不同肠内营养支持方案对2型糖尿病并发急性脑卒中患者肠黏膜屏障功能和Th17/Treg免疫应答平衡的影响

摘要 目的：对比不同肠内营养支持方案对2型糖尿病（T2DM）并发急性脑卒中患者肠黏膜屏障功能和Th17/Treg免疫应答平衡的影响。方法：选取我院2019年3月~2021年2月期间收治的93例T2DM并发急性脑卒中患者,按照信封抽签法将患者分为对照组（46例）和研究组（47例）,对照组接受传统肠内营养支持方案,研究组接受个体化肠内营养支持方案,观察两组血糖水平、营养指标、肠黏膜屏障功能和Th17/Treg免疫应答平衡变化。结果：研究组治疗 1~7 d 后的血糖水平均低于对照组（P<0.05）。治疗7 d后,研究组血清前白蛋白、白蛋白、血红蛋白水平高于对照组（P<0.05）。治疗7 d后,研究组血浆内毒素（ET）、二胺氧化酶（DAO）低于对照组（P<0.05）。治疗7 d后,研究组Th17、Th17/Treg低于对照组;Treg高于对照组（P<0.05）。结论：个体化肠内营养支持方案有利于降低T2DM并发急性脑卒中患者的血糖水平,促进肠粘膜屏障功能的恢复,进而改善机体营养状态及免疫状态。     

艾迪注射液联合XELOX方案对晚期结直肠癌患者肿瘤标志物、血管生成和外周血Th17/Treg平衡的影响

摘要 目的：观察艾迪注射液联合XELOX方案（卡培他滨+奥沙利铂）对晚期结直肠癌患者肿瘤标志物、血管生成和外周血辅助性T细胞17（Th17）/调节性T细胞（Treg）平衡的影响。方法：选择我院2018年2月~2019年8月期间收治的98例经内镜活检证实的晚期结直肠癌患者,根据随机数字表法分为对照组（XELOX方案化疗,49例）和研究组（艾迪注射液联合XELOX化疗方案治疗,49例）。观察两组患者近、远期疗效,对比两组治疗前后的肿瘤标志物、血管生成和Th17/Treg平衡相关指标水平,记录两组用药安全性。结果：与对照组相比,研究组的客观缓解率、疾病控制率明显更高（P<0.05）;研究组的总生存率、无进展生存率高于对照组,但两组间差异无统计学意义（P>0.05）。治疗后,两组癌胚抗原（CEA）、糖链抗原125（CA125）、糖链抗原242（CA242）水平均下降,且研究组低于对照组同期（P<0.05）。治疗后,两组血管内皮生长因子（VEGF）、肿瘤微血管密度（MVD）水平均下降,且研究组低于对照组同期（P<0.05）。治疗后,两组Th17比例、Th17/Treg比值均升高,Treg比例均下降（P<0.05）;研究组治疗后Th17比例、Th17/Treg比值高于对照组同期,Treg比例低于对照组同期（P<0.05）。两组不良反应总发生率对比未见明显差异（P>0.05）。结论：艾迪注射液联合XELOX方案治疗晚期结直肠癌的疗效确切,可能与改善Th17/Treg平衡、抑制血管生成有关。     

