BACKGROUND & AIMS: :The ovarian steroid hormone estrogen (E2) elicits a multiplicity of both systemic and uterotropic responses in vivo. For example, the administration of E2 to ovariectomized (Ovx) and sexually immature rodents leads to uterine-specific inflammatory infiltrates. In this study, we quantitated the number of eosinophils and BM8+, Ia+, and CD4+ cells in uteri obtained from adult Ovx control and E2-treated C57BL/6J, C3H/HeJ, and (C57BL/6J x C3H/HeJ) (B6C3) F1 hybrid mice. All three strains exhibited a significant increase in the number of uterine eosinophils and BM8+ macrophages after E2 treatment. However, C57BL/6J and B6C3 F1 hybrid mice responded with a greater number of infiltrating eosinophils and macrophages as compared with C3H/HeJ. A similar analysis of Ia+ and CD4+ cells showed that E2 treatment either down-regulates or does not affect the number of such cells in all three strains. Genome exclusion mapping using a (C57BL/6J x C3H/HeJ) x C3H/HeJ backcross population localized Est1, the major locus controlling the number of eosinophils infiltrating the uterus after E2 treatment, to chromosome 4. In addition, suggestive linkage to marker loci on chromosomes 10 and 16 was detected and evidence for locus interaction is presented. Our results conclusively demonstrate that E2-regulated/ dependent responses can be genetically controlled, indicating that the phenotypic variation observed in both the normal and pathological effects of E2 may, in part, be due to a genetic component.

背景与目标： ：卵巢类固醇激素雌激素（E2）在体内引起多种全身和子宫促反应。例如，将E2给予去卵巢（Ovx）和性不成熟的啮齿动物会导致子宫特异性炎症浸润。在这项研究中，我们量化了从成年Ovx对照和经E2处理的C57BL / 6J，C3H / HeJ和（C57BL / 6J x C3H / HeJ）（B6C3）获得的子宫中嗜酸性粒细胞和BM8，Ia和CD4细胞的数量F1杂种小鼠。在E2处理后，所有三个菌株均表现出子宫嗜酸性粒细胞和BM8巨噬细胞数量的显着增加。但是，与C3H / HeJ相比，C57BL / 6J和B6C3 F1杂种小鼠反应的浸润性嗜酸性粒细胞和巨噬细胞数量更多。对Ia和CD4细胞的类似分析显示，E2处理在所有三个菌株中均下调或不影响此类细胞的数量。使用（C57BL / 6J x C3H / HeJ）x C3H / HeJ回交群体定位的基因组排斥定位于Est1，Est1是控制E2处理后浸润子宫的嗜酸性粒细胞数量的主要基因座，并指向染色体4。检测到10号和16号染色体上的DNA，并提供了基因座相互作用的证据。我们的结果最终证明，E2调节/依赖性反应可以通过基因控制，这表明在E2的正常和病理效应中观察到的表型变异可能部分归因于遗传成分。

BACKGROUND & AIMS: :Rat basophilic leukemia (RBL 2H3) cells were passively sensitized by exposure to monoclonal anti-trinitrophenol mouse immunoglobulin E (anti-trinitrophenol IgE) (0.5 microgram/ml) and triggered by exposure to a sub-optimal concentration of trinitrophenol ovalbumin conjugate (5 ng/ml). At this concentration, trinitrophenol-ovalbumin increased histamine release from a basal rate of 4.8 +/- 0.5 to 28.5 +/- 4.6% and peptidoleukotrienes from less than 0.1 to 4.2 +/- 1.3 ng/10(6) cells in the activated cells. Ro 19-3704 and Ro 19-1400, platelet activating factor (PAF) antagonists which are structural analogs of PAF, potently inhibited both the IgE-dependent release of histamine (IC50 values of 3.0 and 3.6 microM, respectively) and LT release (IC50 values of 5.0 microM for both compounds) from the cells. These effects appeared to be independent to the ability of the compounds to act as PAF antagonists since PAF on its own had no effect on mediator release, and WEB 2086 and BN 52021, structurally distinct PAF antagonists, were relatively ineffective as inhibitors of mediator release. Ro 19-3704 and Ro 19-1400 were observed to be potent inhibitors of the soluble phospholipase A2 activity in synovial fluid from rheumatoid arthritic patients (IC50 values of 6.5 and 8.4 microM, respectively). In contrast, WEB 2086 and BN 52021 had no effect on this phospholipase A2. Ro 19-3704 significantly inhibited the IgE-dependent formation of inositol phosphates in RBL 2H3 cells (IC50 value of 7.0 microM). These data suggest that the mediator release inhibitory action of these compounds may be related to the ability of these compounds to inhibit phospholipase A2 and/or phospholipase C.

