Development and application of a loop-mediated isothermal amplification method on rapid detection of Pseudomonas aeruginosa strains

We developed and evaluated the specificity and sensitivity of a simple loop-mediated isothermal amplification (LAMP) method for rapid detection of P. aeruginosa strains. The optimal reaction condition was found to be 65°C for 45 min, with the detection limit as 100 fg DNA/tube and 10 CFU/reaction. Application of LAMP assays were performed 426 clinical samples (including 252 P. aeruginosa and 174 non- P. aeruginosa isolates) using a rapid procedure and easy result confirmation. Sensitivity of LAMP and PCR assays was found to be 97.6% (246/252) and 90.5% (228/252), respectively; with a 100% specificity for both assays.     

海洋细菌Agarivorans albus NBRC102603琼胶酶的分离纯化

利用盐析、分子筛和离子交换等方法对海洋细菌Agarivorans albus NBRC102603分泌的琼胶酶粗酶液进行分离纯化,得到琼胶酶A和琼胶酶B.琼胶酶A纯化倍数为17.51倍,酶比活力为881.82 U/mg;琼胶酶B纯化倍数为16.64倍,酶比活力为838.32 U/mg.纯化的琼胶酶经SDS-PAGE检测,显示为单一条带,其相对分子质量分别为酶A 8.36×104和酶B 3.68×104.     

Chromogenic media for MRSA diagnostics

For the past decade, a number of chromogenic media for methicillin-resistance Staphylococcus aureus (MRSA) detection have been developed and applied, including Oxoid Brilliance? MRSA, CHROMagar? MRSA, BBL? CHROMagar? MRSA, MRSASelect and chromID MRSA. The advantages of these chromogenic media offers direct detection of visible staphylococcal colonies, coupled with the use of chromogenic enzymatic substrates that can be hydrolyzed by S. aureus to confirm species or strain identification. BBL? CHROMagar? MRSA and MRSASelect are designed for detection of nasal colonization by MRSA, while CHROMagar? MRSA, Oxoid Brilliance? MRSA and chromID MRSA are readily applied in bacterial screening. This review summarizes the characteristics, principles and capacities of these selective media, and focuses on comparison of different chromogenic media.     

Validation of Respiratory Inductance Plethysmography for Measuring Tidal Volume in Swine

Measuring tidal volume (V_T) in nonintubated swine or swine with leaking breathing circuits is challenging. The aim of this study was to validate respiratory inductance plethysmography (RIP) for measuring V_T in swine that are comparable in size to adult humans. To determine calibration curves, V_T and RIP readings were obtained from anesthetized swine (n = 8; weight, 46–50 kg) during positive-pressure (mechanical) ventilation and spontaneous breathing. For positive-pressure ventilation, 6 pigs were mechanically ventilated by using the pressure-control mode. The 2 pigs in the spontaneously breathing cohort each received a single intravenous bolus dose of propofol to abolish spontaneous breathing; V_T was measured during gradual return of their respiratory drive. A flow–volume sensor was placed between the proximal end of the endotracheal tube and breathing circuit for the recording of inspiratory and expiratory V_T. RIP readings were recorded by using 2 bands, which simultaneously measured ribcage and abdominal excursions. The data revealed that V_T was linearly correlated with the movements of both ribcage and abdomen as measured by using plethysmography over a large range of tidal volume (44 to 1065 mL). In addition, the intercept of the linear equation was small or even negative during spontaneous breathing but increased significantly (maximum, 145 mL, 59.2 ± 35.1 mL) during positive pressure ventilation. Our results indicate that V_T in swine can be calculated by using a simple univariate linear regression equation with RIP readings obtained during either mechanical ventilation or spontaneous breathing.Abbreviations: RIP, respiratory inductance plethysmography; V_T, tidal volumeIn animal care and research, the respiratory function of swine is widely assessed by measuring tidal volume (V_T).⁶ Although V_T is easy to measure in intubated pigs, it is difficult to measure in swine that are either nonintubated or have significant breathing circuit leakage^16,24. Using facemasks attached to minimally restrained swine is currently the method of choice for measuring V_T. This methodology has multiple limitations, including incomplete sealing between the pig''s face and the mask, which is difficult to adapt to these animals. A reliable, minimally invasive method for determining V_T in this species is needed.Respiratory inductance plethysmography (RIP)^11,15 is a viable alternative to measuring V_T. Using RIP, both ribcage and abdominal movements can be recorded. The amplitude changes of the ribcage and abdomen are linearly correlated to the size of the corresponding V_T in humans. This method has been validated in humans^1,13 and is widely used clinically in adults,^14,18 newborns and infants.^5,8 RIP has also been used on animals including sheep⁴ and dogs.¹⁷However, RIP has not yet been validated for use in swine that are comparable in size to adult humans. Considering the profound physiologic and anatomic differences in the respiratory system between swine and humans,^12,20 the validation of plethysmography for use in humans might not extrapolate readily to swine, which are one of the most commonly used animal models in biomedical research. Here we sought to validate the RIP method for calculating V_T in swine during both positive-pressure ventilation and spontaneous breathing.     

