BACKGROUND & AIMS: :Agents to control melanogenesis are in demand for the development of cosmetics to improve pigmentation disorders of skin and hair. In this study, we examined and evaluated the effects of flavonoids on melanogenesis in the melanogenic cells model, murine B16F10 melanoma cells. In the course of this study, we found that incubation of the cells in a medium containing 10 μM of the 4'-O-methylated flavonoids, diosmetin (4'-O-methylluteolin), acacetin (4'-O-methylapigenin) or kaempferide (4'-O-methylkaempferol), increased the melanin contents of the cells 3- to 7-fold higher than the control cells. The concentration-dependence test revealed that 20 μM acacetin showed the highest effect, up to 33-fold higher than the vehicle. On the other hand, the corresponding 4'-OH-type flavonoids, luteolin, apigenin and kaempferol, had a significantly smaller effect. Furthermore, by evaluating the melanogenic proteins, we found that the cells treated with 4'-O-methylated flavonoids showed higher tyrosinase activity, as well as upregulation of tyrosinase expression, preceded by activation of cAMP response element binding protein (CREB) and extracellular signal-regulated kinases types 1 and 2 (ERK1/2). These results indicate that the 4'-O-methyl group of flavonoids plays an important role in the induction of melanogenesis by activating its major signal transduction pathway through the upregulation of phospho-CREB in murine B16F10 melanoma cells.

背景与目标： : 控制黑素生成的药物是化妆品开发的需求，以改善皮肤和头发的色素沉着疾病。在这项研究中，我们检查并评估了类黄酮对黑色素生成细胞模型鼠B16F10黑素瘤细胞中黑色素生成的影响。在这项研究的过程中，我们发现细胞在含有10μm的4 '-O-甲基化类黄酮，地奥美汀 (4'-O-methylluteolin)，acacetin (4 '-O-methyapigenin) 或kaempferide (4'-O-methylameferol)，使细胞的黑色素含量比对照细胞高3至7倍。浓度依赖性测试表明，20μm acacetin的效果最高，比媒介物高33倍。另一方面，相应的4 '-OH型类黄酮木犀草素，芹菜素和山奈酚的作用明显较小。此外，通过评估黑素蛋白，我们发现用4 '-O-甲基化类黄酮处理的细胞显示出更高的酪氨酸酶活性以及酪氨酸酶表达的上调，在激活cAMP反应元件结合蛋白 (CREB) 和细胞外信号调节激酶类型1和2 (ERK1/2) 之前。这些结果表明，类黄酮的4 '-O-甲基通过上调小鼠B16F10黑素瘤细胞的磷酸化CREB激活其主要信号转导途径，在黑素生成的诱导中起重要作用。

BACKGROUND & AIMS: :Utilizing X-ray crystal structure analysis, (3S,5R)-5-[4-(2-chlorophenyl)-2,2-dimethyl-5-oxopiperazin-1-yl]piperidine-3-carboxamides were designed and identified as renin inhibitors. The most potent compound 15 demonstrated favorable pharmacokinetic and pharmacodynamic profiles in rat.

背景与目标： : 利用x射线晶体结构分析，设计了 (3S，5R)-5-[4-(2-氯苯基)-2,2-二甲基-5-氧代哌嗪-1-基] piperidine-3-carboxamides，并鉴定为肾素抑制剂。最有效的化合物15在大鼠中显示出良好的药代动力学和药效学特征。