大柴胡汤对轻症急性胰腺炎患者Th17/Treg免疫平衡和血清水通道蛋白的影响

摘要 目的：观察大柴胡汤对轻症急性胰腺炎（MAP）患者辅助性T细胞17 /调节性T细胞（Th17/Treg）免疫平衡和血清水通道蛋白（AQP）的影响。方法：按照随机数字表法，将天津中医药大学第一附属医院2020年4月~2022年4月期间收治的95例MAP患者分为对照组（西医基础治疗，47例）和研究组（对照组基础上接受大柴胡汤治疗，48例）。对比两组中医证候积分、Th17/Treg免疫平衡、AQP1、AQP5、AQP6和实验室指标。结果：两组治疗7 d后两胁胀痛，矢气则舒，右中上腹痛，恶心呕吐，抑郁易怒，嗳气呃逆，大便不畅评分均下降，且研究组低于对照组（P<0.05）。两组治疗7 d后Treg细胞比例升高，Th17细胞比例、Th17/Treg下降，且研究组的改善幅度大于对照组（P<0.05）。两组治疗7 d后AQP1、AQP5、AQP6升高，且研究组高于对照组（P<0.05）。两组治疗7 d后淀粉酶（AMY）、脂肪酶（LIP）、C反应蛋白（CRP）下降，且研究组低于对照组（P<0.05）。结论：大柴胡汤治疗MAP疗效确切，可以缓解患者的临床症状，有效改善MAP患者Th17/Treg免疫平衡和血清AQP1、AQP5、AQP6水平。     

基于NRS营养风险评估的整蛋白型肠内营养对老年肝硬化患者营养状态、肠道菌群及外周血Th17/Treg平衡的影响

摘要 目的：探讨基于营养风险筛查（NRS）营养风险评估的整蛋白型肠内营养对老年肝硬化患者营养状态、肠道菌群及外周血辅助性T细胞17（Th17）/调节性T细胞（Treg）平衡的影响。方法：按照随机数字表法将2019年12月~2022年8月期间长沙市第一医院收治的103例老年肝硬化患者将患者分为对照组（常规饮食治疗,51例）和研究组（对照组基础上接受基于NRS营养风险评估的整蛋白型肠内营养治疗,52例）。比较两组NRS2002评分、营养状态指标、肠道菌群指标及外周血Th17/Treg平衡变化情况。结果：治疗3个月后,两组NRS2002评分下降,且研究组低于对照组（P<0.05）。治疗3个月后,两组血清总蛋白（TP）、白蛋白（ALB）及前白蛋白（PA）升高,且研究组高于对照组（P<0.05）。治疗3个月后,两组双歧杆菌、乳酸杆菌、肠球菌、肠杆菌升高,且研究组高于对照组（P<0.05）。治疗3个月后,两组酵母样真菌下降,且研究组低于对照组（P<0.05）。治疗3个月后,两组Treg升高,且研究组高于对照组（P<0.05）,治疗3个月后,两组Th17、Th17/Treg下降,且研究组低于对照组（P<0.05）。结论：老年肝硬化患者经基于NRS营养风险评估的整蛋白型肠内营养治疗,有利于降低营养不良发生风险,可能与改善患者的营养状态、肠道菌群及外周血Th17/Treg平衡有关。     

桥本甲状腺炎患者血清miR-326、miR-155与甲状腺相关抗体和Th17/Treg细胞因子失衡的相关性分析

摘要 目的：探讨桥本甲状腺炎（HT）患者血清微小核糖核酸（miR）-326、miR-155与甲状腺相关抗体和辅助性T细胞17（Th17）/调节性T细胞（Treg）细胞因子失衡的相关性。方法：选择2021年1月至2022年10月空军军医大学唐都医院收治的HT患者82例作为研究组，同期接受体检的健康人80例作为对照组，比较两组血清miR-326、miR-155、甲状腺功能参数[促甲状腺激素（TSH）、游离三碘甲状原氨酸（FT₃）、游离甲状腺素（FT₄）]、甲状腺相关抗体[甲状腺过氧化物酶抗体（TPOAb）、甲状腺球蛋白抗体（TGAb）]、Th17、Treg水平及其细胞因子水平，并分析HT患者血清miR-326、miR-155与甲状腺相关抗体和Th17/Treg细胞因子失衡的相关性。结果：研究组TSH、TPOAb、TGAb显著高于对照组，FT₃、FT₄显著低于对照组（P<0.05）。研究组血清miR-326、miR-155表达水平显著高于对照组（P<0.05）。研究组Th17、Th17/Treg、白细胞介素-17（IL-17）、γ干扰素（IFN-γ）显著高于对照组，Treg、白细胞介素-10（IL-10）水平显著低于对照组（P<0.05）。Pearson相关分析显示，HT患者血清miR-326、miR-155分别与TSH、TPOAb、TGAb、Th17、Th17/Treg、IL-17、IFN-γ呈正相关，与FT₃、FT₄、Treg、IL-10呈负相关（P<0.05）。结论：HT患者血清miR-326、miR-155水平异常升高，其水平与TPOAb、TGAb自身抗体及Th17/Treg失衡相关，血清miR-326、miR-155可能通过影响Th17/Treg免疫平衡，促进HT的发生和发展。     

