant Staphylococcus aureus (MRSA) Infection

Nanoemulsion adjuvant vaccines have attracted extensive attention because of their small particle size, high thermal stability, and ability to induce validly immune responses. However, establishing a series of comprehensive protocols to evaluate the immune response of a novel nanoemulsion adjuvant vaccine is vital. Therefore, this article features a rigorous procedure to determine the physicochemical characteristics of a vaccine (by transmission electron microscopy [TEM], atomic force microscopy [AFM], and dynamic light scattering [DLS]), the stability of the vaccine antigen and system (by a high-speed centrifuge test, a thermodynamic stability test, SDS-PAGE, and western blot), and the specific immune response (IgG1, IgG2a, and IgG2b). Using this approach, researchers can evaluate accurately&#160;the protective effect of a novel nanoemulsion adjuvant vaccine in a lethal MRSA252 mouse model. With these protocols, the most promising nanoemulsion vaccine adjuvant in terms of effective adjuvant potential can be identified. In addition, the methods can help optimize novel vaccines for future development.

Evaluating the Immune Response of a Nanoemulsion Adjuvant Vaccine Against Methicillin-Resistant Staphylococcus aureus MRSA Infection

The present protocol prepares and evaluates the physical properties, immune response, and in vivo protective effect of a novel nanoemulsion adjuvant vaccine.

评估纳米乳剂佐剂疫苗对耐甲氧西林 金黄色葡萄球菌 （MRSA）感染的免疫反应

Nanoemulsion adjuvant vaccines have attracted extensive attention because of their small particle size, high thermal stability, and ability to induce ...

In Vivo Mouse Model of Spinal Implant Infection

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The purpose of this method is to describe a novel mouse model of spinal implant infection (SII) that was created to provide an inexpensive, rapid, and accurate in vivo tool to test potential therapeutics and treatment strategies for spinal implant infections.
In this method, we present a model of posterior-approach spinal surgery in which a stainless-steel k-wire is transfixed into the L4 spinous process of 12-week old C57BL/6J wild-type mice and inoculated with 1 x 103 CFU of a bioluminescent strain of Staphylococcus aureus Xen36 bacteria. Mice are then longitudinally imaged for bioluminescence in vivo on post-operative days 0, 1, 3, 5, 7, 10, 14, 18, 21, 25, 28, and 35. Bioluminescence imaging (BLI) signals from a standardized field of view are quantified to measure in vivo bacterial burden.
To quantify bacteria adhering to implants and peri-implant tissue, mice are euthanized and the implant and surrounding soft tissue are harvested. Bacteria are detached from the implant by sonication, cultured overnight and then colony forming units (CFUs) are counted. The results acquired from this method include longitudinal bacterial counts as measured by in vivo S. aureus bioluminescence (mean maximum flux) and CFU counts following euthanasia.
While prior animal models of instrumented spine infection have involved invasive, ex vivo tissue analysis, the mouse model of SII presented in this paper leverages noninvasive, real time in vivo optical imaging of bioluminescent bacteria to replace static tissue study. Applications of the model are broad and may include utilizing alternative bioluminescent bacterial strains, incorporating other types of genetically engineered mice to contemporaneously study host immune response, and evaluating current or investigating new diagnostic and therapeutic modalities such as antibiotics or implant coatings.

The protocol describes a novel in vivo mouse model of spinal implant infection where a stainless-steel k-wire implant is infected with bioluminescent Staphylococcus aureus Xen36. Bacterial burden is monitored longitudinally with bioluminescent imaging and confirmed with colony forming unit counts after euthanasia.

脊柱植入物感染的体内小鼠模型

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The ...

医学

<meta charset="utf8"></head><body> 氧化锌纳米颗粒的合成及其抗菌作用分析

Preparation of Zinc Oxide Nanoparticles and the Evaluation of their Antibacterial Effects

Nosocomial bacterial infections have become increasingly challenging due to their inherent resistance to antibiotics. The emergence of multidrug-resistant bacterial strains in hospitals has been attributed to the extensive and varied use of antibiotics, further exacerbating the problem of antibiotic resistance. Metal nanomaterials have been widely studied as an alternative solution for eradicating antibiotic-resistant bacterial cells. Metallic nanoparticles attack bacterial cells through various mechanisms, such as the release of antibacterial ions, generation of reactive oxygen species, or physical disruption, against which bacteria cannot develop resistance. Among the actively researched antimicrobial metal nanoparticles, zinc oxide nanoparticles, which are FDA-approved, are known for their biocompatibility and antibacterial properties. In this study, we focused on successfully developing a precipitation method for synthesizing zinc oxide nanoparticles, analyzing the properties of these nanoparticles, and conducting antimicrobial tests. Zinc oxide nanoparticles were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), ultraviolet/visible spectroscopy, and X-ray diffraction (XRD). Antibacterial tests were conducted using the broth microdilution test with the multidrug-resistant strains of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. This study demonstrated the potential of zinc oxide nanoparticles in inhibiting the proliferation of antibiotic-resistant bacteria.

<meta charset="utf8"></head><body>作者聚焦：探索氧化锌纳米颗粒在克服抗生素耐药性中的抗菌作用

In this study, zinc oxide nanoparticles were synthesized using a precipitation method. The antibacterial effect of the synthesized particles was tested against multidrug-resistant methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa bacterial strains.

氧化锌纳米颗粒的制备及其抗菌效果评价

Nosocomial bacterial infections have become increasingly challenging due to their inherent resistance to antibiotics. The emergence of multidrug-resistant ...

皮下感染耐甲氧西林金黄色葡萄球菌（MRSA）的

摘要

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此视频中的章节

生物传感器检测抗生素耐药金黄色葡萄球菌细菌

耐甲氧西林实验性心内膜炎模型

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深部真皮注射液作为白色念珠菌皮肤感染模型的组织学分析

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

标准化体外测定，以可视化和量化人中性粒细胞与金黄色葡萄球菌生物膜之间的相互作用

评估纳米乳剂佐剂疫苗对耐甲氧西林金黄色葡萄球菌（MRSA）感染的免疫反应

脊柱植入物感染的体内小鼠模型

氧化锌纳米颗粒的制备及其抗菌效果评价

氧化锌纳米颗粒的制备及其抗菌效果评价

皮下感染耐甲氧西林金黄色葡萄球菌（MRSA）的

摘要

探索更多视频

此视频中的章节

生物传感器检测抗生素耐药金黄色葡萄球菌细菌

耐甲氧西林实验性心内膜炎模型

??PCR?????????????????????MEC??????V

深部真皮注射液作为白色念珠菌皮肤感染模型的组织学分析

一种评价金黄色葡萄球菌感染先天免疫反应的小鼠模型

标准化 体外 测定，以可视化和量化人中性粒细胞与 金黄色葡萄球菌 生物膜之间的相互作用

评估纳米乳剂佐剂疫苗对耐甲氧西林 金黄色葡萄球菌 （MRSA）感染的免疫反应

脊柱植入物感染的体内小鼠模型

氧化锌纳米颗粒的制备及其抗菌效果评价

氧化锌纳米颗粒的制备及其抗菌效果评价

标准化体外测定，以可视化和量化人中性粒细胞与金黄色葡萄球菌生物膜之间的相互作用

评估纳米乳剂佐剂疫苗对耐甲氧西林金黄色葡萄球菌（MRSA）感染的免疫反应