BACKGROUND & AIMS: :The prevalence of IgG antibodies against Mycobacterium leprae recombinant culture filtrate protein-10 (rCFP-10) was investigated in serum samples from 56 leprosy patients, 15 tuberculosis (TB) patients, 14 other skin-diseased patients and 20 healthy subjects. On classifying the patients into bacterial index (BI)-positive and BI-negative groups, the assay showed 83.3 % (15/18) sensitivity for detection of BI-positive leprosy patients. On the other hand, the sensitivity for detection of BI-negative patients was 18.4 % (7/38). None of the 15 TB patients and 14 other skin-diseased patients was positive; however, only one out of 20 healthy individuals was positive, indicating that antibody response to culture filtrate protein-10 (CFP-10) was highly specific (98.0 %; 48/49). Statistically, the performance of the CFP-10-based assay was found to be comparable (P>0.05) with that of an anti-phenolic glycolipid-I (PGL-I) antibody-detecting assay. Thus, M. leprae CFP-10 is potentially a specific antigen for measuring antibody response in BI-positive leprosy patients. Being a secreted antigen, CFP-10 may act as a marker for the viability of M. leprae inside the host, and hence its serological potential is worth exploring for application in monitoring the response of patients with BI-positive leprosy (a highly infectious form) during the course of chemotherapy. When comparing the bacteriological and serological results, an agreement of 82.1 % showed that seropositivity to M. leprae CFP-10 corresponded well with bacteriological criteria. Hence, CFP-10 seems to be a suitable antigen for classification of leprosy patients into BI-positive and BI-negative groups.

背景与目标： ：从56名麻风患者，15名结核病患者，14名其他皮肤病患者和20名健康受试者的血清样本中研究了抗麻风分枝杆菌重组培养物滤液蛋白-10（rCFP-10）IgG抗体的患病率。在将患者分为细菌指数（BI）阳性和BI阴性组后，该测定显示出83.3％（15/18）的灵敏度可检测BI阳性麻风病患者。另一方面，检测BI阴性患者的敏感性为18.4％（7/38）。 15例TB患者和14例其他皮肤病患者均无阳性。然而，在20个健康个体中只有1个是阳性，表明对培养滤液蛋白10（CFP-10）的抗体反应具有高度特异性（98.0％； 48/49）。从统计学上讲，发现基于CFP-10-的检测方法的性能可与抗酚糖脂I（PGL-1）抗体检测方法相媲美（P> 0.05）。因此，麻风分枝杆菌CFP-10潜在地是用于测量BI阳性麻风病患者的抗体应答的特异性抗原。 CFP-10是一种分泌性抗原，可作为宿主体内麻风分枝杆菌生存能力的标志物，因此其血清学潜力值得探索，可用于监测BI阳性麻风病患者（一种高度感染性形式）的反应）在化疗过程中。当比较细菌学和血清学结果时，82.1％的一致性表明，对麻风分枝杆菌CFP-10的血清阳性与细菌学标准非常吻合。因此，CFP-10似乎是将麻风病人分为BI阳性和BI阴性组的合适抗原。

BACKGROUND & AIMS: :Epithelial cells play an important role in orchestrating mucosal immune responses. In allergic-type inflammation, epithelial cells control the recruitment of eosinophils into the mucosa. Th2-type cytokine-driven release of eosinophil-active chemokines from epithelial cells directs eosinophil migration into the mucosal epithelium. CCR3, the main eosinophil chemokine receptor, regulates this process; however, the respective contribution of individual CCR3 ligands in eosinophil transepithelial migration is less well understood. Using an in vitro transepithelial chemotaxis system, we found that eotaxin-3 produced by IL-4-stimulated airway epithelial cells and CCR3 on eosinophils exclusively mediate eosinophil transepithelial migration. Eotaxin-3 protein levels were also increased in the nasal mucosal epithelium recovered from allergic patients as compared to non-allergic patients. Surprisingly, eotaxin-3 in IL-4-stimulated airway epithelial cells was predominantly cell surface bound, and the cell surface form was critical for eosinophil transepithelial migration. Eotaxin-3 cell surface association was partially glycosaminoglycan (GAG) dependent, but was completely protein dependent, suggesting that eotaxin-3 associates with both GAG and cell surface proteins. We thus provide evidence that cell surface-associated eotaxin-3 is the critical IL-4-dependent chemotactic signal mediating eosinophil transepithelial migration in the setting of allergic inflammation.

背景与目标： ：上皮细胞在协调粘膜免疫反应中起重要作用。在过敏型炎症中，上皮细胞控制嗜酸性粒细胞向粘膜的募集。 Th2型细胞因子驱动的嗜酸性粒细胞活性趋化因子从上皮细胞的释放引导嗜酸性粒细胞迁移到粘膜上皮中。 CCR3是主要的嗜酸性粒细胞趋化因子受体，调节这一过程。然而，单个CCR3配体在嗜酸性粒细胞上皮细胞迁移中的各自作用尚不清楚。使用体外经上皮趋化系统，我们发现由IL-4刺激的气道上皮细胞和嗜酸性粒细胞上的CCR3产生的eotaxin-3专门介导嗜酸性粒细胞跨上皮迁移。与非过敏患者相比，从过敏患者中回收的鼻粘膜上皮中的Eotaxin-3蛋白水平也有所增加。出人意料的是，IL-4刺激的气道上皮细胞中的eotaxin-3主要与细胞表面结合，并且细胞表面形式对于嗜酸性粒细胞跨上皮迁移至关重要。 Eotaxin-3细胞表面缔合部分依赖糖胺聚糖（GAG），但完全依赖蛋白质，这表明eotaxin-3与GAG和细胞表面蛋白缔合。因此，我们提供证据，在过敏性炎症的情况下，细胞表面相关的eotaxin-3是介导嗜酸性粒细胞跨上皮迁移的关键IL-4依赖性趋化信号。

