BACKGROUND & AIMS: :Estrogen mediates its effects through multiple cellular receptors. In addition to the classical nuclear estrogen receptors (ERalpha and ERbeta), estrogen also signals through the seven-transmembrane G-protein-coupled receptor (GPCR) GPR30. Although estrogen is a cell-permeable ligand, it is often assumed that all GPCRs function solely as cell surface receptors. Our previous results showed that GPR30 appeared to be expressed predominantly in the endoplasmic reticulum. A critical question that arises is whether this localization represents the site of functional receptor. To address this question, we synthesized a collection of cell-permeable and cell-impermeable estrogen derivatives. We hypothesized that if functional GPR30 were expressed at the cell surface, both permeable and impermeable derivatives would show activity. However, if functional GPR30 were predominantly intracellular, like ERalpha, only the permeable ligands should show activity. Cell permeability was assessed using cells expressing ERalpha as a model intracellular estrogen-binding receptor. Our results reveal that despite exhibiting similar binding affinities for GPR30, only the cell-permeable ligands are capable of stimulating rapid calcium mobilization and phosphoinositide 3-kinase (PI3K) activation. We conclude that GPR30 expressed intracellularly is capable of initiating cellular signaling and that there is insufficient GPR30 expressed on the cell surface to initiate signaling in response to impermeable ligands in the cell lines examined. To our knowledge, this is the first definitive demonstration of a functional intracellular transmembrane estrogen receptor.

背景与目标： 雌激素通过多种细胞受体介导其作用。除了经典的核雌激素受体 (ERalpha和ERbeta) 外，雌激素还通过七跨膜g蛋白偶联受体 (GPCR) gpr30发出信号。尽管雌激素是细胞可渗透的配体，但通常认为所有gpcr仅作为细胞表面受体起作用。我们先前的结果表明，GPR30似乎主要在内质网中表达。出现的一个关键问题是，这种定位是否代表功能受体的位点。为了解决这个问题，我们合成了细胞可渗透和细胞不可渗透的雌激素衍生物的集合。我们假设，如果在细胞表面表达功能性GPR30，则可渗透和不可渗透的衍生物都将显示活性。但是，如果功能性GPR30主要是细胞内的，例如ERalpha，则只有可渗透的配体才应显示活性。使用表达ERalpha的细胞作为模型细胞内雌激素结合受体评估细胞通透性。我们的结果表明，尽管对GPR30表现出相似的结合亲和力，但只有细胞可渗透的配体能够刺激快速的钙动员和磷酸肌醇3激酶 (PI3K) 激活。我们得出的结论是，细胞内表达的GPR30能够启动细胞信号传导，并且在细胞表面表达的GPR30不足以响应所检查细胞系中不可渗透的配体来启动信号传导。据我们所知，这是功能性细胞内跨膜雌激素受体的第一个明确证明。

BACKGROUND & AIMS: :Efficient recombinant expression of N-acyl-L-aminoacylase 1 from pig kidney (pAcy1) was achieved in the prokaryotic host Escherichia coli. An optimized nucleotide sequence (codon adaptation index 0.95 for E. coli), was cloned into vector pET-52(b) yielding an E. coli-expressible pAcy1 gene. Formation of inclusion bodies was alleviated by co-expression of molecular chaperones resulting in 2.7- and 4.2-fold increased recovery of active pAcy1 using trigger factor or GroEL-GroES, respectively. Facile purification was achieved via StrepTag affinity chromatography. Overall, more than 80 mg highly active pAcy1 (94 U/mg) was obtained per liter of cultivation broth. The protein was analyzed for structural and functional identity, and the performances of further described expression and purification systems for pAcy1 were compared.

