BACKGROUND & AIMS: :A comprehensive immunohistochemistry with the isoform-distinguishable antibodies against prostaglandin (PG) F2α and PGE2 biosynthetic enzymes was undertaken to identify the cellular types and enzyme isoforms in rat ovary and uterus around parturition. In general ovarian and uterine cells showed positive immunoreactions for phospholipase A2 groups 4A and 6A, but not group 2A, and cyclooxygenase (COX)-1 rather than COX-2. Their immunoreactions for PGF2α synthase and PGE2 synthase were cell type-dependently variable. The putative PGF2α and PGE2 producing cell types included, as expected, ovarian luteal cells, uterine endometrial epithelium and myometrium, and cervical connective tissue and, unexpectedly, ovarian stromal cells and basal lamina of cervical endometrium. Obtained data indicate the generation of PGF2α and PGE2 by multiple sites, which are entirely the same as established sites of actions, in parturition processes and tissue-dependent differential usage of PG biosynthetic pathway.

背景与目标： : 采用针对前列腺素 (PG) F2α 和PGE2生物合成酶的亚型可区分抗体进行了全面的免疫组织化学检查，以鉴定分娩前后大鼠卵巢和子宫中的细胞类型和酶同工型。一般而言，卵巢和子宫细胞对磷脂酶A2组4A和6A组显示出阳性免疫反应，但对2A组不显示阳性，以及环氧合酶 (COX)-1而不是COX-2。他们对PGF2α 合酶和PGE2合酶的免疫反应是细胞类型依赖性可变的。如预期的那样，推定的PGF2α 和PGE2产生细胞类型包括卵巢黄体细胞，子宫内膜上皮和子宫肌层，宫颈结缔组织，以及出乎意料的卵巢基质细胞和宫颈子宫内膜基底层。获得的数据表明，在分娩过程和PG生物合成途径的组织依赖性差异使用中，多个位点产生了PGF2α 和PGE2，这些位点与已建立的作用位点完全相同。

BACKGROUND & AIMS: :CD4(+) helper T cells are critical for protective immune responses and yet suboptimally primed in response to tumors. Cell-based vaccination strategies are under evaluation in clinical trials but limited by the need to derive antigen-presenting cells (APC) from patients or compatible healthy donors. To overcome these limitations, we developed CD4(+) T cell-targeted synthetic microbead-based artificial APC (aAPC) and used them to activate CD4(+) T lymphocytes specific for a tumor-associated model antigen (Ag) directly from the naive repertoire. In vitro, aAPC specifically primed Ag-specific CD4(+) T cells that were activated to express high levels of CD44, produced mainly interleukin 2, and could differentiate into Th1-like or Th2-like cells in combination with polarizing cytokines. I.v. administration of aAPC led to Ag-specific CD4(+) T-cell activation and proliferation in secondary lymphoid organs, conferred partial protection against subcutaneous tumors, and prevented the establishment of lung metastasis. Taken together, our data support the use of cell-free, synthetic aAPC as a specific and versatile alternative to expand peptide-specific CD4(+) T cells in adoptive and active immunotherapy.

背景与目标： : CD4(+) 辅助性T细胞对保护性免疫反应至关重要，但对肿瘤的反应却不是最佳的。基于细胞的疫苗接种策略正在临床试验中进行评估，但由于需要从患者或相容的健康供体获得抗原呈递细胞 (APC) 的限制。为了克服这些局限性，我们开发了基于CD4 () T细胞靶向的基于合成微珠的人工APC (aAPC)，并使用它们直接从原始库中激活针对肿瘤相关模型抗原 (Ag) 的CD4 () T淋巴细胞。在体外，aAPC特异性引发Ag特异性CD4(+) T细胞，其被激活以表达高水平的CD44，主要产生白介素2，并且可以与极化细胞因子结合分化为Th1-like或Th2-like细胞。I.静脉给药aAPC导致Ag特异性CD4 () T细胞在次级淋巴器官中活化和增殖，对皮下肿瘤提供了部分保护，并阻止了肺转移的建立。综上所述，我们的数据支持使用无细胞，合成的aAPC作为一种特异性和多功能的替代品，以在过继和主动免疫疗法中扩展肽特异性CD4 () T细胞。