BACKGROUND & AIMS: :IL-7 signals are crucial for the survival of naive and memory T cells, and the IL-7R is expressed on the surface of these cells. Following viral infection, the IL-7R is expressed on only a subset of effector CD8 T cells, and has been demonstrated to be important for the survival of these memory precursors. IL-7 message levels remain relatively constant during the T cell response to lymphocytic choriomeningitis virus, but a short-lived burst of GM-CSF is observed soon after infection. Retroviral expression of a chimeric GM-CSF/IL-7R, in which binding of GM-CSF by T cells leads to IL-7 signaling, allows for the delivery of an IL-7 signal in all effector T cells expressing the receptor. In mice infected with lymphocytic choriomeningitis virus, CD8 and CD4 T cells transduced with this chimeric receptor underwent an enhanced proliferative response compared with untransduced populations in the same host. Similarly, TCR transgenic CD8 cells expressing the chimeric receptor produced higher effector numbers during the peak of the T cell response to infection. Surprisingly, the enhanced proliferation did not lead to higher memory numbers, as the subsequent contraction phase was more pronounced in the transduced cell populations. These findings demonstrate that artificial IL-7 signaling during an infection leads to significantly increased Ag-specific effector T cell numbers, but does not result in increased numbers of memory progeny. The extent of contraction may be dictated by intrinsic factors related to the number of prior cell divisions.

背景与目标： ：IL-7信号对于幼稚和记忆T细胞的存活至关重要，而IL-7R在这些细胞的表面表达。病毒感染后，IL-7R仅在效应CD8 T细胞的一部分上表达，并已证明对这些记忆前体的存活很重要。在对淋巴细胞性脉络膜脑膜炎病毒的T细胞应答过程中，IL-7信息水平保持相对恒定，但是感染后不久就观察到了短暂的GM-CSF爆发。嵌合GM-CSF / IL-7R的逆转录病毒表达（其中T细胞与GM-CSF的结合导致IL-7信号传导）允许在表达该受体的所有效应T细胞中传递IL-7信号。在感染了淋巴细胞性脉络膜脑膜炎病毒的小鼠中，用该嵌合受体转导的CD8和CD4 T细胞与同一宿主中未转导的种群相比，具有增强的增殖反应。同样，表达嵌合受体的TCR转基因CD8细胞在感染的T细胞反应高峰期间产生更高的效应子数量。出人意料的是，增强的增殖并未导致更高的记忆数，因为随后的收缩期在转导的细胞群中更为明显。这些发现表明，感染期间的人工IL-7信号转导会导致Ag特异性效应T细胞数量显着增加，但不会导致记忆后代数量增加。收缩的程度可以由与先前细胞分裂数有关的内在因素决定。

BACKGROUND & AIMS: :Immunocompromise after a major injury is presumed to be a predisposing factor for sepsis. Mice subjected to sublethal cecal ligation and puncture (CLP) and challenged 5 days later with Pseudomonas aeruginosa had more bacterial growth in lung tissue, lower serum interferon gamma (IFN-gamma) and interleukin (IL) 12,and higher serum IL-10 when compared with sham CLP mice challenged with Pseudomonas. To test the functional significance of these alterations in cytokine production in the immune response to bacteria, we administered IFN-gamma and anti-IL-10 to post-CLP mice before the Pseudomonas challenge. Administration of IFN-gamma and anti-IL-10 did not improve bacterial clearance or mortality in post-CLP mice. In further studies, we administered IFN-gamma to IL-10 knockout mice before a challenge with P. aeruginosa. Our results showed no significant differences in bacterial clearance or mortality in IL-10 knockout mice with or without IFN-gamma treatment compared with wild-type controls. Finally, because most mortality occurred within 2 to 3 days of the Pseudomonas challenge in the aforementioned studies and was likely associated with a marked proinflammatory response, we investigated the effect of IFN-gamma and anti-IL-10 on clearance of Pseudomonas in C3H/HeJ mice, which do not mount an exaggerated proinflammatory response to endotoxin or Gram-negative bacteria. Neither clearance of the Pseudomonas bacteria nor mortality was improved in C3H/HeJ mice receiving anti-IL-10 and IFN-gamma. These results suggest that the suppressed IFN-gamma and IL-12 responses, in combination with an exaggerated IL-10 response to P. aeruginosa challenge after injury, do not correlate with bacterial clearance or survival.

