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： : 氟西汀 (FLX) 百忧解中的活性药物成分 (API) (®) 是一种广泛使用的精神活性药物，在水生环境中普遍存在，这与废水处理厂 (WWTP) 系统的去除率差有关。该API充当选择性5-羟色胺再摄取抑制剂 (SSRI)，经常报道会在非目标物种中引起破坏作用。这项研究的目的包括在暴露于75 ng L(-1) FLX的两周内对贻贝Mytilus galloprovincialis进行多生物标志物反应评估，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 (GST); 脂质过氧化 (LPO)，乙酰胆碱酯酶 (AChE) 通过碱性不稳定磷酸盐 (ALP) 间接测量卵黄蛋白原样蛋白的神经毒性反应和内分泌破坏。结果显示，短暂的组织特异性酶反应和损伤主要影响贻贝g。然而，在两种性别分化的性腺中，整个过程中明显的ALP水平抑制提供了FLX作为内分泌干扰物而不是氧化或神经毒性诱导剂增强作用的证据。

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) 抑制剂的抗炎潜力被巧合地扩展为对p38α MAPK和磷酸二酯酶4 (PDE4) 的双重抑制，并且两种炎症相关酶的阻断所产生的潜在益处被彻底研究。在对啮齿动物，狗和猴子施用1之后，在体外实验以及体外和体内临床前研究中相继评估了最有希望的化合物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。为了在向健康的人类志愿者施用1时再次确认动物研究的结果，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征的进一步信息外，还证明了p38α MAPK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：

BACKGROUND & AIMS: :An extract from red ginseng [steamed and dried roots of Panax ginseng C.A. Meyer (RGE)] has been shown to have various actions on physiological functions. The mechanisms by which RGE promotes cholesterol metabolism in the liver are unclear, but RGE decreases the plasma levels of cholesterol. We investigated whether RGE affected the mRNA expression of cholesterol metabolism-related proteins such as cytochrome P450 (CYP)7A1 and bile salt export pump (BSEP) in the liver in hypercholesterolemic rats and rat primary hepatocytes. In-vivo studies showed the upregulation of CYP7A1 mRNA in hypercholesterolemic rats treated with RGE. Treatment with RGE exhibited decreased ratios of low-density lipoprotein-cholesterol to high-density lipoprotein-cholesterol compared with hypercholesterolemia without RGE. In-vitro studies also showed the upregulation of CYP7A1 mRNA and protein levels by the addition of RGE to rat primary hepatocytes. The mRNA levels of BSEP exhibited few changes. The sustained levels of the liver X receptor (LXR) in vivo and the increased levels of LXR in vitro on RGE treatment could be involved in the upregulation of CYP7A1. To clarify the effects of 11 ginsenosides including RGE on the mRNA levels of CYP7A1 and BSEP, we performed in-vitro experiments using rat primary hepatocytes. The ginsenosides Ro, Rg3, Re, Rg1, and Rg2 exhibited increased mRNA levels of CYP7A1. These results suggest that several ginsenosides including RGE promoted cholesterol metabolism due to upregulation of CYP7A1.

背景与目标： : 来自红参的提取物 [人参c.A.的蒸干根Meyer (RGE)] 已被证明对生理功能具有多种作用。RGE促进肝脏中胆固醇代谢的机制尚不清楚，但RGE会降低血浆胆固醇水平。我们研究了RGE是否影响高胆固醇血症大鼠和大鼠原代肝细胞肝脏中胆固醇代谢相关蛋白 (例如细胞色素P450 (CYP)7A1和胆盐出口泵 (BSEP)) 的mRNA表达。体内研究表明，用RGE治疗的高胆固醇血症大鼠CYP7A1 mRNA上调。与没有RGE的高胆固醇血症相比，RGE治疗显示低密度脂蛋白胆固醇与高密度脂蛋白胆固醇的比率降低。体外研究还显示，通过向大鼠原代肝细胞添加RGE，CYP7A1 mRNA和蛋白质水平上调。BSEP的mRNA水平几乎没有变化。RGE治疗后体内肝脏X受体 (LXR) 的持续水平和体外LXR水平的升高可能与CYP7A1的上调有关。为了阐明包括RGE在内的11种人参皂甙对CYP7A1和BSEP mRNA水平的影响，我们使用大鼠原代肝细胞进行了体外实验。人参皂苷Ro，Rg3，Re，Rg1和Rg2显示CYP7A1的mRNA水平升高。这些结果表明，由于CYP7A1的上调，包括RGE在内的几种人参皂甙可促进胆固醇代谢。

