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Sample records for amebocyte lysate test

  1. Validation of the limulus amebocyte lysate (LAL) test for routine PET radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Zijlstra, S.; Gerken, P.; Rechin, C.; Wortmann, R.; Notohamiprodjo, G. [Universitatsklinik der Ruhr-Universitat, Bad Oeynhausen (Germany). Herz-und Diabeteszentrum Nordrhein-Westfalen

    1997-01-01

    The kinetic turbidimetric limulus amebocyte lysate test was validated as a method for detecting endotoxins in short-lived radiopharmaceutical samples. Using this method, radiopharmaceuticals can be released for administration to humans after the test, without extensive loss of radioactivity. Inhibition or enhancement on the LAL results by the product samples were examined in more detail and eliminated. (Author).

  2. False-positive result in Limulus test caused by Limulus amebocyte lysate-reactive material in immunoglobulin products.

    OpenAIRE

    Ikemura, K; Ikegami, K.; Shimazu, T; T. Yoshioka; Sugimoto, T.

    1989-01-01

    Limulus amebocyte lysate (LAL)-reactive material other than endotoxin was detected in the plasma and urine of patients after intravenous immunoglobulin therapy. Thirty-seven vials of six different immunoglobulin products were analyzed for the LAL-reactive material by combined use of a conventional chromogenic Limulus test and a chromogenic endotoxin-specific test. The amount of LAL-reactive material in reconstituted immunoglobulin solutions ranged from a mean (standard deviation) of 10.2 (2.1...

  3. The effect of refrigeration and mixing on detection of endotoxin in parenteral drugs using the Limulus Amebocyte Lysate (LAL) test.

    Science.gov (United States)

    Guilfoyle, D E; Yager, J F; Carito, S L

    1989-01-01

    Prior to testing for the presence of bacterial endotoxin, parenteral products are handled and stored in a variety of ways. Two incidents, detected by the U.S. Food and Drug Administration, revealed that differences in product handling and storage may have played a role in causing analytical discrepancies in the testing of identical samples. The testing procedure was the USP Bacterial Endotoxin test using Limulus Amebocyte Lysate (LAL) reagent. Consequently, an evaluation was made at the two principal factors that contributed to the suspected analytical anomaly. The factors were sample storage and the degree of agitation prior to sample analysis. Additional variables such as bacterial growth medium and adsorption potential of endotoxin by rubber stoppers were also evaluated. It was found that neither the medium employed to grow the E. coli endotoxin nor the storage temperature of the spiked solutions were problematic. However, it was shown that 20-40% of the spiked endotoxin was lost due to non-agitation of solution in vials in which the solution was in contact with the rubber stoppers. A suggested remedy for this problem is to store intact product containers in an upright position and to establish a uniform mixing procedure prior to endotoxin assay.

  4. Detection of Bacterial Endotoxins in Propacetamol Hydrochloride for Injection by Tachypleus Amebocyte Lysate Test%鲎试剂法检测注射用盐酸丙帕他莫中细菌内毒素

    Institute of Scientific and Technical Information of China (English)

    罗丽萍; 郭成希; 黄砚青

    2011-01-01

    Objective: To establish a method for detection of bacterial endotoxins in propacetainol hydrochloride for injection. Method: The experiments were carried out according to the bacterial endotoxins test specified in appendix XI E,volume two,Chinese pharmacopoeia 2010 edition. Result: Samples with the concentration of 5.0 mg·ml-1 had inhibitive effect on the bacterial endntoxins test,however the interference was eliminated when the samples were diluted to the concentration of 2. 5 mg·ml-1 ,detected hy tarhypleus ameboeyte lysates with the sensitivity of 0. 25 EU·ml-1. Conclusion: Tachypleus amebocyte lysate test is feasible for detecting bacterial endotoxins in propacetamol hydrochloride for injection.%目的:考察鲎试剂方法检测注射用盐酸丙帕他莫中细菌内毒素的可行性.方法:参照《中国药典》2010年版(二部)附录ⅪE细菌内毒素检查法对注射用盐酸丙帕他莫进行细菌内毒素检测.结果:质量浓度为5.0 mg·ml-1的样品溶液对凝集反应有抑制作用,质量浓度稀释至2.5 mg·ml-1时,用灵敏度λ为0.25 EU· ml-的鲎试剂进行试验,干扰作用消失.结论:注射用盐酸丙帕他莫可用鲎试剂方法进行细菌内毒素检测.

  5. Interference of silica nanoparticles with the traditional Limulus amebocyte lysate gel clot assay.

    Science.gov (United States)

    Kucki, Melanie; Cavelius, Christian; Kraegeloh, Annette

    2014-04-01

    Endotoxin contaminations of engineered nanomaterials can be responsible for observed biological responses, especially for misleading results in in vitro test systems, as well as in vivo studies. Therefore, endotoxin testing of nanomaterials is necessary to benchmark their influence on cells. Here, we tested the traditional Limulus amebocyte lysate gel clot assay for the detection of endotoxins in nanoparticle suspensions with a focus on possible interference of the particles with the test system. We systematically investigated the effects of nanomaterials made of, or covered by, the same material. Different types of bare or PEGylated silica nanoparticles, as well as iron oxide-silica core shell nanoparticles, were tested. Detailed inhibition/enhancement controls revealed enhanced activity in the Limulus coagulation cascade for all particles with bare silica surface. In comparison, PEGylation led to a lower degree of enhancement. These results indicate that the protein-particle interactions are the basis for the observed inhibition and enhancement effects. The enhancement activity of a particle type was positively related to the calculated particle surface area. For most silica particles tested, a dilution of the sample within the maximum valid dilution was sufficient to overcome non-valid enhancement, enabling semi-quantification of the endotoxin contamination. PMID:23884096

  6. Endotoxin recovery using limulus amebocyte lysate (LAL) assay.

    Science.gov (United States)

    Bolden, Jay S; Warburton, Rob E; Phelan, Robert; Murphy, Marie; Smith, Kelly R; De Felippis, Michael R; Chen, Dayue

    2016-09-01

    A phenomenon initially reported by Chen and Vinther in 2013 [1], and now commonly referred to as low endotoxin recovery (LER), has prompted the Food and Drug Administration (FDA) to request specific data demonstrating the capability of the LAL BET method (i.e., USP ) to recover endotoxin from spiked samples over time. The results of these spike/hold recovery studies are expected to be included in the Biologics License Applications (BLA) for review by the Center for Drug Evaluation and Research (CDER) Hughes (2014) and Hughes et al. (2015) [2,3]. Such studies involve spiking a known amount of a surrogate endotoxin, such as purified lipopolysaccharide (LPS), into undiluted biological products and then testing at different time points to determine the recovery over time. We report here the experience and learning gained from conducting spike/hold recovery studies for a monoclonal antibody (Mab) product. Results from initial hold studies spiked with purified LPS showed rapid loss of endotoxin activity in the drug substance (DS) and significant batch-to-batch variation in the drug product (DP). After careful review and examination of the experimental details, it was determined that the study design and execution differed from the routine batch release USP BET method with regard to mixing time and sampling scheme. The hold study design was subsequently revised so that the mixing time and sampling were the same as the verified USP BET method used for routine batch release testing. The spike/hold recovery studies were repeated and the results demonstrated that LPS could be consistently recovered over time. These findings highlight the importance of carefully controlling sample preparation procedures in a spike/hold recovery study in order to demonstrate the suitability of using the LAL BET method for endotoxin detection. PMID:27470947