背景与目标： ：大伤后的免疫功能低下被认为是败血症的诱因。进行半致死盲肠结扎和穿刺（CLP）并在5天后用铜绿假单胞菌攻击的小鼠的肺组织中细菌生长更多，血清干扰素-γ（IFN-γ）和白介素（IL）12更低，而血清IL-10更高。与假单胞菌攻击的假CLP小鼠相比。为了测试这些变化对细菌免疫反应中细胞因子产生的功能意义，我们在假单胞菌攻击之前向CLP后小鼠施用了IFN-γ和抗IL-10。在CLP后小鼠中，IFN-γ和抗IL-10的使用不能改善细菌清除率或死亡率。在进一步的研究中，我们在铜绿假单胞菌攻击之前向IL-10敲除小鼠施用了IFN-γ。我们的结果表明，与野生型对照相比，接受或未接受IFN-γ治疗的IL-10基因敲除小鼠的细菌清除率或死亡率无显着差异。最后，由于在上述研究中大多数死亡发生在假单胞菌攻击后的2到3天内，并且可能与明显的促炎反应有关，因此我们研究了IFN-γ和抗IL-10对C3H / 3中假单胞菌清除的影响HeJ小鼠，对内毒素或革兰氏阴性细菌没有过度的促炎反应。接受抗IL-10和IFN-γ的C3H / HeJ小鼠的假单胞菌细菌清除率和死亡率均未提高。这些结果表明，损伤后抑制的IFN-γ和IL-12反应，加上对损伤后铜绿假单胞菌攻击的过度IL-10反应，与细菌清除率或存活率无关。

BACKGROUND & AIMS: :Accumulating evidence indicates that Toll-like receptor (TLR) signaling adapter protein interactions with Toll/Interleukin-1 Receptor (TIR) domains present in sensory neurons may modulate neuropathic pain states. Following ligand interaction with TLRs, TIR serves to both initiate intracellular signaling and facilitate recruitment of signaling adapter proteins to the intracytoplasmic domain. Although TLR TIR is central to a number of TLR signaling cascades, its role in sensory neurons is poorly understood. In this study we investigated the degree to which TLR TIR decoy peptide modified to include a TAT sequence (Trans-Activator of Transcription gene in HIV; TAT-4BB) affected LPS-induced intracellular calcium flux and excitation in sensory neurons, and behavioral changes due to TLR4 active metabolite, morphine-3-glucuronide (M3G) exposure in vivo. TAT-4BB inhibited LPS-induced calcium changes in a majority of sensory neurons and decreased LPS-dependent neuronal excitability in small diameter neurons. Acute systemic administration of the TAT-4BB reversed M3G-induced tactile allodynia in a dose-dependent manner but did not affect motor activity, anxiety or responses to noxious thermal stimulus. These data suggest that targeting TLR TIR domains may provide novel pharmacological targets to reduce or reverse TLR4-dependent pain behavior in the rodent.

背景与目标： ：越来越多的证据表明，Toll样受体（TLR）信号转接头蛋白与感觉神经元中存在的Toll / Interleukin-1受体（TIR）域相互作用可能会调节神经性疼痛状态。配体与TLR相互作用后，TIR既可以启动细胞内信号传导，也可以促进信号传导衔接子蛋白募集到胞质内域。尽管TLR TIR是许多TLR信号级联的核心，但对其在感觉神经元中的作用知之甚少。在这项研究中，我们调查了TLR TIR诱饵肽修饰为包含TAT序列（HIV转录基因的反式激活因子； TAT-4BB）在多大程度上影响LPS诱导的细胞内钙通量和感觉神经元的兴奋，以及行为改变对TLR4活性代谢产物吗啡-3-葡糖醛酸（M3G）的体内暴露。 TAT-4BB抑制了LPS诱导的大多数感觉神经元中钙的变化，并降低了小直径神经元中LPS依赖性神经元的兴奋性。 TAT-4BB的急性全身给药以剂量依赖的方式逆转了M3G诱导的触觉异常性疼痛，但并未影响运动活动，焦虑或对有害热刺激的反应。这些数据表明，靶向TLR TIR结构域可提供新的药理学靶标，以减少或逆转啮齿动物中TLR4依赖性的疼痛行为。

BACKGROUND & AIMS: BACKGROUND:Interleukin-17 (IL-17), which is secreted by Th17 cells, is a proinflammatory cytokine that is implicated in the pathogenesis of multiple sclerosis (MS) and plays a role in nonresponse of MS patients to interferon-β (IFN-β) therapy. OBJECTIVES:The purpose of this study was to establish a correlation between nonresponders (NR) and IL-17A serum titers and binding antibodies (BAbs) to IFN-β, as well as to find a correlation between IL-17A serum levels and other features of MS patients. METHODS:Our prospective study included 72 inactive relapsing-remitting multiple sclerosis (RRMS) patients that had been treated for at least 18 months with IFN-β and 15 healthy subjects. We determined the serum levels of IL-17A and of BAbs. IL-17A levels were considered elevated (IL-17A+) if the recorded value was greater than 1.6 pg/ml. RESULTS:Twenty-seven patients (37.5%) were NR and had a significantly higher serum IL-17A level compared to the responders group. Nineteen patients (26.4%) were IL-17A+ and had had a significantly higher number of relapses in the previous year and a higher Expanded Disability Status Score. The majority of IL-17A+ patients were NR and had a shorter MS duration. CONCLUSIONS:RRMS patients with high serum IL-17A levels do not respond well to IFN-β therapy and have shorter MS duration compared to patients with low IL-17A levels. This response is not influenced by the presence of BAbs.