背景与目标： ：暴露于单克隆抗三硝基苯酚小鼠免疫球蛋白E（抗三硝基苯酚IgE）（0.5微克/毫升）可以被动致敏大鼠嗜碱性粒细胞白血病（RBL 2H3）细胞，并通过暴露于次适量浓度的三硝基苯酚卵白蛋白缀合物（5 ng / ml）。在此浓度下，三硝基苯酚-卵清蛋白将组胺释放从基础速率的4.8 /-0.5增加到28.5 /-4.6％，而肽白三烯从少于0.1到4.2 /-1.3 ng / 10（6）细胞增加。 Ro 19-3704和Ro 19-1400，血小板活化因子（PAF）拮抗剂，是PAF的结构类似物，可有效抑制IgE依赖的组胺释放（IC50分别为3.0和3.6 microM）和LT释放（IC50）。两种化合物的浓度值均为5.0 microM）。这些作用似乎与化合物充当PAF拮抗剂的能力无关，因为PAF本身对介质释放没有影响，并且结构不同的PAF拮抗剂WEB 2086和BN 52021作为介质释放抑制剂相对无效。观察到Ro 19-3704和Ro 19-1400是类风湿关节炎患者滑液中可溶性磷脂酶A2活性的有效抑制剂（IC50值分别为6.5和8.4 microM）。相反，WEB 2086和BN 52021对这种磷脂酶A2没有影响。 Ro 19-3704显着抑制RBL 2H3细胞中IgE依赖性肌醇磷酸的形成（IC50值为7.0 microM）。这些数据表明，这些化合物的介质释放抑制作用可能与这些化合物抑制磷脂酶A2和/或磷脂酶C的能力有关。

BACKGROUND & AIMS: Many genotoxic agents kill tumor cells by inducing apoptosis; hence, mutations that suppress apoptosis produce resistance to chemotherapy. Although directly activating the apoptotic machinery may bypass these mutations, how to achieve this activation in cancer cells selectively is not clear. In this study, we show that the drug-resistant 293 cell line is unable to activate components of the apoptotic machinery-the ICE-like proteases (caspases)-following treatment with an anticancer drug. Remarkably, extracts from untreated cells spontaneously activate caspases and induce apoptosis in a cell-free system, indicating that drug-resistant cells have not only the apoptotic machinery but also its activator. Comparing extracts from cells with defined genetic differences, we show that this activator is generated by the adenovirus E1A oncogene and is absent from normal cells. We provide preliminary characterization of this oncogene generated activity (OGA) and show that partially purified OGA activates caspases when added to extracts from untransformed cells. We suggest that agents that link OGA to caspases in cells would kill tumor cells otherwise resistant to conventional cancer therapy. As this killing relies on an activity generated by an oncogene, the effect of these agents should be selective for transformed cells.

背景与目标： 许多遗传毒性剂通过诱导细胞凋亡杀死肿瘤细胞。因此，抑制细胞凋亡的突变产生了对化学疗法的抗性。尽管直接激活凋亡机制可能绕过这些突变，但是如何在癌细胞中选择性地实现这种激活尚不清楚。在这项研究中，我们显示抗药性293细胞系无法激活抗凋亡药物ICE样蛋白酶（胱天蛋白酶）的凋亡机制的组成部分。值得注意的是，未经处理的细胞提取物可自发激活胱天蛋白酶并在无细胞系统中诱导凋亡，这表明耐药细胞不仅具有凋亡机制，还具有其激活剂。比较具有确定的遗传差异的细胞提取物，我们表明该激活剂是由腺病毒E1A癌基因产生的，而正常细胞中却不存在。我们提供了此致癌基因产生的活性（OGA）的初步表征，并表明当添加到未转化细胞的提取物中时，部分纯化的OGA会激活胱天蛋白酶。我们建议，将OGA与细胞中的半胱氨酸蛋白酶连接的药剂会杀死肿瘤细胞，否则它们会对常规的癌症治疗产生抵抗力。由于这种杀伤作用依赖于癌基因产生的活性，因此这些药剂对转化细胞的作用应该是选择性的。

BACKGROUND & AIMS: :Kennedy disease, a degenerative disorder characterized by androgen-dependent neuromuscular weakness, is caused by a CAG/glutamine tract expansion in the androgen receptor (Ar) gene. We developed a mouse model of Kennedy disease, using gene targeting to convert mouse androgen receptor (AR) to human sequence while introducing 113 glutamines. AR113Q mice developed hormone and glutamine length-dependent neuromuscular weakness characterized by the early occurrence of myopathic and neurogenic skeletal muscle pathology and by the late development of neuronal intranuclear inclusions in spinal neurons. AR113Q males unexpectedly died at 2-4 months. We show that this androgen-dependent death reflects decreased expression of skeletal muscle chloride channel 1 (CLCN1) and the skeletal muscle sodium channel alpha-subunit, resulting in myotonic discharges in skeletal muscle of the lower urinary tract. AR113Q limb muscles show similar myopathic features and express decreased levels of mRNAs encoding neurotrophin-4 and glial cell line-derived neurotrophic factor. These data define an important myopathic contribution to the Kennedy disease phenotype and suggest a role for muscle in non-cell autonomous toxicity of lower motor neurons.