BACKGROUND & AIMS: :Antitumor agents that bind to tubulin and disrupt microtubule dynamics have attracted considerable attention in the last few years. To extend our knowledge of the thiazole ring as a suitable mimic for the cis-olefin present in combretastatin A-4, we fixed the 3,4,5-trimethoxyphenyl at the C4-position of the thiazole core. We found that the substituents at the C2- and C5-positions had a profound effect on antiproliferative activity. Comparing compounds with the same substituents at the C5-position of the thiazole ring, the moiety at the C2-position influenced antiproliferative activities, with the order of potency being NHCH(3) > Me > N(CH(3))(2). The N-methylamino substituent significantly improved antiproliferative activity on MCF-7 cells with respect to C2-amino counterparts. Increasing steric bulk at the C2-position from N-methylamino to N,N-dimethylamino caused a 1-2 log decrease in activity. The 2-N-methylamino thiazole derivatives 3b, 3d and 3e were the most active compounds as antiproliferative agents, with IC(50) values from low micromolar to single digit nanomolar, and, in addition, they are also active on multidrug-resistant cell lines over-expressing P-glycoprotein. Antiproliferative activity was probably caused by the compounds binding to the colchicines site of tubulin polymerization and disrupting microtubule dynamics. Moreover, the most active compound 3e induced apoptosis through the activation of caspase-2, -3 and -8, but 3e did not cause mitochondrial depolarization.

背景与目标： : 与微管蛋白结合并破坏微管动力学的抗肿瘤剂在过去几年中引起了极大的关注。为了扩展我们对噻唑环作为combretastatin A-4中存在的顺式烯烃的合适模拟物的了解，我们将3,4，5-三甲氧基苯基固定在噻唑核的C4-position。我们发现C2-和C5-positions的取代基对抗增殖活性具有深远的影响。比较噻唑环C5-position具有相同取代基的化合物，C2-position部分影响抗增殖活性，效力顺序为NHCH(3)> Me> N(CH(3))(2)。相对于C2-amino对应物，N-甲基氨基取代基显着提高了MCF-7细胞的抗增殖活性。从N-甲基氨基到N，N-二甲基氨基C2-position的空间体积增加导致活性降低1-2对数。2-n-甲基氨基噻唑衍生物3b，3d和3e是作为抗增殖剂的最具活性的化合物，其IC(50) 值从低微摩尔到个位数纳摩尔，此外，它们在多药耐药细胞系中也具有活性过表达P-糖蛋白。抗增殖活性可能是由化合物与微管蛋白聚合的秋水仙碱位点结合并破坏微管动力学引起的。此外，活性最高的化合物3e通过激活caspase-2，-3和-8诱导细胞凋亡，但3e并未引起线粒体去极化。

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: :Bacterial-derived lipopolysaccharides (LPS) play an essential role in the inflammatory process of inflammatory bowel disease. A defective intestinal tight junction (TJ) barrier is an important pathogenic factor of inflammatory bowel disease and other inflammatory conditions of the gut. Despite its importance in mediating intestinal inflammation, the physiological effects of LPS on the intestinal epithelial barrier remain unclear. The major aims of this study were to determine the effects of physiologically relevant concentrations of LPS (0 to 1 ng/mL) on intestinal barrier function using an in vitro (filter-grown Caco-2 monolayers) and an in vivo (mouse intestinal perfusion) intestinal epithelial model system. LPS, at physiologically relevant concentrations (0 to 1 ng/mL), in the basolateral compartment produced a time-dependent increase in Caco-2 TJ permeability without inducing cell death. Intraperitoneal injection of LPS (0.1 mg/kg), leading to clinically relevant plasma concentrations, also caused a time-dependent increase in intestinal permeability in vivo. The LPS-induced increase in intestinal TJ permeability was mediated by an increase in enterocyte membrane TLR-4 expression and a TLR-4-dependent increase in membrane colocalization of membrane-associated protein CD14. In conclusion, these studies show for the first time that LPS causes an increase in intestinal permeability via an intracellular mechanism involving TLR-4-dependent up-regulation of CD14 membrane expression.

背景与目标： : 细菌来源的脂多糖 (LPS) 在炎症性肠病的炎症过程中起着至关重要的作用。肠紧密连接 (TJ) 屏障缺陷是炎症性肠病和肠道其他炎症状况的重要致病因素。尽管LPS在介导肠道炎症中的重要性，但其对肠上皮屏障的生理作用仍不清楚。这项研究的主要目的是使用体外 (过滤生长的Caco-2单层) 和体内 (小鼠肠灌注) 确定生理相关浓度的LPS (0至1 ng/mL) 对肠屏障功能的影响。) 肠上皮模型系统。LPS在生理相关浓度 (0至1 ng/mL) 下，在基底外侧区室中产生了Caco-2 TJ通透性的时间依赖性增加，而没有诱导细胞死亡。腹膜内注射LPS (0.1 mg/kg) 导致临床上相关的血浆浓度，也引起体内肠通透性的时间依赖性增加。LPS诱导的肠TJ通透性增加是由肠细胞膜TLR-4表达的增加和膜相关蛋白cd14的膜共定位的TLR-4-dependent增加介导的。总之，这些研究首次表明，LPS通过涉及CD14膜表达的TLR-4-dependent上调的细胞内机制引起肠通透性的增加。

