BACKGROUND & AIMS: :Fluoxetine (FLX) the active pharmaceutical ingredient (API) in Prozac(®) is a widely prescribed psychoactive drug which ubiquitous occurrence in the aquatic environment is associated to a poor removal rate in waste-water treatment plant (WWTP) systems. This API acts as a selective serotonin reuptake inhibitor (SSRI) frequently reported to cause disrupting effects in non-target species. The objective of this study includes a multibiomarker response evaluation on mussel Mytilus galloprovincialis during two weeks exposure to 75 ng L(-1) FLX assessing antioxidant enzymes activities--superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); lipid peroxidation (LPO), acetylcholinesterase (AChE) neurotoxic response and endocrine disruption through alkali-labile phosphates (ALP) indirect measurement of vitellogenin-like proteins. Results show transient tissue-specific enzymatic responses and damage affecting mostly mussel gills. However, the clear ALP levels inhibition throughout time in both sex-differentiated gonads gives evidence to FLX reinforced action as an endocrine disruptor rather than an oxidative or neurotoxic inducer.

背景与目标： ：Prozac®中的活性药物成分（API）氟西汀（FLX）是一种广泛使用的精神活性药物，在水生环境中普遍存在，与废水处理厂（WWTP）系统中的去除率差有关。此API作为一种选择性血清素再摄取抑制剂（SSRI）经常报道引起非目标物种干扰作用。这项研究的目的包括在暴露于75 ng L（-1）FLX的两周期间对贻贝贻贝贻贝进行多生物标志物反应评估，以评估抗氧化酶活性-超氧化物歧化酶（SOD），过氧化氢酶（CAT）和谷胱甘肽S-转移酶（消费税）;脂质过氧化（LPO），乙酰胆碱酯酶（AChE）的神经毒性反应和通过碱不稳定的磷酸盐（ALP）间接测量卵黄蛋白原样蛋白的内分泌干扰。结果表明，短暂的组织特异性酶促反应和损害主要影响贻贝g。然而，在两个性别分化的性腺中，随着时间的推移，明显的ALP水平抑制作用为FLX作为内分泌干扰物而不是氧化或神经毒性诱导物的增强作用提供了证据。

BACKGROUND & AIMS: :The anti-inflammatory potential of p38 mitogen-activated protein kinase (MAPK) inhibitors was coincidentally expanded to a dual inhibition of p38α MAPK and phosphodiesterase 4 (PDE4), and the potential benefits arising from the blockage of both inflammation-related enzymes were thoroughly investigated. The most promising compound, CBS-3595 (1), was successively evaluated in in vitro experiments as well as in ex vivo and in vivo preclinical studies after administration of 1 to rodents, dogs, and monkeys. The resulting data clearly indicated a potent suppression of tumor necrosis factor alpha release. For reconfirming the findings of the animal studies when administering 1 to healthy human volunteers, a phase I clinical trial was conducted. Apart from further information regarding the pharmacokinetic and pharmacodynamic characteristics of 1, it was demonstrated that dual inhibition of p38α MAPK and PDE4 is able to synergistically attenuate the excessive anti-inflammatory response.

背景与目标： ：p38丝裂原活化蛋白激酶（MAPK）抑制剂的抗炎潜能同时扩展到了p38αMAPK和磷酸二酯酶4（PDE4）的双重抑制作用，并且两种炎症相关酶均被阻断产生了潜在的益处调查。给啮齿动物，狗和猴子施用1后，在体外实验以及离体和体内临床前研究中相继评估了最有前途的化合物CBS-3595（1）。所得数据清楚地表明有效抑制了肿瘤坏死因子α的释放。为了确认对健康的人类志愿者给药1时动物研究的结果，进行了I期临床试验。除了有关1的药代动力学和药效学特性的进一步信息外，还证明了对p38αMAPK和PDE4的双重抑制能够协同减弱过度的抗炎反应。

BACKGROUND & AIMS: PURPOSE:Many women with hormone receptor-positive breast cancer discontinue effective aromatase inhibitor (AI) treatment due to joint symptoms. METHODS:We conducted a single-arm, open-label, phase II study evaluating glucosamine-sulfate (1,500 mg/day) + chondroitin-sulfate (1,200 mg/day) for 24 weeks to treat joint pain/stiffness in postmenopausal women with early stage breast cancer who developed moderate-to-severe joint pain after initiating AIs. The primary endpoint was improvement in pain/stiffness at week 24 assessed by the Outcome Measure in Rheumatology Clinical Trials and Osteoarthritis Research Society International (OMERACT-OARSI) criteria. Secondary endpoints assessed changes in pain, stiffness, and function using the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index for hips/knees and the Modified Score for the Assessment and Quantification of Chronic Rheumatoid Affections of the Hands (M-SACRAH) for hands/wrists. The Brief Pain Inventory (BPI) assessed pain interference, severity, and worst pain. RESULTS:Of 53 patients enrolled, 39 were evaluable at week 24. From baseline to week 24, 46 % of patients improved according to OMERACT-OARSI criteria. At week 24, there were improvements (all P < 0.05) in pain and function as assessed by WOMAC and M-SACRAH, and in pain interference, severity, and worst pain as assessed by BPI. Estradiol levels did not change from baseline. The most commonly reported side effects were headache (28 %), dyspepsia (15 %), and nausea (17 %). CONCLUSIONS:In this single-arm study, 24 weeks of glucosamine/chondroitin resulted in moderate improvements in AI-induced arthralgias, with minimal side effects, and no changes in estradiol levels. These results suggest a need to evaluate efficacy in a placebo-controlled trial.

