金乌骨通胶囊联合塞来昔布胶囊治疗强直性脊柱炎的临床研究

摘要 目的：观察金乌骨通胶囊联合塞来昔布胶囊治疗强直性脊柱炎（AS）的临床疗效。方法：选取天津中医药大学第一附属医院2017年4月~2020年4月期间收治的126例AS患者。采用双色球法（红色、绿色）将患者分为对照组（红色）和研究组（绿色）,各63例。对照组接受塞来昔布胶囊治疗,研究组接受金乌骨通胶囊联合塞来昔布胶囊治疗,两组均治疗3个月。治疗3个月后,观察两组疗效,记录治疗期间不良反应发生率,对比两组治疗前、治疗3个月后的临床症状缓解情况、生存质量及实验室指标变化。结果：研究组的临床总有效率高于对照组（P<0.05）。治疗3个月后,研究组巴氏强直性脊柱炎疾病活动指数（BASDAI）、巴氏强直性脊柱炎功能指数（BASFI）评分低于对照组,枕墙距小于对照组,腰椎活动度、胸廓活动度大于对照组（P<0.05）。治疗3个月后,研究组世界卫生组织生存质量简表（WHOQOL-BRIEF）各维度评分高于对照组（P<0.05）。治疗3个月后,研究组血沉（ESR）、C-反应蛋白（CRP）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）低于对照组（P<0.05）。两组不良反应发生率组间比较无统计学差异（P>0.05）。结论：金乌骨通胶囊联合塞来昔布胶囊治疗AS患者,可促进其临床症状及生存质量改善,降低ESR及炎性因子水平,是一种安全有效的治疗方案。     

风湿祛痛胶囊联合塞来昔布胶囊治疗强直性脊柱炎的疗效及对血液流变学和血清炎性因子的影响

摘要 目的：探讨强直性脊柱炎（AS）患者经风湿祛痛胶囊联合塞来昔布胶囊治疗后的疗效及对血液流变学和血清炎性因子的影响。方法：选取2014年8月-2021年12月在联勤保障部队第九八三医院治疗的80例AS患者。按照随机数字表法分为对照组（常规治疗及塞来昔布胶囊治疗，40例）和研究组（常规治疗及风湿祛痛胶囊联合塞来昔布胶囊治疗，40例），两组患者均治疗8周。对比两组血液流变学指标、血清炎性因子指标、视觉模拟评分法（VAS）评分、C反应蛋白（CRP）、巴氏强直性脊柱炎疾病活动指数（BASDAI）评分、红细胞沉降率（ESR），同时记录两组治疗期间不良反应发生情况。结果：研究组治疗8周后BASDAI、VAS评分低于对照组（P<0.05）。研究组治疗8周后CRP、ESR低于对照组（P<0.05）。研究组治疗8周后全血高切黏度、血浆黏度、全血低切黏度、红细胞压积低于对照组（P<0.05）。研究组治疗8周后白介素-23（IL-23）、肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）均低于对照组（P<0.05）。两组不良反应发生率对比无差异（P>0.05）。结论：风湿祛痛胶囊联合塞来昔布胶囊治疗AS，可有效缓解患者的临床症状，抑制疾病进展，可能与调节血液流变学、降低炎性因子水平有关。     