  7. Endotoxin detection--from limulus amebocyte lysate to recombinant factor C.

    Science.gov (United States)

    Ding, Jeak Ling; Ho, Bow

    2010-01-01

    Gram negative bacterial endotoxin is a biological pyrogen that causes fever when introduced intravenously. The endotoxin, also known as lipopolysaccharide (LPS), is found in the outer membrane of Gram-negative bacteria. During Gram-negative sepsis, endotoxin stimulates host macrophages to release inflammatory cytokines. However, excessive inflammation causes multiple organ failure and death. Endotoxins, which are ubiquitous pathogenic molecules, are a bane to the pharmaceutical industry and healthcare community. Thus early and sensitive detection of endotoxin is crucial to prevent endotoxaemia. The limulus amebocyte lysate (LAL) has been widely used for ~30 years for the detection of endotoxin in the quality assurance of injectable drugs and medical devices. The LAL constitutes a cascade of serine proteases which are triggered by trace levels of endotoxin, culminating in a gel clot at the end of the reaction. The Factor C, which normally exists as a zymogen, is the primer of this coagulation cascade. In vivo, Factor C is the perfect biosensor, which alerts the horseshoe crab of the presence of a Gram-negative invader. The hemostatic end-point entraps the invader, killing it and limiting further infection. However, as an in vitro endotoxin detection tool, variations in the sensitivity and specificity of LAL to endotoxin, and the dwindling supply of horseshoe crabs are posing increasing challenges to the biotechnology industry. This has necessitated the innovation of an alternative test for endotoxin. Thus, Factor C became the obvious, albeit tricky target for the recombinant technology effort. This chapter documents the backwater of mining the natural blood lysate of the endangered species to the monumental effort of genetic engineering, to produce recombinant Factor C (rFC). The rFC is a 132 kDa molecule, which was produced as a proenzyme inducible by the presence of trace levels of endotoxin. The rFC forms the basis of the "PyroGene" kit, which is a novel micro

  8. Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay[S

    OpenAIRE

    Pais De Barros, Jean-Paul; Gautier, Thomas; Sali, Wahib; Adrie, Christophe; Choubley, Hélène; Charron, Emilie; Lalande, Caroline; Le Guern, Naig; Deckert, Valérie; Monchi, Mehran; Quenot, Jean-Pierre; Lagrost, Laurent

    2015-01-01

    Quantitation of plasma lipopolysaccharides (LPSs) might be used to document Gram-negative bacterial infection. In the present work, LPS-derived 3-hydroxymyristate was extracted from plasma samples with an organic solvent, separated by reversed phase HPLC, and quantitated by MS/MS. This mass assay was combined with the limulus amebocyte lysate (LAL) bioassay to monitor neutralization of LPS activity in biological samples. The described HPLC/MS/MS method is a reliable, practical, accurate, and ...

  9. Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay.

    Science.gov (United States)

    Pais de Barros, Jean-Paul; Gautier, Thomas; Sali, Wahib; Adrie, Christophe; Choubley, Hélène; Charron, Emilie; Lalande, Caroline; Le Guern, Naig; Deckert, Valérie; Monchi, Mehran; Quenot, Jean-Pierre; Lagrost, Laurent

    2015-07-01

    Quantitation of plasma lipopolysaccharides (LPSs) might be used to document Gram-negative bacterial infection. In the present work, LPS-derived 3-hydroxymyristate was extracted from plasma samples with an organic solvent, separated by reversed phase HPLC, and quantitated by MS/MS. This mass assay was combined with the limulus amebocyte lysate (LAL) bioassay to monitor neutralization of LPS activity in biological samples. The described HPLC/MS/MS method is a reliable, practical, accurate, and sensitive tool to quantitate LPS. The combination of the LAL and HPLC/MS/MS analyses provided new evidence for the intrinsic capacity of plasma lipoproteins and phospholipid transfer protein to neutralize the activity of LPS. In a subset of patients with systemic inflammatory response syndrome, with documented infection but with a negative plasma LAL test, significant amounts of LPS were measured by the HPLC/MS/MS method. Patients with the highest plasma LPS concentration were more severely ill. HPLC/MS/MS is a relevant method to quantitate endotoxin in a sample, to assess the efficacy of LPS neutralization, and to evaluate the proinflammatory potential of LPS in vivo. PMID:26023073

  10. A contribution to the study of pyrogenic substances in radiopharmaceutical preparations. Comparison between methods using Rabbit and those using Limulus amebocytes lysate

    International Nuclear Information System (INIS)

    We have studied two methods for pyrogenic substances detection. We used: the hyperthermic action of these substances after injection in Rabbit, and the gelation reaction of a Limulus amebocytes lysate. To apply these two methods of pyrogenic substances detection to the radiopharmaceutical preparations, we have conceived and designed a material allowing their handling in compliance with the radioactive safety norms. We have compared the sensitivity, reliability and reproducibility of these methods, one based on gelation of Limulus amebocytes lysate in presence of endotoxins, the other on the hyperthermic action of these same endotoxins in the rabbit when injected intravenously or through the suboccipital route. The discussion of the results obtained, shows that the method using the Limulus amebocytes lysate is more sensitive, less expansive and less dangerous. This method particulary well adapted to the control of radiopharmaceutical preparations, brings an additional security to the patients for whom these products are destined

  11. Detection of Bacterial Endotoxins in Coenzyme Complex for Injection by Tachypleus Amebocyte Lysate Test%鲎试剂方法检测注射用复合辅酶中细菌内毒素

    Institute of Scientific and Technical Information of China (English)

    王宗春; 姜俊勇; 宋金春

    2011-01-01

    目的:建立注射用复合辅酶细菌内毒素检查法.方法:按照中国药典2010年版(二部)附XIE细菌内毒素检查法进行干扰试验.结果:用灵敏度γ为0.5EU/ml的鲎试剂进行实验,当注射用复合辅酶中辅酶I的浓度为0.8μg/ml时,供试品不干扰鲎试剂与细菌内毒素的凝集反应.结论:使用鲎试剂方法检查注射片用复合辅酶中细菌内毒素是可行的.%Objective:To study the clinical related factors,the distribution and drug-resistance of the pathogen in 2 type diabetic with pulmonary infection,and the supervision for rational use of drugs in clinical practice.Methods:211 in-patients were chosen in recent three years, which were accompanied with diabetes mellitus and pulmonary infection.A retrospective investigation including the related factors(age,curse of disease,symptom) and 124 cases cultivated pathogen was performed.K-B method was used to test the drug-resistant strain and do drug-Sensitivity test.Results: The incidence of diabetes mellitus complicated by pulmonary infection related closely patients, age and curse of disease (P<0.05).The former five bacteria in diabetic with Pulmonary infection were klebsiella pneumonia(23.4%),pseudomonas aeruginose(18.5%),escherichia coli(16.9%),staphyloccocus aureus rosenbach (13.7)and Candida albicans (9.7%).The former five sensitive antibiotics were imipenem,sulperazon,ceftazidime,amikacin,levofloxacin.The former three antibiotics which were sensitive to G+ cocci,were vancomycin,Rifampicin,minocycline. The sensitive antifungal were Itraconazole and Fluconazole. Conclusion: The result of sputum cultivation and drug sensitivity wre useful to clinic.It was important to choose antibiotic under the direction of it.