背景与目标： ：肯尼迪病是一种以雄激素依赖性神经肌肉无力为特征的变性疾病，是由雄激素受体（Ar）基因中的CAG /谷氨酰胺束扩张引起的。我们开发了肯尼迪病的小鼠模型，使用基因靶向技术将小鼠雄激素受体（AR）转化为人类序列，同时引入113种谷氨酰胺。 AR113Q小鼠发展出激素和谷氨酰胺长度依赖性神经肌肉无力，其特征是肌病性和神经源性骨骼肌病理的早期发生以及脊髓神经元中神经元核内包涵体的发育较晚。 AR113Q男性意外死于2-4个月。我们表明，这种雄激素依赖性死亡反映了骨骼肌氯化物通道1（CLCN1）和骨骼肌钠通道α亚基的表达下降，导致下尿路骨骼肌的肌强直放电。 AR113Q肢体肌肉表现出相似的肌病特征，并表达编码神经营养蛋白4和神经胶质细胞源性神经营养因子的mRNA降低水平。这些数据定义了对肯尼迪病表型的重要肌病性贡献，并暗示了肌肉在下运动神经元的非细胞自主毒性中的作用。

BACKGROUND & AIMS: CONTEXT:Chronic pain in patients with advanced cancer poses a serious clinical challenge. The Δ9-tetrahydrocannabinol (THC)/cannabidiol (CBD) oromucosal spray (U.S. Adopted Name, nabiximols; Sativex(®)) is a novel cannabinoid formulation currently undergoing investigation as an adjuvant therapy for this treatment group. OBJECTIVES:This follow-up study investigated the long-term safety and tolerability of THC/CBD spray and THC spray in relieving pain in patients with advanced cancer. METHODS:In total, 43 patients with cancer-related pain experiencing inadequate analgesia despite chronic opioid dosing, who had participated in a previous three-arm (THC/CBD spray, THC spray, or placebo), two-week parent randomized controlled trial, entered this open-label, multicenter, follow-up study. Patients self-titrated THC/CBD spray (n=39) or THC spray (n=4) to symptom relief or maximum dose and were regularly reviewed for safety, tolerability, and evidence of clinical benefit. RESULTS:The efficacy end point of change from baseline in mean Brief Pain Inventory-Short Form scores for "pain severity" and "worst pain" domains showed a decrease (i.e., improvement) at each visit in the THC/CBD spray patients. Similarly, the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 scores showed a decrease (i.e., improvement) from baseline in the domains of insomnia, pain, and fatigue. No new safety concerns associated with the extended use of THC/CBD spray arose from this study. CONCLUSION:This study showed that the long-term use of THC/CBD spray was generally well tolerated, with no evidence of a loss of effect for the relief of cancer-related pain with long-term use. Furthermore, patients who kept using the study medication did not seek to increase their dose of this or other pain-relieving medication over time, suggesting that the adjuvant use of cannabinoids in cancer-related pain could provide useful benefit.

背景与目标： 背景：晚期癌症患者的慢性疼痛提出了严峻的临床挑战。 Δ9-四氢大麻酚（THC）/大麻二酚（CBD）口腔粘膜喷雾剂（美国采用名称，nabiximols;Sativex®）是一种新型大麻素制剂，目前正在接受治疗，作为该治疗组的辅助疗法。
目的：这项随访研究调查了THC / CBD喷雾剂和THC喷雾剂在缓解晚期癌症患者的疼痛方面的长期安全性和耐受性。
方法：共有43例癌症相关疼痛患者尽管使用了阿片类药物长期服用，但仍没有足够的镇痛作用，他们参加了先前的三组试验（THC / CBD喷雾剂，THC喷雾剂或安慰剂），为期两周的父母随机对照试验，参加了这项开放性，多中心的后续研究。患者自行滴定THC / CBD喷雾剂（n = 39）或THC喷雾剂（n = 4）以缓解症状或最大剂量，并定期检查其安全性，耐受性和临床获益证据。
结果：THC / CBD喷剂患者每次就诊时，“疼痛严重程度”和“最严重疼痛”域的平均简短疼痛清单-简表得分的基线变化的功效终点显示出降低（即改善）。同样，欧洲癌症生活质量研究和治疗组织问卷-C30的得分显示，失眠，疼痛和疲劳程度较基线水平有所降低（即有所改善）。这项研究没有引起与THC / CBD喷雾剂的广泛使用相关的新安全隐患。
结论：这项研究表明，长期使用THC / CBD喷雾剂通常具有良好的耐受性，没有证据表明长期使用THC / CBD喷雾剂可减轻癌症相关疼痛。此外，继续使用研究药物的患者并未寻求随时间增加这种药物或其他缓解疼痛药物的剂量，这表明在与癌症相关的疼痛中辅助使用大麻素可提供有益的益处。

BACKGROUND & AIMS: :Strigolactones control many aspects of plant growth and development, but the active form(s) of strigolactones and their mode of action at the molecular level are unknown. A new study provides evidence that an α/β-fold protein plays a central multifunctional role in strigolactone metabolism, perception and signalling.

