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1.
Topical antimicrobial therapy represents an essential part of burn wound care. In order to prevent and treat burn wound infection dressings with antimicrobial properties are applied directly on the wound surface. Not only the infection control but also promotion of healing is very important in burn wound management. It is well known, that a dressing in bactericidal concentration might also delay wound healing. This study was aimed to evaluate the potential toxic effect of topical antimicrobial agents on murine and human dermal cells. For toxicity testing the method by Vittekova et al. was used to evaluate potential toxic effects of 16 agents and 6 control samples on two in vitro cultured cell systems [3T3 cells and dermal fibroblasts] during the first 24 h. Following the 24 h cell culture with the tested agents the live cell counts were evaluated. According to results obtained on both cell systems, the tested samples were divided into three groups—nontoxic, semi-toxic and toxic. Nontoxic samples included Acetic acid 1%, Acticoat®, Dermacyn®, Framykoin®, Silverlon®, gauze, acellular human allodermis and acellular porcine xenodermis. Semi-toxic group included Algivon®Plus, Aquacel®Ag, Betadine®, Nitrofurazone, Octenisept®, Suprasorb® A and a porcine dermal scaffold Xeno-Impl. Finally, the toxic group included Algivon®, Dermazin®, Ialugen®Plus, Prontoderm®, Suprasorb® A Ag and 20% SDS. As the preliminary results of this study have shown, our findings may serve as a potential guide to selection of the most appropriate topical antimicrobial dressings for treatmet of burns. However before they can be translated into clinical practice recommendations, more research on antimicrobial dressings cytotoxicity testing will be necessary.  相似文献   

2.
The development of single nucleotide polymorphism (SNP) markers in maize offers the opportunity to utilize DNA markers in many new areas of population genetics, gene discovery, plant breeding and germplasm identification. However, the steps from sequencing and SNP discovery to SNP marker design and validation are lengthy and expensive. Access to a set of validated SNP markers is a significant advantage to maize researchers who wish to apply SNPs in scientific inquiry. We mined 1,088 loci sequenced across 60 public inbreds that have been used in maize breeding in North America and Europe. We then selected 640 SNPs using generalized marker design criteria that enable utilization with several SNP chemistries. While SNPs were found on average every 43 bases in 1,088 maize gene sequences, SNPs that were amenable to marker design were found on average every 623 bases; representing only 7% of the total SNPs discovered. We also describe the development of a 768 marker multiplex assay for use on the Illumina® BeadArray? platform. SNP markers were mapped on the IBM2 intermated B73 × Mo17 high resolution genetic map using either the IBM2 segregating population, or segregation in multiple parent-progeny triplets. A high degree of colinearity was found with the genetic nested association map. For each SNP presented we give information on map location, polymorphism rates in different heterotic groups and performance on the Illumina® platform.  相似文献   

3.
A simple but novel mixed surfactant system was designed to fabricate a self-nanoemulsifying drug delivery system (SNEDDS) based on hydrophilic–lipophilic balance (HLB) value. The impacts of HLB and molecular structure of surfactants on the formation of SNEDDS were investigated. After screening various oils and surfactants, nifedipine (NDP)-loaded liquid SNEDDS was formulated with Imwitor® 742 as oil and Tween®/Span® or Cremophor®/Span® as mixed surfactant. Droplet size of the emulsions obtained after dispersing SNEDDS containing Tween®/Span® in aqueous medium was independent of the HLB of a mixed surfactant. The use of the Cremophor®/Span® blend gave nanosized emulsion at higher HLB. The structure of the surfactant was found to influence the emulsion droplet size. Solid SNEDDS was then prepared by adsorbing NDP-loaded liquid SNEDDS comprising Cremophor® RH40/Span® 80 onto Aerosil® 200 or Aerosil® R972 as inert solid carrier. Solid SNEDDS formulations using higher amounts (30–50% w/w) of Aerosil® 200 exhibited good flow properties with smooth surface and preserved the self-emulsifying properties of liquid SNEDDS. Differential scanning calorimetry and X-ray diffraction studies of solid SNEDDS revealed the transformation of the crystalline structure of NDP due to its molecular dispersion state. In vitro dissolution study demonstrated higher dissolution of NDP from solid SNEDDS compared with NDP powder.  相似文献   

4.