背景与目标： : 在原核宿主大肠杆菌中实现了猪肾 (pAcy1) 中N-酰基-L-氨基酰化酶1的有效重组表达。将优化的核苷酸序列 (大肠杆菌的密码子适应指数0.95) 克隆到载体pET-52(b) 中，产生大肠杆菌可表达的pAcy1基因。通过分子伴侣的共表达减轻了包涵体的形成，从而分别使用触发因子或GroEL-GroES使活性pAcy1的回收率增加了2.7倍和4.2倍。通过链霉亲和性色谱法实现了简便的纯化。总体而言，每升培养液可获得超过80 mg的高活性pAcy1 (94 U/mg)。分析了该蛋白的结构和功能特性，并比较了进一步描述的pAcy1表达和纯化系统的性能。

BACKGROUND & AIMS: :Acaricides are used by beekeepers in honey bee (Apis mellifera L.) colonies to control parasitic mites, but may also have adverse effects to honey bees. In this study, five commonly used acaricides were tested for their sublethal effects on memory and expression of neural-related genes in honey bees. Memory measured with the proboscis extension reflex (PER) assay was significantly reduced by topical treatment of bees with a single LD05 dose of formic acid at 2 and 24 h post treatment (hpt). However, tau-fluvalinate, amitraz, coumaphos, and formic acid, but not thymol, resulted in memory loss at 48 hpt. The LD05 doses of the acraricides did not affect expression of neuroligin-1, related to memory, or expression of major royal jelly protein-1, related to both memory and development, although expression of both genes was affected at LD50 doses. The LD05 doses of thymol, formic acid, amitraz and coumaphos increased defensin-1 expression, which is related to both memory and immunity. The effect of thymol, however, may have been due to its impact on the immune response rather than memory. This study demonstrates that acaricides vary in their effects on bee's memory, and that the widely used acaricide, formic acid, is particularly damaging.

背景与目标： : 杀螨剂被蜜蜂 (Apis mellifera L.) 菌落中的养蜂人用来控制寄生螨，但也可能对蜜蜂产生不利影响。在这项研究中，测试了五种常用的杀螨剂对蜜蜂的记忆和神经相关基因表达的亚致死作用。通过在治疗后2和24小时 (hpt) 用单次LD05剂量的甲酸局部治疗蜜蜂，用长鼻延伸反射 (PER) 测定法测得的记忆显着降低。然而，在48 hpt时，tau-氟缬氨酸，amitraz，香豆素和甲酸 (而不是百里香酚) 会导致记忆力减退。LD05剂量的杀菌剂不影响与记忆有关的neuroligin-1的表达，也不影响与记忆和发育有关的主要蜂王浆蛋白1的表达，尽管这两个基因的表达在LD50剂量下均受到影响。LD05剂量的百里香酚，甲酸，阿米特拉兹和库玛磷增加了defensin-1的表达，这与记忆和免疫有关。但是，百里香酚的作用可能是由于其对免疫反应而不是记忆的影响。这项研究表明，杀螨剂对蜜蜂记忆的影响各不相同，并且广泛使用的杀螨剂甲酸尤其具有破坏性。

BACKGROUND & AIMS: :Fifteen patients with laparoscopically diagnosed tubal pregnancy and constant or rising plasma beta-hCG levels were treated with prostaglandin F2 alpha and prostaglandin E2. Prostaglandin F2 alpha (5 mgms diluted in 10 cc of isotonic sodium solution) was injected transabdominally with a 22 gauge spinal needle during laparoscopy into the Fallopian tube. Prostaglandin E2 (500 micrograms ms) was given intramuscularly during three consecutive postoperative days. The treatment was defined as successful if plasma beta-hCG levels declined below the lower limit of detection and no further intervention other than prostaglandin application was required. The treatment was successful in eight patients. Six patients underwent laparotomy and salpingotomy because of rising beta-hCG levels. None of the treated patients displayed any adverse reactions following prostaglandin F2 alpha application. One patient underwent explorative laparotomy during the second postoperative day because of lower abdominal pain. During operation, no pathological change could be found. This patient was excluded from the study. In the group treated successfully (n = 8) seven out of eight patients had beta-hCG levels below 2500 mlU/ml preoperatively. In the unsuccessfully treated group (n = 6), four out of six patients had beta-hCG levels above 2500 mlU/ml preoperatively. Mean duration of beta-hCG decline to 10 percent of the maximum preoperative value was 15.8 +/- 8.64 days (mean +/- S.D.). Postoperatively, hysterosalpingography was performed in six out of eight successfully treated patients after three menstrual cycles (one patient had an intrauterine pregnancy, one patient refused written consent). The Fallopian tubes were patent bilaterally in all six patients.