腰痹通胶囊联合塞来昔布胶囊治疗椎间盘源性腰痛的疗效及对血清炎性因子和疼痛介质的影响

摘要 目的：探讨腰痹通胶囊联合塞来昔布胶囊治疗椎间盘源性腰痛（DLBP）的疗效及对血清炎性因子和疼痛介质的影响。方法：选择2017年5月~2022年5月期间我院接收的75例DLBP患者。依据信封抽签法将患者分为对照组（n=37）和研究组（n=38）,对照组接受塞来昔布胶囊治疗,研究组接受腰痹通胶囊联合塞来昔布胶囊治疗。对比两组疗效、临床症状评分、血清炎性因子和疼痛介质的变化情况,同时观察两组用药安全性。结果：研究组痊愈8例,显效10例,有效18例,临床总有效率94.74%,高于对照组的痊愈4例,显效8例,有效17例,临床总有效率78.38%（P<0.05）。治疗1个月后,研究组的视觉疼痛模拟（VAS）评分、Oswestry功能障碍指数（ODI）较对照组更低（P<0.05）,改良日本骨科协会（JOA）评分较对照组更高（P<0.05）。治疗1个月后,研究组的血清白细胞介素-6（IL-6）、C反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）水平较对照组更低（P<0.05）。治疗1个月后,研究组的血清 P物质（SP）、多巴胺（DA）、前列腺素E₂（PGE₂）水平较对照组更低（P<0.05）。两组不良反应发生率组间对比差异不显著（P>0.05）。结论：腰痹通胶囊联合塞来昔布胶囊治疗DLBP,可有效减轻疼痛,恢复腰椎功能,降低血清炎性因子和疼痛介质水平。     

通滞苏润江胶囊联合塞来昔布对早期膝骨关节炎患者血清炎症因子和疼痛介质的影响

摘要 目的：观察通滞苏润江胶囊联合塞来昔布对早期膝骨关节炎（KOA）患者血清炎症因子和疼痛介质的影响。方法：选择2021年6月-2022年12月期间石家庄市人民医院收治的早期KOA患者80例,采用随机数字表法将患者分为对照组（塞来昔布,40例）和观察组（通滞苏润江胶囊联合塞来昔布,40例）。对比两组疗效、疼痛视觉模拟评分（VAS）、西安大略和麦克马斯特大学骨关节炎指数（WOMAC）评分、炎症因子和疼痛介质。结果：观察组临床总有效率高于对照组（P<0.05）。两组治疗1个月后VAS、WOMAC评分下降,且观察组低于对照组同时间点（P<0.05）。两组治疗1个月后肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）下降,且观察组低于对照组同时间点（P<0.05）。两组治疗1个月后前列腺素E₂（PGE₂）、P物质（SP）下降,且观察组低于对照组同时间点（P<0.05）。两组不良反应发生率对比未见统计学差异（P>0.05）。结论：塞来昔布和通滞苏润江胶囊联合治疗早期KOA患者,可缓解疼痛,提高临床治疗效果,改善关节功能,同时还可改善血清炎症因子和疼痛介质水平。     

温针灸联合塞来昔布胶囊对膝骨关节炎患者骨代谢指标和血清IL-6、IL-17、IL-18水平的影响

摘要 目的：观察温针灸联合塞来昔布胶囊对膝骨关节炎（KOA）患者骨代谢指标和血清白介素-6（IL-6）、白介素-17（IL-17）、白介素-18（IL-18）水平的影响。方法：纳入我院2017年4月~2020年12月间针灸科接收的KOA患者80例,将患者采用信封抽签法分为对照组和研究组,各为40例。对照组给予塞来昔布胶囊进行治疗,研究组给予温针灸联合塞来昔布胶囊进行治疗,均连续治疗8周。观察两组疗效、骨代谢指标[骨保护素（OPG）、降钙素（CT）、骨钙素（BGP）]、炎性因子、量表评分[视觉模拟评分法（VAS）、骨关节炎指数评分表（WOMAC）、Lysholm膝关节评分]等情况,记录治疗期间的不良反应发生率。结果：研究组的临床总有效率高于对照组（P<0.05）。研究组治疗8周后VAS、WOMAC较对照组低,Lysholm膝关节评分较对照组高（P<0.05）。研究组治疗8周后血清OPG、BGP较对照组高（P<0.05）。两组血清CT水平治疗8周后对比差异无统计学意义（P>0.05）。研究组治疗8周后血清IL-6、IL-17、IL-18较对照组低（P<0.05）。两组不良反应总发生率组间对比差异无统计学意义（P>0.05）。结论：温针灸联合塞来昔布胶囊治疗KOA患者,可有效减轻疼痛,促进膝关节功能恢复,同时还可抑制炎症因子,改善骨代谢指标,优化治疗效果。     