Background

The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing TaqMan® and SYBR® Green real-time PCR chemistries. In our study four alternative chemistries: Lux?, Plexor?, Cycling Probe Technology and LNA® were extensively evaluated and compared using TaqMan® chemistry as a reference system.

Results

Amplicons were designed on the maize invertase gene and the 5'-junction of inserted transgene and plant genomic DNA in MON 810 event. Real-time assays were subsequently compared for their efficiency in PCR amplification, limits of detection and quantification, repeatability and accuracy to test the performance of the assays. Additionally, the specificity of established assays was checked on various transgenic and non-transgenic plant species. The overall applicability of the designed assays was evaluated, adding practicability and costs issues to the performance characteristics.

Conclusion

Although none of the chemistries significantly outperformed the others, there are certain characteristics that suggest that LNA® technology is an alternative to TaqMan® when designing assays for quantitative analysis. Because LNA® probes are much shorter they might be especially appropriate when high specificity is required and where the design of a common TaqMan® probe is difficult or even impossible due to sequence characteristics. Plexor? on the other hand might be a method of choice for qualitative analysis when sensitivity, low cost and simplicity of use prevail.
  相似文献   

5.
microRNA(miRNA)是一类广泛存在于真核生物中长度为20~24 nt的内源非编码小RNA,它们通过对靶基因mRNA进行切割或翻译抑制,在转录后水平调控靶基因的表达。近期研究表明,miRNA参与植物生长发育与逆境胁迫响应的多个重要生物学过程,对作物的农艺性状也起到重要的调控作用。玉米作为重要的粮食、饲料和工业原料,提高其产量和品质对于保障世界粮食安全至关重要,然而与模式植物拟南芥和水稻相比,玉米中miRNA的研究仍然相对较少,理解miRNA在玉米中的功能和调控机理有助于通过分子育种对关键农艺性状进行遗传改良。本文综述了玉米中miRNA的发现与鉴定,系统总结了参与玉米miRNA代谢途径的关键蛋白DCL、AGO和HEN1的研究进展,重点阐述了在玉米生长发育和非生物胁迫响应过程中已开展功能研究miRNA的调控作用,并对玉米miRNA研究当前存在的问题和未来的发展趋势进行了讨论。  相似文献   

6.

Background

An increased incidence of fungal infections caused by Candida species, especially Candida glabrata and Candida krusei, which are less susceptible to azoles, has been observed. Standardized susceptibility testing is essential for clinical management and for monitoring the epidemiology of resistance.

Aims

We evaluated the performance of two different susceptibility testing commercial methods, Vitek 2® and Sensititre YeastOne®, and compared them with the standard broth microdilution method (CLSI).

Methods

A total of 80 isolates of several Candida species (Candida albicans, Candida parapsilosis complex, Candida tropicalis, C. glabrata and C. krusei) were selected for this study.

Results

We analyzed the categorical agreement (CA) between the methods, stratifying the disagreements. The average CA between the methods was 96.3% for Vitek 2® and 84% for Sensititre YeastOne®. No very major errors were observed. Major errors and minor errors were found for all the isolates tested. With the azoles, both Vitek 2® and Sensititre YeastOne® had good and similar performance levels, except for C. tropicalis and C. krusei (Sensititre YeastOne® showed low CA, 56.2%). With the echinocandins, both methods showed good performance for C. albicans, C. parapsilosis and C. tropicalis. However, we observed important discrepancies for C. krusei with caspofungin: Vitek 2® had 100% CA while Sensititre YeastOne® had only 25%. With amphotericin B, both Vitek 2® and Sensititre YeastOne® had good performance with high CA.

