The role of motility in the in vitro attachment of Pseudomonas putida PaW8 to wheat roots

The attachment of motile and non-motile strains of Pseudomonas putida PaW8 to sterile wheat roots was assessed in both non-competitive and intra-specific competitive assays. The motile strain showed significantly greater attachment to wheat roots than non-motile strains in phosphate buffer. Overall, the motile strain attached better than the non-motile strain at 10(6), 10(7) and 10(8) cfu ml(-1) in competitive assays and at 10(6) and 10(7) cfu ml(-1) in non-competitive assays. When attachment was studied in Luria broth no significant difference between motile and non-motile strains was detected. P. putida PaW8 cells marked with the luxAB genes were used to compare direct detection of attached cells by luminometry with indirect detection by dilution plate counts following extraction from root material. Although direct detection permitted a rapid assessment (60 s) of attachment to surfaces, dilution plate counts provided a more sensitive method for quantification of bacteria. The detection limits were approximately 10 cfu root(-1) using dilution plate counts compared with 1000 cfu root(-1) using luminometry. All results highlighted the importance of motility for the attachment of P. putida to plant roots in simple model systems. To take this work further, studies to assess the role of motility using complex non-sterile systems are needed.     

Efficacy,cost, and aspects to take into account in the treatment of osteoporosis in the elderly

Age is one of the principal risk factors for development of frailty fractures. Age pyramids show a population that is becoming increasingly more elderly, with an increasing incidence of fractures, and the forecasts for the future are truly alarming. Adequate handling of these patients who are especially at risk, at both the preventive and care levels, with a well-defined orthogeriatric model is necessary to respond to this clinical challenge. The objective of this review is to analyze the efficacy of the different strategies for the handling of geriatric patients with fracture risk.     

Development of a Cell-Based Assay to Assess Binding of the proNGF Prodomain to Sortilin

Sortilin was first identified based on its activity as part of intracellular protein sorting machinery. Recently, it was discovered that sortilin also acts as a cell surface receptor for the propeptide form of nerve growth factor (proNGF), progranulin, and neurotensin. The interaction of sortilin to these neurotrophic ligands is linked to diseases of the nervous system that lead to neurodegeneration and neuropathic pain. Blocking of the interaction of sortilin to these ligands may prevent or slow the progress of these nervous system disorders. In vitro screening assays for blocking compounds or peptides are part of the standard set of tools for drug discovery. However, assays for sortilin biology are not readily available to determine if the selected blocking agent inhibits sortilin activity on the surface of cells. We have developed a sortilin specific cell based assay to identify compounds that specifically block interaction between sortilin and proNGF prodomain. The assay system records both the presence of sortilin on the cell surface and the interaction with the pro domain of NGF. Fluorescent images of the sortilin expressing cells are analyzed for the presence of pro domain of NGF. Sortilin-positive and sortilin-negative cells within one well are concomitantly and automatically analyzed. Sortilin—pro domain interaction can be blocked dose dependently by neurotensin and synthetic compounds. The assay will facilitate the discovery of entities interfering with the binding of sortilin to the NGF pro domain. This assay can be modified to screen for inhibitors of the binding of ligands to other complex cell surface receptors.     

The Efficacy of a Live Attenuated TW I-Type Infectious Bronchitis Virus Vaccine Candidate

Virologica Sinica - Infectious bronchitis (IB) is a highly contagious avian disease caused by infection with infectious bronchitis virus (IBV), which seriously affects the development of the global...     

An Insight into the Active Site of Pseudomonas Fluorecens (P. cepacia) Lipase to Define the Stereochemical Demand for the Transesterification in Organic Solvents

The Pseudomonas fluorescens lipase-catalyzed transesterification of 2-methyl alkanols 1 and the 2-substituted oxiranemethanols 2 with vinyl acetate in organic solvents has been studied and the results discussed in terms of steric and electronic demand within the recently postulated models of the lipase active site.     

The Selective Action of 2, 4 -D on the Weed Flora of a Lawn

Increase in recreational demand on the countryside makes the monitoring of resulting change in vegetation of value, especially in areas of recognized ecological importance. Methods are described for an initial survey of trampling pressure and soil compaction, and their effects on both the soil environment and the plant community structure. With reference to a dune slack habitat the response of the vegetation to trampling levels is assessed by linear regression analysis. Variation in diversity is noted together with changes in the ratio between monocotyledons and dicotyledons. Occurrence indices may be used to identify the species resistant to trampling; these, in general, are found to adopt a hemicryptophytic or therophytic growth form. Such studies can make a useful contribution to field investigations for older secondary school and undergraduate students, and provide starting points for further detailed investigations.     

pEG plasmid involved in styrene degradation: molecular dimorphism and integration of a segment into the chromosome

In the Pseudomonas fluorescens strain ST the ability to utilize styrene as the sole carbon source is due to the presence of a plasmid, pEG. In the present report we show that pEG contains two inverted repeat sequences and we present evidence indicating that the catabolic genes are localized in these repeats. The region separating the inverted repeats can assume alternative orientations. In the chromosome of strain ST, a 3 kbp region is homologous to sequences present at one end of the plasmid repeats. This region is present in one copy in the chromosome and could be a specific site for integration of the plasmid. We suggest that this sequence, which is present twice in the pEG plasmid and once in the chromosome, might be a transposon-like element.     