  12. Application of quartz tuning forks for detection of endotoxins and Gram-negative bacterial cells by monitoring of Limulus Amebocyte Lysate coagulation.

    Science.gov (United States)

    Chałupniak, Andrzej; Waszczuk, Karol; Hałubek-Głuchowska, Katarzyna; Piasecki, Tomasz; Gotszalk, Teodor; Rybka, Jacek

    2014-08-15

    Endotoxins, pyrogens of bacterial origin, are a significant threat in many areas of life. Currently, the test most commonly used for endotoxin level determination is LAL (Limulus Amebocyte Lysate) assay. This paper presents application of commercially available low-frequency piezoelectric tuning forks (QTFs) for endotoxin detection. Measurement of the decrease in the QTF oscillation amplitude provides information about the viscosity changes, occurring in the tested sample upon addition of LAL. That method was used to determine the concentrations of endotoxins and bacterial cells (E. coli O157:H19). The relevance of the obtained results was confirmed using a commercially available colorimetric LAL assay. The constructed system can detect bacterial endotoxins in the range of 0.001-5EU/ml and bacterial cells in the range of 10(2)-10(7)CFU/ml. The presented technique requires very simple sample preparation and the sensor response is obtained using compact, portable readout electronics. The single test cost is low compared to commercial endotoxin assays and other novel systems based on micromechanical sensors. PMID:24632139

  13. Detection of bacterial endotoxin in Xingnaojing injection with Tachypleus Amebocyte lysate%鲎试剂检查醒脑静注射液细菌内毒素的探讨

    Institute of Scientific and Technical Information of China (English)

    叶树林; 王晓蕾

    2011-01-01

    Objective To study the method for the detection of bacterial endotoxin in Xingnaojing injection with Tachypleus Amebocyte lysate(TAL). Methods Experiments were performed according to the bacterial endotoxin tests covered in Chinese Pharmacopeia 2005(part 2). Results Xingnaojing injection did not interfere with its gel reaction to the bacterial endotoxin when it was diluted 10-fold. Conclusion Bacterial endotoxin test can be used to detect bacterial endotoxin in Xingnaojing injection.%目的 探讨用鲎试剂检查醒脑静注射液细菌内毒素方法.方法 根据2005年版Ⅱ部收载的细菌内毒素检查法的要求进行实验.结果 醒脑静注射液经10倍稀释时不干扰鲎试剂与细菌内毒素的凝胶反应.结论 细菌内毒素检查法适用于检测醒脑静注射液的内毒素.

  14. Detection of microbial translocation in HIV and SIV infection using the Limulus amebocyte lysate assay is masked by serum and plasma.

    Directory of Open Access Journals (Sweden)

    Ashwin Balagopal

    Full Text Available OBJECTIVE: Microbial translocation (MT is thought to be a major contributor to the pathogenesis of HIV-related immune activation, and circulating lipopolysaccharide (LPS from gram-negative bacteria is the principle measurement of this process. However, related research has been impeded by inconsistent LPS test results. METHODS: Specimens were obtained from HIV-infected adults enrolled in the PEARLS study (ACTG A5175 and HIV-HCV co-infected participants enrolled in a study of liver disease staging using MRI elastography. Pig-tailed macaque specimens were obtained from SIV-infected and -uninfected animals. Samples were tested for LPS using the LAL assay with diazo-coupling modifications to improve sensitive detection. RESULTS: When exogenous LPS was added to macaque plasma, >25% inhibition of LPS detection was found in 10/10 (100% samples at 20% plasma concentration compared to control; in contrast 5/10 (50% samples at 2% plasma concentration (p = 0.07 and 0/10 (0% at 0.1% plasma concentration (p = 0.004 showed >25% inhibition of LPS detection. Similarly, when LPS was added to human serum, >25% inhibition of LPS detection was found in 5/12 (42% of samples at 2% serum concentration compared to control, while 0/12 (0% of samples in 0.1% serum showed >25% inhibition of LPS detection (p = 0.07. Likewise, LPS detection in human sera without exogenous LPS was improved by dilution: LPS was detected in 2/12 (17% human samples in 2% serum, ranging from 3,436-4,736 pg/mL, compared to 9/12 (75% samples in 0.1% serum, ranging from 123 pg/mL -60,131 pg/mL (p = 0.016. In a separate validation cohort of HIV-HCV co-infected participants sampled at two different times on the same day, LPS measured in 0.2% plasma and with diazo-coupling was closely correlated between the first and second samples (R = 0.66, p<0.05. CONCLUSIONS: Undiluted serum and plasma mask LPS detection. The extent of MT may be substantially underestimated.

  15. AN OVERVIEW OF LIMULUS AMOEBOCYTE LYSATE (LAL TEST

    Directory of Open Access Journals (Sweden)

    Khale Anubha

    2011-04-01

    Full Text Available Limulus Amoebocyte Lysate (LAL test is in vitro test to detect and measure bacterial endotoxins. It is the most critical quality control test required by FDA for all parenteral drugs in their final stages of formulation. Endotoxins are products of metabolism associated with outer membrane of gram negative bacteria especially Pseudomonas, E. coli and Serratia and are most significant source of pyrogens. They are ubiquitous and cause profound physiological changes when administered parenterally. Also they play an important role in occurrence and development of many diseases. Their detection is very important in manufacturing of parental drugs, biological products and medical devices. There is specification of endotoxin limit provided in monograph of each pharmacopoeia. The convential rabbit test has several disadvantages. LAL test is recently developed quantitative test has overcome rabbit test due to many reasons. The article will discuss about the basic principle, development, different methods, comparison with rabbit test and applications of the test.

  16. Detection of free endotoxin in cerebrospinal fluid by the Limulus lysate test.

    OpenAIRE

    Munford, R S; Hall, C L; Grimm, L.

    1984-01-01

    We used a rabbit model of Escherichia coli meningitis to study the basis for positive Limulus lysate tests in infected cerebrospinal fluid. The results indicated that positive Limulus tests are due to endotoxins in cerebrospinal fluid and not to leukocyte proteases or other possible activators of the Limulus clotting system. The results also suggest that bacteria-free endotoxin may be present in localized gram-negative bacterial infections.

  17. A comparative study of Mono Mac 6 cells, isolated mononuclear cells and Limulus amoebocyte lysate assay in pyrogen testing

    DEFF Research Database (Denmark)

    Moesby, Lise; Jensen, S; Hansen, E W;

    1999-01-01

    Pyrogen induced secretion of interleukin 6 (IL-6) in Mono Mac 6 (MM6) cells was measured. The ability of the MM6 cell culture to detect pyrogens was compared to the Limulus amoebocyte lysate (LAL) test and isolated mononuclear cells (MNC). The detection limit of MM6 for lipopolysaccharide (LPS...

  18. Automation of the Limulus amoebocyte lysate test by using the Abbott MS-2 microbiology system.

    OpenAIRE

    Jorgensen, J H; Alexander, G A

    1981-01-01

    A rapid, automated method for the performance of the Limulus amoebocyte lysate endotoxin assay has been developed by using the Abbott MS-2 Microbiology System. This instrument automatically determines sequential changes in the optical density of up to 176 samples at 1- or 5-min increments during a 1-h assay period. Graphic representation of optical density changes can be viewed on a cathode-ray tube or reproduced by using a hard-copy printer. Limulus amoebocyte lysate preparations that were o...