背景与目标： ：Strigolactones控制植物生长和发育的许多方面，但是strigolactones的活性形式及其在分子水平上的作用方式尚不清楚。一项新的研究提供了证据，表明α/β折叠蛋白在松果内酯的代谢，感知和信号传导中起着重要的多功能作用。

BACKGROUND & AIMS: Importance:Opioid use disorder (OUD) frequently begins in adolescence and young adulthood. Intervening early with pharmacotherapy is recommended by major professional organizations. No prior national studies have examined the extent to which adolescents and young adults (collectively termed youth) with OUD receive pharmacotherapy. Objective:To identify time trends and disparities in receipt of buprenorphine and naltrexone among youth with OUD in the United States. Design, Setting, and Participants:A retrospective cohort study was conducted using deidentified data from a national commercial insurance database. Enrollment and complete health insurance claims of 9.7 million youth, aged 13 to 25 years were analyzed, identifying individuals who received a diagnosis of OUD between January 1, 2001, and June 30, 2014, with final follow-up date December 31, 2014. Analysis was conducted from April 25 to December 31, 2016. Time trends were identified and multivariable logistic regression was used to determine sociodemographic factors associated with medication receipt. Exposures:Sex, age, race/ethnicity, neighborhood education and poverty levels, geographic region, census region, and year of diagnosis. Main Outcomes and Measures:Dispensing of a medication (buprenorphine or naltrexone) within 6 months of first receiving an OUD diagnosis. Results:Among 20 822 youth diagnosed with OUD (0.2% of the 9.7 million sample), 13 698 (65.8%) were male and 17 119 (82.2%) were non-Hispanic white. Mean (SD) age was 21.0 (2.5) years at the first observed diagnosis. The diagnosis rate of OUD increased nearly 6-fold from 2001 to 2014 (from 0.26 per 100 000 person-years to 1.51 per 100 000 person-years). Overall, 5580 (26.8%) youth were dispensed a medication within 6 months of diagnosis, with 4976 (89.2%) of medication-treated youth receiving buprenorphine and 604 (10.8%) receiving naltrexone. Medication receipt increased more than 10-fold, from 3.0% in 2002 (when buprenorphine was introduced) to 31.8% in 2009, but declined in subsequent years (27.5% in 2014). In multivariable analyses, younger individuals were less likely to receive medications, with adjusted probability for age 13 to 15 years, 1.4% (95% CI, 0.4%-2.3%); 16 to 17 years, 9.7% (95% CI, 8.4%-11.1%); 18 to 20 years, 22.0% (95% CI, 21.0%-23.0%); and 21 to 25 years, 30.5% (95% CI, 30.0%-31.5%) (P < .001 for difference). Females (7124 [20.3%]) were less likely than males (13 698 [24.4%]) to receive medications (P < .001), as were non-Hispanic black (105 [14.8%]) and Hispanic (1165 [20.0%]) youth compared with non-Hispanic white (17 119 [23.1%]) youth (P < .001). Conclusions and Relevance:In this first national study of buprenorphine and naltrexone receipt among youth, dispensing increased over time. Nonetheless, only 1 in 4 commercially insured youth with OUD received pharmacotherapy, and disparities based on sex, age, and race/ethnicity were observed.

背景与目标： 重要性：阿片类药物使用障碍（OUD）经常在青春期和成年期开始。主要专业组织建议尽早进行药物治疗。以前的国家研究都没有研究OUD的青少年和年轻人（统称为青年）接受药物治疗的程度。
目的：确定美国OUD青年中丁丙诺啡和纳曲酮的接受时间趋势和差异。
设计，地点和参与者：使用来自国家商业保险数据库的身份不明数据进行回顾性队列研究。分析了970万年龄在13至25岁之间的970万青年的入学和完全健康保险理赔，确定了在2001年1月1日至2014年6月30日期间被诊断为OUD的个人，最终随访日期为2014年12月31日。分析于2016年4月25日至12月31日进行。确定了时间趋势，并使用多变量logistic回归确定与用药相关的社会人口统计学因素。
暴露：性别，年龄，种族/民族，邻里教育和贫困程度，地理区域，人口普查区域和诊断年份。
主要结果和措施：在首次接受OUD诊断后的6个月内分发药物（丁丙诺啡或纳曲酮）。
结果：在20822名被诊断为OUD的青年中（占970万样本的0.2％），男性中有13698名（65.8％），非西班牙裔白人中有17119名（82.2％）。在首次观察到的诊断中，平均（SD）年龄为21.0（2.5）岁。从2001年到2014年，OUD的诊断率增长了近6倍（从每100000人年0.26增至每100000人年1.51）。总体而言，在诊断后的6个月内分配了5580（26.8％）名年轻人用药，其中接受丁丙诺啡的4976（89.2％）名接受药物治疗的青年人和接受纳曲酮的604名（10.8％）。药物治疗的收入增加了十倍以上，从2002年的3.0％（引入丁丙诺啡时）到2009年的31.8％，但随后几年却下降了（2014年为27.5％）。在多变量分析中，年龄较小的个体接受药物治疗的可能性较小，年龄调整为13至15岁的概率为1.4％（95％CI，0.4％-2.3％）； 16至17年，9.7％（95％CI，8.4％-11.1％）； 18至20年，22.0％（95％CI，21.0％-23.0％）；和21至25年，分别为30.5％（95％CI，30.0％-31.5％）（差异P <<。001）。与非西班牙裔黑人（105名[14.8％]）和西班牙裔（1165名[20.0]）相比，女性（7124名[20.3％]）与男性（13698名[24.4％]）接受药物治疗的可能性较小（P <.001）。 ％]的年轻人与非西班牙裔白人（17119 [23.1％]）的年轻人相比（P <）。001）。
结论和相关性：在这项关于丁丙诺啡和纳曲酮的年轻人接受的首次国家研究中，随着时间的流逝，配药量增加了。尽管如此，只有四分之一的有OUD的商业保险青年接受了药物治疗，并且观察到了基于性别，年龄和种族/民族的差异。