BACKGROUND & AIMS: :The effects of bread consumption change over time on anthropometric measures have been scarcely studied. We analysed 2213 participants at high risk for CVD from the PREvención con DIeta MEDiterránea (PREDIMED) trial to assess the association between changes in the consumption of bread and weight and waist circumference gain over time. Dietary habits were assessed with validated FFQ at baseline and repeatedly every year during 4 years of follow-up. Using multivariate models to adjust for covariates, long-term weight and waist circumference changes according to quartiles of change in energy-adjusted white and whole-grain bread consumption were calculated. The present results showed that over 4 years, participants in the highest quartile of change in white bread intake gained 0·76 kg more than those in the lowest quartile (P for trend = 0·003) and 1·28 cm more than those in the lowest quartile (P for trend < 0·001). No significant dose-response relationships were observed for change in whole-bread consumption and anthropometric measures. Gaining weight (>2 kg) and gaining waist circumference (>2 cm) during follow-up was not associated with increase in bread consumption, but participants in the highest quartile of changes in white bread intake had a reduction of 33 % in the odds of losing weight (>2 kg) and a reduction of 36 % in the odds of losing waist circumference (>2 cm). The present results suggest that reducing white bread, but not whole-grain bread consumption, within a Mediterranean-style food pattern setting is associated with lower gains in weight and abdominal fat.

背景与目标： : 面包消费随时间变化对人体测量的影响很少被研究。我们分析了来自prevenci ó n con DIeta meditr á nea (predmed) 试验的2213名CVD高危参与者，以评估随时间推移面包和体重变化与腰围增加之间的关联。在基线时使用经过验证的FFQ评估饮食习惯，并在随访的4年中每年重复进行。使用多变量模型对协变量进行调整，根据能量调整后的白色和全麦面包消费量变化的四分位数，计算了长期体重和腰围的变化。目前的结果表明，在过去的4年里，白面包摄入量变化最高四分位数的参与者比最低四分位数的参与者增加了0·76千克 (趋势P = 0·003)，比最低四分位数的参与者增加了1·28厘米 (趋势P <0·001)。对于全面包消费和人体测量的变化，未观察到明显的剂量反应关系。随访期间体重增加 (> 2千克) 和腰围增加 (> 2厘米) 与面包消耗量增加无关，但是，白面包摄入量变化最高四分位数的参与者的减肥几率降低了33% (> 2千克)，而腰围下降的几率降低了36% (> 2厘米)。目前的结果表明，在地中海风格的食物模式环境中，减少白面包而不是全麦面包的消费与体重和腹部脂肪的减少有关。

BACKGROUND & AIMS: :Traumatic brain injury (TBI) remains the leading cause of injury-related death and disability. Brain edema, one of the most major complications of TBI, contributes to elevated intracranial pressure, and poor prognosis following TBI. Nerve growth factor (NGF) appears to be a viable strategy to treat brain edema and TBI. Unfortunately, due to its poor blood-brain barrier (BBB) permeability, the clinical application of NGF has been greatly limited. We previously demonstrated that intranasal NGF could bypass the BBB and distribute throughout the brain. Here we further studied whether intranasal NGF could attenuate TBI-induced brain edema and its putative mechanisms. TBI was produced by a modified weight-drop model. We found that intranasal administration of NGF (5μg/d) attenuated the brain edema, as assayed by hemisphere water content, at 12h, 24h and 72h after TBI induction. This attenuation was associated with a prominent decrease of the content of aquaporin-4, which plays a pivotal role in the formation of brain edema. By the use of RT-PCR and ELISA, we showed that intranasal NGF markedly inhibited the transcription and expression of pro-inflammatory cytokines including IL-1β and TNF-α. An electrophoretic mobility shift assay (EMSA) displayed a significant activation of nuclear factor-κB following TBI, which was, however, much lowered in the NGF-treated rats. Furthermore, upon intranasal NGF supplementation, mitochondria-mediated apoptosis following TBI was minimized, as indicated by upregulation of Bcl-2 and downregulation of caspase-3. Collectively, our findings suggested that intranasal NGF may be a promising strategy to treat brain edema and TBI.