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： : 纤溶酶原激活物抑制剂-1 (PAI-1) 是丝氨酸蛋白酶抑制剂 (serpin) 超家族的成员，其使组织纤溶酶原激活物 (tPA) 失活; 因此，PAI-1抗原水平的升高抵消了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 (如肥胖和糖尿病) 的特征相关。内脏脂肪组织比皮下脂肪组织产生更多的PAI-1: 内脏脂肪组织产生的PAI-1增加是内脏肥胖和心血管疾病之间的重要联系。除了内脏脂肪组织外，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，并负责炎症部位常见的疼痛和肿胀。非甾体抗炎药 (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。然而，NSAID治疗的主要副作用之一是胃溃疡。通过5-脂氧合酶 (5-LO) 酶途径抑制前列腺素的产生和白三烯形成的相关增加可能导致吸引炎症细胞，引起局部炎症部位并产生溃疡。为了确定5-LO抑制对该假设的影响，在大鼠中进行了研究，以评估tepoxalin (一种双重CO/LO抑制剂) 对胃粘膜中白三烯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吲哚美辛 (在4天内每天2 mg/kg) 导致40% 死亡率，并在第四剂药物后24小时评估时伴有肠粘连和穿孔。此外，肠系膜小静脉中的中性粒细胞粘附增加，肠系膜间质中的细胞浸润明显。在每天吲哚美辛治疗前30分钟，在单独的大鼠组中抑制了这些胃肠道副作用。进行了进一步的研究以确定tepoxalin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，脂质过氧化物生成和LTB4生成。给药90分钟后，吲哚美辛 (100 mg/kg，p.o.) 在大鼠胃粘膜中产生升高的LTB4水平。此外，在该剂量下以及使用另一种NSAID萘普生时，肠系膜小静脉中的中性粒细胞粘附增加。此时，胃粘膜中没有明显的脂质过氧化物生成。Tepoxalin (最高400 mg/kg，p.o.) 对胃粘膜LTB4的生成和脂质过氧化物水平没有影响。在最高剂量下观察到中性粒细胞粘附减少。在另一项研究中，用tepoxalin (ED50 = 7.5 mg/kg，p.o.) 或选择性5-LO抑制剂zileuton (100 mg/kg，p.o.) 进行预处理可防止胃粘膜LTB4水平升高和中性粒细胞粘附由吲哚美辛 (100 mg/kg，p、o.)。这些数据表明，LO抑制作用可能在预防NSAID诱导的胃部炎症中起着至关重要的作用，从而深入了解tepoxalin缺乏溃疡性以及可能预防胃副作用的抗炎治疗的新方法。