BACKGROUND & AIMS: :Intracellular supply of dTTP is a highly regulated process and has been a key target for chemotherapeutic drug development. Thymidylate kinase (TMPK) is the key enzyme for dTTP formation in both de novo and salvage pathways. In this study, we used lentiviral-based small hairpin RNA to silence TMPK expression in p53(+/+) and p53(-/-) HCT-116 colon cancer cells. This approach was sufficient to decrease the dTTP pool gradually without affecting p53 expression and generating cytotoxicity. TMPK knockdown significantly increased doxorubicin sensitivity dramatically in p53-proficient, p53-null HCT-116, and LoVo colon cancer cells. The decrease in the dTTP pool using this approach augmented the DNA damage response and enhanced apoptotic induction after exposure to low-dose doxorubicin, leading to cell death. In contrast, silencing of thymidylate synthase which blocks the de novo pathway was incapable of sensitizing p53-null HCT-116 cells to doxorubicin-induced apoptosis because of the compensation by the salvage pathway. Our results suggest the lentiviral delivery of small hairpin RNA targeting TMPK in combination with a low dose of doxorubicin as a new approach to kill colon cancer cells regardless of p53 status.

背景与目标： : dTTP的细胞内供应是一个高度调节的过程，并且一直是化学治疗药物开发的关键目标。胸苷酸激酶 (TMPK) 是从头和挽救途径中dTTP形成的关键酶。在这项研究中，我们使用基于慢病毒的小发夹RNA来沉默结肠癌细胞中p53 (/) 和p53(-/-) HCT-116 TMPK的表达。这种方法足以逐渐减少dTTP库，而不会影响p53表达并产生细胞毒性。TMPK敲低显着提高了p53-proficient，p53-null HCT-116和LoVo结肠癌细胞中阿霉素的敏感性。使用这种方法减少的dTTP库增加了DNA损伤反应，并增强了低剂量阿霉素暴露后的凋亡诱导，导致细胞死亡。相反，由于挽救途径的补偿，阻断从头途径的胸苷酸合酶的沉默不能使p53-null HCT-116细胞对阿霉素诱导的凋亡敏感。我们的结果表明，靶向TMPK的小发夹RNA的慢病毒递送与低剂量的阿霉素相结合，是一种杀死结肠癌细胞的新方法，而与p53状态无关。

BACKGROUND & AIMS: PURPOSE:To investigate the possible role of endogenous prostaglandins in the development of form deprivation myopia, as well as the effects of exogenous prostaglandins using atropine as a positive control. METHODS:Monocular form deprivation was accomplished by mounting a translucent occluder on one eye of 2-3 day old chicks for 1-4 weeks. Ocular occlusion for 1-2 weeks was used for pharmacological blocking experiments. The axial length of the eye was measured by ultrasonography. RESULTS:Indomethacin, administered intramuscularly, subconjunctivally or intravitreally had no significant effect on myopia development. Exogenous PGE2, PGF2alpha and latanoprost acid administered subconjunctivally, or topically as isopropyl ester eyedrops had no statistically significant effect on the myopia development. However, PGF2alpha significantly (p<0.01) attenuated the development of myopia after intravitreal injection. The other two prostaglandins had no statistically significant effect. CONCLUSIONS:Endogenous prostaglandins are unlikely to play a significant role in the development of form deprivation myopia in the chick. However, PGF2alpha suprisingly seems to retard the development of form deprivation myopia, but only when administered intravitreally. Whether the mechanism of the myopia retardation is direct or indirect remains unknown.

背景与目标：

BACKGROUND & AIMS: :A facile test system based on the accumulation of benzo[c]phenanthridine alkaloids in Eschscholzia californica cell suspension culture (an indicator of defense gene activation) has been used to analyze a series of synthetic compounds for elicitor-like activity. Of the 200 jasmonic acid and coronatine analogs tested with this system, representative results obtained with 49 of them are presented here. The following can be summarized concerning structure-activity relationships: there is a large degree of plasticity allowed at the C-3 of jasmonic acid in the activation of defense genes. The carbonyl moiety is not strictly required, but exocyclic double bond character appears necessary. The pentenyl side chain at C-2 cannot tolerate bulky groups at the terminal carbon and still be biologically active. Substitutions to the C-1' position are tolerated if they can potentially undergo beta-oxidation. Either an alkanoic acid or methyl ester is required at C-1, or a side chain that can be shortened by beta-oxidation or by peptidase hydrolysis. Coronatine and various derivatives thereof are not as effective as jasmonic acid, and derivatives in inducing benzo[c]phenanthridine alkaloid accumulation. Jasmonic acid rather than the octadecanoic precursors is therefore considered to be a likely signal transducer of defense gene activation in planta.