背景与目标： : 创伤性脑损伤 (TBI) 仍然是与伤害相关的死亡和残疾的主要原因。脑水肿是TBI的主要并发症之一，可导致颅内压升高和TBI后预后不良。神经生长因子 (NGF) 似乎是治疗脑水肿和TBI的可行策略。不幸的是，由于其血脑屏障 (BBB) 渗透性差，NGF的临床应用受到了极大的限制。我们先前证明鼻内NGF可以绕过BBB并分布在整个大脑中。在这里，我们进一步研究了鼻内NGF是否可以减轻TBI诱导的脑水肿及其推测的机制。TBI是通过改进的重量下降模型产生的。我们发现，经TBI诱导后12小时，24小时和72小时，鼻内施用NGF (5 μ g/d) 可减轻脑水肿，如半球含水量所测定。这种衰减与aquaporin-4含量的显着降低有关，这在脑水肿的形成中起着关键作用。通过rt-pcr和ELISA，我们发现鼻内NGF显着抑制促炎细胞因子 (包括IL-1β 和TNF-α) 的转录和表达。电泳迁移率变化测定 (EMSA) 显示TBI后核因子-κ b的显着激活，但是，在NGF处理的大鼠中，这一激活大大降低。此外，在鼻内补充NGF时，TBI后线粒体介导的凋亡被最小化，如Bcl-2上调和caspase-3下调所表明的。总的来说，我们的发现表明鼻内NGF可能是治疗脑水肿和TBI的有希望的策略。