沙参麦冬汤联合硫酸羟氯喹治疗气阴两虚型原发性干燥综合征的疗效及对Th17/Treg免疫失衡的影响

摘要 目的：观察沙参麦冬汤联合硫酸羟氯喹治疗气阴两虚型原发性干燥综合征（pSS）的疗效及对辅助性T细胞（Th17）/调节性T细胞（Treg）免疫失衡的影响。方法：选择2020年3月~2021年5月期间我院收治的pSS患者80例作为研究对象,采用随机数字表法进行分组,将患者分为对照组和研究组,各为40例。对照组接受硫酸羟氯喹治疗,研究组接受沙参麦冬汤联合硫酸羟氯喹治疗。观察两组疗效、中医证候积分、症状相关指标及Th17/Treg免疫失衡指标,评价两组用药安全性。结果：研究组的临床总有效率较对照组高（P<0.05）。治疗后,两组口干咽燥、两目干涩、唇舌紫暗、关节疼痛、皮肤干涩证候积分下降,且研究组低于对照组（P<0.05）。治疗后,两组红细胞沉降率（ESR）、欧洲抗风湿病联盟干燥综合征疾病活动指数（ESSDAI）评分、免疫球蛋白G（IgG）、类风湿因子（RF）、欧洲抗风湿病联盟干燥综合征患者报告指数（ESSPRI）评分下降,且研究组低于对照组（P<0.05）,基础泪液分泌Ⅰ试验（SIt）、唾液流率升高,且研究组高于对照组（P<0.05）。治疗后,两组白介素-17（IL-17）、Th17下降,白介素-10（IL-10）、Treg升高（P<0.05）,研究组治疗后IL-17、Th17低于对照组,IL-10、Treg高于对照组（P<0.05）。两组不良反应发生率对比无显著性差异（P>0.05）。结论：沙参麦冬汤联合硫酸羟氯喹治疗气阴两虚型pSS疗效显著,可有效改善患者临床症状,阻止病情进展,调节机体Th17/Treg免疫失衡,具有一定的临床应用价值。     

不明原因复发性流产再次妊娠患者血清1,25(OH)2D3、sTim-3与Th17/Treg免疫失衡和妊娠结局的关系

摘要 目的：探讨不明原因复发性流产（URSA）再次妊娠患者血清1,25-二羟维生素D3[1,25（OH)）₂D₃]、可溶性T细胞免疫球蛋白黏蛋白分子3（sTim-3）与辅助性T细胞17（Th17）/调节性T细胞（Treg）免疫失衡和妊娠结局的关系。方法：选择于湖南省妇幼保健院2020年1月~2022年1月就诊的62例URSA再次妊娠患者作为研究组,另选择同期进行孕检的正常早孕妇女30例作为对照组。比较两组孕早期血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平、Th17/Treg比值。Pearson法分析URSA再次妊娠患者血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值平的相关性。根据URSA再次妊娠患者妊娠结局的不同分为妊娠成功分娩组和妊娠再次流产组,比较两组孕早期血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值。受试者工作特征（ROC）曲线分析血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值对妊娠结局的预测价值。结果：研究组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于对照组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于对照组（P<0.05）。Pearson相关分析显示,URSA再次妊娠患者血清1,25(OH) ₂D₃与血清sTim-3、外周血Th17细胞水平、Th17/Treg比值呈负相关,与Treg细胞水平呈正相关（P<0.05）;血清sTim-3与外周血Treg细胞水平呈负相关,与Th17细胞水平、Th17/Treg比值呈正相关（P<0.05）。妊娠再次流产组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于妊娠成功分娩组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于妊娠成功分娩组（P<0.05）。ROC曲线分析显示,血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平及Th17/Treg比值均可预测URSA再次妊娠患者妊娠再次流产的发生风险,且上述指标联合检测的预测效能更高。结论：血清1,25(OH) ₂D₃水平异常降低、sTim-3水平异常升高可导致Th17/Treg免疫失衡,导致URSA再次妊娠患者再次发生流产。上述指标联合检测对URSA再次妊娠患者妊娠再次流产的预测效能更高。     