鳖甲煎丸联合恩替卡韦对慢性乙型肝炎肝纤维化患者肝功能、肝纤维化指标和外周血TGF-β1/ smad信号通路的影响

摘要 目的：探讨鳖甲煎丸辅助恩替卡韦治疗慢性乙型肝炎肝纤维化患者肝功能、肝纤维化指标和外周血转化生长因子β1（TGF-β1）/smad信号通路的影响。方法：选取2021年01月~2022年01月中国中医科学院望京医院收治的132例慢性乙型肝炎肝纤维化患者分为对照组和观察组，各66例。对照组以恩替卡韦治疗，观察组增加鳖甲煎丸治疗。比较两组临床疗效、肝功能指标、肝脾B超指标、中医证候积分、肝纤维化指标、外周血TGF-β1/smad信号通路、不良反应发生率。结果：观察组总有效率为90.91%高于对照组74.24%（P<0.05）。治疗后，观察组肝门静脉主干内径、脾门厚度均小于对照组，中医证候积分低于对照组（P<0.05）。治疗后，观察组直接胆红素（DBIL）、谷草转氨酶（AST）、谷丙转氨酶（ALT）水平均低于对照组（P<0.05）。治疗后，观察组的Ⅲ型前胶原(PC III)、IV型胶原（IV-C）、透明质酸酶（HAase）、层粘连蛋白（LN）水平均低于对照组（P<0.05）。治疗后，观察组外周血Smad同源物7（Smad7）水平高于对照组，TGF-β1、Smad同源物3（Smad3）水平低于对照组（P<0.05）。两组患者不良反应发生率组间对比无统计学意义（P>0.05）。结论：鳖甲煎丸结合恩替卡韦对慢性乙型肝炎肝纤维化患者可提升治疗效果，降低中医证候积分，改善肝功能和肝纤维化损伤状态，其疗效机制可能与调控TGF-β1/smad信号通路有关。     

扶正方对Lewis肺癌小鼠免疫功能、PI3K/AKT信号通路和外周血IL-2、IL-6、INF-γ的影响

摘要 目的：观察扶正方对Lewis肺癌小鼠免疫功能、磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路和外周血白细胞介素（IL）-2、IL-6、γ干扰素（INF-γ）的影响。方法：将40只Lewis肺癌小鼠随机分为模型组（M组）、扶正方低剂量组（A组）、扶正方高剂量组（B组）、顺铂组（S组）,每组10只,A组、B组分别给予扶正方0.4 mL/20 g、0.8 mL/20 g灌胃,M组给予生理盐水0.4 mL/20 g灌胃,S组给予顺铂1 mg/mL,0.4 mL灌胃,连续14d,比较各组小鼠一般情况、肿瘤重量,胸腺指数、脾脏指数、脾脏CD3⁺细胞、CD4⁺细胞、CD8⁺细胞比例细胞百分比,鼠肿瘤组织PI3K/AKT信号通路蛋白表达水平及外周血IL-2、IL-6、INF-γ水平。结果：A组、B组、S组小鼠肿瘤重量低于M组,S组小鼠肿瘤重量低于A组、B组（P<0.05）,治疗前各组小鼠体重比较无统计学差异（P>0.05）,治疗后A组、B组小鼠体重高于S组、M组（P<0.05）。A组、B组小鼠胸腺指数显著高于M组、S组（P<0.05）。A组、B组CD3⁺、CD4⁺、CD4⁺/CD8⁺显著高于M组、S组,CD8⁺显著低于M组、S组（P<0.05）,B组CD3⁺、CD4⁺、CD4⁺/CD8⁺显著高于A组,CD8⁺低于A组（P<0.05）。A组、B组、S组小鼠肿瘤组织PI3K蛋白、AKT蛋白表达水平显著低于M组（P<0.05）。A组、B组、S组小鼠外周血IL-2、INF-γ水平显著高于M组,IL-6水平显著低于M组（P<0.05）。结论：扶正方可以提升Lewis肺癌小鼠免疫功能,调节IL-2、IL-6、INF-γ细胞因子水平,抑制PI3K/AKT信号通路起到抗肺癌的作用。     