背景与目标： : 用前列腺素f2α 和前列腺素e2治疗15例经腹腔镜诊断为输卵管妊娠且血浆 β-hCG水平持续或升高的患者。在腹腔镜检查期间，将前列腺素f2α (在10 cc的等渗钠溶液中稀释的5 mgms) 用22号脊柱针经腹注射到输卵管中。在术后连续三天肌肉注射前列腺素E2 (500微克ms)。如果血浆 β-hCG水平下降到检测下限以下，并且除前列腺素应用外不需要进一步干预，则治疗被定义为成功。八名患者的治疗成功。由于 β-hCG水平升高，六名患者接受了剖腹手术和输卵管切开术。在应用前列腺素f2α 后，治疗的患者均未显示任何不良反应。由于腹痛较低，一名患者在术后第二天接受了探索性剖腹手术。术中未见病理改变。该患者被排除在研究之外。在成功治疗的组中 (n = 8)，八名患者中有七名术前 β-hCG水平低于2500 mlU/ml。在未成功治疗组 (n = 6) 中，六名患者中有四名术前 β-hCG水平高于2500 mlU/ml。Β-hCG下降至最大术前值10% 的平均持续时间为15.8 +/- 8.64天 (平均值 +/-s.d.)。术后，在三个月经周期后，八名成功治疗的患者中有六名进行了子宫输卵管造影 (一名患者宫内妊娠，一名患者拒绝书面同意)。所有六名患者的输卵管均双侧未闭。

BACKGROUND & AIMS: :Non-nucleosidic phosphoramidite linker units suitable for use on commercial DNA synthesis machines have been designed for the direct incorporation of biotin and a new reporter group, phosphotyrosine, at multiple sites on synthetic oligonucleotides. The units are based on a 3-carbon glyceryl backbone where the reporter group is attached to the 2-O-position through a 3-aminopropyl spacer. 17-mer oligonucleotides were synthesized carrying at the 5'-end 1, 2, 4 or 8 biotinyl units or 1, 2, 4 or 8 phosphotyrosinyl units respectively and used for the detection of DNA on nitrocellulose filters by hybridization. Subsequent incubation of the filters with a monoclonal antibody to the reporter group followed by secondary detection using enhanced chemiluminescence (ECL) resulted in amplification of signal strengths as the number of reporter groups was increased. The results were quantitated by use of a charge couple device (CCD) camera. Spacing of biotin moieties by thymidyl residues resulted in further improvements in signal strengths, whereas similar spacing of phosphotyrosinyl units did not.

背景与目标： : 适用于商业DNA合成机器的非核苷亚磷酰胺接头单元已设计用于在合成寡核苷酸的多个位点上直接掺入生物素和新的报告基团磷酸酪氨酸。单元基于3-碳甘油基骨架，其中报告基团通过3-氨基丙基间隔体连接到2-o位。合成了17-聚体寡核苷酸，分别在5 '端携带1、2、4或8个生物素基单元或1、2、4或8个磷酸基酪氨酸基单元，并用于通过杂交检测硝酸纤维素过滤器上的DNA。随后将滤镜与单克隆抗体孵育到报告组，然后使用增强的化学发光 (ECL) 进行二次检测，随着报告组数量的增加，信号强度得到了放大。通过使用电荷耦合器件 (CCD) 相机对结果进行定量。胸苷基残基对生物素部分的间距导致信号强度的进一步提高，而磷酸基酪氨酸单元的间距却没有。