水稻上部节间长度等数量性状基因的定位及其遗传效应分析

对水稻籼粳杂交（窄叶青８号×京系１７）Ｆ＿１的花药进行离体培养,建立一个含１３３个ＤＨ系的作图群体,通过构建分子连锁图谱,对水稻上部节间长度、株高和抽穗期的ＱＴＬ进行区间作图,定位了影响上部节间长度的１２个ＱＴＬ、株高的４个ＱＴＬ和抽穗期的１个ＱＴＬ。对这些ＱＴＬ的遗传效应分析的结果表明,控制抽穗期的１个ＱＴＬ即Ｈｄ８ａ为主效基因,其余的１６个ＱＴＬ为微效基因。控制上部节间长度单个的ＱＴＬ对表型的贡献率介于８－１８％,其加性效应可使所控制的节间长度增加大约１．６－３．６ｃｍ。值得注意的是,一些控制相关性状的、作用方向相同的ＱＴＬ定位于同一染色体的相同或邻近区段上。这一结果揭示了这些性状相关的遗传基础,在水稻育种中运用这些ＱＴＬ将有助于对株高进行精细的遗传调控。     

Determination of phenformin hydrochloride using molecular imprinting technology coupled with flow-injection chemiluminescence

A novel method was developed using molecular imprinting technology (MIT) coupled with flow‐injection chemiluminescence (FI‐CL) for highly sensitive detection of phenformin hydrochloride (PH). The phenformin imprinted polymer was synthesized with methacrylic acid (MAA) as a functional monomer and ethylene glycol dimethacrylate (EGDMA) as a cross‐linker. Newly synthesized molecularly imprinted polymer (MIP) particles were packed into a column as a selective recognition element for determination of PH. A CL method for the determination of PH was developed based on the CL reaction of PH with N‐bromosuccinimide sensitized by eosin Y in basic media. The optimization of detection conditions was investigated. The CL intensity responded linearly to the concentration of PH in the range 0.09–2.0 µg/mL, with a correlation coefficient of 0.9920. The detection limit was 0.031 µg/mL. The relative standard deviation for the determination of 1.0 µg/mL PH solution was 1.0% (n = 11). The method was applied to the determination of PH in urine samples, with satisfactory results. Copyright © 2011 John Wiley & Sons, Ltd.     

莱州湾鱼类群落优势种生态位

根据2011年5月、8月、10月和12月莱州湾底拖网调查资料,对该海域鱼类群落优势种的生态位进行了研究。结果表明,全年调查共捕获鱼类49种,各季节优势种种类数分别为春季3种,夏季5种,秋季3种,冬季4种。夏、冬季优势种的丛生指数较低,春、秋季较高;冬、春季优势种的平均拥挤度较低,夏、秋季较高。基于欧氏距离的优势种丰度聚类结果同优势度排序结果吻合。主成分分析(PCA)表明,青鳞小沙丁鱼(Sardinella zunasi)和赤鼻棱鳀(Thryssa kammalensis)分别是影响第一轴和第二轴的主要种类。绯鱼衔(Callionymus beniteguri)、鲬(Platycephalus indicus)、髭缟虾虎鱼(Tridentiger barbatus)和短吻红舌鳎(Cynoglossus joyneri)是时空二维生态位宽度最高的种类(2)。时空生态位显著重叠(0.6)的种类有7组,其中银姑鱼(Pennahia argentata)和皮氏叫姑鱼(Johnius belengerii)重叠值最高(0.798)。δ~(13)C值变幅(CR)最大的种类为斑鱼祭(Konosirus punctatus),δ~(15)N值变幅(NR)最大的种类为青鳞小沙丁鱼;青鳞小沙丁鱼和鱼祭生态位总面积超过20,皮氏叫姑鱼生态位总面积最小(1.38)且与其他优势种营养生态位重叠较高。等级聚类、排序、PCA和优势种排序结果较一致,而与时空生态位宽度分析结果差异较大,表明莱州湾鱼类群落结构受洄游鱼类的影响较大。时空生态位宽度较高的种类主要为集群特征不明显的周年定居种(绯鱼衔、鲬、短吻红舌鳎等底层鱼类),而季节洄游种(青鳞小沙丁鱼、鱼祭、赤鼻棱鳀等中上层鱼类)因时间生态位宽度较低导致时空生态位宽度较低。时空生态位和营养生态位分析表明,生态位重叠导致的资源利用性竞争并不是导致莱州湾鱼类群落结构现状的决定性因素,而更多可能是人为干扰形成的。     

莱州湾春季鱼卵仔稚鱼群落结构和物种多样性

于2007—2010年春季(5月)利用大型浮游生物网对莱州湾海域位置相同的12个站位进行4个航次拖网调查,研究分析了该海域鱼卵仔稚鱼群落组成、优势种、空间分布及其多样性特征。结果表明:4个航次共获得鱼卵、仔稚鱼26种,隶属于6目16科24属,1种衔科鱼卵鉴定到科,另有1种鱼卵未能鉴定种类。鱼卵、仔稚鱼分布不均匀,莱州湾西部还湾底海域分布较多。斑鰶(Konosiruspunctatus)为该海域鱼卵的第一优势种;鳀(Engraulisjaponicus)在2007年和2008年作为仔稚鱼的第一优势种出现,而2009年和2010年仔稚鱼的第一优势种则是虾虎鱼。调查期间该海域鱼卵、仔稚鱼的均匀度指数和生物多样性指数变化趋势一致,均表现为从2007年开始随年代递减的趋势;种类数和丰富度指数则从2008年开始呈现随年代递减的趋势。2007—2010年4a鱼卵仔稚鱼聚类结果显示,2007年和2008年鱼卵仔稚鱼群落结构相似,与2009年和2010年均有差异。与1982年山东省近岸调查结果相比,莱州湾海域鱼卵仔稚鱼群落结构发生明显变化,斑鰶取代鳀成为第一优势种,小黄鱼等经济鱼类数量逐渐降低。