Chiral assay methods for lifibrol and metabolites in plasma and the observation of unidirectional chiral inversion following administration of the enantiomers to dogs

Lifibrol, a new drug for the treatment of hypercholesterolemia, contains a stereogenic center bearing a secondary alcohol group. A normal-phase achiral–chiral HPLC separation of the enantiomers of lifibrol and two of its metabolites was developed and validated for quantitation in dog plasma. A silica and a Chiralcel OD-H column were operated in series and all six enantiomeric components and internal standard were directly separated. An initial solid-phase extraction (phenyl) clean-up step and a column-switching step to eliminate late-eluting compounds were also utilized. The solid-phase extraction step was automated using a robotic system. Assay development, validation, and application of the method to a bioavailability study of the racemate and enantiomers of lifibrol in dogs are described. The lower limit of quantitation was 0.0125 μg/ml for each enantiomer of lifibrol using 200 μl of dog plasma with UV detection (255 nm). In dog plasma following oral or intravenous administration of the racemate, the (R)/(S) ratio of the enantiomers of lifibrol was greater than one and increased with time. Following administration of the individual enantiomers, chiral inversion of the (S)-enantiomer but not the (R)-enantiomer was observed. © 1994 Wiley-Liss, Inc.     

Role of the ErbB-4 carboxyl terminus in gamma-secretase cleavage

The ErbB-4 receptor tyrosine kinase has a PDZ domain recognition motif at its carboxyl terminus. The first step in ErbB-4 proteolytic processing is a metalloprotease-dependent cleavage of the receptor ectodomain, which is not influenced by deletion of this motif. Metalloprotease cleavage of ErbB-4 produces a membrane-associated 80-kDa fragment that is a substrate for subsequent gamma-secretase cleavage, which releases the cytoplasmic domain from the membrane and allows nuclear translocation of this fragment. Deletion of the PDZ domain recognition motif does abrogate the gamma-secretase cleavage of ErbB-4. The wild-type 80-kDa ErbB-4 fragment forms an association complex with presenilin, thought to be the catalytic moiety of gamma-secretase activity. However, this association is significantly impaired by loss of the PDZ domain recognition motif from ErbB-4. Deletion of this ErbB-4 motif prevents the nuclear localization of the ErbB-4 cytoplasmic domain. Data also show that the basal cleavage of wild-type ErbB-4 by this proteolytic system can produce a sufficient level of ErbB-4 processing to negatively influence cell growth and that loss of the PDZ domain recognition motif abrogates this response.     

Conditional replication of a recombinant adenovirus studied using neomycin as a selective marker

An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.     

辽宁省辣椒疫病菌多态性及致病力分化研究初探

【目的】明确辽宁省辣椒疫病菌多态性及致病力分化与区域性关系。【方法】利用SRAP技术对辽宁省25个辣椒疫病菌菌株进行了PCR扩增及NTSYS-PC聚类分析,用灌根法进行致病力分化试验并对试验结果进行SPSS 11.5分层聚类分析。【结果】利用筛选出的27组引物对25个菌株进行扩增,得到578条条带,每对引物多态性比率在84%?100%之间,多态性丰富;供试菌株间遗传相似性较高,相似系数0.56?0.91,以相似系数0.68为阈值划分,25个菌株可聚为4组。试验菌株80%为中等致病力,聚类结果较为分散。【结论】供试菌株没有表现出明显的区域性特征,菌株致病力强弱分化区域特征性规律不明显。     

西藏东南部棘蛙属一新种记述(两栖纲：蛙科)

对产于西藏东南部察隅和墨脱的察隅棘蛙通过形态特征和数值分类作了进一步比较研究,结果认为：产于察隅的察隅棘蛙是一个有效种,而产于墨脱的标本应为一个新种,即墨脱棘蛙Paa medogensis sp.nov.。     

镇海棘螈早期个体发育研究

根据野外观察和室内实验 ,对镇海棘螈的早期个体发育过程进行了研究。结果显示 ,镇海棘螈早期发育过程与琉球棘螈的个体发育过程非常相似 ,它们的平衡枝均不发达且在前肢芽晚期迅速消失。温度和湿度对胚胎发育具有显著影响。室温下 ,从受精卵到孵化需 2 9d左右。孵化幼体体长呈配合对数曲线增长 ,其体全长 (S)与日龄 (T)的关系可表示为S =1 2 974 9+6 2 398lnT(Sig.F <0 0 1 )。水生生活阶段为 58～ 88d。室内幼体的水生发育时间和变态个体大小与野外的情况有较大的差异 ,可能与营养条件和自然条件下的持续干旱有关。变态前的外鳃萎缩期是镇海棘螈早期个体发育过程中的死亡高峰期。     

黑刺粉虱危害对黄金芽茶叶生化成分含量及抗氧化酶活力的影响

【目的】明确黑刺粉虱Aleurocanthus spiniferus为害对茶叶生化物质含量及抗氧化酶活性的影响,为精准、安全、高效、绿色防治黑刺粉虱,提高茶叶品质提供参考。【方法】测定黑刺粉虱为害不同级别下黄金芽芽下第3叶中生化成分(茶多酚、可溶性糖、游离氨基酸、咖啡碱、儿茶素)含量及3种抗氧化酶(SOD, POD和CAT)活性的变化。【结果】随着黑刺粉虱为害等级的提高,黄金芽芽下第3叶中茶多酚的含量显著性降低,且以为害级别为Ⅳ级时最低,为18.82%±0.21%,而游离氨基酸、咖啡碱、可溶性糖和儿茶素的含量在不同的为害级别间均无显著性变化。黄金芽芽下第3叶中POD和CAT活力随着黑刺粉虱为害等级的增加先显著升高后很快降低,均以为害等级Ⅰ时活性最高,分别为57.14±3.98和28.05±0.40 U/mg pro。SOD活力呈显著下降趋势,且以为害等级Ⅳ时SOD活力最低,为442.73±10.54 U/mg pro。【结论】黑刺粉虱为害对黄金芽茶叶中与抗性提高相关的生化物质咖啡碱、儿茶素含量均没有显著性影响,与抗逆补偿相关的游离氨基酸和可溶性糖含量也没有显著性变化,但茶多酚含量显著降低,3种抗氧化酶活性随为害等级升高呈现先升高后显著降低的趋势,说明黄金芽对黑刺粉虱的抵抗力较弱,需要加强黄金芽上黑刺粉虱发生的科学管理。     

A new species of the genus Amolops (Anura: Ranidae) from high-altitude Sichuan,southwestern China,with a discussion on the taxonomic status of Amolops kangtingensis

A new species of the genus Amolops Cope,1865 is described from Xinduqiao,Kangding,Sichuan.It was previously identified as Amolops kangtingensis,which is synonymized to Amolops mantzorum in this study.The new species,Amolops xinduqiao sp.nov.,is distinguished from all other congeners by the following combination of characters:(1) medium body size,adult males SVL 41.2-47.5 mm (n=15,average 43.9 mm),adult females SVL 48.5-56.6 mm (n=15,average 52.5 mm);(2) head length equal to width or slightly wider than long;(3) tympanum small,but distinct;(4) vomerine teeth in two tiny rows,separated by a space about one vomerine teeth row;(5) bony projections on lower jaw absent;(6) dorsolateral folds usually absent;(7) tarsal folds or glands on tarsus absent;(8) circummarginal groove on disc of finger Ⅰ absent;(9) tibiotarsal articulation reaching nostril or beyond;(10) webs of toe Ⅳ reaching to distal articulation,other toes fully webbed to disc;and (11) vocal sac absent in males.