背景与目标： 目的：许多患有激素受体阳性乳腺癌的妇女由于关节症状而停止有效的芳香化酶抑制剂（AI）治疗。
方法：我们进行了单臂，开放标签，II期研究，评估硫酸氨基葡萄糖（1,500毫克/天）硫酸软骨素（1200毫克/天）持续24周，以治疗绝经后早期女性的关节痛/僵硬启动AI后出现中度至重度关节痛的乳腺癌患者。主要终点是通过风湿病临床试验和国际骨关节炎研究协会（OMERACT-OARSI）标准中的结果评估标准评估的第24周疼痛/僵硬的改善。次要终点是使用西安大略省和麦克马斯特大学骨关节炎（WOMAC）指数评估臀部/膝盖的疼痛，僵硬和功能的变化，以及评估手的慢性类风湿病影响和量化的改良分数（M-SACRAH） / wrists。简短疼痛清单（BPI）评估了疼痛的干扰，严重程度和最严重的疼痛。
结果：在入组的53例患者中，有39例在第24周得到了评估。从基线到第24周，根据OMERACT-OARSI标准，有46％的患者得到了改善。在第24周时，WOMAC和M-SACRAH评估的疼痛和功能以及BPI评估的疼痛干扰，严重程度和最严重疼痛得到改善（所有P <0.05）。雌二醇水平与基线相比没有变化。最常见的副作用是头痛（28％），消化不良（15％）和恶心（17％）。
结论：在这项单臂研究中，葡萄糖胺/软骨素的24周可导致AI引起的关节痛适度改善，副作用最小，且雌二醇水平无变化。这些结果表明需要在安慰剂对照试验中评估疗效。

BACKGROUND & AIMS: :Plasminogen activator inhibitor-1 (PAI-1) is a member of the serine protease inhibitor (serpin) superfamily, which inactivates tissue plasminogen activator (tPA); therefore, increased level of PAI-1 antigen counteracts the anticoagulant effect of tPA and facilitates the fibrin clot formation. Plasma PAI-1 antigen and activity levels are associated with increased body mass index and with features of the insulin resistance syndrome like obesity and diabetes. Visceral adipose tissue produces more PAI-1 than subcutaneous adipose tissue: This increased production of PAI-1 from the visceral adipose tissue is one important link between visceral obesity and cardiovascular disease. Besides visceral adipose tissue, there is mounting evidence that epicardial adipose tissue may be an important source of PAI-1, especially in patients with type 2 diabetes.

背景与目标： ：Plasminogen activator inhibitor-1（PAI-1）是丝氨酸蛋白酶抑制剂（serpin）超家族的一员，它可以使组织纤溶酶原激活物（tPA）失活。因此，PAI-1抗原水平的增加抵消了tPA的抗凝作用，并促进了纤维蛋白凝块的形成。血浆PAI-1抗原和活性水平与体重指数增加以及胰岛素抵抗综合征（如肥胖症和糖尿病）的特征有关。内脏脂肪组织比皮下脂肪组织产生更多的PAI-1：内脏脂肪组织增加的PAI-1产生是内脏肥胖与心血管疾病之间的重要联系。除内脏脂肪组织外，越来越多的证据表明心外膜脂肪组织可能是PAI-1的重要来源，尤其是在2型糖尿病患者中。

BACKGROUND & AIMS: Prostaglandins and thromboxanes are products of arachidonic acid metabolism via the cyclooxygenase (CO) enzyme and are responsible for the pain and swelling common to sites of inflammation. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the production of these substances and are used in the treatment of inflammatory diseases such as arthritis. However, one of the major side-effects of NSAID therapy is gastric ulceration. It is possible that inhibition of prostaglandin production and a related increase in the formation of leukotrienes via the 5-lipoxygenase (5-LO) enzymatic pathway are responsible for attracting inflammatory cells, causing local sites of inflammation and producing ulceration. To determine the effects of 5-LO inhibition on this hypothesis, studies were performed in rats to evaluate the effects of tepoxalin, a dual CO/LO inhibitor on leukotriene B4 levels in gastric mucosa and neutrophil adhesion in mesenteric venules. In rats, chronic oral administration of an NSAID, indomethacin (2 mg/kg daily over 4 days), resulted in 40% mortality, accompanied by intestinal adhesions and perforations when evaluated 24 h after the fourth dose of drug. Additionally, neutrophil adhesion was increased in the mesenteric venules and cell infiltration was evident in the mesenteric interstitium. These gastrointestinal side-effects were inhibited in a separate group of rats administered tepoxalin (20 mg/kg, p.o) 30 min prior to each daily indomethacin treatment. Further studies were performed to determine tepoxalin's effects on early events associated with NSAID-induced gastrointestinal inflammation, including neutrophil adhesion, lipid peroxide generation and LTB4 production. Indomethacin (100 mg/kg, p.o.) produced elevated levels of LTB4 in rat gastric mucosa 90 min after administration. Additionally, neutrophil adhesion in mesenteric venules was increased at this dose and with the administration of another NSAID, naproxen. No generation of lipid peroxides was evident in the gastric mucosa at this timepoint. Tepoxalin (up to 400 mg/kg, p.o.) did not have an effects on gastric mucosal LTB4 generation and lipid peroxide levels. A decrease in neutrophil adhesion was observed at the highest dose. In another study, pretreatment with tepoxalin (ED50=7.5 mg/kg, p.o.) or the selective 5-LO inhibitor zileuton (100 mg/kg, p.o.) prevented the increases in gastric mucosal LTB4 levels and neutrophil adhesion induced by indomethacin (100 mg/kg, p.o.). These data suggest that LO inhibition may play a vital role in the prevention of NSAID-induced gastric inflammation, providing insight into the lack of ulcerogenicity with tepoxalin and new approaches to anti-inflammatory therapy which may prevent gastric side effects.