背景与目标： : 基于苯并 [c] 菲吡啶生物碱在加利福尼亚Eschscholzia细胞悬浮培养物 (防御基因激活的指标) 中的积累的简便测试系统已用于分析一系列具有激发子样活性的合成化合物。在用该系统测试的200茉莉酸和冠冕素类似物中，用其中的49个获得的代表性结果在这里给出。关于结构-活性关系，可以总结以下内容: 在防御基因的激活中，茉莉酸的C-3允许有很大程度的可塑性。羰基部分不是严格要求的，但是环外双键特征似乎是必需的。在C-2处的戊烯基侧链不能耐受末端碳处的大体积基团并且仍然具有生物活性。如果它们可能经历 β-氧化，则对C-1位置的取代是可以容忍的。C-1需要链烷酸或甲酯，或者可以通过 β-氧化或肽酶水解缩短的侧链。Coronatine及其各种衍生物在诱导苯并 [c] 菲吡啶生物碱积累方面不如茉莉酸有效。因此，茉莉酸而不是十八碳前体被认为是植物中防御基因激活的可能信号转导子。

BACKGROUND & AIMS: BACKGROUND:Autologous monocyte-derived dendritic cells (DCs) electroporated with synthetic messenger RNA (mRNA) encoding a CD40 ligand, a constitutively active Toll-like receptor 4 and CD70, together with mRNA encoding fusion proteins of a human leukocyte antigen (HLA)-class II targeting signal (DC-LAMP) and a melanoma-associated antigen (MAA); either MAGE-A3, MAGE-C2, tyrosinase or gp100) (TriMixDC-MEL) are superiorly immunogenic. PATIENTS AND METHODS:In this phase IB clinical trial, 24 million viable DCs were administered by four biweekly combined intradermal (id) and intravenous (iv) administrations, and a fifth administration on week 16. The number of iv-administered DCs was escalated in four sequentially treated cohorts. Immune responses were assessed by analysis of antigen specificity of blood-derived T-cells and skin infiltrating lymphocytes (SKILs). RESULTS:Fifteen patients with pretreated advanced melanoma tolerated administration of TriMixDC-MEL well. Two patients achieved a complete response and two patients a partial response. All objective responders are progression-free after a follow-up of, respectively, 24+, 28+, 33+, and 34+ months. Post-therapy antigen-specific SKILs were documented in 6 of 12 patients, and antigen-specific CD8(+) T-cells were detected in the blood of 4 of 5 patients. CONCLUSIONS:Cellular immunotherapy with TriMixDC-MEL is safe and immunogenic. Antitumor activity with durable disease control is observed across the investigated iv-dose levels. CLINICALTRIALSGOV IDENTIFIER:NCT01066390.

背景与目标：

BACKGROUND & AIMS: :There is a clear need for low-cost, self-applied, long-lasting approaches to prevent human immunodeficiency virus (HIV) infection in both men and women, even with the advent of pre-exposure prophylaxis (PrEP). Broadly neutralizing antibodies represent an option to improve HIV prophylaxis, but intravenous delivery, cold-chain stability requirements, low cervicovaginal concentrations, and cost may preclude their use. Here, we present an approach to express the anti-GP120 broadly neutralizing antibody PGT121 in the primary site of inoculation, the female reproductive tract, using synthetic mRNA. Expression is achieved through aerosol delivery of unformulated mRNA in water. We demonstrated high levels of antibody expression for over 28 days with a single mRNA administration in the reproductive tract of sheep. In rhesus macaques, neutralizing antibody titers in secretions developed within 4 h and simian-HIV (SHIV) infection of ex vivo explants was prevented. Persistence of PGT121 in vaginal secretions and epithelium was achieved through the incorporation of a glycosylphosphatidylinositol (GPI) anchor into the heavy chain of the antibody. Overall, we present a new paradigm to deliver neutralizing antibodies to the female reproductive tract for the prevention of HIV infections.

背景与目标： : 显然需要低成本，自我应用，持久的方法来预防男性和女性的人类免疫缺陷病病毒 (HIV) 感染，即使出现了接触前预防 (PrEP)。广泛中和抗体是改善HIV预防的一种选择，但是静脉内给药，冷链稳定性要求，低宫颈阴道浓度和成本可能会阻止其使用。在这里，我们提出了一种使用合成mRNA在接种的主要部位女性生殖道中表达anti-GP120的广泛中和抗体PGT121的方法。通过在水中气溶胶递送未配制的mRNA来实现表达。我们在绵羊的生殖道中单次施用mRNA证明了超过28天的高水平抗体表达。在恒河猴中，分泌物中的中和抗体滴度在4小时内产生，并防止了离体外植体的猿猴HIV (SHIV) 感染。PGT121在阴道分泌物和上皮中的持久性是通过将糖基磷脂酰肌醇 (GPI) 锚入抗体的重链中实现的。总体而言，我们提出了一种向女性生殖道提供中和抗体以预防HIV感染的新范式。

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。