  19. In vitro test for pyrogenes in radiopharmaceuticals

    International Nuclear Information System (INIS)

    Procedure and results of determination of pyrogenic substances in radiopharmaceutical preparations by an in vitro method based on the reaction between bacterial endotoxine and Limulus Amebocyte Lysate are presented. The advantage of this method as compared to the test in experimental animals performed so far has also been analyzed and proved by the fact that it enables avoidance of introduction of radioactive materials in experimental animals and of radiation effects on the results obtained in efficiency studies. The in vitro method is a quick one and requires only small quantities of the radiopharmaceutical preparation to be examined. (author)

  20. Evaluation of the Limulus amoebocyte lysate test in conjunction with a gram negative bacterial plate count for detecting irradiation of chicken

    International Nuclear Information System (INIS)

    A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjunction with a Gram negative bacterial (GNB) plate count for detecting the irradiation of chicken is described. Preliminary studies demonstrated that chickens irradiated at an absorbed dose of 2.5 kGy could be differentiated from unirradiated birds by measuring levels of endotoxin and of numbers of GNB on chicken skin. Irradiated birds were found to have endotoxin levels similar to those found in unirradiated birds but significantly lower numbers of GNB. In a limited study the test was found to be applicable to birds from different processors. The effect of temperature abuse on the microbiological profile, and thus the efficacy of the test, was also investigated. After temperature abuse, the irradiated birds were identifiable at worst up to 3 days after irradiation treatment at the 2.5 kGy level and at best some 13 days after irradiation. Temperature abuse at 150C resulted in rapid recovery of surviving micro-organisms which made differentiation of irradiated and unirradiated birds using this test unreliable. The microbiological quality of the bird prior to irradiation treatment also affected the test as large numbers of GNB present on the bird prior to irradiation treatment resulted in larger numbers of survivors. In addition, monitoring the developing flora after irradiation treatment amd during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeast and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds. (author)

  1. Evaluation of the limulus amoebocyte lysate test in conjunction with a gram negative bacterial plate count for detecting irradiation of chicken

    Science.gov (United States)

    Scotter, Susan L.; Wood, Roger; McWeeny, David J.

    A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjuction with a Gram negative bacteria (GNB) plate count for detecting the irradiation of chicken is described. Preliminary studies demonstrated that chickens irradiated at an absorbed dose of 2.5 kGy could be differentiated from unirradiated birds by measuring levels of endotoxin and of numbers of GNB on chicken skin. Irradiated birds were found to have endotoxin levels similar to those found in unirradiated birds but significantly lower numbers of GNB. In a limited study the test was found to be applicable to birds from different processors. The effect of temperature abuse on the microbiological profile, and thus the efficacy of the test, was also investigated. After temperature abuse, the irradiated birds were identifiable at worst up to 3 days after irradiation treatment at the 2.5 kGy level and at best some 13 days after irradiation. Temperature abuse at 15°C resulted in rapid recovery of surviving micro-organisms which made differentiation of irradiated and unirradiated birds using this test unreliable. The microbiological quality of the bird prior to irradiation treatment also affected the test as large numbers of GNB present on the bird prior to irradiation treatment resulted in larger numbers of survivors. In addition, monitoring the developing flora after irradiation treatment and during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeasts and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds.

  2. Evaluation of the Limulus amoebocyte lysate test in conjunction with a gram negative bacterial plate count for detecting irradiation of chicken

    Energy Technology Data Exchange (ETDEWEB)

    Scotter, S.L.; Wood, R.; McWeeny, D.J. (Ministry of Agriculture, Fisheries and Food, Norwich (UK). Food Science Lab.)

    1990-01-01

    A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjunction with a Gram negative bacterial (GNB) plate count for detecting the irradiation of chicken is described. Preliminary studies demonstrated that chickens irradiated at an absorbed dose of 2.5 kGy could be differentiated from unirradiated birds by measuring levels of endotoxin and of numbers of GNB on chicken skin. Irradiated birds were found to have endotoxin levels similar to those found in unirradiated birds but significantly lower numbers of GNB. In a limited study the test was found to be applicable to birds from different processors. The effect of temperature abuse on the microbiological profile, and thus the efficacy of the test, was also investigated. After temperature abuse, the irradiated birds were identifiable at worst up to 3 days after irradiation treatment at the 2.5 kGy level and at best some 13 days after irradiation. Temperature abuse at 15{sup 0}C resulted in rapid recovery of surviving micro-organisms which made differentiation of irradiated and unirradiated birds using this test unreliable. The microbiological quality of the bird prior to irradiation treatment also affected the test as large numbers of GNB present on the bird prior to irradiation treatment resulted in larger numbers of survivors. In addition, monitoring the developing flora after irradiation treatment amd during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeast and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds. (author).

  3. Airborne Endotoxin from Indoor and Outdoor Environments:Effect of Sample Dilution on the Kinetic Limulus Amebocyte Lysate (LAL) Assay

    Science.gov (United States)

    Airborne endotoxin in occupational environments are a potential respiratory hazard to individuals. In this study, total and inhalable airborne endotoxin samples were collected via filtration from inside animal housing units and downwind from agricultural production sites and a wastewater treatment ...

  4. Comparison of the Limulus amoebocyte lysate test with plate counts and chemical analyses for assessment of the quality of lean fish.

    OpenAIRE

    Sullivan, J. D.; Ellis, P C; Lee, R. G.; Combs, W S; Watson, S. W.

    1983-01-01

    The quality of lean fish was assessed simply and rapidly with Limulus amoebocyte lysate. The endotoxin levels agreed with aerobic plate counts and chemical indices of spoilage. Correlation between level of endotoxin and level of total volatile bases was found to be highly significant (r = 0.8579; P less than 0.001).

  5. Short time bacterial endotoxins test for positron emission tomography by means of positively charged filters

    International Nuclear Information System (INIS)

    Positron emission tomography (PET) radiotracers have very short physical half-lives. It is hard to complete a bacterial endotoxins test prior to release from medical institutes. For endotoxin quantitative determination, limulus amebocyte lysate (LAL) reagent and kinetic-turbidimetry system were previously developed. We investigated the possibility of a short time test by means of positively charged filters. As a result of this study, the effects of positively charged filters on endotoxin removal were over 99.5% for [18F]FDG and [18F]NaF, which were contaminated with the indicated concentration of endotoxin. Combining this filter and the kinetic-turbidimetric method, it was possible to complete a bacterial endotoxins test in 5 min prior to the patient's administration. This test should be required prior to release for PET radiopharmaceutical quality control. It has been suggested that this combination is a good method for this purpose. (author)

  6. Influence of radiation on endotoxin test using the PTSTM for 18-FDG radiopharmaceutical

    Energy Technology Data Exchange (ETDEWEB)

    Santos-Oliveira, Ralph, E-mail: roliveira@ien.gov.br [Instituto de Engenharia Nuclear (IEN/CNEN-RJ), Rio de Janeiro, RJ (Brazil). Div. de Radiofarmacia