BACKGROUND & AIMS: :The treatment of glioblastoma remains a major challenge in the field of neuro-oncology. There is emerging evidence that glioblastomas consist of heterogeneous cell populations with a small subset of cells with stem cell-like properties which might be resistant to conventional therapy and are thus crucial for tumor recurrence. These glioma-initiating cells (GICs) are therefore an attractive therapeutic target. Death receptor activation is one promising approach of cancer therapy. The synthetic hexameric cluster of differentiation 95 (CD95) agonist APO010 exhibits strong antiglioma activity towards human glioma cell lines, as well as in cell cultures of primary glioblastoma. Here, we investigated the ability of APO010 to induce cell death in a panel of previously well-defined GIC lines. The GIC lines and their derived differentiated cultures expressed CD95 on the cell surface and were sensitive towards APO010-mediated cell death to a variable extent. Temozolomide enhanced sensitivity of GICs to APO010. APO010 warrants being further evaluated as a tool to target GICs.

背景与目标： : 胶质母细胞瘤的治疗仍然是神经肿瘤学领域的主要挑战。有新的证据表明，胶质母细胞瘤由异质细胞群组成，其中一小部分具有干细胞样特性的细胞亚群可能对常规疗法具有抵抗力，因此对肿瘤复发至关重要。因此，这些神经胶质瘤起始细胞 (gic) 是有吸引力的治疗靶标。死亡受体激活是一种有前途的癌症治疗方法。合成的六聚体分化簇95 (CD95) 激动剂APO010对人神经胶质瘤细胞系以及原发性胶质母细胞瘤的细胞培养物表现出很强的抗神经胶质瘤活性。在这里，我们研究了APO010在一组先前定义明确的GIC系中诱导细胞死亡的能力。GIC系及其衍生的分化培养物在细胞表面表达CD95，并在不同程度上对APO010-mediated细胞死亡敏感。替莫唑胺增强了GICs对apo010的敏感性。APO010认股权证被进一步评估为针对gic的工具。

BACKGROUND & AIMS: :A screening program was conducted to find microorganisms that modify the synthetic cannabinoid nabilone. After purification, the products from three cultures were analyzed by spectral methods to determine their chemical structures. An optically active 9S-hydroxy-6aR,10aR-trans cannabinoid was isolated from a culture of an unidentified soil bacterium designated A24007. From Bacillus cereus cultures were isolated a 9S,6'-dihydroxy-6aR,10aR-trans cannabinoid, a 9S-hydroxy-6'-keto-6aR,10aR-trans cannabinoid, a 9-keto-6'-hydroxy-6aS,10aS-trans cannabinoid, and a 6',9-diketo-6aS,10aS-trans cannabinoid. All of these products were optically active, as was a 9S-hydroxy-6aS,10AS-trans cannabinoid also isolated from B. cereus cultures. A series of acidic products were isolated from cultures of Nocardia salmonicolor. All of these products contained a carboxylic acid group at the terminal end of three-position alkyl side chains having varying numbers of carbon atoms. Two of the acidic products contained a 9-keto group, whereas all other carboxylic acid products were 9-hydroxy cannabinoids. The array of products obtained from incubation of nabilone indicates the usefulness of microbial transformations in the preparation of new cannabinoids.

背景与目标： : 进行了一项筛选程序，以发现修饰合成大麻素纳比酮的微生物。纯化后，通过光谱法分析三种培养物的产物以确定其化学结构。从鉴定为a24007的未鉴定土壤细菌的培养物中分离出光学活性的9s-羟基-6ar，10ar-反式大麻素。从蜡样芽孢杆菌培养物中分离出9S，6 '-dihydroxy-6aR，10ar-反式大麻素，9s-羟基-6'-keto-6aR，10ar-反式大麻素，9-酮-6 '-hydroxy-6aS，10as-反式大麻素和6'，9-二酮-6as，10as-反式大麻素。所有这些产品都是光学活性的，从蜡状芽孢杆菌培养物中也分离出9s-羟基-6as，10as-反式大麻素也是如此。从鲑鱼诺卡氏菌培养物中分离出一系列酸性产物。所有这些产物在具有不同碳原子数的三位烷基侧链的末端均包含羧酸基团。其中两个酸性产物含有9-酮基团，而所有其他羧酸产物都是9-羟基大麻素。从纳比酮的孵育中获得的一系列产品表明微生物转化在制备新大麻素中的有用性。