BACKGROUND & AIMS: BACKGROUND CONTEXT:Although autogenous bone is still considered to be the gold standard graft material for promoting spinal fusion, other bone graft substitutes have been developed in an attempt to improve arthrodesis rates and avoid the complications associated with the procurement of autograft. The bone morphogenetic proteins (BMPs) represent a family of osteoinductive growth factors that are known to stimulate the osteoblastic differentiation of stem cells. Osteogenic protein-1 (OP-1) Putty is a commercially available BMP preparation that is already approved for use in humans. Previous clinical studies involving patients with degenerative spondylolisthesis have reported that the efficacy and safety of OP-1 Putty is comparable to that of autograft at both 1- and 2-year follow-up. PURPOSE:The purpose of this study was to evaluate the intermediate-term efficacy and safety of OP-1 Putty as an alternative to autogenous bone by comparing the 4-year radiographic, clinical, and safety data of these same patients who underwent decompression and uninstrumented fusion with either OP-1 Putty or iliac crest autograft. STUDY DESIGN/SETTING:A prospective, randomized, controlled, multicenter clinical pilot study. PATIENT SAMPLE:Thirty-six patients undergoing decompressive laminectomy and single-level uninstrumented fusion for degenerative spondylolisthesis and symptomatic spinal stenosis were randomized in a 2:1 fashion to receive either OP-1 Putty (24 patients) or autogenous iliac crest bone graft (12 patients). OUTCOME MEASURES:Patient-reported outcome measures consisting of Oswestry Disability Index and Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) scores were used to evaluate clinical efficacy. Perioperative data including operative time, estimated blood loss, and duration of hospital stay were also recorded for each surgery. Postoperatively, a neurological examination and an assessment of donor-site pain (if applicable) were performed at every follow-up visit. Radiographic fusion success was defined as the presence of continuous bridging bone formation between the transverse processes at the level of the spondylolisthesis with minimal motion evident on dynamic lateral x-ray films. The primary efficacy endpoint was the overall success rate, a composite measure derived from both radiographic and clinical parameters. The safety of OP-1 Putty was confirmed by comparing the nature and frequency of all adverse events and complications that were prospectively observed in either of the groups. METHODS:Thirty-six patients with degenerative spondylolisthesis and symptoms of neurogenic claudication underwent decompressive laminectomy and single-level uninstrumented fusion with either OP-1 Putty or autograft. All patients were evaluated at 6 weeks and 3, 6, 9, 12, and 24 months, after which time they were instructed to return on a yearly basis. Multiple neuroradiologists blinded to the assigned treatment reviewed static and dynamic X-ray films with digital calipers to assess fusion status according to the presence of continuous bridging bone across the transverse processes as well as the amount of residual motion evident at the level of interest. Oswestry Disability Index surveys and SF-36 questionnaires were used to assess clinical outcomes. RESULTS:At the 48-month time point, complete radiographic and clinical data were available for 22 of 36 patients (16 OP-1 Putty and 6 autograft) and 25 of 36 patients (18 OP-1 Putty and 7 autograft), respectively. Radiographic evidence of a solid arthrodesis was present in 11 of 16 OP-1 Putty patients (68.8%) and 3 of 6 autograft patients (50%). Clinically successful outcomes defined as at least a 20% improvement in preoperative Oswestry scores were experienced by 14 of 19 OP-1 Putty patients (73.7%) and 4 of 7 autograft patients (57.1%); these clinical findings were corroborated by similar increases in SF-36 scores. The respective overall success rates of the OP-1 Putty and autograft group were 62.5% and 33.3%. In this study, there were no incidents of local or systemic toxicity, ectopic bone production, or other adverse events directly related to the use of OP-1 Putty. CONCLUSION:Despite the challenges associated with obtaining a solid uninstrumented fusion in patients with degenerative spondylolisthesis, the rates of radiographic fusion, clinical improvement, and overall success associated with the use of OP-1 Putty were at least comparable to that of the autograft controls for at least 48 months after surgery. These results appear to validate the short-term results previously reported for OP-1 Putty and suggest that this material may potentially represent a viable bone graft substitute for certain fusion applications.

背景与目标：

BACKGROUND & AIMS: :Cigarette smoking is the predominant risk factor for bladder cancer. Aromatic amines such as 4-aminobiphenyl (ABP) is the major carcinogens found in tobacco smoke. Although it is generally accepted that ABP is metabolically activated via N-hydroxylation by CYP1A2 in human liver, previous studies using Cyp1a2-null mice indicated the involvement of other enzyme(s). Here we found that CYP2A13 can metabolically activate ABP to show genotoxicity by Umu assay. The K(m) and V(max) values for ABP N-hydroxylation by recombinant CYP2A13 in E. coli were 38.5 +/- 0.6 microM and 7.8 +/- 0.0 pmol/min/pmol CYP, respectively. The K(m) and V(max) values by recombinant CYP1A2 were 9.9 +/- 0.9 microM and 39.6 +/- 0.9 pmol/min/pmol CYP, respectively, showing 20-fold higher intrinsic clearance than CYP2A13. In human bladder, CYP2A13 mRNA, but not CYP1A2, is expressed at a relatively high level. Human bladder microsomes showed ABP N-hydroxylase activity (K(m) = 34.9 +/- 4.7 microM and V(max) = 57.5 +/- 1.9 pmol/min/mg protein), although the intrinsic clearance was 5-fold lower than that in human liver microsomes (K(m) = 33.2 +/- 2.0 microM and V(max) = 293.9 +/- 5.8 pmol/min/mg protein). The activity in human bladder microsomes was prominently inhibited by 8-methoxypsoralen, but not by fluvoxamine, anti-CYP1A2 or anti-CYP2A6 antibodies. CYP2S1, which is expressed in human bladder and has relatively high amino acid identities with CYP2As, did not show detectable ABP N-hydroxylase activity. In conclusion, although the enzyme responsible for ABP N-hydroxylation in human bladder microsomes could not be determined, we found that CYP2A13 metabolically activates ABP.