Efficacy of a Comfrey root extract ointment in comparison to a Diclo-fenac gel in the treatment of ankle distortions: Results of an observer-blind, randomized, multicenter study

In the treatment of minor blunt injuries several topical drugs are known to have anti-inflammatory and analgesic properties. They represent, however, two fundamentally different major pharmacological therapy approaches: the “chemical-synthetical” and the “phytotherapeutical” approach. The main objective of this trial (CODEC_2004) was to compare the efficacy and tolerability of an ointment of Comfrey extract (Extr. Rad. Symphyti) with that of a Diclofenac gel in the treatment of acute unilateral ankle sprain (distortion). In a single-blind, controlled, randomized, parallel-group, multicenter and confirmatory clinical trial outpatients with acute unilateral ankle sprains (n=164, mean age 29.0 years, 47.6% female) received either a 6 cm long ointment layer of Kytta-Salbe^® f (Comfrey extract) (n=82) or of Diclofenac gel containing 1.16 g of diclofenac diethylamine salt (n=82) for 7±1 days, four times a day.

Primary variable was the area-under-the-curve (AUC) of the pain reaction to pressure on the injured area measured by a calibrated caliper (tonometer). Secondary variables were the circumference of the joint (swelling, figure-of-eight method), the individual spontaneous pain sensation at rest and at movement according to a Visual Analogue Scale (VAS), the judgment of impaired movements of the injured joint by the method of “neutral-zero”, consumption of rescue medication (paracetamol), as well as the global efficacy evaluation and the global assessment of tolerability (both by physician and patient, 4 ranks). In this study the primary variable was also to be validated prospectively.

It was confirmatorily shown that Comfrey extract is non-inferior to diclofenac. The 95% confidence interval for the AUC (Comfrey extract minus Diclofenac gel) was 19.01–103.09 h*N/cm² and was completely above the margin of non-inferiority. Moreover, the results of the primary and secondary variables indicate that Comfrey extract may be superior to Diclofenac gel.     

Development of a Loop‐Mediated Isothermal Amplification Assay to Detect Fusarium oxysporum

We report the development of a loop‐mediated isothermal amplification (LAMP) assay targeting the CYP51C element for visual detection of F. oxysporum which caused Fusarium wilt in soybean. The CYP51C‐LAMP assay efficiently amplified the target gene in 60 min at 62°C. And specificity was evaluated against F. oxysporum, Fusarium spp. and other fungal species. The detection limit of the CYP51C‐specific LAMP assay for F. oxysporum was four conidia per gram soil. The assay also detected F. oxysporum from inoculated soybean tissues and residues. These results suggest that this CYP51C‐LAMP assay can be used to detect residues on plants in the field.     

依达拉奉联合银杏达莫治疗2型糖尿病合并急性脑梗塞的临床效果分析

目的:本研究旨在探究2型糖尿病合并急性脑梗死的临床疗效。方法:对我市医院门诊及住院部在2014年1月至2015年9月间收录的358例2型糖尿病合并急性脑梗塞患者的治疗情况进行,依照患者治疗方法的差异,将本组患者分成两组,分别是对照组和观察组,对照组实施常规糖尿病及脑梗塞治疗,观察组在其基础上加用依达拉奉联合银杏达莫治疗,持续治疗3周后,评估治疗效果。结果:治疗三周后,两组患者的椎基底动脉血流速度均有明显改善(P0.05),观察组的改善程度较对照组更为显著(P0.05);观察组患者的神经功能缺损评分为(10.96±2.4)分,对照组为(16.21±3.1)分,则观察组患者神经功能显著由于对照组(P0.05);观察组患者临床治疗有效率为95.35%,对照组为72.03%,则观察组治疗有效率显著高于对照组,且具有统计学意义(P0.05);对照组的平均住院时间为(4.2±1.3)周,观察组的平均住院时间为(2.8±0.9)周。结论:对于2型糖尿病合并急性脑梗塞患者,在临床综合治疗的基础上加用依达拉奉联合银杏达莫治疗效果显著,能够显著提升患者椎基底动脉的血流速度,改善神经功能缺损,提高临床治疗效果,具有较大推广应用优势及价值。     

Development of a PCR-based Assay for the Detection of Plasmodiophora brassicae in Soil

