Isolation and phylogenetic analysis of arcobacter spp. in ground chicken meat and environmental water in Japan and Thailand

Prevalence of Arcobacter spp. in chicken meat samples and environmental water samples in Japan and Thailand was investigated. Arcobacter was isolated from 48% of chicken meat samples (20/41) and 23% of river water samples (4/17) from Japan, and 100% of chicken meat samples (10/10) and 100% of canal water samples (7/7) from Thailand. A. butzleri was among the species isolated from all positive samples. About 10% genetic diversity was seen in the rpoB-rpoC in Arcobacters, and phylogenetic trees were divided into two clusters. In both countries, the results suggested that chicken and environmental water were highly contaminated with a genetically diverse population of Arcobacter.     

Determination of WR-1065 in human blood by high-performance liquid chromatography following fluorescent derivatization by a maleimide reagent ThioGlo3

In order to improve the sensitivity and stability of human blood samples containing WR-1065 (i.e., active metabolite of the cytoprotective agent amifostine), a high-performance liquid chromatographic method was developed and validated using fluorescent derivatization with ThioGlo3. Using a sample volume of only 100 microl, the method was specific, sensitive (limit of quantitation=10 nM in deproteinized blood or 20 nM in whole blood), accurate (error < or = 3.2%) and reproducible (CV < or = 8.7%). In addition, the stability of WR-1065 in deproteinized and derivatized blood samples was assured for at least four weeks at -20 degrees C. This method should be particularly valuable in translating the kinetic-dynamic relationship of WR-1065 in preclinical models to that in cancer patients.     

On-column preconcentration of glutathione and glutathione disulfide using pH-mediated base stacking for the analysis of microdialysis samples by capillary electrophoresis

Capillary electrophoresis (CE) has become a useful analytical tool for the analysis of microdialysis samples. However, CE with UV detection (CE-UV) does not provide detection limits sufficient to quantify glutathione (GSH) and glutathione disulfide (GSSG) in biological samples such as liver microdialysates, because of the small optical path length in the capillary. To overcome this limitation, an on-column preconcentration technique, pH-mediated base stacking, was used in this study to improve the sensitivity of CE-UV. This stacking technique allowed large volumes of high ionic strength sample injection without deterioration of the separation efficiency and resolution. A 26-fold increase in sensitivity was achieved for both GSH and GSSG using the pH-mediated base stacking, relative to normal injection without stacking. The limit of detection for GSH and GSSG was found to be 0.75 microM (S/N=6) and 0.25 microM (S/N=6), respectively. The developed method was used to analyze GSH and GSSG in liver microdialysates of anesthetized Sprague Dawley male rats. The basal concentrations of GSH and GSSG in the liver microdialysates of male rats were found to be 4.73+/-2.08 microM (n=7) and 5.52+/-3.66 microM (n=7), respectively.     

Solid-phase extraction or liquid chromatography coupled on-line with gas chromatography in the analysis of biological samples

This review provides an overview of the on-line coupling of solid-phase extraction or liquid chromatography with gas chromatography for the analysis of biological samples. Principles relevant to techniques are briefly presented and selected applications are described. Benefits of the coupled systems are discussed.     

Re-caching of acorns by rodents: Cache management in eastern deciduous forests of North America

Scatter-hoarding rodents such as tree squirrels selectively cache seeds for subsequent use in widely-spaced caches placed below the ground surface. This behavior has important implications for seed dispersal, seedling establishment, and tree regeneration. Hoarders manage these caches by recovering and eating some seeds, and moving and re-caching others. This process of re-caching, however, is poorly understood. Here, we use radio-telemetry to evaluate re-caching behavior for the management of acorn caches by rodents in eastern deciduous forests. We also test the hypothesis that as seeds are re-cached, the distance from the source increases. Radio transmitters were implanted in Northern red oak (Quercus rubra) acorns and presented to rodents in a natural setting over 3 seasons. We used radio-telemetry to track and document evidence of recovery and re-caching. We tracked a total of 102 acorns. Of the 39 radio-tagged acorns initially cached, 19 (49%) were cached on two or more occasions; one acorn was cached four times. The hypothesis that rodents move seeds to progressively greater distances from the source is not well-supported, suggesting that acorns are being moved within an individual's home range. Given the species of rodents in the study area, gray squirrels (Sciurus carolinensis) are the most likely to be responsible for the caching and re-caching events. Gray squirrels appear to engage in extensive re-caching during periods of long-term food storage, which has important implications for understanding how caching behavior influences acorn dispersal and oak regeneration.     