    2010-07-15

    F-18 FDG (2-[18-F] fluoro-2-deoxy-D-glucose) is the most frequently used radiopharmaceutical for PET and PET CT imaging exams. The FDA recently approved the use of the PTS{sup TM} (Portable Test System) as an alternative to the standard test proposed by the United States Pharmacopeia using the LAL (Limulus Amebocyte Lysates), that takes longer to perform (about 1h) than the PTS{sup TM} (15 min). Recent studies have demonstrated that radiation could interfere with the PTS{sup TM} test. In order to study the effects of radiation on the PTS{sup TM} test and/or equipment, 27 batches of F-18 FDG produced in the Nuclear Engineering Institute were analyzed. The results showed that no direct correlation with radiation was found in any of the cases. (author)

  7. Short time bacterial endotoxins test for positron emission tomography by means of positively charged filters

    Energy Technology Data Exchange (ETDEWEB)

    Nakazawa, Nobuhiro; Wakita, Kazuo [Nishijin Hospital, Kyoto (Japan); Mineura, Katsuyoshi [Kyoto Prefectural Univ. of Medicine (Japan)] (and others)

    2002-11-01

    Positron emission tomography (PET) radiotracers have very short physical half-lives. It is hard to complete a bacterial endotoxins test prior to release from medical institutes. For endotoxin quantitative determination, limulus amebocyte lysate (LAL) reagent and kinetic-turbidimetry system were previously developed. We investigated the possibility of a short time test by means of positively charged filters. As a result of this study, the effects of positively charged filters on endotoxin removal were over 99.5% for [{sup 18}F]FDG and [{sup 18}F]NaF, which were contaminated with the indicated concentration of endotoxin. Combining this filter and the kinetic-turbidimetric method, it was possible to complete a bacterial endotoxins test in 5 min prior to the patient's administration. This test should be required prior to release for PET radiopharmaceutical quality control. It has been suggested that this combination is a good method for this purpose. (author)

  8. Quantitative determination of endotoxin in human peripheral venous blood with limulus lysate test%偶氮显色鲎试验定量测定人体外周静脉血微量内毒素的研究

    Institute of Scientific and Technical Information of China (English)

    徐鹏远; 蒋朱明; 舒红; 刘跃武; 梁存河; 杨乃发

    2000-01-01

    Objective AIM to set up a high sencitive, good reproducible method to detect trace endotoxin and endotoxin in human peripheral venous blood. Methods The method of quantitative determination of endotoxin was established by using Limulus Lysate teat with chromogenie substrate, and the endotoxin levee of peripheral venous blood was determined from 50 healthy subjects and 51 patients. Results 1. Improved Limulus Lysate test with ehromogenic substrate could betect trace endotoxin (10~2Eu/ml) and the CV was 2.29% (n=5). 2. Endotoxin levels of healthy Chinese in peripheral venous blood were below 0. 033Eu/ml. 3. Endotoxin levels were higher in patients sith some major operations, Which were correlated with the severity of stress (R2=0. 665). Conclusions This method is an effective measure ment for the Gram-negative bactexiemio and studies of endotoxemia.%目的建立偶氮显色鲎试验并测定健康人和临床病人外周静脉血内毒素.方法利用偶氮显色法原理建立检测微量内毒素技术,测定50例健康成人及51例不同创伤病人外周静脉血的内毒素并对测定方法进行探讨.结果1.此法检测内毒素可达微量(10-2Eu/ml),自身变异系数为2.29%(n=5).2.健康成人外周静脉血内毒素正常值为<0.0330Eu/ml,与国际水准一致.3.病人外周静脉血内毒素水平与创伤程度有关,其血浓度随着创伤程度增加而增加,两者之间呈正相关,R2=0.667.结论本法灵敏度高,重复性好并可精确定量.

  9. Bacterial challenge of NISSHO ultrafilter ETF 609: results of in vitro testing.

    Science.gov (United States)

    Krautzig, S; Lonnemann, G; Shaldon, S; Koch, K M

    1996-07-01

    In hemodialysis, a certain degree of bacterial contamination on the dialysate side is a regular finding. Concern has been growing that this contamination may lead to a chronic inflammatory response in the patient. Ultrafiltration of dialysate can be used to reduce bacterial content and levels of cytokine-inducing substances upstream of the patient's dialyzer. The aim of this study was to test in vitro the rejection capacity of a polysulfone hollow-fiber ultrafilter (ETF 609, NISSHO Co., Osaka, Japan) challenged with bacterial filtrates derived from Pseudomonas aeruginosa PA103. Results showed a reduction of interleukin-1 beta-inducing activity (measured on peripheral blood mononuclear cells) from 5,035 +/- 394 pg/ml prefilter to nondetectable levels postfilter and endotoxin levels (limulus amebocyte lysate assay) of 4,167 +/- 1,079 versus 12 +/- 2 pg/ml, respectively. In conclusion, ultrafiltration of dialysate with the polysulfone ultrafilter ETF 609 leads to a potent reduction of cytokine-inducing activity.

  10. Rapid detection of significant bacteriuria by use of an automated Limulus amoebocyte lysate assay.

    OpenAIRE

    Jorgensen, J H; Alexander, G A

    1982-01-01

    Previous studies have demonstrated that significant gram-negative bacteriuria can be detected by using the Limulus amoebocyte lysate test. A series of 580 urine specimens were tested in parallel with the automated MS-2 (Abbott Laboratories) assay and with quantitative urine bacterial cultures. The overall ability of the MS-2 Limulus amoebocyte lysate test to correctly classify urine specimens as containing either greater than or equal to 10(5) organisms or less than 10(5) organisms per ml dur...

  11. Assessing endotoxins in equine-derived snake antivenoms: Comparison of the USP pyrogen test and the Limulus Amoebocyte Lysate assay (LAL).

    Science.gov (United States)

    Solano, Gabriela; Gómez, Aarón; León, Guillermo

    2015-10-01

    Snake antivenoms are parenterally administered; therefore, endotoxin content must be strictly controlled. Following international indications to calculate endotoxin limits, it was determined that antivenom doses between 20 mL and 120 mL should not exceed 17.5 Endotoxin Units per milliliter (EU/mL) and 2.9 EU/mL, respectively. The rabbit pyrogen test (RPT) has been used to evaluate endotoxin contamination in antivenoms, but some laboratories have recently implemented the LAL assay. We compared the capability of both tests to evaluate endotoxin contamination in antivenoms, and we found that both methods can detect all endotoxin concentrations in the range of the antivenom specifications. The acceptance criteria of RPT and LAL must be harmonized by calculating the endotoxin limit as the quotient of the threshold pyrogenic dose and the therapeutic dose and the dose administered to rabbits as the quotient of the threshold pyrogenic dose and the endotoxin limit. Since endotoxins from Gram-negative bacteria exert different pyrogenicity, if contamination occurred, antivenom batches that induce pyrogenic reactions may be found in spite of passing LAL specifications. Although LAL assay can be used to assess endotoxin content throughout the antivenom manufacturing process, we recommend that the release of final products be based on the results of both methods. PMID:26325294

  12. Study of endotoxin release by Salmonella thyphi by antibiotics in Chromogenic Limulus Amebocyte Lyssate Method

    Directory of Open Access Journals (Sweden)