沙库巴曲缬沙坦钠对老年慢性心力衰竭合并2型糖尿病患者肾功能、神经内分泌激素和TGF-β1/Smad3信号通路的影响

摘要 目的：探讨沙库巴曲缬沙坦钠对老年慢性心力衰竭（CHF）合并2型糖尿病（T2DM）患者肾功能、神经内分泌激素和转化生长因子-β1（TGF-β1）/Smad3信号通路的影响。方法：选择我院2020年5月~2021年5月期间收治的92例老年CHF合并T2DM患者,按照双色球法将患者分为对照组（n=46,接受常规治疗）和研究组（n=46,对照组基础上接受沙库巴曲缬沙坦钠治疗）。对比两组心功能、肾功能、神经内分泌激素、TGF-β1/Smad3信号通路相关指标和不良反应发生情况。结果：治疗12个月后,研究组心输出量（CO）、左心室射血分数（LVEF）高于对照组,左心室舒张末期内径（LVEDD）低于对照组（P<0.05）。治疗12个月后,研究组血肌酐（Scr）、血尿素氮（BUN）、血尿酸（UA）、中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、胱抑素C（Cys-C）低于对照组（P<0.05）。治疗12个月后,研究组去甲肾上腺素（NE）、醛固酮（ALD）和血管紧张素II（AngII）低于对照组（P<0.05）。治疗12个月后,研究组Smad7 mRNA高于对照组,Smad2 mRNA、TGF-β1 mRNA、Smad3 mRNA低于对照组（P<0.05）。两组不良反应发生率组间比较差异无统计学意义（P>0.05）。结论：沙库巴曲缬沙坦钠用于治疗老年CHF合并T2DM患者,可有效改善患者的心功能、肾功能、神经内分泌激素和TGF-β1/Smad3信号通路相关指标,安全性较好。     

基于JAK2/STAT3信号通路探讨头针结合艾灸对缺血缺氧脑瘫幼鼠模型的影响及作用机制

摘要 目的：基于JAK2/STAT3信号通路探讨头针结合艾灸对缺血缺氧脑瘫幼鼠模型的影响及作用机制。方法：选取40只7 d龄的清洁级SD幼鼠,按随机数字表法分为A组、B组、C组、D组、E组,每组8只。E组不予以任何处理,其余各组幼鼠均以改良的缺氧缺血性脑病造模方法构建缺血缺氧脑瘫模型。造模成功后A组、B组、C组分别给予头针结合艾灸治疗、头针治疗、艾灸治疗,D组、E组均给予50 ?滋L生理盐水。对比各组幼鼠脑组织Janus蛋白酪氨酸激酶2（JAK2）及转录活化因子3（STAT3）蛋白表达量、血清炎症因子[白介素-6（IL-6）、肿瘤坏死因子--α（TNF-α）、白介素-1（IL-1）]水平、神经递质[去甲肾上腺素（NE）、5-羟色胺（5-HT）]水平、记忆功能。结果：A组脑组织JAK2、STAT3 蛋白表达量低于B、C、D组,但高于E组（P<0.05）;A组血清IL-6、IL-1和TNF-α水平低于B、C、D组,但高于E组（P<0.05）;A组血清NE、5-HT水平低于B、C、D组,但高于E组（P<0.05）;A组Y迷宫实验正确次数多于B、C、D组（P<0.05）,而少于E组（P<0.05）。结论：头针结合艾灸可通过靶向抑制JAK2/STAT3信号通路相关蛋白的表达下调机体炎症反应水平,从而降低缺血缺氧脑瘫幼鼠脑组织损伤,促进其记忆功能恢复。     