背景与目标： 前列腺素和血栓烷是花生四烯酸通过环氧合酶（CO）酶代谢的产物，它们是炎症部位常见的疼痛和肿胀的原因。非甾体类抗炎药（NSAIDs）抑制这些物质的产生，并用于治疗炎症性疾病，例如关节炎。但是，NSAID治疗的主要副作用之一是胃溃疡。通过5-脂氧合酶（5-LO）酶促途径抑制前列腺素的产生以及白三烯形成的相关增加可能是吸引炎症细胞，引起局部炎症和产生溃疡的原因。为了确定5-LO抑制对这一假设的影响，在大鼠中进行了研究以评估替泊沙林（一种双重CO / LO抑制剂）对胃粘膜白三烯B4水平和肠系膜小静脉中性粒细胞粘附的影响。在大鼠中，长期口服NSAID，消炎痛（2天每天2 mg / kg，共4天）可导致40％的死亡率，并在第四剂药物后24小时进行评估，并伴有肠粘连和穿孔。另外，在肠系膜小静脉中嗜中性粒细胞的粘附增加，并且在肠系膜间质中细胞浸润明显。在每天进行消炎痛治疗前30分钟，给另一组大鼠服用tepoxalin（20 mg / kg，p.o），抑制了这些胃肠道副作用。进行了进一步的研究以确定替泊沙林对与NSAID诱导的胃肠道炎症相关的早期事件的影响，包括中性粒细胞粘附，脂质过氧化物的产生和LTB4的产生。吲哚美辛（100 mg / kg，p.o.）给药90分钟后，大鼠胃黏膜中LTB4的水平升高。另外，在该剂量下并通过使用另一种NSAID萘普生，肠系膜小静脉中的中性粒细胞粘附增加。在这个时间点在胃粘膜中没有明显的脂质过氧化物的产生。 Tepoxalin（最高400 mg / kg，p.o.）对胃粘膜LTB4的产生和脂质过氧化物的水平没有影响。在最高剂量下观察到嗜中性粒细胞粘附性降低。在另一项研究中，用替泊沙林（ED50 = 7.5 mg / kg，口服）或选择性5-LO抑制剂齐留通（100 mg / kg，口服）预处理可防止由吲哚美辛（100 mg / min）引起的胃粘膜LTB4水平增加和嗜中性白细胞粘附/ kg，po）。这些数据表明，LO抑制可能在预防NSAID引起的胃部炎症中起着至关重要的作用，从而提供了对特泊沙林缺乏致溃疡性的认识，并提供了可预防胃部副作用的新的抗炎治疗方法。

BACKGROUND & AIMS: 1. Selective 5-hydroxytryptamine (5-HT; serotonin) reuptake inhibitors (SSRIs) cause a greater increase in extracellular 5-HT in the forebrain when the somatodendritic 5-HT1A autoreceptor is blocked. Here, we investigated whether blockade of the terminal 5-HT1B autoreceptor influences a selective 5-HT reuptake inhibitor in the same way, and whether there is an additional effect of blocking both the 5-HT1A and 5-HT1B autoreceptors. 2. Extracellular 5-HT was measured in frontal cortex of the anaesthetized rat by use of brain microdialysis. In vivo extracellular recordings of 5-HT neuronal activity in the dorsal raphe nucleus (DRN) were also carried out. 3. The selective 5-HT reuptake inhibitor, paroxetine (0.8 mg kg-1, i.v.), increased extracellular 5-HT about 2 fold in rats pretreated with the 5-HT1A receptor antagonist, WAY100635. When administered alone neither paroxetine (0.8 mg kg-1, i.v.) nor WAY100635 (0.1 mg kg-1, i.v.) altered extracellular 5-HT levels. 4. Paroxetine (0.8 mg kg-1, i.v.) did not increase 5-HT in rats pretreated with the 5-HT1B/D receptor antagonist, GR127935 (1 mg kg-1, i.v.). GR127935 (1 and 5 mg kg-1, i.v.) had no effect on extracellular 5-HT when administered alone. 5. Interestingly, paroxetine (0.8 mg kg-1, i.v.) caused the greatest increase in 5-HT (up to 5 fold) when GR127935 (1 or 5 mg kg-1, i.v.) was administered in combination with WAY100635 (0.1 mg kg-1, i.v.). Administration of GR127935 (5 mg kg-1, i.v.) plus WAY100635 (0.1 mg kg-1, i.v.) without paroxetine, had no effect on extracellular 5-HT in the frontal cortex. 6. Despite the lack of effect of GR127935 on 5-HT under basal conditions, when 5-HT output was elevated about 3 fold (by adding 1 microM paroxetine to the perfusion medium), the drug caused a dose-related (1 and 5 mg kg-1, i.v.) increase in 5-HT. 7. By itself, GR127935 slightly but significantly decreased 5-HT cell firing in the DRN at higher doses (2.0-5.0 mg kg-1, i.v.), but did not prevent the inhibition of 5-HT cell firing induced by paroxetine. 8. In summary, our results suggest that selective 5-HT reuptake inhibitors may cause a large increase in 5-HT in the frontal cortex when 5-HT autoreceptors on both the somatodendrites (5-HT1A) and nerve terminals (5-HT1B) are blocked. This increase is greater than when either set of autoreceptors are blocked separately. The failure of a 5-HT1B receptor antagonist alone to enhance the effect of the selective 5-HT reuptake inhibitor in our experiments may be related to a lack of tone on the terminal 5-HT1B autoreceptor due to a continued inhibition of 5-HT cell firing. These results are discussed in relation to the use of 5-HT autoreceptor antagonists to augment the antidepressant effect of selective 5-HT reuptake inhibitors.