    Shagari G

    1997-07-01

    Full Text Available It seems, rapid destruction of gram negative bacteria by antibiotics contribute to the clinical deterioration of some patients with lethal endotoxemia. In this research we evaluated LPS (lipopolysaccharide release during antibiotic killing of salmonella typhi (Ty2-5536. The organism was incubated in the presence of Chloramphenicol, Ampicillin and Co-trimoxazole at concentrations that killed >99.9% of the organisms as determined by quantitative culture techniques. After incubation the antibiotic-bacterial cultures were centrifuged and the supernatants were filtered and collected for "in vivo" and "in vitro" studies. Injection of 1 ml/kg of filtrates in rabbits raised normal temperature of the animals by 1.2°C that it showes the presence of LPS in the filtrates. Quantitative chromogenic limulus amebocyte lyssate (L.A.L assay was used to determine the amount of LPS in the filtrates. The amount of LPS was 86.67±2.53 Pg/ml for chloramphenicol, 113.33±8.07 Pg/ml for Ampicillin and 134.18±11.59 Pg/ml for Co-trimoxazole. According to our investigation chloramphenicol is the best antibiotic against S.typhi because it decrease the induced-pathological effects of LPS in gram negative infection

  13. Avaliação de pirogênios em produtos de uso veterinário pelos testes da hipertermia em coelhos e do lisado de amebócitos do Limulus Pyrogens in veterinary products by the rabbit pyrogen test and the Limulus amoebocyte lysate test

    Directory of Open Access Journals (Sweden)

    Thiago Barth

    2007-02-01

    Full Text Available Realizou-se a avaliação de pirogênios em produtos veterinários de uso parenteral, pelo método da hipertermia em coelhos, calculando-se, para o teste das amostras, doses com concentrações de três a sete vezes superiores à terapêutica. Preconizou-se como resposta positiva o aumento de temperatura de 0,6°C. Utilizou-se também o ensaio do lisado de amebócitos do Limulus (LAL por geleificação, semiquantitativo, executando o teste de interferentes, validando o procedimento e estabelecendo a máxima diluição válida para a análise de cada produto. Paralelamente, efetuou-se avaliação comparativa de amostras com o método do LAL cromogênico, quantitativo, demonstrando correlação e reprodutibilidade dos resultados. Avaliaram-se vinte e oito produtos de diferentes classes farmacológicas, observando-se que dois não cumpriram as especificações, sendo reprovados. Sugere-se que as especificações estudadas sejam adotadas, contribuindo para aprimorar o controle de contaminantes, garantindo a qualidade e a segurança dos produtos veterinários.The rabbit pyrogen test was used to evaluate veterinary products, suggesting the temperature rise of 0.6°C as the ending point for the positive results. The test doses were calculated based on the therapeutic dose increased from three to seven times. The semi-quantitative Limulus amoebocyte lysate (LAL gel clot test was performed and compared to the LAL spectrophotometric chromogenic, quantitative assay. The comparative evaluation of the samples showed correlation and reproducibility of the results. The interference test was carried out, the procedure validated and the maximum valid dilution established for the analysis of the products without Pharmacopoeial specifications. Two of the twenty-eight products of different pharmacological groups evaluated didn't meet the requirements and were reproved. The specifications investigated are suggested to be used for the purity evaluation of the

  14. Red blood cell lysate modulates the expression of extracellular matrix proteins in dermal fibroblasts.

    Science.gov (United States)

    Akbari, Amir; Li, Yunyuan; Kilani, Ruhangiz T; Ghahary, Aziz

    2012-11-01

    During the early stage of wound healing process, blood clots can be served as a temporary extracellular matrix (ECM) to let skin cell migration and proliferation. The red blood cells are generally thought as inert bystanders in the early and inflammatory phase of wound healing. Here, we provide evidence that red blood cells (RBC) also play an important role in modulation of key ECM components such as type-I collagen, α-smooth muscle actin, fibronectin, and matrix metalloproteinases (MMPs). In this study, we used western blot analysis and showed a significant increase in the level of MMP-1, 2, 3. Furthermore, we found that RBC lysate significantly down-regulates type-I collagen and α-smooth muscle actin while up-regulates fibronectin expression in dermal fibroblasts. To further explore the mechanism by which RBC lysate modulates MMP-1 expression, the effect of inhibitors for three MAPK signaling pathways on RBC inducing MMP-1 expression by dermal fibroblasts were tested. The result showed that the inhibitor of ERK1/2 could abrogate the stimulatory effect of RBC lysate on MMP-1 expression in dermal fibroblasts. Consistently, RBC treatment results in an increase of ERK1/2 phosphorylation in dermal fibroblast. In conclusion, these findings suggest that RBC lysate can modulate the expression of MMPs and key ECM components which are important in healing process.

  15. Bifidobacterium longum lysate, a new ingredient for reactive skin.

    Science.gov (United States)

    Guéniche, Audrey; Bastien, Philippe; Ovigne, Jean Marc; Kermici, Michel; Courchay, Guy; Chevalier, Veronique; Breton, Lionel; Castiel-Higounenc, Isabelle

    2010-08-01

    Reactive skin is characterized by marked sensitivity to physical (heat, cold, wind) or chemical (topically applied products) stimuli and by the impairment of the skin barrier's ability to repair itself. Several lines of evidence suggest that beyond their capacity to positively influence the composition of intestinal microbiota, some probiotic bacteria can modulate the immune system both at local and systemic levels, thereby improving immune defense mechanisms and/or down-regulating immune disorders such as allergies and intestinal inflammation. Several recent human clinical trials clearly suggest that probiotic supplementation might be beneficial to the skin. Using a probiotic lysate, Bifidobacterium longum sp. extract (BL), we demonstrated first in vitro, and then in a clinical trial, that this non-replicating bacteria form applied to the skin was able to improve sensitive skin. The effect of BL were evaluated first on two different models. Using ex vivo human skin explant model we found a statistically significant improvement versus placebo in various parameters associated with inflammation such as a decrease in vasodilation, oedema, mast cell degranulation and TNF-alpha release. Moreover, using nerve cell cultures in vitro, we showed that after 6 h of incubation in culture medium (0.3-1%), the probiotic lysate significantly inhibited capsaicin-induced CGRP release by neurones. Then, a topical cream containing the active extract was tested in a randomized, double-blind, placebo-controlled trial. Sixty-six female volunteers with reactive skin were randomly given either the cream with the bacterial extract at 10% (n = 33) or the control cream (n = 33). The volunteers applied the cream to the face, arms and legs twice a day for two months. Skin sensitivity was assessed by stinging test (lactic acid) and skin barrier recovery was evaluated by measuring trans-epidermal water loss following barrier disruption induced by repeated tape-stripping at D1, D29 and D57. The

  16. Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination.

    Directory of Open Access Journals (Sweden)

    Iris Bosschem

    Full Text Available Helicobacter suis (H. suis is a widespread porcine gastric pathogen, which is also of zoonotic importance. The first goal of this study was to investigate the efficacy of several vaccine adjuvants (CpG-DNA, Curdlan, Freund's Complete and Incomplete, Cholera toxin, administered either subcutaneously or intranasally along with H. suis whole-cell lysate, to protect against subsequent H. suis challenge in a BALB/c infection model. Subcutaneous immunization with Freund's complete (FC/lysate and intranasal immunization with Cholera toxin (CT/lysate were shown to be the best options for vaccination against H. suis, as determined by the amount of colonizing H. suis bacteria in the stomach, although adverse effects such as post-immunization gastritis/pseudo-pyloric metaplasia and increased mortality were observed, respectively. Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects. A CCR4 antagonist that transiently inhibits the migration of regulatory T cells was also included as a new adjuvant in this second study. Results confirmed that immunization with CT (intranasally or sublingually is among the most effective vaccination protocols, but increased mortality was still observed. In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed. Compared to the FC/lysate immunized group, gastric pseudo-pyloric metaplasia was less severe or even absent in the CCR4 antagonist/lysate immunized group. In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

  17. Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination.

