Peptide autoinducers of staphylococcal pathogenicity

葡萄球菌致病性肽自诱导剂

• 批准号: 8669288
• 负责人: Richard P. Novick
• 金额: $ 15.12万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-31 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8669288
• 关键词: Active Sites; Affect; Agonist; Air; Amino Acids; Attenuated; Bacteria; Bacterial Proteins; Binding; Binding Sites; Blood Circulation; Body Weight decreased; C-terminal; Cellular biology; Complex; Detection; Distal; Floods; Genus staphylococcus; Goals; Grant; Histidine; Hydrolysis; Image; Infection; Ligand Binding; Ligands; Luciferases; Maps; Medical center; Membrane; Modeling; Molecular; Monitor; Morbidity - disease rate; Mus; Outcome; Pathogenicity; Peptides; Phosphorylation; Phosphotransferases; Production; Prokaryotic Cells; Protein Secretion; Proteins; Protomer; Receptor Inhibition; Reporter; Running; Side; Signal Transduction; Site; Solutions; Specificity; Structure; System; Temperature; Testing; Therapeutic; Time; Treatment Efficacy; Virulence; Virulent; Water; Work; cost; design; dimer; kidney infection; mortality; mutant; novel; protein-histidine kinase; receptor; receptor binding; receptor function; renal abscess; research study; subcutaneous; transmission process

The devastating impact of Super Storm Sandy on the NYU Medical center was caused by the total flooding of the lower levels of the entire Medical Center, including the Smilow Vivarium, which was under 15’ of water. This caused not only the loss of electric power, air circulation, running water, etc., but also the destruction of our IVIS imaging camera and the loss of all of our mice, 30 of which were at that time being used for an experiment on the effect of the AIP on kidney infections following the IV inoculation of virulent staphylococci. We had been using bacteria tagged with a luciferase reporter to monitor the fate of the infecting bacteria by imaging in the IVIS. Although our luciferase reporters worked well for the subcutaneous infection model, we found, during a subsequent experiment that our luciferase constructs did not provide a sufficiently strong signal for the detection of bacteria in the kidney abscesses which develop following IV inoculation of staphylococci and so we had to use other parameters, such as weight loss, morbidity, and mortality to evaluate the results. Consequently, we spent a considerable amount of time developing new reporters with higher luciferase activities or with fluorescent proteins (E2- crimson and iRFP) that are excited and emit in the near infra-red. The newer luciferase constructs were being used in the experiment that was in progress at the time of the storm. Since these strains were still being tested, they were stored temporarily on plates in the cold room and in a -20 freezer. Among the consequences of the storm, described in more detail in the accompanying application for a for-cost extension of our other grant, R01-AI22159, were the loss of power for these freezers and the cold room, resulting in the loss of all these temporary cultures, and in our entire supply of AIP’s, which were stored in solution in a +4 refrigerator, whose temperature reached 50C during the power outage, causing hydrolysis of the thiolactone bond essential for AIP activity.

超级风暴桑迪对纽约大学医疗中心的破坏性影响是由总 洪水淹没了整个医疗中心的下层，包括Smilow Vivarium， 在15“水中。这不仅造成电力、空气流通、自来水等的损失， 但也破坏了我们的IVIS成像相机，我们所有的老鼠都死了，其中30只是 当时用于IV给药后AIP对肾脏感染的影响的实验， 接种烈性葡萄球菌。 我们一直在使用荧光素酶报告标记的细菌来监测感染者的命运。 通过在IVIS中成像来检测细菌。尽管我们的荧光素酶报告基因在皮下 感染模型，我们发现，在随后的实验中，我们的荧光素酶结构没有 提供足够强的信号用于检测肾脓肿中的细菌 因此，我们必须使用其他参数，如体重 损失率、发病率和死亡率来评估结果。因此，我们花了相当多的钱 开发具有更高荧光素酶活性或荧光蛋白（E2- 深红色和iRFP），它们被激发并发出近红外线。新的荧光素酶构建体 在风暴发生时正在进行的实验中使用。由于这些菌株 仍在测试时，将它们暂时储存在冷藏室和-20 ℃冰箱中的平板上。 在风暴的后果中，在所附的申请中有更详细的描述， 我们的另一项赠款R01-AI22159的成本扩展是这些冷冻机的电力损失， 冷藏室，导致所有这些临时培养物的损失，以及我们所有AIP的供应， 将其储存在+4冰箱中的溶液中，其温度在功率期间达到50 C 中断，导致AIP活性所必需的硫代内酯键水解。