基于TGF-β1/Smads信号通路探讨扶正化瘀胶囊联合恩替卡韦分散片治疗慢性乙型肝炎肝纤维化的疗效机制

摘要 目的：基于转化生长因子-β1（TGF-β1）/Smads信号通路探讨恩替卡韦分散片联合扶正化瘀胶囊治疗慢性乙型肝炎肝纤维化的疗效机制。方法：纳入新疆医科大学第一附属医院2020年4月~2022年1月期间收治的慢性乙型肝炎肝纤维化患者104例,将患者根据随机数字表法分为对照组和观察组,例数各为52例。在常规治疗的基础上,对照组接受恩替卡韦分散片治疗,观察组接受扶正化瘀胶囊联合恩替卡韦分散片治疗。对比两组肝功能指标、肝纤维化指标、TGF-β1/Smads信号通路相关指标和影像学指标变化情况。结果：治疗24周后,两组丙氨酸氨基转移酶 （ALT）、天门冬氨酸氨基转移酶（AST）水平均下降,且观察组的下降程度较对照组更大（P<0.05）。治疗24周后,两组Ⅲ型前胶原（PCⅢ）、层粘连蛋白（LN）、Ⅳ型胶原（Ⅳ-C）、透明质酸（HA）水平均下降,且观察组的下降程度较对照组更大（P<0.05）。治疗24周后,两组TGF-β1mRNA、Smad2mRNA、Smad3mRNA水平下降, Smad7mRNA水平升高,且观察组的改善程度较对照组更大（P<0.05）。治疗24周后,两组门静脉内径、脾脏厚度均下降,且观察组较对照组更低（P<0.05）。结论：恩替卡韦分散片联合扶正化瘀胶囊治疗慢性乙型肝炎肝纤维化,可改善肝功能,对肝纤维化具有明显的抑制作用,可能与调控TGF-β1/Smads信号通路有关。     

Alkylation of isolated chromatin with N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea

When isolated chromatin is incubated with the carcinogens N-methyl-N-nitrosourea (MeNU) and N-ethyl-N-nitrosourea (EtNU), DNA and chromosomal proteins become alkylated to increasingly greater extents as the carcinogen concentrations increase. With either MeNU or EtNU, the core and linker DNA of chromatin are alkylated to essentially identical extents. Alkylation of chromatin DNA as well as free DNA is drastically reduced at physiological ionic strengths (e.g. 0.15 M NaCl). The presence of 0.15 M NaCl, on the other hand, enhances alkylation of chromosomal proteins. While EtNU is much less reactive to DNA than MeNU, alkylation of chromosomal proteins relative to that of chromatin DNA has been found to be markedly greater with EtNU than with MeNU. Such a difference in their relative reactivities toward DNA and proteins may be related to the known difference of carcinogenic potency between these N-nitroso compounds.     

Association of the eukaryotic V1VO ATPase subunits a with d and d with A

Owing to the complex nature of V₁V_O ATPases, identification of neighboring subunits is essential for mechanistic understanding of this enzyme. Here, we describe the links between the V₁ headpiece and the V_O-domain of the yeast V₁V_O ATPase via subunit A and d as well as the V_O subunits a and d using surface plasmon resonance and fluorescence correlation spectroscopy. Binding constants of about 60 and 200 nM have been determined for the a-d and d-A assembly, respectively. The data are discussed in light of subunit a and d forming a peripheral stalk, connecting the catalytic A₃B₃ hexamer with V_O.