    Science.gov (United States)

    Bosschem, Iris; Bayry, Jagadeesh; De Bruyne, Ellen; Van Deun, Kim; Smet, Annemieke; Vercauteren, Griet; Ducatelle, Richard; Haesebrouck, Freddy; Flahou, Bram

    2015-01-01

    Helicobacter suis (H. suis) is a widespread porcine gastric pathogen, which is also of zoonotic importance. The first goal of this study was to investigate the efficacy of several vaccine adjuvants (CpG-DNA, Curdlan, Freund's Complete and Incomplete, Cholera toxin), administered either subcutaneously or intranasally along with H. suis whole-cell lysate, to protect against subsequent H. suis challenge in a BALB/c infection model. Subcutaneous immunization with Freund's complete (FC)/lysate and intranasal immunization with Cholera toxin (CT)/lysate were shown to be the best options for vaccination against H. suis, as determined by the amount of colonizing H. suis bacteria in the stomach, although adverse effects such as post-immunization gastritis/pseudo-pyloric metaplasia and increased mortality were observed, respectively. Therefore, we decided to test alternative strategies, including sublingual vaccine administration, to reduce the unwanted side-effects. A CCR4 antagonist that transiently inhibits the migration of regulatory T cells was also included as a new adjuvant in this second study. Results confirmed that immunization with CT (intranasally or sublingually) is among the most effective vaccination protocols, but increased mortality was still observed. In the groups immunized subcutaneously with FC/lysate and CCR4 antagonist/lysate, a significant protection was observed. Compared to the FC/lysate immunized group, gastric pseudo-pyloric metaplasia was less severe or even absent in the CCR4 antagonist/lysate immunized group. In general, an inverse correlation was observed between IFN-γ, IL-4, IL-17, KC, MIP-2 and LIX mRNA expression and H. suis colonization density, whereas lower IL-10 expression levels were observed in partially protected animals.

  18. Combinations of SPR and MS for characterization of native and recombinant proteins in cell lysates

    DEFF Research Database (Denmark)

    Borch, Jonas; Roepstorff, Peter

    2006-01-01

    Surface plasmon resonance and mass spectrometry (SPR-MS) has been combined for quality check of recombinant 6xHis-tagged 14-3-3 proteins expressed in Escherichia coli. Lysates were injected over an SPR sensorchip with immobilized Ni2+ for SPR analysis of the specific Ni2+ binding response...... of 14-3-3 proteins in order to test their activity. Specific binding of recombinant and native 14-3-3 proteins in complex mixtures to immobilized phosphopeptides and subsequent elution was also tested by SPR-MS. Ammonium sulfate precipitate fractions from lysates of E. coli expressing 14-3-3 protein...... and of cauliflower were investigated for specific binding to the phosphopeptide ligands immobilized on a sensorchip by SPR. Subsequently, the bound protein was eluted and analyzed by MS for characterization of intact mass and peptide mass mapping....

  19. DETERMINATION OF ENDOTOXIN IN SMALL-VOLUME PARENTERALS (SVPs: A COMPARISON OF LAL GEL-CLOT METHOD WITH U.S.P. RABBIT PYROGEN TEST

    Directory of Open Access Journals (Sweden)

    Morteza Rafiee-Tehrani M.H. Djamshidi

    1994-08-01

    Full Text Available The simplicity and sensitivity of the Limulus Amebocyte Lysate (LAL gel-clot method in detection of endotoxin in Small Volume Parenterals (SVPs were studied and compared with rabbit pyrogen test."nAll formulations were artificially spiked with 0.25, 0.5 and 5EU/ml of Escherichia coli 0111:B4 endotoxin (Iambda=0.5EU/ml. The Threshold Pyrogenic Dose (TPD for E.coli 0111 :B4 endotoxin was found to be more than lng per kg of body weight (0.5EU/ml. However, the sensitivity of LAL test was found to be 0.25 Eu/ml for water for injection and 0.5 EU/ml for other formulations. The observed inhibitory effect of electrolyte containing solutions was deleted by some degree of dilution prior to test. However, none of the samples showed inhibitory at the Maximum Valid Dilution (MVD. The U.S.P. rabbit pyrogen test was insignificant for all the solutions tested. These results suggest, LAL gel-clot test to be useful for determination of bacterial endotoxin as final release test in SVPs. A comparison of the pyrogenicity assays also indicates, that LAL is more sensitive and simple and less time-consuming than the conventional rabbit test.

  20. Contamination of nanoparticles by endotoxin: evaluation of different test methods

    Directory of Open Access Journals (Sweden)

    Smulders Stijn

    2012-11-01

    Full Text Available Abstract Background Nanomaterials can be contaminated with endotoxin (lipopolysaccharides, LPS during production or handling. In this study, we searched for a convenient in vitro method to evaluate endotoxin contamination in nanoparticle samples. We assessed the reliability of the commonly used limulus amebocyte lysate (LAL assay and an alternative method based on toll-like receptor (TLR 4 reporter cells when applied with particles (TiO2, Ag, CaCO3 and SiO2, or after extraction of the endotoxin as described in the ISO norm 29701. Results Our results indicate that the gel clot LAL assay is easily disturbed in the presence of nanoparticles; and that the endotoxin extraction protocol is not suitable at high particle concentrations. The chromogenic-based LAL endotoxin detection systems (chromogenic LAL assay and Endosafe-PTS, and the TLR4 reporter cells were not significantly perturbed. Conclusion We demonstrated that nanoparticles can interfere with endotoxin detection systems indicating that a convenient test method must be chosen before assessing endotoxin contamination in nanoparticle samples.

  1. Evaluation of a portable test system for assessing endotoxin activity in raw milk.

    Science.gov (United States)

    Suzuki, Yohko; Suzuki, Kazuyuki; Shimamori, Toshio; Tsuchiya, Masakazu; Niehaus, Andrew; Lakritz, Jeffrey

    2016-01-01

    The aim of the present study was to compare endotoxin activities detected in raw milk samples obtained from cattle by a commercially available portable test system (PTS) and traditional microplate limulus amebocyte lysate (LAL)-based assay, which determined activities using a kinetic turbidimetric (KT) assay. Raw milk samples were obtained from 53 and 12 dairy cattle without and with clinical mastitis, respectively. Comparison between the KT and PTS was performed by the Friedman test. The Pearson product moment correlation coefficients were calculated to evaluate associations between any two continuous variables. Linear regression model analysis was also performed to obtain the equation describing the relationship between PTS and KT assay. The endotoxin activities detected in 200- or 400-fold diluted milk samples were similar between PTS and KT assay, whereas a significant difference was observed in 100-fold diluted milk (P<0.001). The results obtained from 200- (r(2)=0.778, P<0.001) and 400-fold diluted milk samples (r(2)=0.945, P<0.001) using PTS correlated with those using KT assay. The median milk endotoxin activities in Gram-positive and Gram-negative clinical mastitis cows were 0.655 and 11,523.5 EU/ml, respectively. The results of the present study suggest that PTS as a simple and easy test to assess endotoxin activity in raw milk is efficient, simple and reproducible. PMID:26279135