Conclusions

Despite the limited isolates tested, we concluded that both methods have good performance and are reliable for antifungal susceptibility testing. However, caspofungin activity against C. krusei and C. glabrata should be interpreted carefully when using Sensititre YeastOne® because we observed a low CA.  相似文献   

7.
Reduced rates of lung cancer have been observed in several occupational groups exposed to high levels of organic dusts contaminated by endotoxin. The underlying anti-neoplastic mechanism of endotoxin may be an increased secretion of endogenous anti-neoplastic mediators and activation of the toll-like receptors (TLR). A detoxified endotoxin derivative, Monophosphoryl Lipid A (MPL®) is marketed in Europe since 1999 as part of the adjuvant systems in allergy vaccines for treatment of allergic rhino-conjunctivitis and allergic asthma. Over 200,000 patients have used them to date (nearly 70% in Germany). Since detailed exposure (MPL® dose and timing of administration) and individual data are potentially available, an observational follow-up study could be conducted in Germany to investigate the protective effect of MPL® against cancer, comparing cancer incidence in two groups of patients with allergic rhinitis: those treated with allergoids plus MPL® and those treated with a vaccine including the same allergoids but not MPL®. The protective effect of MPL® could be quantified in ever and never smokers. If this proposed observational study provides evidence of protective effects, MPL® could be immediately used as a chemo-preventive agent since it is already in use as adjuvant in human vaccines against cancer.  相似文献   

8.
Aims: The development and evaluation of a sensitive and specific TaqMan® real-time polymerase chain reaction (PCR) for the detection and identification of Pantoea stewartii on maize. Methods and Results: A TaqMan®-based real-time PCR assay targeting the cpsD gene enabling specific detection of P. stewartii in maize leaves and seeds was developed. Under optimal conditions, the selected primers and probe were specific for the detection of all 14 reference P. stewartii strains by real-time PCR. The 32 non-Panteoa and eight other Pantoea strains tested negative. The TaqMan® PCR assay detected 1 pg of purified DNA and 104P. stewartii colony forming units per millilitre (10 cells per reaction) in pure cultures consisting of 92·0% intact (viable) cells. Direct processing of leaf lesions and seeds by the real-time PCR detected 10 and 50 P. stewartii cells per reaction respectively. TaqMan® real-time PCR results were validated by dilution plating of macerates and PCR-based subcloning followed by DNA sequencing. Conclusions: The real-time PCR assay described is a rapid, reliable and more sensitive tool for the detection of P. stewartii. Significance and Impact of the study: This real-time PCR assay would avoid false-negative results and reduce the time required for certifying maize seed shipments.  相似文献   

9.
Analytical methods based on light microscopy, 90° light-scattering and surface plasmon resonance (SPR) allowed the characterization of aggregation that can occur when antibodies are mixed with human plasma. Light microscopy showed that aggregates formed when human plasma was mixed with 5% dextrose solutions of Herceptin® (trastuzumab) or Avastin® (bevacizumab) but not Remicade® (infliximab). The aggregates in the plasma-Herceptin®-5% dextrose solution were globular, size range 0.5–9 μm, with a mean diameter of 4 μm. The aggregates in the plasma-Avastin®-5% dextrose samples had a mean size of 2 μm. No aggregation was observed when 0.9% NaCl solutions of Herceptin®, Avastin® and Remicade® were mixed with human plasma. 90° light-scattering measurements showed that aggregates were still present 2.5 h after mixing Herceptin® or Avastin® with 5% dextrose-plasma solution. A SPR method was utilized to qualitatively describe the extent of interactions of surface-bound antibodies with undiluted human serum. Increased binding was observed in the case of Erbitux® (cetuximab), whereas no binding was measured for Humira® (adalimumab). The binding of sera components to 13 monoclonal antibodies was measured and correlated with known serum binding properties of the antibodies. The data presented in this paper provide analytical methods to study the intrinsic and buffer-dependent aggregation tendencies of therapeutic proteins when mixed with human plasma and serum.  相似文献   

10.
Barley (Hordeum vulgare L.) is one of the founder crops of agriculture, and today it is the fourth most important cereal grain worldwide. Barley is used as malt in brewing and distilling industry, as an additive for animal feed, and as a component of various food and bread for human consumption. Progress in stable genetic transformation of barley ensures a potential for improvement of its agronomic performance or use of barley in various biotechnological and industrial applications. Recently, barley grain has been successfully used in molecular farming as a promising bioreactor adapted for production of human therapeutic proteins or animal vaccines. In addition to development of reliable transformation technologies, an extensive amount of various barley genetic resources and tools such as sequence data, microarrays, genetic maps, and databases has been generated. Current status on barley transformation technologies including gene transfer techniques, targets, and progeny stabilization, recent trials for improvement of agricultural traits and performance of barley, especially in relation to increased biotic and abiotic stress tolerance, and potential use of barley grain as a protein production platform have been reviewed in this study. Overall, barley represents a promising tool for both agricultural and biotechnological transgenic approaches, and is considered an ancient but rediscovered crop as a model industrial platform for molecular farming.  相似文献   