Structured summary

MINT-7012054: d (uniprotkb:P32366) binds (MI:0407) to A (uniprotkb:P17255) by fluorescence correlation spectroscopy (MI:0052)MINT-7012041: d (uniprotkb:P32366) binds (MI:0407) to A (uniprotkb:P17255) by surface plasmon resonance (MI:0107)MINT-7012028: d (uniprotkb:P32366) binds (MI:0407) to a (uniprotkb:P32563) by surface plasmon resonance (MI:0107)     

Association of upregulated Ras/Raf/ERK1/2 signaling with autism

Autism is a neurodevelopmental disorder characterized by impairments in social interaction, verbal communication and repetitive behaviors. BTBR mouse is currently used as a model for understanding mechanisms that may be responsible for the pathogenesis of autism. Growing evidence suggests that Ras/Raf/ERK1/2 signaling plays death-promoting apoptotic roles in neural cells. Recent studies showed a possible association between neural cell death and autism. In addition, two studies reported that a deletion of a locus on chromosome 16, which includes the MAPK3 gene that encodes ERK1, is associated with autism. We thus hypothesized that Ras/Raf/ERK1/2 signaling could be abnormally regulated in the brain of BTBR mice that models autism. In this study, we show that expression of Ras protein was significantly elevated in frontal cortex and cerebellum of BTBR mice as compared with B6 mice. The phosphorylations of A-Raf, B-Raf and C-Raf were all significantly increased in frontal cortex of BTBR mice. However, only C-Raf phosphorylation was increased in the cerebellum of BTBR mice. In addition, we further detected that the activities of both MEK1/2 and ERK1/2, which are the downstream kinases of Ras/Raf signaling, were significantly enhanced in the frontal cortex. We also detected that ERK1/2 is significantly over-expressed in frontal cortex of autistic subjects. Our results indicate that Ras/Raf/ERK1/2 signaling is upregulated in the frontal cortex of BTBR mice that model autism. These findings, together with the enhanced ERK1/2 expression in autistic frontal cortex, imply that Ras/Raf/ERK1/2 signaling activities could be increased in autistic brain and involved in the pathogenesis of autism.     

Infection of Promecotheca papuana with Synnematium jonesii

The fungus Synnematium jonesii was isolated from naturally infected adults of Promecotheca papuana and cultured in vitro. Caged beetles were successfully inoculated using an aqueous suspension of conidia and also a dust prepared from dried sclerotia. The implications of using this fungus for the control of P. papuana outbreaks is discussed.     

板蓝根颗粒对甲型流感病毒小鼠的作用

目的测定板蓝根颗粒抗流感病毒的药效作用。方法 A/California/7/2009（CA7）病毒滴鼻感染BALB/c小鼠观察14d,观察板蓝根对甲型H1N1流感病毒感染小鼠的保护作用,计算小鼠存活率、存活天数以及延长生命率。感染的小鼠第5天每组小鼠处死一半,取肺组织,观察板蓝根对甲型H1N1流感病毒感染小鼠肺组织的保护作用。结果板蓝根可明显延长甲型H1N1流感病毒感染小鼠的存活天数并提高存活率,病理结果显示板蓝根对甲型H1N1流感病毒感染的小鼠的肺组织有一定程度的保护作用,与模型组比较差异显著（P〈0.05）。结论板蓝根颗粒对甲型H1N1流感病毒感染的小鼠有较好的保护作用。     

Hebeloma species associated with Cistus

The genus Hebeloma has a number of species highly specific to Cistus and others that occur with several host genera. This paper discusses the species of Hebeloma that appear to be ectomycorrhizal with Cistus, judging from their occurrence when Cistus is the only available host. The previously unknown species H. plesiocistum spec. nov. is described. We also provide a key to the known Hebeloma associates of Cistus. Molecular analyses based on ITS sequence data further illustrate the distinctness of the newly described species and difficulties in the species delimitation with view to H. erumpens. Specific associations with Cistus may have evolved more than once within the genus Hebeloma.     