背景与目标： 背景：Th17细胞分泌的白细胞介素17（IL-17）是一种促炎细胞因子，与多发性硬化症（MS）的发病机制有关，并且在MS患者对干扰素-β（IFN- β）疗法。
目的：本研究的目的是建立无应答者（NR）与IL-17A血清滴度和针对IFN-β的结合抗体（BAbs）之间的相关性，以及找出IL-17A血清水平与其他特征之间的相关性MS患者。
方法：我们的前瞻性研究包括72例接受IFN-β治疗至少18个月的非活动性复发缓解型多发性硬化症（RRMS）患者和15名健康受试者。我们确定了IL-17A和BAbs的血清水平。如果记录值大于1.6 pg / ml，则认为IL-17A水平升高（IL-17A）。
结果：27名患者（37.5％）为NR，与应答者组相比，血清IL-17A水平显着更高。 19名患者（26.4％）为IL-17A，在前一年中复发率明显更高，而“扩展残疾状态评分”更高。大多数IL-17A患者为NR，MS病程较短。
结论：与低IL-17A水平的患者相比，血清IL-17A水平高的RRMS患者对IFN-β治疗的反应不佳，MS病程较短。该反应不受BAbs的存在的影响。

BACKGROUND & AIMS: OBJECTIVE:To identify early clinical factors that are correlated with death or severe disability in paediatric patients who have sustained an injury by hanging or strangulation. METHODS:A retrospective review of all patient records from January 1, 1997, to September 30, 2007, was conducted. Patient records were identified by International Classification of Diseases and Related Health Problems, Tenth Revision, Canada diagnostic codes for asphyxia, strangulation, hypoxic-ischemic encephalopathy, hanging, hypoxemia, hypoxia or anoxia. RESULTS:A total of 109 records were identified. Of these, 41 met the inclusion criteria for the study. Of 19 (46%) children who were pulse-less and received cardiopulmonary resuscitation, 16 died and the survivors were severely disabled. Of the 22 (54%) children who were found with a pulse, 18 made a full recovery. CONCLUSIONS:Children who are pulseless at discovery for hanging injuries are at high risk of death or severe disability. Early clinical and neurophysiological indicators should be applied systematically to best guide clinicians and parents in their decision making.

背景与目标： 目的：确定与因吊死或绞窄而受伤的小儿患者的死亡或严重残疾相关的早期临床因素。
方法：对1997年1月1日至2007年9月30日的所有患者记录进行回顾性回顾。通过国际疾病分类和相关健康问题分类（第十次修订版，加拿大）对窒息，窒息，缺氧缺血性脑病，悬挂，低氧血症，缺氧或缺氧的诊断代码确定了患者的记录。
结果：共鉴定到109条记录。其中有41个符合研究的纳入标准。在19名（46％）无脉搏并接受心肺复苏的儿童中，有16人死亡，幸存者严重残疾。在发现有脉搏的22名儿童中（54％），其中18名完全康复。
结论：发现悬吊伤害而无脉搏的儿童极有可能导致死亡或严重残疾。应系统地应用早期临床和神经生理指标，以最好地指导临床医生和父母进行决策。

BACKGROUND & AIMS: :IL-23 and IL-17 are pro-inflammatory cytokines. IL-23 is secreted by activated macrophages and dendritic cells, while IL-17 by Th17 cells. Serum IL-23 and IL-17 are known to be elevated in numerous inflammatory diseases including neurodegenerative diseases. The role of serum IL-23 and IL-17 in aneurysmal subarachnoid hemorrhage (aSAH) has still not been investigated. The present work investigates the serum IL-23 and IL-17 levels and their association with post hemorrhagic complications and clinical outcome in patients with aSAH. METHODS:In this study, 80 patients with aSAH (Hunt and Hess grade I-V) were prospectively recruited. We enrolled 24 control patients with lumbar spinal stenosis. Peripheral venous blood was withdrawn from controls and from aSAH patients at day 1 and day 7, allowed to clot and centrifuged to obtain serum. Enzyme linked immunoassay kits were employed to quantify the serum levels of IL-23 and IL-17 by applying 50µL of serum samples. Post hemorrhagic complications and clinical outcome were documented prospectively from patient's hospital record. RESULTS:Serum IL-23 and IL-17 levels were significantly elevated in aSAH patients at day 1 and day 7 (n=80) as compared to control patients (n=24). Further analysis after dichotomy of patients who suffered from post hemorrhagic complications including cerebral vasospasm, chronic hydrocephalus, seizures, cerebral ischemia, delayed neurological deficits showed differential correlations with different post hemorrhagic complications (Table 1). Serum IL-23 and IL-17 levels did not correlate with clinical outcome. CONCLUSION:Serum IL-23 and IL-17 levels were elevated in patients with aSAH showing upregulation of IL-23/IL-17 inflammatory axis after aSAH. Serum IL-23 and IL-17 showed differential correlations with post hemorrhagic complications and no correlation with clinical outcome.

背景与目标： ：IL-23和IL-17是促炎性细胞因子。 IL-23由活化的巨噬细胞和树突状细胞分泌，而IL-17由Th17细胞分泌。已知血清IL-23和IL-17在包括神经退行性疾病在内的许多炎性疾病中均升高。血清IL-23和IL-17在动脉瘤性蛛网膜下腔出血（aSAH）中的作用尚未进行研究。本工作调查了aSAH患者的血清IL-23和IL-17水平及其与出血后并发症和临床结局的关系。
方法：在这项研究中，前瞻性招募了80例aSAH患者（Hunt和Hess I-V级）。我们招募了24名腰椎管狭窄的对照患者。在第1天和第7天从对照和aSAH患者中抽取外周静脉血，使其凝结并离心以获得血清。采用酶联免疫分析试剂盒，通过施加50µL血清样品定量IL-23和IL-17的血清水平。从患者的住院记录中前瞻性地记录了出血后并发症和临床结局。
结果：与对照组患者（n = 24）相比，aSAH患者在第1天和第7天的血清IL-23和IL-17水平显着升高（n = 80）。二分法对患有出血后并发症（包括脑血管痉挛，慢性脑积水，癫痫发作，脑缺血，迟发性神经功能缺损）的患者进行二分法手术后的进一步分析显示，不同出血后并发症的相关性不同（表1）。血清IL-23和IL-17水平与临床结果无关。
结论：aSAH患者血清IL-23和IL-17水平升高，显示aSAH后IL-23 / IL-17炎症轴上调。血清IL-23和IL-17与出血后并发症呈差异相关，与临床结局无相关性。