11.
P. K. Wright, J. Marshall and M. Desai Comparison of SurePath ® and ThinPrep ® liquid‐based cervical cytology using positive predictive value, atypical predictive value and total predictive value as performance indicators Objective: Two liquid‐based cytology (LBC) systems are in widespread use in the UK: ThinPrep® and SurePath®. A number of studies have now compared LBC with conventional cytology in cervical screening. However, to date, we are aware of no studies that have compared ThinPrep® with SurePath® LBC. As the selection and use of specific diagnostic systems in a laboratory has significant clinical and economic implications, there is a clear need to compare directly existing LBC technology. The objective of this study was to compare ThinPrep® with SurePath® LBC in a single cytology laboratory using performance indicators. Methods: Data were collected for all cervical cytology samples processed at Manchester Cytology Centre over a 1‐year period. ThinPrep® LBC was compared with SurePath® LBC using positive predictive value (PPV), atypical predictive value (APV) and total predictive value (TPV), reflecting outcome of cervical intraepithelial neoplasia (CIN) grade 2 or worse for high‐grade dyskaryosis (PPV), low‐grade dyskaryosis or borderline (atypical) cytology (APV) and all (total) abnormal cytology (TPV). Results: 2287 (out of 56 467) (ThinPrep®) and 586 (out of 22 824) (SurePath®) samples showed borderline or worse cytology after exclusion criteria. PPV, APV and TPV were within acceptable ranges for both ThinPrep® and SurePath®. Conclusions: ThinPrep® and SurePath® were equivalent based on three performance indicators. We suggest that APV and TPV should be used as an adjunct to PPV and other methods of quality assurance for cervical screening.  相似文献   

12.
《Autophagy》2013,9(8):883-891
Autophagic flux can be measured by determining the declining abundance of autophagic substrates such as sequestosome 1 (SQSTM1, better known as p62), which is sequestered in autophagosomes upon its direct interaction with LC3. However, the total amount of p62 results from two opposed processes, namely its synthesis (which can be modulated by some cellular stressors including autophagy inducers) and its degradation. To avoid this problem, we generated a stable cell line expressing a chimeric protein composed by p62 and the HaloTag® protein, which serves as a receptor for fluorescent HaloTag® ligands. Upon labeling with HaloTag® ligands (which form covalent, near-to-undissociable bonds with the Halotag® receptor) and washing, the resulting fluorescent labeling is not influenced by de novo protein synthesis, therefore allowing for the specific monitoring of the fusion protein decline without any interference by protein synthesis. We demonstrate that a HaloTag®-p62 fusion protein stably expressed in suitable cell lines can be used to monitor autophagy by flow cytometry and automated fluorescence microscopy. We surmise that this system could be adapted to high-throughput applications.  相似文献   

13.
Chlorella ellipsoidea is a single-celled eukaryotic green microalgae with high nutritional value. Its value may be further increased if a simple, reliable and cost-effective transformation method for C. ellipsoidea can be developed. In this paper, we describe a novel transformation method for C. ellipsoidea . This system is based on treatment of C. ellipsoidea cells with cellulolytic enzymes to weaken their cell walls, making them become competent to take up foreign DNA. To demonstrate the usefulness and effectiveness of this method, we treated C. ellipsoidea cells with a cell wall-degrading enzyme, cellulase, followed by transformation with plasmid pSP-Ubi-GUS harbouring both the zeocin resistance gene and the beta-glucuronidase (GUS) reporter gene that serve as selective makers for transformation. Transformants were readily obtained on zeocin selection medium, reaching transformation efficiency of 2.25 × 103 transformants/μg of plasmid DNA. PCR analysis has also demonstrated the presence of the GUS reporter gene in the zeocin-resistant transformants. Histochemical assays further showed the expression of the GUS activity in both primary transformants and transformants after long-term growth (10 months) with antibiotic selection on and off. Availability of a simple and efficient transformation system for C. ellipsoidea will accelerate the exploration of this microalga for a broader range of biotechnological applications, including its use as a biologic factory for the production of high-value human therapeutic proteins.  相似文献   