BACKGROUND & AIMS: :Cystic fibrosis (CF), an inherited disease characterized by defective epithelial Cl- transport, damages lungs via chronic inflammation and oxidative stress. Glutathione, a major antioxidant in the epithelial lung lining fluid, is decreased in the apical fluid of CF airway epithelia due to reduced glutathione efflux (Gao L, Kim KJ, Yankaskas JR, and Forman HJ. Am J Physiol Lung Cell Mol Physiol 277: L113-L118, 1999). The present study examined the question of whether restoration of chloride transport would also restore glutathione secretion. We found that a Cl- channel-forming peptide (N-K4-M2GlyR) and a K+ channel activator (chlorzoxazone) increased Cl- secretion, measured as bumetanide-sensitive short-circuit current, and glutathione efflux, measured by high-performance liquid chromatography, in a human CF airway epithelial cell line (CFT1). Addition of the peptide alone increased glutathione secretion (181 +/- 8% of the control value), whereas chlorzoxazone alone did not significantly affect glutathione efflux; however, chlorzoxazone potentiated the effect of the peptide on glutathione release (359 +/- 16% of the control value). These studies demonstrate that glutathione efflux is associated with apical chloride secretion, not with the CF transmembrane conductance regulator per se, and the defect of glutathione efflux in CF can be overcome pharmacologically.

背景与目标： 囊性纤维化 (CF) 是一种以上皮细胞Cl-转运缺陷为特征的遗传性疾病，通过慢性炎症和氧化应激损害肺部。谷胱甘肽是上皮肺衬里液中的主要抗氧化剂，由于谷胱甘肽外排减少 (Gao L，Kim KJ，Yankaskas JR和Forman HJ. Am J Physiol肺细胞Mol Physiol 277: L113-L118，1999)，在CF气道上皮的顶端液中减少。本研究研究了恢复氯化物运输是否也会恢复谷胱甘肽分泌的问题。我们发现形成Cl通道的肽 (N-K4-M2GlyR) 和K通道激活剂 (氯唑沙宗) 增加了Cl分泌，以布美他尼敏感的短路电流测量，并通过高效液相色谱法测量谷胱甘肽外排，在人CF气道上皮细胞系 (CFT1) 中。单独添加肽增加谷胱甘肽分泌 (对照值的181 +/- 8%)，而单独的氯唑沙宗不显著影响谷胱甘肽外排; 然而，氯唑沙宗增强了肽对谷胱甘肽释放的作用 (对照值的359 +/- 16%)。这些研究表明，谷胱甘肽的外排与顶端氯化物的分泌有关，而与CF跨膜电导调节剂本身无关，并且可以从药理学上克服CF中谷胱甘肽外排的缺陷。

BACKGROUND & AIMS: :The present study was undertaken to investigate the involvement of prostaglandins in the secretagogue action, observed after intraduodenal administration of rhein anthrone and rhein in rats. After intraduodenal administration of rhein anthrone (50 mg/kg), the active metabolite of sennosides, a very marked increase of secretion was observed compared to control. The amount of secretion was calculated by dividing the total weight of the small intestine, obtained 30 minutes after administration of the drug, by the total length. The effect seen with rhein (50 mg/kg) is far less pronounced than that with rhein anthrone and is not significant when compared with control. Pretreating the animals with indomethacin (15 mg/kg, p.o., 1 hour in advance) or with ibuprofen (15 mg/kg, p.o., 1 hour in advance) largely prevents the secretagogue effect of rhein anthrone, suggesting that prostaglandins play an important role in the observed pharmacological action. This idea is reinforced by the observation that pretreatment with hydrocortisone (50 mg/kg, p.o., 6 hours in advance) is also able to counteract the effect of rhein anthrone. After administration of rhein anthrone, an almost tenfold increase of the tissue content of prostaglandin E2 was observed. Here again, the results with rhein were far less pronounced. It is concluded that prostaglandins play an important role in the secretagogue action of rhein anthrone.