BACKGROUND & AIMS: :An effective immune response to antigen requires professional antigen-presenting cell (APC), which not only present antigen, but also provide costimulation and cytokines (eg, IL-12) that drive T cell differentiation down the appropriate effector pathway (Tc1/TH1). For T cell-based immunotherapy protocols, the availability of large numbers of autologous professional APC is a major limitation because professional APC do not proliferate in vitro. T cells themselves can proliferate exponentially in vitro and have the ability to present antigen. They can also express costimulatory molecules after activation. Therefore, we hypothesized that if activated T cells were genetically modified to express proinflammatory cytokines required to polarize T cells toward a Tc1 response, they could fulfill the requirements for an abundant, autologous APC. To test this potential, T cells were activated by CD3/CD28 antibodies and pulsed with model HLA-A2+ peptides derived from CMVpp65, MAGE-3, and MART-1. Activated T-APC readily reactivated CD8 pp65 memory T cells from healthy CMV seropositive donors; however, the activation of MAGE-3 and MART-1-specific CD8 T cells required both IL-7 and IL-12, which could be provided either exogenously or by genetic modification of the T-APC. Responder T cells could be expanded to large numbers with subsequent stimulations using activated, peptide-pulsed T-APC and IL-2. Tumor antigen-specific T cell lines killed both peptide-pulsed target cells and tumor cell lines. Thus, T cells provide a platform for the generation of autologous APC that can be customized to express both antigens and therapeutic molecules for the induction of antigen-specific T cell immunity.

背景与目标： ：对抗原的有效免疫反应需要专业的抗原呈递细胞（APC），它不仅呈递抗原，而且还提供共刺激和细胞因子（例如IL-12），以驱动T细胞向适当的效应子途径（Tc1 / TH1）分化）。对于基于T细胞的免疫疗法方案，大量的自体专业APC的可用性是一个主要限制，因为专业APC不会在体外增殖。 T细胞本身可以在体外成倍增殖，并具有呈递抗原的能力。它们还可以在激活后表达共刺激分子。因此，我们假设，如果对活化的T细胞进行基因修饰以表达使T细胞朝Tc1反应极化所需的促炎细胞因子，那么它们就可以满足丰富的自体APC的要求。为了测试这种潜力，T细胞被CD3 / CD28抗体激活，并用衍生自CMVpp65，MAGE-3和MART-1的模型HLA-A2肽脉冲。激活的T-APC可以很容易地激活来自健康CMV血清反应阳性供体的CD8 pp65记忆T细胞。然而，MAGE-3和MART-1特异性CD8 T细胞的激活需要IL-7和IL-12，这可以通过外源或通过T-APC的遗传修饰来提供。可以使用激活的，肽脉冲的T-APC和IL-2在随后的刺激下将应答剂T细胞扩增为大量。肿瘤抗原特异性T细胞系杀死了肽脉冲的靶细胞和肿瘤细胞系。因此，T细胞提供了用于产生自体APC的平台，该平台可以被定制以表达抗原和治疗性分子以诱导抗原特异性T细胞免疫。

BACKGROUND & AIMS: OBJECTIVES:To characterize the naturally occurring expanded-spectrum beta-lactamase from an Escherichia coli clinical isolate and to compare it with a wild-type beta-lactamase. METHODS:The chromosome-borne ampC genes from E. coli BER and E. coli EC2 were PCR amplified, sequenced and cloned into an expression vector. Antimicrobial susceptibilities of the parental isolate and the recombinant strains were determined by agar dilution methods. Kinetic parameters were determined from purified AmpC BER and AmpC EC2. RESULTS:AmpC BER was overexpressed in its original clinical isolate because of mutations in the promoter region of its gene at positions -42 and -18. The analysis of the ampC coding sequence revealed a 6 bp insertion when compared with the wild-type sequence leading to the tandem duplication of two alanine residues inside the H-10 helix. AmpC BER-producing recombinants were resistant to ceftazidime, had reduced susceptibility to other oxyiminocephalosporins (cefotaxime and cefepime), but had a greater susceptibility to cefoxitin when compared with the recombinant expressing the wild-type beta-lactamase AmpC EC2. The affinity of AmpC BER for cephalosporins and imipenem was increased, whereas the hydrolysis rate was decreased for all these compounds. In addition, the IC50 values of clavulanic acid and tazobactam for AmpC BER were increased. CONCLUSIONS:This work sheds new light on structure-function relationships of expanded-spectrum AmpC beta-lactamases.