BACKGROUND & AIMS: :Repetitive transcranial magnetic stimulation (rTMS), a non-invasive form of brain stimulation, has shown experimental and clinical efficacy in a range of neuromodulatory models, even when delivered at low intensity (i.e. subthreshold for action potential generation). After central nervous system (CNS) injury, studies suggest that reactive astrocytes and microglia can have detrimental but also beneficial effects; thus modulating glial activity, for example through application of rTMS, could potentially be a useful therapeutic tool following neurotrauma. Immunohistochemistry was used to measure the effect of low intensity rTMS (LI-rTMS) on GFAP (astrocyte), IBA1 (microglial), and CS56 (proteoglycan) expression in a unilateral penetrating cortical stab injury model of glial scarring in young adult and aged male and female C57BL6/J mice. Mice received contralateral low frequency, ipsilateral low frequency, ipsilateral high frequency or sham LI-rTMS (4-5mT intensity), for two weeks following injury. There was no significant difference in the overall volume of tissue containing GFAP positive (+) astrocytes, IBA1+ microglia, or proteoglycan expression, between sham and LI-rTMS-treated mice of all ages and sex. Importantly however, the density of GFAP+ astrocytes and IBA1+ microglia immediately adjacent to the injury was significantly reduced following ipsilateral low and high frequency stimulation in adult and aged females (p≤0.05), but was significantly increased in adult and aged males (p≤0.05). LI-rTMS effects were generally of greater magnitude in aged mice compared to young adult mice. These results suggest that sex differences need to be factored into therapeutic rTMS protocols. In particular, more work analyzing frequency and intensity specific effects, especially in relation to age and sex, is required to determine how rTMS can best be used to modify glial reactivity and phenotype following neurotrauma.

背景与目标： ：反复经颅磁刺激（rTMS）是一种非侵入性的脑刺激形式，即使在低强度下（即产生动作电位的阈值下），也已在一系列神经调节模型中显示出实验和临床功效。中枢神经系统（CNS）损伤后，研究表明反应性星形胶质细胞和小胶质细胞可能具有有害作用，但也有有益作用。因此，例如通过应用rTMS调节神经胶质活动可能是神经创伤后的一种有用的治疗工具。免疫组织化学方法用于测量低强度rTMS（LI-rTMS）对单侧穿透性胶质瘢痕形成的成年男性和老年男性的单侧穿透性皮质刺伤模型中GFAP（星形胶质细胞），IBA1（小胶质细胞）和CS56（蛋白聚糖）表达的影响。和雌性C57BL6 / J小鼠。小鼠在受伤后两周接受对侧低频，同侧低频，同侧高频或假LI-rTMS（4-5mT强度）。在假手术和LI-rTMS处理的所有年龄和性别的小鼠之间，含有GFAP阳性（）星形胶质细胞，IBA1小胶质细胞或蛋白聚糖表达的组织总体积均无显着差异。然而重要的是，成年和老年女性在同侧低频和高频刺激后，紧邻损伤的GFAP星形胶质细胞和IBA1小胶质细胞的密度显着降低（p≤0.05），而成年和老年男性显着增加（p≤0.05） ）。与成年小鼠相比，老年小鼠的LI-rTMS效应通常更大。这些结果表明，性别差异需要纳入治疗性rTMS方案中。特别是，需要做更多的工作来分析频率和强度的具体影响，尤其是与年龄和性别相关的影响，以确定rTMS如何最好地用于改变神经外伤后的神经胶质反应性和表型。

BACKGROUND & AIMS: :In order to understand the effect of pH on the formation of electrostatic complexes between lysozyme and low methoxyl (LM) pectin, mixtures were prepared at a fixed lysozyme concentration (0.714g.L-1) by progressive addition of LM pectin (from 0 to 4g.L-1). Turbidity analysis allowed to determine specific conditions of pH and lysozyme/LM pectin ratio for optimal complex aggregation. The intrinsic fluorescence enhancement observed upon binding of LM pectin to lysozyme was correlated with the formation of intermolecular aggregates. Conversely, the intrinsic fluorescence decrease observed at higher LM pectin amounts was correlated with the dissociation of intermolecular aggregates. UV absorption spectroscopy showed modifications in lysozyme conformation during both the aggregation phase and the dissociation phase. The role of electrostatic interactions in the formation of lysozyme/LM pectin complexes is discussed in relation to the overall structure and the charge density profile of the two biopolymers.