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.选择性5-羟色胺 (5-HT; 5-羟色胺) 再摄取抑制剂 (SSRIs) 在体细胞肌腱5-HT1A自身受体受阻时，会导致前脑细胞外5-HT的增加。在这里，我们研究了末端5-HT1B自身受体的阻断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自身受体的附加作用。2.使用脑微透析在麻醉的大鼠额叶皮层中测量细胞外5-HT。还进行了中缝背核 (DRN) 中5-HT神经元活性的体内细胞外记录。3.在用5-HT1A受体拮抗剂way100635预处理的大鼠中，选择性5-HT再摄取抑制剂帕罗西汀 (0.8 mg kg-1，i.v.) 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，静脉注射) 和WAY100635 (0.1 mg kg-1，静脉注射) 都不会改变细胞外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.) 不增加5-HT。GR127935 (1和5 mg kg-1，静脉注射) 单独给药时对细胞外5-HT没有影响。5.有趣的是，当GR127935 (1或5 mg kg-1，静脉注射) 与WAY100635 (0.1 mg kg-1，静脉注射) 联合给药时，帕罗西汀 (0.8 mg kg-1，静脉注射) 引起5-HT的最大增加 (高达5倍)。i.v.)。施用GR127935 (5 mg kg-1，i.v.) 加WAY100635 (0.1 mg kg-1，i.v.) 而不使用帕罗西汀，对额叶皮层的细胞外5-HT没有影响。6.尽管在基础条件下GR127935对5-HT缺乏作用，但当5-HT输出量增加约3倍 (通过向灌注培养基中添加1 microM帕罗西汀) 时，药物导致5-HT剂量相关 (1和5 mg kg-1，静脉注射) 增加。7.就其本身而言，GR127935在较高剂量 (2.0-5.0 mg kg-1，i.v.) 下略微但显著降低DRN中的5-HT细胞放电，但没有阻止帕罗西汀诱导的5-HT细胞放电的抑制。8.总之，我们的结果表明，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，当体突肌 (5-HT1A) 和神经末梢 (5-HT1B) 上的5-HT自身受体被阻断。此增加大于单独阻塞任一组自身感受器时的增加。在我们的实验中，单独的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，这可能与由于持续抑制5-HT1B自身受体而导致末端5-HT1B自身受体缺乏音调有关。ht细胞放电。讨论了这些结果，这些结果与使用5-HT自身受体拮抗剂来增强选择性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对预防静脉血栓栓塞有效。出血并发症很少见，但在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，证明了直接凝血酶抑制剂melagatran诱导全血 (WB) 凝血谱的剂量依赖性异常，如最近描述的改良血栓弹力图模型所记录，并且rFVIIa或APCC能够改善止血能力。使用来自30名健康男性的WB进行了实验。将melagatran添加到WB中的体外滴定实验 (n = 10)，对应于血浆浓度范围为0至5.0微米 (12步)，结果显示凝块起始的剂量依赖性延长和最大凝块传播速率的特征性降低。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完成了体外干预研究 (n = 20)。在所有测试的melagatran浓度下，rFVIIa均显着缩短了melagatran诱导的长时间凝块起始，但仅诱导了减少的凝块繁殖的微小改善。相反，APCC显着且剂量依赖性地缩短了凝块的起始并加速了凝块的传播。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的影响，但尤其是APCC显着改善了wb凝块形成。APCC的明显稳定作用可能是由其凝血酶原和活化的凝血因子的含量引起的。

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： : 血浆蛋白酶是参与宿主细胞血红蛋白降解的天冬氨酸蛋白酶，被疟原虫用作食物来源。血浆蛋白酶作为药物靶标非常有希望，尤其是与抑制也参与血红蛋白分解代谢的falcipains结合使用时。在这篇综述中，鉴于过渡态模拟物作为潜在的铅开发化合物的兴趣，我们讨论了血浆蛋白酶i-iv的机理。总结了针对plasmepsin II的抑制剂开发以及相关的晶体结构，以概述该领域。计算技术的应用，尤其是通过线性相互作用能方法进行的结合亲和力预测，在疟疾血浆蛋白酶抑制剂的开发中非常成功，并进行了详细讨论。同源性建模和分子对接在当前的抑制剂设计项目中很有用，并且分析了此类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：