背景与目标： 1.当抑制体树突状5-HT1A自体受体时，选择性5-羟色胺（5-HT； 5-羟色胺）再摄取抑制剂（SSRIs）导致前脑细胞外5-HT的增加。在这里，我们调查了终端5-HT1B自身受体的阻断作用是否以相同的方式影响选择性5-HT再摄取抑制剂，以及是否存在阻断5-HT1A和5-HT1B自身受体的其他作用。 2.通过使用脑微透析在麻醉的大鼠的额皮质中测量细胞外5-HT。还进行了背缝核（DRN）中5-HT神经元活性的体内细胞外记录。 3.选择性5-HT再摄取抑制剂帕罗西汀（0.8mg kg-1，静脉内）在用5-HT1A受体拮抗剂WAY100635预处理的大鼠中使细胞外5-HT增加约2倍。当单独给药时，帕罗西汀（0.8 mg kg-1，i.v.）和WAY100635（0.1 mg kg-1，i.v.）均未改变细胞外5-HT水平。 4.在用5-HT1B / D受体拮抗剂GR127935（1 mg kg-1，i.v.）预处理的大鼠中，帕罗西汀（0.8 mg kg-1，i.v.）不会增加5-HT。单独给药时，GR127935（1和5 mg kg-1，静脉内）对细胞外5-HT无效。 5.有趣的是，当GR127935（1或5 mg kg-1，iv）与WAY100635（0.1 mg）联合使用时，帕罗西汀（0.8 mg kg-1，iv）引起5-HT的最大增加（最多5倍）。千克-1，iv）。不含帕罗西汀的GR127935（5 mg kg-1，i.v.）加上WAY100635（0.1 mg kg-1，i.v.）的给药对额皮质中的细胞外5-HT没有影响。 6.尽管在基本条件下GR127935对5-HT缺乏影响，但当5-HT产量提高了约3倍时（通过向灌注培养基中添加1 microM帕罗西汀），该药物引起了剂量相关（1和5 mg kg-1，iv）5-HT升高。 7.单独使用时，GR127935在较高剂量（2.0-5.0 mg kg-1，i.v。）下可轻微但显着降低DRN中5-HT细胞的发射，但不能阻止帕罗西汀诱导的5-HT细胞发射的抑制。 8.总而言之，我们的研究结果表明，当在肢体树突（5-HT1A）和神经末梢（5-HT1B）上都存在5-HT自身受体时，选择性5-HT再摄取抑制剂可能会导致额叶皮层5-HT大量增加。被阻止。该增加量大于分别封闭任一组自动受体时的增加量。在我们的实验中，单独使用5-HT1B受体拮抗剂未能增强选择性5-HT再摄取抑制剂的作用可能与由于持续抑制5-HT细胞而导致末端5-HT1B自体受体缺乏音调有关射击。这些结果与使用5-HT自身受体拮抗剂增强选择性5-HT再摄取抑制剂的抗抑郁作用有关。

BACKGROUND & AIMS: :Direct thrombin inhibitors have proven efficacious in prevention of venous thromboembolism. Bleeding complications are rare, but in case of acute serious bleeding, an effective and instant haemostatic intervention may be required. In the present study it was demonstrated that the direct thrombin inhibitor melagatran induces dose-dependent abnormalities in whole blood (WB) clotting profiles as recorded by a recently described modified thrombelastographic model, and that rFVIIa or APCC are capable of improving the haemostatic capacity. Experiments were performed using WB from 30 healthy males. In-vitro titration experiments (n = 10) with addition of melagatran to WB corresponding to plasma concentrations ranging from 0 to 5.0 microM (12 steps) showed a dose-dependent prolongation of the clot initiation and characteristic decrease of the maximum rate of clot propagation. In-vitro intervention studies (n = 20) were completed with four different concentrations of melagatran as well as addition of four different levels of rFVIIa or APCC. At all tested concentrations of melagatran, rFVIIa significantly shortened the melagatran-induced prolonged clot initiation but induced only minor improvements of the reduced clot propagation. In contrast, APCC significantly and dose-dependently shortened the clot initiation and accelerated the clot propagation. In conclusion, our thrombelastographic model appears useful for evaluating the effect of direct thrombin inhibitors on dynamicWB clot formation and rFVIIa, but especially APCC significantly improved theWB clot formation. The pronounced stabilizing effect of APCC may be caused by its content of prothrombin and activated coagulation factors.

背景与目标： ：直接凝血酶抑制剂已被证明可有效预防静脉血栓栓塞。出血并发症很少见，但在发生急性严重出血的情况下，可能需要有效且即时的止血干预。在本研究中，正如最近描述的改良血栓弹力图模型所记录的那样，已证明直接凝血酶抑制剂美拉加群在全血（WB）凝血曲线中诱导了剂量依赖性异常，并且rFVIIa或APCC能够提高止血能力。使用来自30名健康男性的WB进行实验。体外滴定实验（n = 10），向血浆中添加美拉加群，对应血浆浓度范围为0至5.0 microM（12步），显示了血块起始的剂量依赖性延长和最大血块繁殖速率的特征性降低。体外干预研究（n = 20）用四种不同浓度的美拉加群以及添加四种不同水平的rFVIIa或APCC完成。在所有测试浓度的黑加仑中，rFVIIa显着缩短了由黑加仑引起的血凝块起始时间的延长，但仅引起血凝块减少的轻微改善。相比之下，APCC显着且剂量依赖性地缩短了血凝块起始并加速了血凝块繁殖。总之，我们的血栓弹力图模型似乎可用于评估直接凝血酶抑制剂对动态WB凝块形成和rFVIIa的作用，但尤其是APCC可以显着改善WB凝块的形成。 APCC明显的稳定作用可能是由于其凝血酶原含量和活化的凝血因子引起的。