背景与目标： 目的：鉴定来自大肠杆菌临床分离株的天然存在的广谱β-内酰胺酶，并将其与野生型β-内酰胺酶进行比较。
方法：PCR扩增来自大肠杆菌BER和大肠杆菌EC2的染色体传播的ampC基因，测序并克隆到表达载体中。通过琼脂稀释法测定亲本分离株和重组菌株的抗药性。动力学参数由纯化的AmpC BER和AmpC EC2确定。
结果：AmpC BER在其原始临床分离株中过表达，因为其基因的启动子区域位于-42和-18位。 ampC编码序列的分析显示，与野生型序列相比，插入了6 bp，导致H-10螺旋内部两个丙氨酸残基串联重复。与表达野生型β-内酰胺酶AmpC EC2的重组体相比，产生AmpC BER的重组体对头孢他啶具有抗性，对其他氧亚氨基头孢菌素（头孢噻肟和头孢吡肟）的敏感性降低，但对头孢西丁的敏感性更高。 AmpC BER对头孢菌素和亚胺培南的亲和力增加，而所有这些化合物的水解速率均降低。此外，棒酸和他唑巴坦对AmpC BER的IC50值增加。
结论：这项工作为广谱AmpCβ-内酰胺酶的结构-功能关系提供了新的思路。

BACKGROUND & AIMS: :Germinal centers (GCs) support high-affinity, long-lived humoral immunity. How memory B cells develop in GCs is not clear. Through the use of a cell-cycle-reporting system, we identified GC-derived memory precursor cells (GC-MP cells) that had quit cycling and reached G0 phase while in the GC, exhibited memory-associated phenotypes with signs of affinity maturation and localized toward the GC border. After being transferred into adoptive hosts, GC-MP cells reconstituted a secondary response like genuine memory B cells. GC-MP cells expressed the interleukin 9 (IL-9) receptor and responded to IL-9. Acute treatment with IL-9 or antibody to IL-9 accelerated or retarded the positioning of GC-MP cells toward the GC edge and exit from the GC, and enhanced or inhibited the development of memory B cells, which required B cell-intrinsic responsiveness to IL-9. Follicular helper T cells (TFH cells) produced IL-9, and deletion of IL-9 from T cells or, more specifically, from GC TFH cells led to impaired memory formation of B cells. Therefore, the GC development of memory B cells is promoted by TFH cell-derived IL-9.

背景与目标： ：生殖器中心（GC）支持高亲和力，长寿命的体液免疫。尚不清楚GC中记忆B细胞如何发育。通过使用细胞周期报告系统，我们确定了GC衍生的记忆前体细胞（GC-MP细胞）已经退出循环并达到G0期，而在GC中则表现出与记忆相关的表型，并伴有亲和力成熟和定位于GC边界。在被转移到过继宿主中之后，GC-MP细胞像真正的记忆B细胞一样重新构成了次级反应。 GC-MP细胞表达白介素9（IL-9）受体并对IL-9作出反应。用IL-9或IL-9抗体进行的急性治疗可加速或延迟GC-MP细胞向GC边缘定位并从GC退出，并增强或抑制记忆B细胞的发育，这需要B细胞内在的响应能力IL-9。卵泡辅助性T细胞（TFH细胞）产生IL-9，T细胞或更具体地从GC TFH细胞中删除IL-9导致B细胞记忆形成受损。因此，源自TFH细胞的IL-9促进了记忆B细胞的GC发育。

BACKGROUND & AIMS: BACKGROUND:Time trade-off (TTO) exercises typically present respondents with a limited time horizon, for example 10 years, thus implicitly considerably reducing remaining life expectancy for the average respondent. It is unclear how this affects health state valuations. AIM:The aim of the study is to investigate how awareness of the reduced life span implied by a 10-year TTO affects health state valuations, using an experimental design. METHODS:Two Web-based questionnaires (Q1 and Q2) were administered in a sample representative of the Dutch population. Both questionnaires contained three 10-year TTO exercises valuing three distinct health states, specified using the EQ-5D. Q1 used a TTO instruction not explicitly emphasizing the fact that remaining life expectancy was reduced to 10 years, while in Q2 respondents were explicitly made aware of this fact by emphasizing their implied age of death. Respondents answering Q1 were asked retrospectively whether they had been aware of their reduced life span due to the 10-year TTO. RESULTS:In total, 656 respondents completed the questionnaires (Q1: 339 and Q2: 317). The average age of the respondents was 43 years and 51 % of respondents were male. The average numbers of years traded off for the respondents of Q1 were for TTO1 0.443, TTO2 0.552, and TTO3 2.083 years. For the respondents of Q2, these averages were lower, i.e., TTO1 0.401 (p = 0.085 vs. Q1), TTO2: 0.546 (p = 0.036 vs. Q1), and TTO3: 1.467 years (p = 0.000 vs. Q1). Fifty-seven percent of respondents in Q1 confirmed that they were aware of the reduced life span. This spontaneous awareness had a limited and mixed influence on results. The generalized negative binomial regression analysis, explaining the time traded off showed that age, subjective life expectancy, and questionnaire Q2 (vs. Q1) were negatively associated with the years traded off, whereas education and worse health states in the TTO exercise had a significant positive impact on the years traded off. The probit model investigating the impact on the willingness to trade showed that age (-), education (+), subjective life expectancy (-), questionnaire Q2 versus Q1 (-), the interaction between Q2 and male gender (+), and worse health states in the TTO exercise (+) had a significant impact on the willingness to trade. CONCLUSION:These findings emphasize the importance of expected and implied life expectancy in TTOs.