Prosthecium species with Stegonsporium anamorphs on Acer

Data from microscopic morphology, single-spore cultures, and DNA analyses of teleomorphs and anamorphs support the recognition of five species of Prosthecium with Stegonsporium anamorphs on Acer: P. acerinum sp. nov., the teleomorph of S. acerinum; P. acerophilum comb. nov., formerly known as Dictyoporthe acerophila; P. galeatum comb. nov., originally described as Massaria galeata; P. opalus sp. nov.; and P. pyriforme sp. nov., the teleomorph of S. pyriforme s. str. The morphology of both type specimens and freshly collected material was investigated. The teleomorphs have brown ellipsoidal ascospores with five distosepta and often a longitudinal distoseptum. The anamorphs of all species described here belong to Stegonsporium; their connection to the Prosthecium teleomorphs was demonstrated by morphology and DNA sequences of single spore cultures derived from both ascospores and conidia. The anamorphs and teleomorphs of all five Prosthecium species are described and illustrated by LM images, and a key to these species is provided. As perceived from this work, S. pyriforme is restricted to Europe and does not occur in North America, whereas S. acerinum is restricted to North America, not found in Europe. The host associations given in the literature are revised and evidence is provided that only A. opalus, A. pseudoplatanus, and A. saccharum are confirmed hosts of Prosthecium with Stegonsporium anamorphs. Molecular phylogenetic analyses of tef1, ITS rDNA, and partial nuLSU rDNA sequences confirm that the species with Stegonsporium anamorphs are closely related to P. ellipsosporum, the generic type species. Stilbospora macrosperma is confirmed as the anamorph of P. ellipsosporum by DNA data of single spore isolates obtained from both ascospores and conidia.     

Interaction of anthelmintic benzimidazoles with AscarisSuum embryonic tubulin

The ability of mebendazole and fenbendazole to bind to tubulin in cytosolic fractions from 8-day Ascaris suum embryos was determined by inhibition studies with [³H]colchicine. Colchicine binding in the presence of 1·10^?6 M mebendazole was completely inhibited during a 6 h incubation period at 37°C. Inhibition of colchicine binding to A. suum embryonic tubulin by mebendazole and fenbendazole appeared to be noncompetative. The inhibition constants of mebendazole and fenbendazole for A. suum embryonic tubulin were 1.9·10^?8 M and 6.5·10^?8 M, respectively. Mebendazole and fenbendazole appeared to be competitive inhibitors of colchicine binding to bovine brain tubulin. The inhibition constants of mebendazole and fenbendazole for bovine brain tubulin were 7.3·10^?6 M and 1.7·10^?5 M, respectively. These values are 250–400 times greater than the inhibition constants of fenbendazole and mebendazole for A. suum embryonic tubulin. Differential binding affinities between nematode tubulin and mammalian tubulin for benzimidazoles may explain the selective toxicity. The importance of tubulin as a receptor for anthelmintic benzimidazoles in animal parasitic nematodes is discussed.     

Association of Cdc42/N-WASP/Arp2/3 signaling pathway with Golgi membranes

Recent findings indicate that Cdc42 regulates Golgi-to-ER (endoplasmic reticulum) protein transport through N-WASP and Arp2/3 (Luna et al. 2002, Mol. Biol. Cell, 13:866-879). To analyse the components of the Cdc42-governed signaling pathway in the secretory pathway, we localized Cdc42, N-WASP and Arp2/3 in the Golgi complex by cryoimmunoelectron microscopy. Cdc42 is found throughout the Golgi stack, particularly in cis/middle cisternae, whereas N-WASP and Arp3 (a component of the Arp2/3 complex) are restricted to cis cisternae. Arp3 also colocalized in peri-Golgi tubulovesicular structures with either KDEL receptor or GM130. Even though Arp3 is not found in TGN46-positive cisternal elements, a small fraction of Arp3-labeled tubulo-vesicular elements showed TGN46 labeling. Active Cdc42 (GTP-bound form) induced relocation of N-WASP and Arp3 to the lateral rims of Golgi cisternae. These results show that the actin nucleation and polymerization signaling pathway governed by Cdc42/N-WASP/Arp operates in the Golgi complex of mammalian cells, further implicating actin dynamics in Golgi-associated membrane trafficking.