BACKGROUND & AIMS: :Plasmepsins are aspartic proteases involved in the degradation of the host cell hemoglobin that is used as a food source by the malaria parasite. Plasmepsins are highly promising as drug targets, especially when combined with the inhibition of falcipains that are also involved in hemoglobin catabolism. In this review, we discuss the mechanism of plasmepsins I-IV in view of the interest in transition state mimetics as potential compounds for lead development. Inhibitor development against plasmepsin II as well as relevant crystal structures are summarized in order to give an overview of the field. Application of computational techniques, especially binding affinity prediction by the linear interaction energy method, in the development of malarial plasmepsin inhibitors has been highly successful and is discussed in detail. Homology modeling and molecular docking have been useful in the current inhibitor design project, and the combination of such methods with binding free energy calculations is analyzed.

背景与目标： ：胃蛋白酶是天冬氨酸蛋白酶，参与宿主细胞血红蛋白的降解，被疟原虫用作食物来源。纤溶酶作为药物靶标非常有前途，特别是与抑制也与血红蛋白分解代谢有关的falcipains结合使用时。在这篇综述中，我们鉴于对过渡态模拟物作为潜在的潜在铅化合物的发展感兴趣，讨论了纤溶酶I-IV的机制。概述了针对纤溶酶II的抑制剂的开发以及相关的晶体结构，以便对该领域进行概述。计算技术的应用，特别是通过线性相互作用能法的结合亲和力预测，在疟疾纤溶酶抑制剂的开发中已经取得了很大的成功，并进行了详细的讨论。均相建模和分子对接在当前的抑制剂设计项目中非常有用，并且分析了这些方法与结合自由能计算的结合。

BACKGROUND & AIMS: PURPOSE:Steroid sulfatase (STS) inhibitors that can decrease or prevent the biosynthesis of estrogenic steroids via the sulfatase route may play an important role in the treatment of breast cancer. We compare the in vivo efficacy of two potent STS inhibitors, STX64 and STX213, in a xenograft breast cancer model. EXPERIMENTAL DESIGN:MCF-7 cells stably expressing STS cDNA (MCF-7STS) were generated. Ovariectomized MF-1 female nude mice receiving s.c. injections of estradiol sulfate (E2S) and bearing both MCF-7STS and wild-type MCF-7 (MCF-7WT) tumors were orally treated with STX64 and STX213. Treatment was given for 49 days followed by a recovery period of 35 days in which animals received only E2S. Mice were weighed, and tumor measurements were taken weekly. RESULTS:STX64 and STX213 exhibited potent STS inhibition in vivo. However, STX213 showed a greater duration of activity. In vehicle-treated nude mice receiving E2S, tumor volumes increased 5.5-fold for MCF-7WT and 3.8-fold for MCF-7STS after 49 days compared with day 0. MCF-7WT tumor growth was reduced by 56% by STX213 over the dosing period, and subsequent growth was retarded during the recovery period. All treatments fully inhibited growth of MCF-7STS tumors, and recovery of these tumors was significantly retarded (P<0.01). All compounds completely inhibited liver and tumor STS activity. Additionally, STS mRNA expression in the MCF-7STS tumors directly correlated with the corresponding STS enzyme activity. CONCLUSIONS:This study indicates that STS inhibitors attenuate hormone-dependent human breast cancer growth and therefore offer a potentially novel treatment for this condition.

背景与目标： 用途：类固醇硫酸酯酶（STS）抑制剂可通过硫酸酯酶途径减少或阻止雌激素类固醇的生物合成，可能在乳腺癌的治疗中起重要作用。我们比较了两种有效的STS抑制剂STX64和STX213在异种移植乳腺癌模型中的体内疗效。
实验设计：产生稳定表达STS cDNA（MCF-7STS）的MCF-7细胞。接受皮下切除的卵巢切除的MF-1雌性裸鼠注射雌二醇硫酸盐（E2S）并同时携带MCF-7STS和野生型MCF-7（MCF-7WT）肿瘤的患者，应使用STX64和STX213口服治疗。给予49天的治疗，然后恢复35天，其中动物仅接受E2S。称重小鼠，每周进行一次肿瘤测量。
结果：STX64和STX213在体内表现出有效的STS抑制作用。但是，STX213显示了更长的活动时间。在接受E2S的媒介物治疗的裸鼠中，与第0天相比，49天后MCF-7WT的肿瘤体积增加了5.5倍，MCF-7STS的肿瘤体积增加了3.8倍。STX213使MCF-7WT的肿瘤生长比给药减少了56％期间，随后的生长在恢复期受到阻碍。所有治疗均完全抑制MCF-7STS肿瘤的生长，并且这些肿瘤的恢复显着受阻（P <0.01）。所有化合物均完全抑制肝脏和肿瘤STS活性。此外，MCF-7STS肿瘤中的STS mRNA表达与相应的STS酶活性直接相关。
结论：这项研究表明，STS抑制剂可减弱激素依赖性人类乳腺癌的生长，因此可为这种情况提供潜在的新颖治疗方法。

BACKGROUND & AIMS: :Plasminogen activator inhibitor-1 (PAI-1) is an inhibitor of fibrinolysis. Increased plasma PAI-1 levels play an essential role in the pathogenesis of cardiovascular risk and other diseases associated with thrombosis. The 4G/5G polymorphism of the PAI-1 promoter region has been extensively studied in different populations. We studied 160 healthy unrelated Lebanese individuals using a reverse hybridization PCR assay to detect the 5G/5G, 4G/5G and, 4G/4G genotypes of the PAI-1 gene and the frequencies of the 4G and 5G alleles. We found that 4G/5G genotype was the most prevalent (45.6%) followed by 5G/5G (36.9%) and 4G/4G (17.5%). The frequencies of the 4G and 5G alleles were calculated to be 0.403 and 0.597, respectively. Compared to other ethnic communities, the Lebanese population was found to harbour a relatively high prevalence of the rare 4G allele. This, in turn, may predispose this population to develop cardiovascular diseases and other thrombotic clinical conditions. This study aids to enhance our understanding of the genetic features of the Lebanese population.