背景与目标： 背景：时间权衡（TTO）练习通常为受访者提供有限的时间范围（例如10年），从而隐含地大大降低了平均受访者的剩余预期寿命。目前尚不清楚这如何影响健康状况评估。
目的：该研究的目的是使用实验设计，调查对10年TTO所隐含的寿命缩短的认识如何影响健康状况评估。
方法：在代表荷兰人口的样本中进行了两份基于Web的问卷（Q1和Q2）。两份问卷均包含三个为期10年的TTO演习，评估了使用EQ-5D指定的三种不同的健康状态。第一季度使用的是TTO指令，没有明确强调预期寿命会缩短至10年，而第二季度的受访者则通过强调其隐性死亡年龄来明确意识到这一事实。回顾性地回答了回答第一季度的受访者，他们是否知道由于10年的TTO而缩短了寿命。
结果：共有656名受访者完成了问卷（第一季度：339；第二季度：317）。受访者的平均年龄为43岁，而51％的受访者为男性。第一季度受访者需要权衡的平均年数为TTO1 0.443，TTO2 0.552和TTO3 2.083年。对于第二季度的受访者来说，这些平均值较低，即TTO1 0.401（p = 0.085 vs.Q1），TTO2：0.546（p = 0.036 vs.Q1）和TTO3：1.467年（p = 0.000 vs.Q1）。第一季度有57％的受访者确认他们知道寿命缩短了。这种自发的意识对结果的影响有限而又复杂。广义负二项式回归分析解释了折衷的时间，表明年龄，主观预期寿命和问卷Q2（相对于Q1）与折衷的年负相关，而TTO锻炼中的教育程度和健康状况较差对折中年的积极影响。概率模型对贸易意愿的影响进行了调查，结果显示年龄（-），教育程度（），主观预期寿命（-），调查问卷Q2与Q1（-），Q2与男性之间的相互作用（）和健康状况较差TTO演习中的州（）对贸易意愿产生了重大影响。
结论：这些发现强调了预期和隐含的预期寿命在TTO中的重要性。

BACKGROUND & AIMS: :hCAP-18/LL-37 is an antimicrobial peptide that is mainly expressed in epithelial cells. Gingival epithelial cells play pivotal roles in antimicrobial defense by expressing hCAP-18/LL-37. Porphyromonas gingivalis is a primary pathogen for chronic periodontitis and produces cysteine proteinase gingipains, which induce proinflammatory cytokines production, leading to enhance inflammatory responses. In contrast, gingipains attenuate immune responses, leading to induce anti-inflammatory responses. In this study, we investigated the ability of gingipains to attenuate P. gingivalis-induced hCAP-18/LL-37 production by human gingival epithelial Ca9-22 cells. The expression of LL-37 mRNA was increased by the infection of Ca9-22 cells with a P. gingivalis gingipains-null mutant KDP136 compared with P. gingivalis wild-type strain ATCC 33277. Interleukin (IL)-33 is involved in the development of chronic inflammatory diseases, and P. gingivalis infection increases IL-33 production by human gingival epithelial cells. P. gingivalis-induced LL-37 mRNA expression was augmented in IL-33 small interfering RNA-transfected Ca9-22 cells. Maxacalcitol (22-oxacalcitriol: OCT) is a biologically active metabolite of vitamin D3 analog, and OCT increases hCAP-18/LL-37 production by human gingival epithelial cells. The increasing expression of LL-37 mRNA by OCT was down-regulated by infection of the cells with P. gingivalis ATCC 33277 in Ca9-22 cells. Furthermore, P. gingivalis infection induced IL-33 mRNA expression in Ca9-22 cells; therefore, P. gingivalis-induced endogenous IL-33 down-regulated hCAP-18/LL-37 production by the bacterium. These findings suggested that endogenous IL-33 down-regulates the induction of hCAP-18/LL-37 production in human gingival epithelial cells.

背景与目标： ：hCAP-18 / LL-37是一种抗菌肽，主要在上皮细胞中表达。牙龈上皮细胞通过表达hCAP-18 / LL-37在抗菌防御中起关键作用。牙龈卟啉单胞菌是慢性牙周炎的主要病原体，并产生半胱氨酸蛋白酶齿龈蛋白酶，从而诱导促炎细胞因子的产生，从而增强炎症反应。相反，姜黄素减弱免疫反应，导致诱导抗炎反应。在这项研究中，我们调查了牙龈蛋白酶减弱人牙龈上皮Ca9-22细胞对牙龈卟啉单胞菌诱导的hCAP-18 / LL-37产生的能力。与齿龈假单胞菌野生型菌株ATCC 33277相比，齿龈假单胞菌齿龈蛋白酶空突变体KDP136感染Ca9-22细胞可增加LL-37 mRNA的表达。白介素（IL）-33参与了发育慢性炎症性疾病和牙龈卟啉单胞菌感染会增加人牙龈上皮细胞产生IL-33的数量。在IL-33小干扰RNA转染的Ca9-22细胞中，牙龈卟啉单胞菌诱导的LL-37 mRNA表达增加。 Maxacalcitol（22-oxacalcitriol：OCT）是维生素D3类似物的生物活性代谢产物，OCT可增加人牙龈上皮细胞产生的hCAP-18 / LL-37。通过OCT LL-37 mRNA的表达增加被Ca9-22细胞中牙龈卟啉单胞菌ATCC 33277感染的细胞下调。此外，牙龈卟啉单胞菌感染诱导Ca9-22细胞中IL-33 mRNA表达。因此，牙龈卟啉单胞菌诱导的内源性IL-33下调了细菌的hCAP-18 / LL-37产生。这些发现表明内源性IL-33下调人牙龈上皮细胞中hCAP-18 / LL-37产生的诱导。