背景与目标： 吸烟是膀胱癌的主要危险因素。芳香胺 (例如4-氨基联苯 (ABP)) 是烟草烟雾中发现的主要致癌物。尽管人们普遍认为ABP通过CYP1A2在人肝脏中通过N-羟基化代谢激活，但先前使用Cyp1a2-null小鼠的研究表明其他酶的参与。在这里，我们发现CYP2A13可以通过Umu测定代谢激活ABP以显示遗传毒性。重组CYP2A13在大肠杆菌中abpn-羟基化的K(m) 和V(max) 值分别为38.5 +/- 0.6 microM和7.8 +/- 0.0 pmol/min/pmol CYP。重组CYP1A2的K(m) 和V(max) 值分别为9.9 +/- 0.9 microM和39.6 +/- 0.9 pmol/min/pmol CYP，显示出比CYP2A13高20倍的固有清除率。在人膀胱中，CYP2A13 mRNA而不是CYP1A2以相对较高的水平表达。人膀胱微粒体显示ABP N-羟化酶活性 (K(m) = 34.9 +/- 4.7微米和V(max) = 57.5 +/- 1.9 pmol/min/mg蛋白)，虽然内在清除率比人肝微粒体低5倍 (K(m) = 33.2 +/- 2.0微米和V(max) = 293.9 +/- 5.8 pmol/min/mg蛋白)。8-甲氧基补骨脂素显着抑制人膀胱微粒体的活性，但氟伏沙明，anti-CYP1A2或anti-CYP2A6抗体不抑制人膀胱微粒体的活性。CYP2S1在人膀胱中表达，与CYP2As具有较高的氨基酸同一性，但未显示可检测到的ABP N-羟化酶活性。总之，尽管无法确定负责人膀胱微粒体中ABP N-羟基化的酶，但我们发现CYP2A13代谢激活ABP。

BACKGROUND & AIMS: :A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.

背景与目标： : 整联蛋白家族的跨膜细胞外基质受体 α6β4是半染色体的组成部分，半染色体是上皮细胞-结缔组织附着中重要的粘附复合物 (Stepp，m.a.，S. Spurr-Michaud，A. Tisdale，J. Elwell，和I. K. Gipson。1990. Proc. Natl. Acad. Sci.美国。87:8970-8974; Jones，j.c.R.，M.Kurpakus，H. M. Cooper和V. Quaranta。1991.细胞调控。2:427-438)。半桥粒的胞质组分包括大疱性类天疱疮 (BP) 抗原，而细胞外组分包括锚定细丝 (CAF) 的125-kD组分和含VII型胶原的锚定原纤维。我们已经在体外伤口愈合外植体模型中监测了 α6β4整合素在形成半桥体中的掺入。在最近从裸露的结缔组织上未受伤部位的边缘迁移的上皮细胞中，α6β4首先出现在整个细胞表面。在这个阶段，这些细胞含有很少或没有胞质半染色体成分，至少可以通过使用BP自身抗体的免疫荧光检测到，而它们已经对层粘连蛋白和CAF呈阳性。在稍后的阶段，随着细胞对胞质半桥粒成分 (例如BP抗原) 以及VII型胶原呈阳性，α6β4沿着上皮细胞的基极集中，并与外植体的结缔组织邻接。针对 β4的多克隆抗体不会干扰外植体中上皮细胞的迁移。但是，它们阻止了半桥粒复合体的组装，并抑制了在伤口区域迁移的细胞中VII型胶原的表达。此外，它们还会诱导外植体未受伤区域的非迁移细胞中已建立的半桥粒的破坏。针对 α6的单克隆抗体具有更显着的作用，因为它们完全脱离了外植体未受伤区域的上皮细胞。针对CAF的抗体也可以通过诱导基底膜区的上皮和结缔组织之间的分离而分离未受伤区域的上皮细胞。这些结果提出了一个模型，通过该模型，α6β4极化到细胞的基底表面，可能是由假定的锚固丝相关配体诱导的，触发了半桥粒斑块的组装。