背景与目标： ：为了了解pH值对溶菌酶和低甲氧基（LM）果胶之间形成静电复合物的影响，通过逐步添加LM果胶（0至4g）以固定的溶菌酶浓度（0.714gL-1）制备混合物.L-1）。浊度分析可以确定特定的pH值条件和溶菌酶/ LM果胶比例，以实现最佳的复合物聚集。 LM果胶与溶菌酶结合后观察到的固有荧光增强与分子间聚集体的形成相关。相反，在较高LM果胶量下观察到的固有荧光下降与分子间聚集体的解离相关。紫外吸收光谱显示在聚集阶段和解离阶段的溶菌酶构象都发生了改变。关于两种生物聚合物的整体结构和电荷密度分布，讨论了静电相互作用在溶菌酶/ LM果胶复合物形成中的作用。

BACKGROUND & AIMS: :Motor imagery tasks are well established procedures in brain computer interfaces, but are also used in the assessment of patients with disorders of consciousness. For testing awareness in unresponsive patients it is necessary to know the natural variance of brain responses to motor imagery in healthy subjects. We examined 22 healthy subjects using EEG in three conditions: movement of both hands, imagery of the same movement, and an instruction to hold both hands still. Single-subject non-parametric statistics were applied to the fast-Fourier transformed data. Most effects were found in the α- and β-frequency ranges over central electrodes, that is, in the μ-rhythm. We found significant power changes in 18 subjects during movement and in 11 subjects during motor imagery. In 8 subjects these changes were consistent over both conditions. The significant power changes during movement were a decrease of μ-rhythm. There were 2 subjects with an increase and 9 subjects with a decrease of μ-rhythm during imagery. α and β are the most responsive frequency ranges, but there is a minor number of subjects who show a synchronization instead of the more common desynchronization during motor imagery. A (de)synchronization of μ-rhythm can be considered to be a normal response.

背景与目标： ：运动图像任务是大脑计算机界面中公认的程序，但也用于评估意识障碍患者。为了测试反应迟钝的患者的意识，有必要了解健康受试者中大脑对运动图像反应的自然变化。我们在以下三种情况下使用脑电图检查了22名健康受试者：双手运动，同一动作的图像以及保持双手静止的指令。将单对象非参数统计信息应用于快速傅立叶变换后的数据。在中央电极的α频率和β频率范围内，即在μ节奏中，发现了大多数影响。我们发现运动过程中的18个对象和运动图像中的11个对象的功率发生了显着变化。在8位受试者中，这些变化在两种情况下均一致。运动过程中功率的显着变化是μ节律的降低。在成像过程中，有2位受试者的μ节律增加，而9位受试者的μ节律减小。 α和β是最敏感的频率范围，但是在电机成像过程中，有少数被摄对象显示出同步，而不是更常见的不同步。 μ节奏的（去）同步可以被认为是正常反应。

BACKGROUND & AIMS: :Spontaneous intracranial hemorrhage is a debilitating form of stroke, often leading to death or permanent cognitive impairment. Many of the causative genes and the underlying mechanisms implicated in developmental cerebral-vascular malformations are unknown. Recent in vitro and in vivo studies in mice have shown inhibition of the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) pathway to be effective in stabilizing cranial vessels. Using a combination of pharmacological and genetic approaches to specifically inhibit the HMGCR pathway in zebrafish (Danio rerio), we demonstrate a requirement for this metabolic pathway in developmental vascular stability. Here we report that inhibition of HMGCR function perturbs cerebral-vascular stability, resulting in progressive dilation of blood vessels, followed by vessel rupture, mimicking cerebral cavernous malformation (CCM)-like lesions in humans and murine models. The hemorrhages in the brain are rescued by prior exogenous supplementation with geranylgeranyl pyrophosphate (GGPP), a 20-carbon metabolite of the HMGCR pathway, required for the membrane localization and activation of Rho GTPases. Consistent with this observation, morpholino-induced depletion of the β-subunit of geranylgeranyltransferase I (GGTase I), an enzyme that facilitates the post-translational transfer of the GGPP moiety to the C-terminus of Rho family of GTPases, mimics the cerebral hemorrhaging induced by the pharmacological and genetic ablation of HMGCR. In embryos with cerebral hemorrhage, the endothelial-specific expression of cdc42, a Rho GTPase involved in the regulation of vascular permeability, was significantly reduced. Taken together, our data reveal a metabolic contribution to the stabilization of nascent cranial vessels, requiring protein geranylgeranylation acting downstream of the HMGCR pathway.

背景与目标： ：自发性颅内出血是中风的一种虚弱形式，通常会导致死亡或永久性认知障碍。涉及发展性脑血管畸形的许​​多致病基因和潜在机制尚不清楚。最近在小鼠中进行的体外和体内研究表明，抑制3-羟基-3-甲基戊二酰辅酶A还原酶（HMGCR）途径可有效稳定颅骨血管。使用药理学和遗传学方法的组合来特异性抑制斑马鱼（Danio rerio）中的HMGCR途径，我们证明了这种代谢途径在发育性血管稳定性中的需求。在这里我们报道抑制HMGCR功能扰乱脑血管稳定性，导致血管进行性扩张，继而血管破裂，在人和鼠模型中模仿脑海绵状畸形（CCM）样病变。可以通过事先外源补充geranylgeranyl焦磷酸（GGPP）（HMGCR途径的20个碳代谢物）进行膜定位和激活Rho GTPases来挽救脑部出血。与此观察结果一致，吗啉代诱导的香叶基香叶基转移酶I（GGTase I）的β亚基耗竭，该酶有助于将GGPP部分翻译后转移至GTPases Rho家族的C末端，从而模拟脑出血由HMGCR的药理和遗传消融诱导。在患有脑出血的胚胎中，参与调节血管通透性的Rho GTPase cdc42的内皮特异性表达显着降低。综上所述，我们的数据揭示了新陈代谢对稳定新生颅血管的作用，需要在HMGCR途径下游作用的蛋白质香叶基香叶基化作用。