背景与目标： ：Plasminogen activator inhibitor-1（PAI-1）是纤维蛋白溶解抑制剂。血浆PAI-1水平升高在心血管风险和其他与血栓形成有关的疾病的发病机理中起着至关重要的作用。 PAI-1启动子区域的4G / 5G多态性已在不同人群中进行了广泛研究。我们使用反向杂交PCR方法研究了160名健康的黎巴嫩无关个体，以检测PAI-1基因的5G / 5G，4G / 5G和4G / 4G基因型以及4G和5G等位基因的频率。我们发现4G / 5G基因型是最普遍的（45.6％），其次是5G / 5G（36.9％）和4G / 4G（17.5％）。计算得出4G和5G等位基因的频率分别为0.403和0.597。与其他种族社区相比，发现黎巴嫩人口中稀有4G等位基因的患病率较高。反过来，这可能使该人群容易患上心血管疾病和其他血栓形成性临床状况。这项研究有助于增进我们对黎巴嫩人口遗传特征的了解。

BACKGROUND & AIMS: :The recent progress in chemotherapy for malignant gliomas is attributable to the introduction of the DNA-methylating agent temozolomide (TMZ); however, drug resistance remains a major issue. Previous studies have shown that TMZ induces prolonged arrest of human glioma cells in the G2/M phase of the cell cycle followed by a senescence-like phenomenon or mitotic catastrophe. These findings suggest that the G2 checkpoint is linked to DNA repair mechanisms. We investigated the effect of a cyclin-dependent kinase (Cdk) inhibitor flavopiridol (FP) that inhibits the action of Cdc2, a key protein in the G2 checkpoint pathway, on TMZ-treated glioma cells. Colony formation efficiency revealed that FP potentiated the cytotoxicity of TMZ in glioma cells in a p53-independent manner. This effect was clearly associated with the suppression of key proteins at the G2-M transition, accumulation of the cells exclusively at the G2 phase, and increase in a double-stranded DNA break marker (seen on performing immunoblotting). TMZ-resistant clones showed activation of the G2 checkpoint in response to TMZ, while FP treatment resensitized these clones to TMZ. FP also enhanced the cytotoxicity of TMZ in U87MG-AktER cells. Moreover, administration of TMZ and/or FP to nude mice with xenografted U87MG cells revealed that FP sensitized xenografted U87MG cells to TMZ in these mice. Our findings suggest that TMZ resistance could be promoted by enhanced DNA repair activity in the G2-M transition and that a Cdk inhibitor could suppress this activity, leading to potentiation of TMZ action on glioma cells.

背景与目标： ：恶性神经胶质瘤化学疗法的最新进展归因于DNA甲基化剂替莫唑胺（TMZ）的引入；但是，耐药性仍然是一个主要问题。先前的研究表明，TMZ诱导人神经胶质瘤细胞在细胞周期的G2 / M期延长停滞，随后出现衰老样现象或有丝分裂灾难。这些发现表明，G2检查点与DNA修复机制有关。我们研究了细胞周期蛋白依赖性激酶（Cdk）抑制剂flavopiridol（FP）抑制TMZ处理的神经胶质瘤细胞对Cdc2（G2检查点途径中的关键蛋白）的作用。集落形成效率表明，FP以非p53依赖性方式增强了胶质瘤细胞中TMZ的细胞毒性。这种作用显然与关键蛋白在G2-M转换中的抑制，细胞仅在G2期的蓄积以及双链DNA断裂标记的增加（见于进行免疫印迹）有关。耐TMZ的克隆显示出对TMZ的响应，激活了G2检查点，而FP处理使这些克隆对TMZ再次敏感。 FP还增强了U87MG-AktER细胞中TMZ的细胞毒性。此外，对具有异种移植的U87MG细胞的裸鼠施用TMZ和/或FP显示在这些小鼠中FP使异种移植的U87MG细胞对TMZ敏感。我们的发现表明，TMZ耐药性可以通过在G2-M转换中增强DNA修复活性来促进，而Cdk抑制剂可以抑制该活性，从而增强TMZ对神经胶质瘤细胞的作用。