背景与目标： : 本研究旨在研究前列腺素在大鼠十二指肠内给予大黄酸和大黄酸后观察到的促分泌作用。十二指肠内给药大黄酸蒽酮 (50 mg/kg) 后，与对照组相比，观察到分泌明显增加。分泌量是通过将施用药物后30分钟获得的小肠的总重量除以总长度来计算的。大黄酸 (50 mg/kg) 的作用远不如大黄酸蒽酮明显，与对照组相比也不明显。用吲哚美辛 (15 mg/kg，p.o.，提前1小时) 或布洛芬 (15 mg/kg，p.o.，提前1小时) 预处理动物，很大程度上阻止了大黄酸蒽酮的促分泌作用，提示前列腺素在观察到的药理作用中起重要作用。观察到氢化可的松预处理 (50 mg/kg，p.o.，提前6小时) 也能够抵消大黄酸蒽酮的作用，从而强化了这一想法。给予大黄酸蒽酮后，观察到前列腺素E2的组织含量几乎增加了十倍。同样，使用rhein的结果远没有那么明显。结论前列腺素在大黄酸蒽酮的促分泌作用中起重要作用。

BACKGROUND & AIMS: :We report the first synthetic peptide vaccine eliciting strong CD8(+) and CD4(+) T lymphocyte responses in humans. The vaccine, representing the C-terminal region of the circumsporozoite protein of Plasmodium falciparum (amino acids 282-383) was well tolerated and strong sporozoite-specific antibodies were elicited. In addition, robust lymphocyte proliferation responses were equally elicited with concomitant in vitro production of IFN-gamma, crucial in the elimination of the parasite. Most importantly, we also observed the development of CD8(+) T lymphocyte responses decisive in the immunity to malaria. The latter finding opens new, possibly safer, avenues for vaccination strategies when a CD8(+) T cell response is needed.

背景与目标： : 我们报告了第一个合成肽疫苗，在人类中引起强烈的CD8 () 和CD4 () T淋巴细胞反应。代表恶性疟原虫环子孢子蛋白 (氨基酸282-383) 的C末端区域的疫苗具有良好的耐受性，并产生了强的子孢子特异性抗体。此外，伴随IFN-γ 的体外产生同样引起了强大的淋巴细胞增殖反应，这对于消除寄生虫至关重要。最重要的是，我们还观察到CD8 () T淋巴细胞反应的发展对疟疾的免疫力起决定性作用。当需要CD8(+) T细胞应答时，后者的发现为疫苗接种策略开辟了新的，可能更安全的途径。

BACKGROUND & AIMS: :In a previous study, we developed newly synthesized arylthio derivatives of cyclopentenone prostaglandins (GIF-0642, GIF-0643, GIF-0644, GIF-0745 and GIF-0747), which are neuroprotective against both manganese toxicity in PC12 cells and glutamate toxicity in HT22 cells. In the present study, we showed that these compounds and their lead compound, NEPP11, are potent inducers of glial cell line-derived neurotrophic factor (GDNF) expression in C6 glioma cells and primary astrocytes. These neuroprotective cyclopentenone prostaglandins also induced the gene expression of nerve growth factor and, to a lesser extent, brain-derived neurotrophic factor. The induction of GDNF mRNA was transcription-dependent, and the overexpression of dominant-negative Nrf2 attenuated the ability of the (arylthio)cyclopentenone prostaglandins to stimulate GDNF gene expression. These results suggest that (arylthio)cyclopentenone prostaglandins increase GDNF gene expression partly via the Keap1/Nrf2 pathway. A growing number of reports demonstrate the importance of increasing the amounts of neurotrophic factors, especially GDNF, in neuropathological states. Although the precise mechanisms by which the GIF compounds inhibit cell death are under investigation, an increase in neurotrophic factors may contribute to the diverse pharmacological properties of (arylthio)cyclopentenone prostaglandins in vivo and will make them potentially valuable in the treatment of neurodegenerative disorders.