BACKGROUND & AIMS: :The possibility that endocrine factors may influence the clinical course of malignant melanoma is suggested by the superior survival data of women. In preclinical models we observed a higher rate of colony formation by human melanoma cells in male compared to female SCID mice, but only in the case of the liver and not in other organs. The gender difference could be seen at an early phase of colony formation. On the other hand, in our human melanoma cell lines we failed to detect steroid receptor protein expression, and treatment with sex hormones did not considerably influence their in vitro behavior. Investigating the possible contribution of host cells to the observed gender difference, we performed in vivo blocking experiments applying pretreatment of the animals with Kupffer cell inhibitor gadolinium chloride and the NK cell inhibitor anti-asialo GM1 antibody. While Kupffer cell blockade enhanced melanoma liver colonization equally in the two sexes, a more prominent increase was observed in female than in male mice in the case of NK cell inhibition. Further supporting the importance of NK cells in the lower liver colonization efficiency of melanoma cells in females, gender difference in colony formation was lost in NSG mice lacking NK activity. Although in humans no organ selectivity of gender difference in melanoma progression has been observed according to data in the literature, our results possibly indicate a contribution of natural host defense mechanisms to gender difference in survival of patients with melanoma or other tumor types as well.

背景与目标： 女性优越的生存率数据提示内分泌因素可能影响恶性黑色素瘤的临床进程。在临床前模型中，我们观察到，与雌性SCID小鼠相比，雄性人黑素瘤细胞在雄性中的集落形成率更高，但仅在肝脏而不是在其他器官中。可以在菌落形成的早期阶段看到性别差异。另一方面，在我们的人类黑素瘤细胞系中，我们未能检测到类固醇受体蛋白的表达，而性激素的治疗​​并未显着影响其体外行为。为了研究宿主细胞对观察到的性别差异的可能贡献，我们进行了体内阻断实验，该实验应用了用Kupffer细胞抑制剂氯化chloride和NK细胞抑制剂抗亚洲GM1抗体对动物进行预处理的实验。虽然库普弗细胞阻断剂在两个性别中均能平等地增强黑色素瘤的肝脏定植，但在抑制NK细胞的情况下，雌性小鼠比雄性小鼠观察到更明显的增加。进一步证实了NK细胞在降低雌性黑素瘤细胞肝脏定植效率中的重要性，在缺乏NK活性的NSG小鼠中，丧失了克隆形成的性别差异。尽管在人类中，根据文献中的数据，没有观察到器官对黑素瘤进展中性别差异的选择性，但我们的结果可能表明，天然宿主防御机制对黑素瘤或其他肿瘤类型患者的生存中的性别差异也有贡献。

BACKGROUND & AIMS: OBJECTIVE:Postoperative ileus (POI), the most frequent complication after intestinal surgery, depends on dendritic cells (DCs) and macrophages. Here, we have investigated the mechanism that activates these cells and the contribution of the intestinal microbiota for POI induction. DESIGN:POI was induced by manipulating the intestine of mice, which selectively lack DCs, monocytes or macrophages. The disease severity in the small and large intestine was analysed by determining the distribution of orally applied fluorescein isothiocyanate-dextran and by measuring the excretion time of a retrogradely inserted glass ball. The impact of the microbiota on intestinal peristalsis was evaluated after oral antibiotic treatment. RESULTS:We found that Cd11c-Cre+ Irf4flox/flox mice lack CD103+CD11b+ DCs, a DC subset unique to the intestine whose function is poorly understood. Their absence in the intestinal muscularis reduced pathogenic inducible nitric oxide synthase (iNOS) production by monocytes and macrophages and ameliorated POI. Pathogenic iNOS was produced in the jejunum by resident Ly6C- macrophages and infiltrating chemokine receptor 2-dependent Ly6C+ monocytes, but in the colon only by the latter demonstrating differential tolerance mechanisms along the intestinal tract. Consistently, depletion of both cell subsets reduced small intestinal POI, whereas the depletion of Ly6C+ monocytes alone was sufficient to prevent large intestinal POI. The differential role of monocytes and macrophages in small and large intestinal POI suggested a potential role of the intestinal microbiota. Indeed, antibiotic treatment reduced iNOS levels and ameliorated POI. CONCLUSIONS:Our findings reveal that CD103+CD11b+ DCs and the intestinal microbiome are a prerequisite for the activation of intestinal monocytes and macrophages and for dysregulating intestinal motility in POI.