BACKGROUND & AIMS: The present study examined the differences of self-ratings of 4 h sleep in three statesL-WE, where the percentage of waking time and urinary epinephrine are low (< 20% waking time); H-W, where the percentage of waking time and epinephrine levels increase along the basal regression line as determined by a previous study (20-100% waking time and < 7 ng/min); H-E, where epinephrine levels increase more than expected from the basal regression line for the two parameters (> 7 ng/min). Eight healthy male subjects participated twice in a 4 h polysomnograph experiment with four types of sleep onset (total of 64 observations). In group L-WE (52 observations for eight subjects), there were no excessively negative feelings on sleep latency, sleep depth, and feelings of sleep compared with usual sleep according to the questionnaire. Subjective sleep diagrams in group L-WE were similar to polysomnographic findings. Thus, group L-WE was thought objectively and subjectively to have a good sleep state. Groups H-W (eight observations for four subjects) and H-E (four observations for two subjects) had negative feelings regarding sleep depth and feelings of sleep compared with usual sleep. Approximately half the group H-W underrated their sleep compared with objective diagrams, while all cases in group H-E remarkably underrated their sleep in the subjective diagrams. The state of remarkable adrenal medullary secretory activity seen in group H-E and that of the slightly increased activity shown in group H-W were included in poor sleep states objectively and subjectively.

背景与目标： 本研究检查了三种状态下4 h睡眠自我评分的差异，其中清醒时间和尿肾上腺素的百分比较低 (< 20% 清醒时间); H-w，其中清醒时间和肾上腺素水平的百分比沿基础回归线增加，如先前的研究 (20-100% 清醒时间和 <7 ng/min); H-e，其中肾上腺素水平增加超过两个参数的基础回归线的预期 (> 7 ng/min)。八名健康的男性受试者两次参加了4小时多导睡眠图实验，其中有四种睡眠发作类型 (总共64个观察结果)。根据问卷，在l-we组 (八名受试者的52项观察结果) 中，与通常的睡眠相比，在睡眠潜伏期，睡眠深度和睡眠感觉方面没有过度负面的感觉。L组的主观睡眠图-我们与多导睡眠图的发现相似。因此，客观和主观地认为L-WE组具有良好的睡眠状态。与通常的睡眠相比，h-w组 (四名受试者的八次观察) 和h-e组 (两名受试者的四次观察) 对睡眠深度和睡眠感觉有负面感觉。与客观图相比，大约一半的h-w组低估了他们的睡眠，而h-e组的所有病例在主观图中都明显低估了他们的睡眠。客观和主观地将h-e组中明显的肾上腺髓质分泌活动状态和h-w组中显示的轻度活动状态包括在不良的睡眠状态中。

BACKGROUND & AIMS: :In addition to our recent report on a series of rationally designed benzylindolyldiketo acids acting as potent HIV-1 integrase strand transfer inhibitors, we disclose the results obtained with novel compounds chemically modified on the diketo acid moiety in order to investigate its influence on the biological activity and cytotoxicity. The activity of designed and synthesized 4-[(1-benzyl-1H-indol-3-yl)carbonyl]-3-hydroxyfuran-2(5H)-one derivatives lies in the micromolar range with regard to HIV IN enzymatic activity. The microwave-assisted synthesis was employed in some steps of the chemical procedures.

背景与目标： : 除了我们最近关于一系列合理设计的苄基吲哚二酮酸作为有效的HIV-1整合酶链转移抑制剂的报道之外，我们公开了用在二酮酸部分上化学修饰的新型化合物获得的结果，以研究其对生物活性和细胞毒性的影响。设计和合成的4-[(1-苄基-1H-吲哚-3-基) 羰基]-3-羟基呋喃-2(5H)-一衍生物的酶活性在微摩尔范围内。微波辅助合成用于化学程序的某些步骤。