背景与目标： : 在先前的研究中，我们开发了新合成的环戊烯酮前列腺素的芳基硫代衍生物 (GIF-0642，GIF-0643，GIF-0644，GIF-0745和GIF-0747)，它们对PC12细胞中的锰毒性和HT22细胞中的谷氨酸毒性均具有神经保护作用。在本研究中，我们表明这些化合物及其铅化合物NEPP11是C6神经胶质瘤细胞和原代星形胶质细胞中胶质细胞源性神经营养因子 (GDNF) 表达的有效诱导剂。这些神经保护性环戊烯酮前列腺素还诱导了神经生长因子的基因表达，并在较小程度上诱导了脑源性神经营养因子的基因表达。GDNF mRNA的诱导是转录依赖性的，显性负Nrf2的过度表达减弱了 (芳基硫代) 环戊烯酮前列腺素刺激GDNF基因表达的能力。这些结果表明 (芳基硫代) 环戊烯酮前列腺素部分通过Keap1/Nrf2途径增加GDNF基因表达。越来越多的报道证明了增加神经营养因子量的重要性,尤其是GDNF，在神经病理状态下，尽管GIF化合物抑制细胞死亡的确切机制正在研究中，但神经营养因子的增加可能有助于 (芳基硫代) 环戊烯酮前列腺素在体内的多种药理特性，并将使其在治疗神经退行性疾病中具有潜在的价值。

BACKGROUND & AIMS: :UDP-glucose:glycoprotein glucosyltransferase plays a key role in glycoprotein quality control in the endoplasmic reticulum, by virtue of its ability to discriminate folding states. Although lines of evidence have clarified the ability of UGGT to recognize a partially unfolded protein, its mechanistic rationale has been obscure. In this study, the substrate recognition mechanism of UGGT was studied using synthetic substrate of UGGT. Although UGGT has high extent of surface hydrophobicity, it clearly lacks property of typical molecular chaperones. Furthermore, it was revealed that the addition of the substrate caused secondary structure change of UGGT in a dose-dependent manner, resulting that the K(d) value of the UGGT-substrate interaction was estimated from theoretical formula based on 1:1 complexation between UGGT and the acceptor substrate. Moreover, the kinetic analysis of glucosyltransferase activity of UGGT elucidated Michaelis constant K(m) correctly.

背景与目标： : UDP-葡萄糖: 糖蛋白葡萄糖基转移酶凭借其区分折叠状态的能力，在内质网的糖蛋白质量控制中起着关键作用。尽管有大量证据阐明了UGGT识别部分未折叠蛋白质的能力，但其机理原理尚不清楚。在这项研究中，使用UGGT的合成底物研究了UGGT的底物识别机制。尽管UGGT具有很高的表面疏水性，但显然缺乏典型分子伴侣的特性。此外，揭示了底物的添加以剂量依赖性方式引起UGGT的二级结构变化，导致UGGT-底物相互作用的K(d) 值是根据基于UGGT与受体底物之间的1:1络合的理论公式估算的。此外，UGGT的葡萄糖基转移酶活性的动力学分析正确阐明了米氏常数K(m)。

BACKGROUND & AIMS: :Staphylococcus aureus is a major human pathogen producing different types of toxins. Enterotoxin A (SEA) is the most common type among clinical and food-related strains. The aim of the present study was to estimate functional regions of SEA that are responsible for emetic and superantigenic activities using synthetic peptides. A series of 13 synthetic peptides corresponding to specific regions of SEA were synthesized, and the effect of these peptides on superantigenic activity of SEA including interferon γ (IFN-γ) production in mouse spleen cells, SEA-induced lethal shock in mice, spleen cell proliferation in house musk shrew, and emetic activity in shrews were assessed. Pre-treatment of spleen cells with synthetic peptides corresponding to the regions 21-40, 35-50, 81-100, or 161-180 of SEA significantly inhibited SEA-induced IFN-γ production and cell proliferation. These peptides also inhibited SEA-induced lethal shock. Interestingly, peptides corresponding to regions 21-40, 35-50 and 81-100 significantly inhibited SEA-induced emesis in house musk shrews, but region 161-180 did not. These findings indicated that regions 21-50 and 81-100 of SEA are important for both superantigenic and emetic activities of SEA molecule while region 161-180 is involved in superantigenic activity but not emetic activity of SEA. These regions could be important targets for therapeutic intervention against SEA exposure.