BACKGROUND & AIMS: BACKGROUND:Hyperpigmentary disorders like melasma, actinic and senile lentigines are a major cosmetic concern. Therefore, many topical products are available, containing various active ingredients aiming to reduce melanin production and distribution. The most prominent target for inhibitors of hyperpigmentation is tyrosinase, the key regulator of melanin production. Many inhibitors of tyrosinase are described in the literature; however, most of them lack clinical efficacy. METHODS:We were interested in evaluating the inhibition of skin pigmentation by well-known compounds with skin-whitening activity like hydroquinone, arbutin, kojic acid and 4-n-butylresorcinol. We compared the inhibition of human tyrosinase activity in a biochemical assay as well as inhibition of melanin production in MelanoDerm skin model culture. For some compounds, the in vivo efficacy was tested in clinical studies. RESULTS:Arbutin and hydroquinone only weakly inhibit human tyrosinase with a half maximal inhibitory concentration (IC(50)) in the millimolar range. Kojic acid is 10 times more potent with an IC(50) of approximately 500 μmol/L. However, by far the most potent inhibitor of human tyrosinase is 4-n-butylresorcinol with an IC(50) of 21 μmol/L. In artificial skin models, arbutin was least active with an IC(50) for inhibition of melanin production > 5000 μmol/L. Kojic acid inhibited with an IC(50) > 400 μmol/L. Interestingly, hydroquinone inhibited melanin production in MelanoDerms with an IC(50) below 40 μmol/L, probably due to a mechanism different from tyrosinase inhibition. Again, 4-n-butylresorcinol was the most potent inhibitor with an IC(50) of 13.5 μmol/L. In vivo efficacy of 4-n-butyl-resorcinol was confirmed in clinical studies. Subjects with age spots on the forearm treated twice daily two age spots with a formula containing 4-n-butylresorcinol and two control age spots with the corresponding vehicle. Within 8 weeks, 4-n-butylresorcinol reduced visibly the appearance of age spots, while the control spots showed no improvement. A second study showed that 4-butylresorcinol was more effective than 4-hexylresorcinol and 4-phenylethylresorcinol. CONCLUSION:The present in vitro and in vivo data prove the high inhibitory capacity of 4-n-butylresorcinol on human tyrosinase activity, exceeding by far the potency of hydroquinone, arbutin and kojic acid. The resulting clinical improvement of skin hyperpigmentations reveals 4-n-butylresorcinol as a very valuable active compound for the management of pigmentation disorders.

背景与目标： 背景：诸如色素沉着，光化性和老年性小扁豆的色素沉着过度症是主要的美容问题。因此，可获得许多外用产品，其包含旨在减少黑色素产生和分布的各种活性成分。色素沉着过度抑制剂的最主要目标是酪氨酸酶，它是黑色素生成的关键调节剂。文献中描述了许多酪氨酸酶抑制剂。然而，它们大多数缺乏临床功效。
方法：我们对评价具有美白活性的对苯二酚，熊果苷，曲酸和4-正丁基间苯二酚等具有美白活性的化合物对皮肤色素沉着的抑制作用感兴趣。我们在生化分析中比较了对人酪氨酸酶活性的抑制作用以及对MelanoDerm皮肤模型培养物中黑色素生成的抑制作用。对于某些化合物，在临床研究中测试了体内功效。
结果：熊果苷和对苯二酚仅能微弱地抑制人酪氨酸酶，最大抑制浓度（IC（50））在毫摩尔范围内。曲酸的效价高10倍，IC（50）约为500μmol/ L。但是，到目前为止，最有效的人酪氨酸酶抑制剂是4-正丁基间苯二酚，IC（50）为21μmol/ L。在人造皮肤模型中，熊果苷对IC（50）的抑制作用最小，可抑制> 5000μmol/ L的黑色素生成。曲酸被IC（50）> 400μmol/ L抑制。有趣的是，对苯二酚抑制了MelanoDerms中黑色素的生成，IC（50）低于40μmol/ L，这可能是由于其与酪氨酸酶抑制作用不同的机制所致。同样，4-正丁基间苯二酚是最有效的抑制剂，IC（50）为13.5μmol/ L。在临床研究中证实了4-正丁基间苯二酚的体内功效。前臂上有老年斑的受试者每天两次用含4-正丁基间苯二酚的配方治疗两个老年斑，并用相应的媒介物治疗两个对照老年斑。在8周内，4-正丁基间苯二酚明显减少了老年斑的出现，而对照斑则没有改善。第二项研究表明4-丁基间苯二酚比4-己基间苯二酚和4-苯基乙基间苯二酚更有效。
结论：目前的体内和体外数据证明4-正丁基间苯二酚对人酪氨酸酶活性具有很高的抑制能力，远远超过了对苯二酚，熊果苷和曲酸的效力。导致皮肤色素沉着过度的临床改善表明，4-正丁基间苯二酚是用于治疗色素沉着疾病的非常有价值的活性化合物。

BACKGROUND & AIMS: :Complement C3, when its cDNA was transfected into COS-1 cells, was synthesized as a precursor, pro-C3, which was intracellularly processed into the alpha and beta subunits, although not completely. A cDNA for rat alpha 1-protease inhibitor (alpha 1-PI) was mutated in vitro to encode its variant with the modified active site (Met352----Arg). In cells co-transfected with the mutant alpha 1-PI cDNA and the C3 cDNA, pro-C3 expressed was secreted without being processed into the subunits. Co-transfection of the mutant alpha 1-PI cDNA and the albumin cDNA also resulted in the inhibition of intracellular conversion of proalbumin into serum-type albumin. No inhibition of the processing of each preform was observed in cells co-transfected with the normal alpha 1-PI cDNA. Taken together, the results indicate that the alpha 1-PI variant (Met352----Arg) expressed inhibits specifically an intracellular enzyme which is involved in the proteolytic processing of both pro-C3 and proalbumin.