BACKGROUND & AIMS: :This study was designed to measure symptoms, IL-4, IL-5, IFN-gamma, and eosinophilic cationic protein (ECP) in nasal secretions from subjects experiencing an artificial allergy season and to look for evidence of priming. Clinically relevant allergen was administered intranasally out of season to 12 asymptomatic individuals with seasonal allergic rhinitis. These individuals were then randomized to receive allergen or saline daily for the next 7 days. Nasal secretions and scrapings of nasal epithelium were obtained at baseline (day 1), 24 hours after the initial allergen administration (day 2), and 24 hours after the last instillation of allergen or saline (day 9). Nasal symptom scores (p < 0.0002), IL-5 mRNA (p = 0.03), and ECP (p < 0.02) increased after receiving the first challenge (day 2 compared with day 1). In the six subjects randomized to receive seven sequential daily challenges with allergen, symptom scores remained elevated (p < 0.02), IL-5 protein increased (p = 0.02), and IFN-gamma (p = 0.02) levels decreased (day 9 compared with day 1). In the six subjects randomized to receive seven sequential daily challenges with placebo, symptom scores, IL-5, and IFN-gamma levels were not significantly different (day 9 compared with day 1). Compared with the findings at day 2 (n = 12), the treated subjects (n = 6) had no further increase in symptoms but did show a further increase in IL-5 (p = 0.01) and a decrease in IFN-gamma (p = 0.02) at day 9. Daily instillation of moderate doses of allergen intranasally is characterized by persistent symptoms, elevation of IL-5, and reduced levels of IFN-gamma.

背景与目标： ：本研究旨在测量经历了人工过敏季节的受试者的鼻分泌物中的症状，IL-4，IL-5，IFN-γ和嗜酸性阳离子蛋白（ECP），并寻找引发的证据。临床相关的变应原是在季节外鼻内给予12例季节性变应性鼻炎的无症状个体。然后将这些个体随机分组，在接下来的7天中每天接受过敏原或生理盐水。在基线（第1天），首次施用过敏原后24小时（第2天）和最后滴入过敏原或生理盐水后第24小时（第9天）获得鼻腔分泌物和鼻上皮的刮痕。接受首次攻击后（第2天与第1天相比），鼻症状评分（p <0.0002），IL-5 mRNA（p = 0.03）和ECP（p <0.02）升高。在随机接受每日7次连续性过敏原攻击的6名受试者中，症状评分保持升高（p <0.02），IL-5蛋白升高（p = 0.02），IFN-γ（p = 0.02）水平降低（比较第9天）与第1天）。在随机接受安慰剂的七个连续每日挑战的六名受试者中，症状评分，IL-5和IFN-γ水平无显着差异（第9天与第1天相比）。与第2天的结果（n = 12）相比，接受治疗的受试者（n = 6）的症状没有进一步增加，但确实表现出IL-5进一步增加（p = 0.01）和IFN-γ降低（ p = 0.02）在第9天。鼻内每日滴入中等剂量的过敏原的特征是持续症状，IL-5升高和IFN-γ水平降低。

BACKGROUND & AIMS: :The key component in the so-called EPRI effective dose equivalent (EDE) methodology is an algorithm that utilizes two dosimeters (instead of multiple dosimeters) to predict the EDE for external photon exposures. The exposure scenarios that were previously studied in deriving the algorithm include parallel photon beams and point sources 33 cm from the body surface. The motivation for this study was the need to investigate source locations within 33 cm from the body so the method is more widely applicable. The ORNL stylized mathematical human phantoms and the MCNP code were used to calculate organ doses in this study. This paper presents the EDE data for point gamma sources at 0.3, 1.0, and 1.5 MeV, respectively, which are located at 10 cm from the surface of the body. The results and analyses show that the locations ranging from the overhead to the foot have resulted in conservative ratios except for two general regions near the front upper thigh and directly overhead. If all locations considered in this study were averaged for each photon energy, the overall ratio is on the conservative side. These data suggest that the EPRI EDE methodology is still valid for sources located 10 cm from the body, although the chance for resulting in a non-conservative estimate of the EDE has increased in comparison with the sources located at 30 cm from the body. Finally, this paper provides recommendations on how to apply the EPRI EDE methodology.

背景与目标： ：所谓的EPRI有效剂量当量（EDE）方法中的关键组件是一种算法，该算法利用两个剂量计（而不是多个剂量计）来预测外部光子暴露的EDE。先前在推导算法时曾研究过的曝光场景包括平行光子束和距体表33 cm的点光源。这项研究的动机是需要调查距人体33厘米以内的放射源位置，因此该方法更广泛地适用。在本研究中，使用了ORNL程式化的数学人体模型和MCNP代码来计算器官剂量。本文介绍了分别位于距人体表面10 cm处的0.3 MeV，1.0 MeV和1.5 MeV的点伽马源的EDE数据。结果和分析表明，从头顶到脚的位置范围导致了保守的比率，除了大腿前上方和直接头顶附近的两个一般区域。如果在本研究中考虑的所有位置均针对每种光子能量求平均值，则总体比率处于保守的一面。这些数据表明，EPRI EDE方法对于距离人体10厘米处的放射源仍然有效，尽管与位于人体30厘米处的放射源相比，导致EDE非保守估计的机会有所增加。最后，本文提供了有关如何应用EPRI EDE方法的建议。