背景与目标： 目的：术后肠梗阻（POI）是小肠手术后最常见的并发症，取决于树突状细胞（DC）和巨噬细胞。在这里，我们研究了激活这些细胞的机制以及肠道菌群对POI诱导的贡献。
设计：POI是通过操纵小鼠肠道诱导的，小鼠的肠道选择性地缺乏DC，单核细胞或巨噬细胞。通过确定口服荧光素异硫氰酸酯-右旋糖酐的分布并测量逆行插入的玻璃球的排泄时间来分析小肠和大肠的疾病严重程度。口服抗生素治疗后，评估了微生物群对肠道蠕动的影响。
结果：我们发现Cd11c-Cre Irf4flox / flox小鼠缺少CD103 CD11b DC，这是肠道功能独特的DC子集，其功能尚不清楚。它们在肠道肌层中的缺失减少了单核细胞和巨噬细胞以及改善的POI的致病性诱导型一氧化氮合酶（iNOS）的产生。致病性iNOS是由常驻Ly6C-巨噬细胞和浸润趋化因子受体2依赖性Ly6C单核细胞在空肠中产生，但仅在结肠中表现出沿肠道的不同耐受机制，而在结肠中产生。一致地，两个细胞亚群的消耗减少了小肠POI，而仅Ly6C单核细胞的消耗就足以防止大肠POI。单核细胞和巨噬细胞在小肠和大肠POI中的不同作用提示了肠道菌群的潜在作用。确实，抗生素治疗降低了iNOS水平并改善了POI。
结论：我们的发现表明，CD103 CD11b DC和肠道微生物组是激活肠道单核细胞和巨噬细胞以及POI肠道运动失调的先决条件。

BACKGROUND & AIMS: :Previously reported propellane derivative KNT-42 preferred the κ receptor and functioned as a message part in the message-address concept, but its affinity for the κ receptor was not high. To improve affinity, we synthesized five pentacyclic propellane derivatives designed for the purpose of fixing the conformation of KNT-42. The etheno- and ethano-bridged derivatives SYK-347 and SYK-393 exhibited high affinity and selectivity for the κ receptor, whereas the other derivatives did not. These results would be due to the different ranges of movement of the basic nitrogens and less basicity of the nitrogens due to the electron withdrawing effect of the introduced hydroxy or keto group. SYK-347 and SYK-393 preferring the κ receptor were expected to be useful for designing selective ligands for opioid receptor types, especially the κ receptor.

背景与目标： ：以前报道的螺ella衍生物KNT-42更喜欢κ受体，并在消息地址概念中起消息部分的作用，但它对κ受体的亲和力不高。为了提高亲和力，我们合成了五种五环丙烷衍生物，旨在固定KNT-42的构象。乙炔和乙桥接的衍生物SYK-347和SYK-393对κ受体表现出高亲和力和选择性，而其他衍生物则没有。这些结果将归因于碱性氮的移动范围不同以及由于引入的羟基或酮基的电子吸收作用而使氮的碱性降低。期望κ受体的SYK-347和SYK-393可用于设计针对阿片样受体类型，特别是κ受体的选择性配体。

BACKGROUND & AIMS: :Little is known about molecular recognition of acetylated N termini, despite prevalence of this modification among eukaryotic cytosolic proteins. We report that the family of human DCN-like (DCNL) co-E3s, which promote ligation of the ubiquitin-like protein NEDD8 to cullin targets, recognizes acetylated N termini of the E2 enzymes UBC12 and UBE2F. Systematic biochemical and biophysical analyses reveal 40- and 10-fold variations in affinities among different DCNL-cullin and DCNL-E2 complexes, contributing to varying efficiencies of different NEDD8 ligation cascades. Structures of DCNL2 and DCNL3 complexes with N-terminally acetylated peptides from UBC12 and UBE2F illuminate a common mechanism by which DCNL proteins recognize N-terminally acetylated E2s and how selectivity for interactions dependent on N-acetyl-methionine are established through side chains recognizing distal residues. Distinct preferences of UBC12 and UBE2F peptides for inhibiting different DCNLs, including the oncogenic DCNL1 protein, suggest it may be possible to develop small molecules blocking specific N-acetyl-methionine-dependent protein interactions.

背景与目标： ：尽管在真核细胞溶质蛋白中普遍存在这种修饰，但对乙酰化N末端的分子识别知之甚少。我们报告说，人类DCN样（DCNL）co-E3s的家族，促进泛素样蛋白NEDD8与cullin目标的连接，识别E2酶UBC12和UBE2F的乙酰化N末端。系统的生化和生物物理分析表明，不同的DCNL-cullin和DCNL-E2复合物之间的亲和力变化分别为40倍和10倍，从而导致了不同NEDD8连接级联反应效率的变化。具有来自UBC12和UBE2F的N末端乙酰化肽的DCNL2和DCNL3配合物的结构阐明了DCNL蛋白质识别N末端乙酰化E2的共同机制以及如何通过识别远端残基的侧链建立依赖N-乙酰甲硫氨酸的相互作用的选择性。 UBC12和UBE2F肽对于抑制不同的DCNL（包括致癌DCNL1蛋白）的不同偏好，表明可能开发出阻断特定N-乙酰基-蛋氨酸依赖性蛋白相互作用的小分子。