背景与目标： 补体C3，当其cDNA转染到COS-1细胞中时，被合成为前体pro-C3，pro-C3在细胞内被加工成α和β亚基，尽管不完全。大鼠α1-蛋白酶抑制剂（α1-P​​I）的cDNA在体外发生突变，以编码其具有修饰的活性位点的变体（Met352-Arg）。在用突变体α1-PI cDNA和C3 cDNA共转染的细胞中，表达的pro-C3被分泌而没有被加工成亚基。突变体α1-PI cDNA和白蛋白cDNA的共转染也导致抑制了原白蛋白向血清型白蛋白的细胞内转化。在用正常的α1-PI cDNA共转染的细胞中未观察到每种预型体的加工受到抑制。两者合计，结果表明表达的α1-PI变体（Met352 ---- Arg）特异性抑制细胞内酶，该酶参与pro-C3和proalbumin的蛋白水解过程。

BACKGROUND & AIMS: :We recently reported the discovery of octahydropyrrolo[1,2-a]pyrazine A as a lead compound for an inhibitor of apoptosis proteins (IAP) antagonist. To develop IAP antagonists with favorable PK profiles, we designed novel tri-cyclic compounds, octahydro-1H-cyclopropa[4,5]pyrrolo[1,2-a]pyrazines 1 and 2 based on co-crystal structural analysis of A with cellular IAP-1 (cIAP-1). The additional cyclopropane moiety was used to block the predicted metabolic site of compound A without detriment to the binding affinity for cIAP. Compounds 1 and 2 were stereoselectively synthesized via intermediates 4a and 5b', which were obtained by Simmons-Smith cyclopropanation of ethylester 3a and silyl ether 3b'. Compounds 1 and 2 showed strong growth inhibition in MDA-MB-231 breast cancer cells and improved metabolic stability in comparison to A. Compound 2 exhibited significant in vivo PD effects to increase tumor necrosis factor-alpha mRNA in a dose dependent manner.

背景与目标： ：我们最近报道了八氢吡咯并[1,2-a]吡嗪A作为凋亡蛋白（IAP）拮抗剂抑制剂的先导化合物的发现。为了开发具有良好PK谱的IAP拮抗剂，我们基于A与细胞的共晶体结构分析，设计了新颖的三环化合物，八氢-1H-环丙[4,5]吡咯并[1,2-a]吡嗪1和2。 IAP-1（cIAP-1）。额外的环丙烷部分用于封闭化合物A的预测代谢位点，而不会损害对cIAP的结合亲和力。经由中间体4a和5b'立体选择性地合成化合物1和2，所述中间体通过乙酯3a和甲硅烷基醚3b'的Simmons-Smith环丙烷化获得。与A相比，化合物1和2在MDA-MB-231乳腺癌细胞中显示出强大的生长抑制作用，并改善了代谢稳定性。化合物2在剂量方面具有显着的体内PD效应，从而增加了肿瘤坏死因子-αmRNA的表达。

BACKGROUND & AIMS: PURPOSE:To elucidate the intraocular pressure (IOP)-lowering effects and associated characteristics of Y-39983, a selective Rho-associated coiled coil-forming protein kinase (ROCK) inhibitor derived from Y-27632, in animal eyes. METHODS:Y-39983 was compared with Y-27632 for selectivity of ROCK inhibition by biochemical assay. The IOP was monitored by pneumatonometer in albino rabbits and cynomolgus monkeys that were given topically administered Y-39983. The total outflow facility and uveoscleral outflow were measured by two-level constant-pressure perfusion and perfusion technique using fluorescein isothiocyanate-dextran, respectively, at 2 hours after topical administration of Y-39983 in albino rabbits. The ocular toxicologic effects of topical administration of Y-39983 were observed in albino rabbits and cynomolgus monkeys. RESULTS:A biochemical assay showed that Y-39983 inhibited ROCK more potently than Y-27632. In rabbits, topical administration of Y-39983 significantly increased conventional outflow by 65.5%, followed by significant, dose-dependent reduction in IOP. Maximum IOP reduction was 13.2 +/- 0.6 mm Hg (mean +/- SE) at 0.1% Y-39983 in rabbits. In monkeys, at 3 hours after topical administration of 0.05% Y-39983, maximum reduction of IOP was 2.5 +/- 0.8 mm Hg. No serious side effects were observed in ocular tissues except sporadic punctate subconjunctival hemorrhage during long-term topical administration of Y-39983 four times a day (at 2-hour intervals) in rabbits or monkeys. However, punctate subconjunctival hemorrhage was not observed with administration twice daily (at a 6-hour interval) or three times a day (at 5-hour intervals). CONCLUSIONS:Y-39983 causes increased outflow facility followed by IOP reduction. Y-39983 ophthalmic solution may be a candidate drug for lowering of IOP, since it increases conventional outflow and produces relatively few side effects.

背景与目标： 目的：为了阐明动物眼中Y-39983（一种选自R-27632的选择性Rho相关的卷曲螺旋形成蛋白激酶（ROCK）抑制剂）Y-39983的降低眼内压（IOP）的作用和相关特征。
方法：通过生化分析比较Y-39983和Y-27632对ROCK的选择性。通过肺气量计在局部给予Y-39983的白化兔和食蟹猴中监测IOP。在白化兔中局部施用Y-39983后2小时，分别通过两级恒压灌注和灌注技术使用异硫氰酸荧光素-右旋糖酐测量总流出设施和葡萄膜巩膜流出。在白化病兔和食蟹猴中观察到局部施用Y-39983的眼部毒理作用。
结果：生化分析表明，Y-39983比Y-27632对ROCK的抑制作用更强。在兔子中，局部施用Y-39983可将常规流出量显着增加65.5％，然后是IOP的剂量依赖性显着降低。在0.1％的Y-39983中，兔子的最大IOP降低为13.2 /-0.6毫米汞柱（平均/-SE）。在猴子中，局部给药0.05％Y-39983后3小时，IOP的最大降低为2.5±0.8 mm Hg。在兔子或猴子中，一天四次（以2小时为间隔）每天四次Y-39983长期局部给药期间，偶发性点状结膜下出血除外，在眼组织中未观察到严重的副作用。但是，每日两次（每隔6小时）或每天三次（每隔5小时）给药未观察到点状结膜下出血。
结论：Y-39983引起流出设施增加，随后IOP降低。 Y-39983眼用溶液剂可能是降低IOP的候选药物，因为它增加了常规流出量并且产生了相对较少的副作用。