Effective mosquito and arbovirus surveillance using metabarcoding

Effective vector and arbovirus surveillance requires timely and accurate screening techniques that can be easily upscaled. Next‐generation sequencing (NGS) is a high‐throughput technology that has the potential to modernize vector surveillance. When combined with DNA barcoding, it is termed ‘metabarcoding.’ The aim of our study was to establish a metabarcoding protocol to characterize pools of mosquitoes and screen them for virus. Pools contained 100 morphologically identified individuals, including one Ross River virus (RRV) infected mosquito, with three species present at different proportions: 1, 5, 94%. Nucleic acid extracted from both crude homogenate and supernatant was used to amplify a 269‐bp section of the mitochondrial cytochrome c oxidase subunit I (COI) locus. Additionally, a 67‐bp region of the RRV E2 gene was amplified from synthesized cDNA to screen for RRV. Amplicon sequencing was performed using an Illumina MiSeq, and bioinformatic analysis was performed using a DNA barcode database of Victorian mosquitoes. Metabarcoding successfully detected all mosquito species and RRV in every positive sample tested. The limits of species detection were also examined by screening a pool of 1000 individuals, successfully identifying the species and RRV from a single mosquito. The primers used for amplification, number of PCR cycles and total number of individuals present all have effects on the quantification of species in mixed bulk samples. Based on the results, a number of recommendations for future metabarcoding studies are presented. Overall, metabarcoding shows great promise for providing a new alternative approach to screening large insect surveillance trap catches.     

超声乳化术与小切口囊外摘除术治疗白内障患者的疗效及对角膜内皮细胞及生存质量的影响

目的:探讨超声乳化术与小切口囊外摘除术治疗白内障患者的疗效及对角膜内皮细胞及生存质量的影响。方法:选取2015年3月到2017年6月在我院接受治疗的白内障患者203例,根据随机数字表法将患者分为对照组100例和观察组103例。对照组患者给予小切口囊外摘除术进行治疗,观察组给予超声乳化术进行治疗。于术前、术后1周、术后1个月、术后3个月对所有患者的视力进行检测。于术前、术后2周检测患者的眼角膜内皮细胞数量,并计算细胞密度和六角形细胞比率。于术前和术后1个月采用生活质量评定量表对患者的生活质量进行评测,记录患者术后出现的并发症。结果:术前、术后1个月、术后3个月,两组患者的视力比较差异无统计学意义(P0.05),术后1周观察组患者的视力明显高于对照组(P0.05);术后2周两组患者的细胞密度、六角形细胞比率均降低,对照组的细胞密度低于观察组(P0.05)。术前、术后1个月两组患者的自理能力、活动能力、社交能力、心理情绪得分比较差异无统计学意义(P0.05),术后1个月两组患者的自理能力、活动能力、社交能力、心理情绪得分均高于术前(P0.05)。两组患者的并发症发生率比较差异无统计学意义(P0.05)。结论:与小切口囊外摘除术相比,超声乳化术能在短期内更快的提升患者视力,且对患者的眼角膜内皮细胞损伤较小,但两种手术近期疗效和并发症情况基本一致,且对患者生活质量的影响无差异。     

经膝下途径导管直接溶栓治疗混合型下肢深静脉血栓形成

目的:评价经膝下(Below the Knee, BTK)途径导管直接溶栓(Catheter-Directed Thrombolysis, CDT)治疗混合型下肢深静脉血栓形成(Deep Vein Thrombosis, DVT)的安全性和有效性。方法:回顾性分析2013年1月至2017年10月于徐州市肿瘤医院介入科接受经BTK途径CDT治疗的38例急性混合型DVT患者临床资料,其中男性患者26例,女性患者12例,年龄54±11岁。结果:技术成功率为100%。经小隐静脉穿刺途径28例,小隐静脉切开途径4例,胫后静脉切开途径4例,胫后静脉穿刺途径2例。行CDT前,所有患者均接受临时可回收滤器置入,并于术后2周内取出。36名患者DVT血栓溶解成功(Ⅱ和Ⅲ级)。2名未获得血栓溶解成功的患者,从症状发生到CDT时间均大于10天。对13例患者(左侧10例,右侧3例)进行髂静脉球囊扩张和支架植入。围手术期出现切口渗血4例,切开部位麻木5例,没有新发肺栓塞和大出血发等主要并发症。平均随访时间为2年(1个月-4年),28例患者超过1.5年。2例患者由于未服用华法林而出现反复髂股DVT。随访期间通畅率和PTS率分别为81.6%(31/38)和31.6%(12/38)。结论:采用经BTK途径行CDT治疗混合型DVT是安全有效的,围手术期及随访期间结果满意,是一种值得推广的方法。     

Small molecule modulators of PCSK9 – A literature and patent overview

Proprotein convertase subtilisin kexin like type 9 (PCSK9) has since its discovery been a key protein target for the modulation of LDL cholesterol. The interest in PCSK9 has grown even more with the positive clinical trial outcomes in cardiovascular disease recently reported for two PCSK9 antibodies. Currently, there are no PCSK9 small molecule programs active in clinical development. However, there has been a steady increase in publications and patent applications within the PCSK9 small molecule field. This digest will provide a summary of small molecule and peptide PCSK9 modulators reported both in scientific journals and in patent applications, most of them originating from the last 3–4?years. As such, this digest will serve as an introduction to the field and assist further identification and discovery of small molecule PCSK9 modulators.