14.
Aim: To determine the performance of the Ridascreen® SET Total kit, after sample extraction and concentration by dialysis, with regard to its use in official controls for staphylococcal enterotoxins under European Regulation (EC) No. 2073/2005 modified. This study was conducted on naturally contaminated cheese samples and compared with the results of the previously validated Vidas® SET2 kit. Methods and Results: The effectiveness of the Ridascreen® SET Total kit on naturally contaminated cheeses was compared to that of the Vidas® SET2 kit by applying the EN ISO 16140 standard. Sensitivity and specificity were also compared using spiked buffer solutions and cheese samples with SEA to SEE toxins. Conclusions: This study showed that the Ridascreen® SET Total kit is as effective as the Vidas® SET2 kit. Significance and Impact of the Study: The Ridascreen® SET Total kit was found to specifically detect SEA to SEE in cheeses. The Ridascreen® SET Total can therefore be used to check the staphylococcal enterotoxin content and ensure consumer protection.  相似文献   

15.
In this article, we explore the intellectual property (IP) landscape relevant to the production and commercialization of Carolight?, a transgenic multivitamin corn variety created on humanitarian grounds to address micronutrient deficiencies in low‐and‐middle‐income countries. The successful production of this variety requires IP rights risk management because there is a strong protection on inventions and processes via patent portfolios in both developing and industrialized countries. The IP framework is complex, and specialist patent lawyers are usually employed to perform such analysis, but the costs cannot always be met by small, publicly funded projects. We report an alternative strategy, a do‐it‐yourself patent analysis, to produce a review with limited legal value that can nevertheless lay the foundations for a subsequent more in‐depth professional freedom‐to‐operate opinion.  相似文献   

16.
The protective effect of a commercial preparation (STW 5, Iberogast®), containing the extracts of bitter candy tuft, lemon balm leaf, chamomile flower, caraway fruit, peppermint leaf, liquorice root, Angelica root, milk thistle fruit and greater celandine herb, against the development of gastric ulcers was previously reported in an earlier publication (Khayyal et al., 2001). All extracts produced a dose dependent anti-ulcerogenic effect associated with a reduced acid output, an increased mucin secretion, an increase in prostaglandin E2 release and a decrease in leukotrienes. The effect on pepsin content was not uniform and did not seem to bear a relationship with the anti-ulcerogenic activity. The best effects were observed with the combined formulation, STW 5. Furthermore, the effect of the latter in protecting against the development of rebound gastric acidity was examined experimentally in rats and compared with the effect of some commercial antacid preparations (Rennie®, Talcid® and Maaloxan®). A model of testing rebound acidity was developed by inducing a marginal increase in gastric acidity through the administration of indomethacin, in such a way that it could be easily neutralized, allowing any eventual secondary increase in acidity to be measured within a few hours of administration. In addition, the serum gastrin level was measured after drug treatment to establish any correlation between it and any rebound acidity. The results obtained demonstrated that STW 5 did not only lower the gastric acidity as effectively as the commercial antacid, but it was more effective in inhibiting the secondary hyperacidity. Moreover, STW 5 was capable of inhibiting the serum gastrin level in rats, an effect which ran parallel to its lowering effect on gastric acid production.  相似文献   

17.
To analyze the suitability of Gateway® vectors for transformation of chloroplasts, we converted a standard plastid transformation vector into a Gateway® destination vector containing the necessary recombination sites attR1 and attR2. Insertion of the green fluorescent protein (GFP) coding sequence with associated T7g10 ribosome binding site into this destination vector created the expression vector for transformation of tobacco chloroplasts with the biolistic method. Correct integration of the transgene into the plastid genome was verified by PCR and the homoplasmic nature of the transformed plants was confirmed by Southern Blot analysis. Expression of the GFP reporter protein was monitored by confocal laser scanning microscopy (CLSM) and quantification by western blot analysis showed a GFP accumulation level of 3 % total soluble protein (TSP). The presented results clearly demonstrate that the Gateway® recombination sites are compatible with all steps of plastid transformation, from generation of transplastomic plants to expression of GFP. This is the first report of a plastid transformation vector made by the Gateway® recombinant cloning technology, which proves the suitability of this system for use in chloroplasts.  相似文献   