火针联合外用5%米诺地尔酊对肝肾不足型斑秃患者外周血Th17细胞、Treg细胞和心理状态的影响

摘要 目的：观察火针联合外用5%米诺地尔酊对肝肾不足型斑秃患者外周血辅助性T细胞17（Th17）、调节性T细胞（Treg）和心理状态的影响。方法：选取2019年3月~2021年4月期间我院收治的102例肝肾不足型斑秃患者,按随机数字表法分为对照组和研究组,各为51例。对照组患者接受5%米诺地尔酊外用治疗,研究组患者接受火针联合外用5%米诺地尔酊治疗,治疗3个月后,观察两组疗效,对比两组外周血Th17细胞、Treg细胞占CD4⁺T淋巴细胞比例及相关细胞因子水平、心理状态、斑块面积的变化,记录两组毳毛长出时间、毳毛变黑时间以及治疗期间不良反应情况。结果：与对照组相比,研究组的临床总有效率明显更高（P<0.05）。两组治疗后Th17细胞比例、白介素-6（IL-6）、白介素-23（IL-23）水平较治疗前下降,Treg细胞比例较治疗前升高,且研究组变化程度大于对照组（P<0.05）。两组治疗后斑块面积均缩小,且研究组小于对照组（P<0.05）;研究组的毳毛长出时间、毳毛变黑时间均短于对照组（P<0.05）。两组治疗后焦虑自评量表（SAS）、抑郁自评量表（SDS）评分较治疗前下降,且研究组低于对照组（P<0.05）。两组不良反应发生率比较无统计学差异（P<0.05）。结论：火针联合外用5%米诺地尔酊治疗肝肾不足型斑秃,可有效改善患者临床症状,调节外周血Th17细胞、Treg细胞比例及相关细胞因子水平,有效改善患者焦虑抑郁情绪,安全可靠。     

枯草杆菌二联活菌肠溶胶囊联合泮托拉唑对UC患者炎性因子、肠黏膜功能及外周血Th17、CD4+CD25+Treg细胞表达的影响

摘要 目的：观察枯草杆菌二联活菌肠溶胶囊联合泮托拉唑对溃疡性结肠炎（UC）患者炎性因子、肠黏膜功能及外周血Th17、CD4⁺CD25⁺Treg细胞表达的影响。方法：研究对象选择2014年7月~2018年9月期间来我院香山路门诊部接受诊治的80例UC患者,随机分为联合组（枯草杆菌二联活菌肠溶胶囊联合泮托拉唑治疗）、对照组（泮托拉唑治疗）,各40例。对比两组的疗效、炎性因子、肠黏膜功能及外周血中Th17及CD4⁺CD25⁺Treg细胞表达 。记录两组治疗期间不良反应发生情况。结果：联合组的临床总有效率为92.50%（37/40）,对照组为70.00%（28/40）,两组比较差异有统计学意义（P<0.05）。两组不良反应发生率对比无差异（P>0.05）。联合组治疗6个月后血清白介素-8（IL-8）、肿瘤坏死因子-α（TNF-α）、C反应蛋白（CRP）水平均明显比对照组低（P<0.05）。联合组治疗6个月后血清D-乳酸含量、二胺氧化酶（DAO）水平均明显比对照组低（P<0.05）。联合组治疗6个月后外周血中Th17细胞表达比对照组低,CD4⁺CD25⁺Treg细胞表达比对照组高（P<0.05）。结论：枯草杆菌二联活菌肠溶胶囊联合泮托拉唑治疗UC患者,可有效改善肠道环境,使外周血中的 Th17 细胞表达降低,CD4⁺CD25⁺Treg细胞表达增加,并缓解炎症状态,临床效果满意且安全性好。     