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： : 纤溶酶原激活物抑制剂-1 (PAI-1) 是纤维蛋白溶解的抑制剂。血浆PAI-1水平的升高在心血管风险和与血栓形成相关的其他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测定法研究了160个健康的无关黎巴嫩个体，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基因型最普遍 (45.6%)，其次是5G/5G (36.9%) 和4G/4G (17.5%)。4g和5g等位基因的频率分别计算为0.403和0.597。与其他种族社区相比，发现黎巴嫩人口的稀有4g等位基因患病率相对较高。反过来，这可能使该人群易患心血管疾病和其他血栓性临床疾病。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: :Retinal proteins function as photoreceptors and ion pumps. Xanthorhodopsin of Salinibacter ruber is a recent addition to this diverse family. Its novel and distinctive feature is a second chromophore, a carotenoid, which serves as light-harvesting antenna. Here we discuss the properties of this carotenoid/retinal complex most relevant to its function (such as the specific binding site controlled by the retinal) and its relationship to other retinal proteins (bacteriorhodopsin, archaerhodopsin, proteorhodopsin and retinal photoreceptors of archaea and eukaryotes). Antenna addition to a retinal protein has not been observed among the archaea and emerged in bacteria apparently in response to environmental conditions where light-harvesting becomes a limiting factor in retinal protein functioning.

背景与目标： : 视网膜蛋白作为光感受器和离子泵的功能。萨氏杆菌的黄hodopsin是这个多样化家族的最新成员。它新颖而独特的特征是第二个发色团，一种类胡萝卜素，可作为集光天线。在这里，我们讨论了这种类胡萝卜素/视网膜复合物与其功能最相关的特性 (例如由视网膜控制的特异性结合位点) 及其与其他视网膜蛋白 (细菌视紫红质，古细菌视紫红质，蛋白视紫红质和古细菌和真核生物的视网膜光感受器) 的关系。在古细菌中尚未观察到天线添加到视网膜蛋白中，并且在细菌中出现显然是对环境条件的响应，在环境条件下，光收集成为视网膜蛋白功能的限制因素。

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： : 恶性神经胶质瘤化疗的最新进展归因于DNA甲基化剂替莫唑胺 (TMZ) 的引入; 然而，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期长期停滞，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Cdc2 (G2检查点途径中的关键蛋白) 对TMZ处理的神经胶质瘤细胞的作用。集落形成效率表明，FP以p53-independent的方式增强了神经胶质瘤细胞中TMZ的细胞毒性。这种作用显然与G2-M转变时关键蛋白的抑制，仅在G2期细胞的积累以及双链DNA断裂标记物的增加 (在进行免疫印迹时可见) 有关。TMZ抗性克隆显示出响应TMZ的G2检查点激活，而FP处理使这些克隆对TMZ敏感。FP还增强了TMZ在U87MG-AktER细胞中的细胞毒性。此外，向具有异种移植的U87MG细胞的裸鼠施用TMZ和/或FP显示，在这些小鼠中，FP致敏异种移植的U87MG细胞至TMZ。我们的发现表明，TMZ抗性可以通过在G2-M转变中增强DNA修复活性来促进，并且Cdk抑制剂可以抑制这种活性，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：

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被转染到COS-1细胞中时，被合成为前体pro-C3，其在细胞内加工成 α 和 β 亚基，尽管不完全。大鼠 α1-蛋白酶抑制剂 (α1-pi) 的cDNA在体外进行突变，以用修饰的活性位点 (Met352 ---- Arg) 编码其变体。在用突变体 α1-picdna和c3cdna共转染的细胞中，表达的pro-C3被分泌而不被加工成亚基。突变体 α1-pi cDNA和白蛋白cDNA的共转染也导致抑制前白蛋白向血清型白蛋白的细胞内转化。在用正常 α1-PI cDNA共转染的细胞中，未观察到每种预成型体的加工抑制。总之，结果表明表达的 α1-pi变体 (Met352 ---- Arg) 特异性抑制了参与pro-C3和前白蛋白的蛋白水解加工的胞内酶。