A single-tube nested polymerase chain reaction (STN PCR) method was developed for detecting the causal agent of clubroot disease, Plasmodiophora brassicae. Outer primer PBTZS-2 (5′-CCGAATTCGCGTCAGCGTGA-3′) to amplify a 1457 bp-fragment from P. brassicae DNA and nested primers, PBTZS-3 (5′-CCACGTCGATCACGTTGCAAT-3′) and PBTZS-4 (5′-GCTGGCGTTGATGTACTGGAA-TT-3′), to amplify a 398 bp-fragment internal of the 1457 bp-fragment were used for the STN PCR. The 398 bp-fragment was amplified from as little as 1 fg of P. brassicae DNA with the STN PCR. A protocol for extracting P. brassicae DNA directly from soil was developed. By using the protocol, DNA was extracted from artificially infested soil containing various numbers of P. brassicae resting spores and the resulting DNA was used as template for the STN PCR. As little as one resting spore of P. brassicae per g of soil was detectable with the STN PCR. The STN PCR was applied to naturally infested soil from 3 fields and one canal bed. The 398 bp-fragment was amplified from soil of 2 fields and the canal bed. To improve the detection of P. brassicae, the STN PCR products were subjected to second PCR amplification (double PCR) using the nested primers PBTZS-3 and PBTZS-4. The double PCR amplification generated a single 398 bp-DNA band which was visualized clearly on the agarose gel for all the 4 soil samples tested. A combination of the STN PCR and the double PCR appears a useful assay method for detecting P. brassicae resting spores in field soil.     

The Application of Adsorption Modeling and Fourier Transform Infrared Spectroscopy to the Comparison of Two Species of Plant Growth–Promoting Rhizobacteria as Biosorbents of Cadmium in Different pH Solutions

Batch experiments were performed to determine the cadmium absorption capacity of two plant growth–promoting rhizobacteria at different pH levels and in different cadmium concentrations. Comparison of the mean metal removal from two species of bacteria studied showed that Pseudomonas florescence is the superior species for removing cadmium at all cadmium concentrations. The maximum cadmium absorption by P. florescence and P. putida were at 5 mg/L of cadmium concentration in pH 6 and 7, respectively. The applicability of the Langmuir and Freundlich isotherm models was surveyed. Comparison of two isotherm parameters (Q _m and a) further confirmed that P. fluorescence was better at binding cadmium ions (52.6 and 7.7 mg/g, respectively). Adsorption reaction also was considered by Fourier transform infrared (FTIR) spectroscopy. The FTIR analysis implied that the principal functional sites in the bacterial cell walls were phosphoryl and hydroxyl, carboxyl, amide I, amide II, and amine groups.     

多价免疫原用绿脓杆菌CMCC(B)-10117株替换株的初选

从临床分离绿脓杆菌菌株中,筛选与CMCC(B)-10117株抗原高度相关的菌株PA02010,PA02015,PA02033,PA02037和PA02048进行抗原成分分析。在此基础上,选择了PA02048进行家兔抗原性试验和小鼠保护性试验。家兔抗原性试验证明,肌肉注射的效价为1：320～1：640;静脉注射产生抗体能力强于肌肉注射,效价为1：2560～1：5120。从动物保护性试验看,免疫动物均能很好耐受5个LD50的本菌攻击,在10个LD50的本菌攻击时,免疫动物的存活率能达到90％;而对照组动物没有一个存活。所有试验表明。PA02048菌株是替代CMCC(B)-10117菌株的较为理想的菌株。     

Development of a PCR Assay for the Molecular Detection of Phytophthora boehmeriae in Infected Cotton

Cotton blight, caused by the oomycete Phytophthora boehmeriae, is a serious disease of cotton in China. In wet weather conditions, P. boehmeriae is usually the primary pathogen, followed by many saprophytic fungi and pathogens such as Pythium spp., Fusarium spp., Rhizoctonia and others. As P. boehmeriae grows much slower than other pathogens, it is difficult to isolate and identify. A rapid and accurate method for its specific identification is necessary for the detection of blight in infected cotton tissue. The internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) from three isolates of P. boehmeriae were amplified using the polymerase chain reaction (PCR) with the universal primers DC6 and ITS4. PCR products were cloned and sequenced. The sequences were aligned with those published of 50 other Phytophthora species, and a region specific to P. boehmeriae was used to construct the specific PCR primers PB1 and PB2. Over 106 isolates of 14 Phytophthora species and at least 20 other fungal species were used to check the specificity of the primers. PCR amplification with primers PB1 and PB2 resulted in the amplification of a product of approximately 750 bp only from isolates of P. boehmeriae. Using primers PB1 and PB2, detection sensitivity was approximately 10 fg DNA/μl. In inoculated plant material, P. boehmeriae could be detected in tissue 1 day after inoculation, prior to the appearance of symptoms. The PB primer‐based PCR assay provides an accurate and sensitive method for detecting P. boehmeriae in cotton tissue.