BACKGROUND & AIMS: :The promotion potential of phenobarbital (PB) and 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) on liver carcinogenesis after initiation with various doses of diethylnitrosamine (DEN) was assessed using an in vivo short term system. Male F344 rats were pretreated with a single intraperitoneal injection of varying doses of DEN (0, 6, 12, 25, 50, 100 or 200 mg/kg body wt), and 2 weeks later were treated with 0.05% PB or 0.06% 3'-Me-DAB for 6 weeks. All animals were subjected to partial hepatectomy 3 weeks after the DEN treatment. Quantitation of gamma-glutamyltranspeptidase-positive (gamma-GT+) foci revealed a DEN dose-dependent response. Magnitude of promotion by PB and more pronounced by 3'-Me-DAB was, in contrast, strongest at the lower doses of DEN. The results suggest that quantitative differences with regard to initiation level may exist, influencing the promotability of initiated cells.

背景与目标： : 使用体内短期系统评估了苯巴比妥 (PB) 和3 '-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) 在各种剂量的二乙基亚硝胺 (DEN) 引发肝癌后的促进潜力。用单次腹腔注射不同剂量的DEN (0、6、12、25、50、100或200 mg/kg体重) 预处理雄性F344大鼠，2周后用0.05% PB或0.06% 3 '-Me-DAB治疗6周。所有动物在DEN治疗后3周接受部分肝切除术。Γ-谷氨酰转肽酶阳性 (γ-GT) 病灶的定量显示出DEN剂量依赖性反应。相比之下，在较低剂量的DEN下，PB和3 '-Me-DAB的促进程度最高。结果表明，在起始水平方面可能存在定量差异，从而影响起始细胞的启动子性。

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: :Tumor suppressor PAR-4 acts in part by modulating sensitivity to apoptosis, but the basis for its activity is not fully understood. In this study, we describe a novel mechanism of antiapoptosis by NF-κB, revealing that it can block PAR-4-mediated apoptosis by downregulating trafficking of the PAR-4 receptor GRP78 from the endoplasmic reticulum to the cell surface. Mechanistic investigations revealed that NF-κB mediated this antiapoptotic mechanism by upregulating expression of UACA, a proinflammatory protein in certain disease settings. In clinical specimens of cancer, a strong correlation existed between NF-κB activity and UACA expression, relative to normal tissues. UACA bound to intracellular PAR-4 in diverse cancer cells, where it prevented translocation of GRP78 from the endoplasmic reticulum to the cell surface. This pathway of antiapoptosis could be inhibited by suppressing levels of NF-κB or UACA expression, which enhanced endoplasmic reticulum stress and restored GRP78 trafficking to the cell surface, thereby sensitizing cancer cells to apoptosis by extracellular PAR-4 or GRP78 agonistic antibody. In summary, our results identify a novel intracellular pathway of apoptosis mediated by NF-κB through UACA elevation, which by attenuating endoplasmic reticulum stress and GRP78 translocation to the cell surface can blunt the sensitivity of cancer cells to apoptosis.

背景与目标： : 肿瘤抑制因子PAR-4部分通过调节对凋亡的敏感性起作用，但其活性的基础尚不完全清楚。在这项研究中，我们描述了NF-κ b抗凋亡的新机制，揭示了它可以通过下调PAR-4受体GRP78从内质网到细胞表面的运输来阻断PAR-4介导的凋亡。机理研究表明，NF-κ b通过上调UACA (在某些疾病环境中是一种促炎蛋白) 的表达来介导这种抗凋亡机制。在癌症的临床标本中，相对于正常组织，NF-κ b活性与UACA表达之间存在很强的相关性。UACA与多种癌细胞中的细胞内PAR-4结合，阻止了GRP78从内质网转移到细胞表面。抑制NF-κ b或UACA表达的水平可以抑制这种抗凋亡途径，从而增强内质网应激并恢复GRP78向细胞表面的运输，从而使癌细胞对细胞外PAR-4或GRP78激动抗体的凋亡敏感。总之，我们的结果确定了由NF-κ b通过UACA升高介导的新的细胞内凋亡途径，该途径通过减弱内质网应激和GRP78易位到细胞表面可以减弱癌细胞对凋亡的敏感性。