疏肝健脾活血方对溃疡性结肠炎患者肠粘膜屏障功能、氧化应激反应及Th1/Th2细胞因子的影响

摘要 目的：探讨疏肝健脾活血方对溃疡性结肠炎（UC）患者肠粘膜屏障功能、氧化应激反应及辅助性T细胞亚群（Th1/Th2）细胞因子的影响。方法：选取2020年4月~2022年3月期间我院收治的UC患者80例,按照随机数字表法,将80例UC患者分为对照组（n=40,美沙拉嗪肠溶片治疗）和研究组（n=40,美沙拉嗪肠溶片联合疏肝健脾活血方治疗）,观察两组肠粘膜屏障功能、疗效、氧化应激反应相关指标、中医证候积分、Th1/Th2细胞因子的变化情况。结果：研究组的临床总有效率高于对照组（P<0.05）。两组治疗2个月后痛泻、胸胁胀闷、神疲懒言、嗳气评分均下降,且研究组低于对照组（P<0.05）。两组治疗2个月后二胺氧化酶（DAO）、D-乳酸、内毒素均下降,且研究组低于对照组（P<0.05）。研究组治疗2个月后超氧化物歧化酶（SOD）高于对照组,丙二醛（MDA）低于对照组（P<0.05）。研究组治疗2个月后白介素-4（IL-4）高于对照组,干扰素-γ（INF-γ）、INF-γ/IL-4低于对照组（P<0.05）。两组不良反应发生率组间对比无明显差异（P>0.05）。结论：疏肝健脾活血方辅助治疗UC患者,可调节Th1/Th2免疫平衡,改善机体氧化应激反应及肠粘膜屏障功能,促进临床症状改善。     

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.     

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.     

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。     

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.     

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.     

Biological monitoring of bisphenol A with HLPC/FLD and LC/MS/MS assays

Biological monitoring is a necessary process for risk assessment of endocrine disrupting chemicals (EDCs), particularly, bisphenol A (BPA), in breast milk, because its human risks are not clear yet, and infants, who feed on breast milk, are highly susceptible for EDCs. Concerning biological monitoring of BPA, the HPLC/FLD has been widely used before the LC/MS/MS. However, there was no report, which simultaneously evaluated the two methods in real analyses. Therefore, we analyzed BPA with LC/MS/MS and HPLC/FLD in human breast milk and conducted comparison of two methods in analyzed BPA levels. After establishing optimal condition, e.g. linearity, recovery, reproducibility and free BPA system, we analyzed BPA levels in human breast milk samples (N = 100). The LOQs were similar in the two methods, i.e. 1.8 and 1.3 ng/mL for the HPLC/FLD and LC/MS/MS assays, respectively. There were strong associations between total BPA levels with the two methods (R² = 0.40, p < 0.01), however, only 11% of them were analyzed as similar levels with 15% CVs. In addition, the detection range of BPA was broader in the HPLC method than the LC/MS/MS method. However, the BPA levels in the HPLC/FLD analysis were lower than those in the LC/MS/MS analysis (p < 0.01). Thus, the differences in BPA levels between the two methods may come from mainly over-estimation with the LC/MS/MS method in low BPA samples and some of poor resolution with the HPLC/FLD in high BPA samples.