背景与目标： : 金黄色葡萄球菌是产生不同类型毒素的主要人类病原体。肠毒素A (SEA) 是临床和食品相关菌株中最常见的类型。本研究的目的是使用合成肽估算负责催吐和超抗原活性的SEA功能区域。合成了一系列与SEA特定区域相对应的13种合成肽，并研究了这些肽对SEA超抗原活性的影响，包括小鼠脾细胞中干扰素 γ (IFN-γ) 的产生，小鼠SEA诱导的致死性休克，家常麝香sh的脾细胞增殖，并评估了鼩的催吐活性。用对应于SEA区域21-40、35-50、81-100或161-180的合成肽预处理脾细胞显著抑制了SEA诱导的IFN-γ 的产生和细胞增殖。这些肽还抑制了SEA诱导的致命性休克。有趣的是，对应于21-40、35-50和81-100区的肽显著抑制了家式麝香鼩的海诱导呕吐，但161-180区没有。这些发现表明，SEA的21-50和81-100区域对于SEA分子的超抗原和催吐活性都很重要，而161-180区域则参与了SEA的超抗原活性，但不参与SEA的催吐活性。这些区域可能是针对海洋暴露进行治疗干预的重要目标。

BACKGROUND & AIMS: Objectives:To determine the effect of medical treatment on work disability in patients with active PsA in a real-world setting. Methods:Four hundred patients with active PsA commencing or switching to anti-TNF or conventional synthetic DMARD (csDMARD) were recruited to a multicentre UK prospective observational cohort study. Work disability was measured using the work productivity and activity-specific health problem instrument and peripheral joint activity was measured with the disease activity in PsA composite measure. Results:Four hundred patients were recruited, of whom 229 (57.25%) were working (of any age). Sixty-two patients of working age (24%) were unemployed. At 6 months there was a 10% improvement in presenteeism ( P = 0.007) and a 15% improvement in work productivity ( P = 0.001) among working patients commenced on csDMARDs ( n = 164) vs a larger and more rapid 30% improvement in presenteeism ( P < 0.001) and 40% improvement in work productivity ( P < 0.001) among those commenced on anti-TNF therapy ( n = 65). Clinical response was poor among patients commenced on a csDMARD ( n = 272), with an 8.4 point improvement in disease activity in PsA ( P < 0.001) vs those commenced on anti-TNF therapy ( n = 121), who had a 36.8 point improvement ( P < 0.001). Conclusion:We report significant and clinically meaningful improvements in both work disability and clinical outcomes after commencement of anti-TNF therapy in a real-world setting. Improvements in all outcomes among those commencing csDMARDs were slower and of a smaller magnitude.

背景与目标：

BACKGROUND & AIMS: :Essential genes are absolutely required for the survival of any living entity. Investigation of essential genes is therefore expected to advance tremendously our understanding of the universal principles of life. Determination of a minimal set of essential genes needed to sustain life also plays an important role in the emerging field of synthetic biology, whose goals include creation of a stringently controlled minimal cell with predesigned phenotypic traits. In addition, due to their indispensability for survival of bacteria, genes encoding essential cellular functions have great potential in medicine as promising targets for the development of novel antimicrobials. Here, we review recent advances in the investigation of essential genes, with emphasis on the practical applications in medicine and synthetic biology.

背景与目标： : 任何生命实体的生存都绝对需要必需的基因。因此，对基本基因的研究有望极大地促进我们对生命普遍原理的理解。确定维持生命所需的最小必需基因集在合成生物学的新兴领域中也起着重要作用，其目标包括创建具有预先设计的表型特征的严格控制的最小细胞。此外，由于其对细菌生存的不可或缺性，编码必需细胞功能的基因在医学上具有巨大的潜力，可作为开发新型抗菌剂的有希望的靶标。在这里，我们回顾了必需基因研究的最新进展，重点是在医学和合成生物学中的实际应用。