18.
We describe the haemodynamic and pathological characteristics of four types of impervious arterial prostheses, two alloplastic (Milrathane® and Gore-Tex®), and two chemically processed bovine heterografts (Solcograft® and Solco P®). They were implanted in the thoracic aortae of dogs for durations of 24 hours, 48 hours, one weeks, two weeks, one month, three months and six months. Haemodynamic analyses showed no relation between the shear rate index, I·Y, and compliance, CD. The observed shear rates are 6.5 times lower than those likely to damage the endothelial cell layer. Macroscopic and microscopic observations of explanted grafts showed the presence of obstructive thrombi at the anastomoses of Mitrathane® grafts as early as one week. Gore-Tex® grafts develop in the area of anastomoses parietal-thrombi which reorganize and become covered with pseudo-endothelial cells. The bovine heterografts show a similar behaviour. However, whereas Solcograft® has an irregular thin wall, Solco P® had improved characteristics except in the graft implanted for three months which demonstrated some manufacturing weaknesses. Both types showed the development of anastomotic pannus covered with endothelial-like cells. All grafts, whether alloplastic or chemically processed, suffered from an absence of healing of the middle part of the prosthesis. The cause of this problem will be found in the analysis of the biochemical and enzymatic reactations between the material used and its physiological surrounding.  相似文献   

19.
Improving foreign protein accumulation is crucial for enhancing the commercial success of plant-based production systems since product yields have a major influence on process economics. Cereal grain evolved to store large amounts of proteins in tightly organized aggregates. In maize, γ-Zein is the major storage protein synthesized by the rough endoplasmic reticulum (ER) and stored in specialized organelles called protein bodies (PB). Zera® (γ-Zein ER-accumulating domain) is the N-terminal proline-rich domain of γ-zein that is sufficient to induce the assembly of PB formation. Fusion of the Zera® domain to proteins of interest results in assembly of dense PB-like, ER-derived organelles, containing high concentration of recombinant protein. Our main goal was to increase recombinant protein accumulation in plants in order to enhance the efficiency of orally-delivered plant-made vaccines. It is well known that oral vaccination requires substantially higher doses than parental formulations. As a part of a project to develop a plant-made plague vaccine, we expressed our model antigen, the Yersinia pestis F1-V antigen fusion protein, with and without a fused Zera® domain. We demonstrated that Zera®-F1-V protein accumulation was at least 3× higher than F1-V alone when expressed in three different host plant systems: Ncotiana benthamiana, Medicago sativa (alfalfa) and Nicotiana tabacum NT1 cells. We confirmed the feasibility of using Zera® technology to induce protein body formation in non-seed tissues. Zera® expression and accumulation did not affect plant development and growth. These results confirmed the potential exploitation of Zera® technology to substantially increase the accumulation of value-added proteins in plants.  相似文献   

20.
Silicates have the potential to induce disease resistance in plants. Induction of nematode resistance usually results from paralysis of nurse cell development or activation of the hypersensitivity response. This study aimed to evaluate the effects of silicon (Si) treatment on the penetration and development of Meloidogyne javanica in various crops. The experiment was set up in a randomized (3 × 4) + 1 factorial design, with 3 Si sources (Silifort®, Rocksil® and wollastonite), 4 crops (maize, rice, common bean and soybean) and 1 treatment control (distilled water). The Si treatments included adding wollastonite to the soil 10 days prior to seedling transplantation, or spraying with solutions of Silifort® or Rocksil®, 2 days after seedlings transplantation. Twelve days after transplantation, the plants were inoculated with 1000 eggs and eventual second‐stage juveniles (J2) of M. javanica. At 3, 8, 13 and 18 days after inoculation (DAI), the plants were harvested and nematode penetration evaluated by optical microscopy. All Si treatments adversely affected development of M. javanica in soybean, common bean and rice and reduced nematode penetration of rice roots. Silifort® and wollastonite reduced nematode penetration in common bean and soybean roots, respectively. However, none of the Si treatments influenced the variables analysed in maize. The results of this study illustrate the potential of Si treatment to control M. javanica parasitism in plants.  相似文献   

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