  2. 枸橼酸钠注射液细菌内毒素检查方法的考察%Studies of Bacterial Endotoxins Test of Sodium Citrate Injection

    Institute of Scientific and Technical Information of China (English)

    何进; 史国兵; 高军; 陈金旺

    2011-01-01

    Objective To establish a method of bacterial endotoxin test of sodium citrate injection by gel clot.Methods The method was based on the bacterial endotoxin test in the appendix of Chp 2005.A series of tests were performed, such as the sensitivity test of tachypleus amebocyte lysate, first interference test and interference tests.Results No interference was found after the sodium citrate injection was diluted 6 times in the bacterial endotoxin interference factors test.Conclusion The present method can replace the pyrogen test with rabbits for examination of bacterial endotoxin of sodium citrate injection.%目的 建立枸橼酸钠注射液细菌内毒素检查方法.方法 按2005年版附录细菌内毒素检查法,分别进行了鲎试剂灵敏度复核试验、细菌内毒素干扰试验.结果 枸橼酸钠注射液稀释6倍后,不干扰鲎试剂与细菌内毒素的凝胶反应.结论 可用细菌内毒素检查法替代家兔法检查枸橼酸钠注射液中的细菌内毒素.

  3. Serodiagnosis of Lyme disease by kinetic enzyme-linked immunosorbent assay using recombinant VlsE1 or peptide antigens of Borrelia burgdorferi compared with 2-tiered testing using whole-cell lysates.

    Science.gov (United States)

    Bacon, Rendi Murphree; Biggerstaff, Brad J; Schriefer, Martin E; Gilmore, Robert D; Philipp, Mario T; Steere, Allen C; Wormser, Gary P; Marques, Adriana R; Johnson, Barbara J B

    2003-04-15

    In a study of US patients with Lyme disease, immunoglobulin (Ig) G and IgM antibody responses to recombinant Borrelia burgdorferi antigen VlsE1 (rVlsE1), IgG responses to a synthetic peptide homologous to a conserved internal sequence of VlsE (C6), and IgM responses to a synthetic peptide comprising the C-terminal 10 amino acid residues of a B. burgdorferi outer-surface protein C (pepC10) were evaluated by kinetic enzyme-linked immunoassay. At 99% specificity, the overall sensitivities for detecting IgG antibody to rVlsE1 or C6 in samples from patients with diverse manifestations of Lyme disease were equivalent to that of 2-tiered testing. When data were considered in parallel, 2 combinations (IgG responses to either rVlsE1 or C6 in parallel with IgM responses to pepC10) maintained high specificity (98%) and were significantly more sensitive than 2-tiered analysis in detecting antibodies to B. burgdorferi in patients with acute erythema migrans. In later stages of Lyme disease, the sensitivities of the in parallel tests and 2-tiered testing were high and statistically equivalent.

  4. Application of bacterial endotoxin test to the safety evaluation of clinical use of hydrocolloid dressing%细菌内毒素检查法在水胶体敷料临床使用安全性评价中的应用

    Institute of Scientific and Technical Information of China (English)

    许雷鸣; 顾倩; 武谷; 堵伟锋; 李翔; 林园园

    2014-01-01

    目的:建立水胶体敷料的细菌内毒素检查方法,为评价临床使用的安全性提供依据。方法按水胶体敷料表面积每1 cm2加入4 mL细菌内毒素检查用水,37℃浸提2 h,用2个不同厂家的鲎试剂进行干扰试验研究并采用凝胶半定量试验测定产品中的细菌内毒素含量。结果根据临床实际应用情况,确定水胶体敷料的内毒素限值L=0.5 EU·cm-2;在本实验条件下,可用灵敏度为0.125 EU·mL-1及0.125 EU·mL-1以上的鲎试剂检测水胶体敷料中的细菌内毒素。结论本试验建立的细菌内毒素检查方法可用于水胶体敷料的细菌内毒素检查,保障其临床使用的安全性。%Objective To establish a bacterial endotoxin test method for hydrocolloid dressing and to provide the experimental basis forthe safety evaluation of clinical use.Methods Each 1 cm2 total surface area of sample was added with 4 mL water for bacterial endo-toxin test and was extracted by heating at 37℃ for 2 hours.Tachypleus amebocyte lysate from two manufacturers were used for interfer-ence test and semi-quantitative test was used in the endotoxin assay of hydrocolloid dressing.Results According to the dose of clinicalapplication,the most suitable limit of bacterial endotoxin for hydrocolloid dressing was 0.5 EU·cm -2 .Under this experimental condi-tion,it was effective that bacterial endotoxin was tested by tachypleus amebocyte lysate with the sensitivity of 0.125 EU·mL -1 or high-er sensitivity.Conclusions This method can be used in bacterial endotoxin test of hydrocolloid dressing and can be applied to securethe safety of clinical use.

  5. Establishment of Bacterial Endotoxin Test of Iodine [131 I]Actuximab Injection%碘[131I]爱克妥昔单抗注射液细菌内毒素检查方法建立

    Institute of Scientific and Technical Information of China (English)

    孙祥敏; 张云; 张先; 杨薇薇; 蔡海燕

    2015-01-01

    为建立碘[131 I]爱克妥昔单抗注射液的细菌内毒素检查方法,采用《中国药典》二部细菌内毒素检查法进行实验。结果表明,将供试品稀释10倍、20倍、40倍进行细菌内毒素检查均存在干扰;对供试品稀释80倍进行检查或采用抗增液复溶鲎试剂,再将供试品稀释20倍进行检查,均无干扰。供试品的细菌内毒素检查可采用将供试品至少稀释80倍进行检验的方法;也可采用抗增液复溶鲎试剂,将供试品稀释至少20倍进行检查。结果表明,直接稀释法为产品检验的首选方法。%To establish a method for the bacterial endotoxin test of Iodine [1 3 1 I]Actux-imab Injection,The experiment was carried out according to the appendix ⅪE in Vol.Ⅱof the 2010 version of Chinese Pharmacopoeia.The results demonstrated that the prepa-ration interfered with the test at 10 times,20 times and 40 times dilution,and the inter-ference can be eliminated after the sample was diluted to 80 times or the sample was diluted to 20 times with the endotoxin-specific buffer dissolving the Tachypleus Amebo-cyte Lysate.Two methods can be used to test the bacterial endotoxin of Iodine [1 3 1 I] Actuximab Injection.One is the sample should be diluted to atleast 80 times,the other is the sample was diluted to 20 times with the endotoxin-specific buffer dissolving the Tachypleus Amebocyte Lysate.The dilution method is the preferred method for the quality control.

  6. 注射用丹参多酚酸盐细菌内毒素检查%Feasibility Study on Bacterial Endotoxin Test of Salvianolate for Injection

    Institute of Scientific and Technical Information of China (English)

    汪伟

    2012-01-01

    Objective: To establish a method for the bacterial endotoxin test of salvianolate for injection. Method: The bacterial endotoxin test was carried out according to the requirements in the appendix XI E of Chinese Pharmacopeia( edition 2010, Vol II ). Result: The salvianolate for injection at the concentration not greater than 0. 8 mg ? Ml-1 showed no interference on tachypiens amebocyte lysate teat. Conclusion: Bacterial endotoxin test ( gel process ) is applicable in the bacterial endotoxin limit test of salvianolate for injection.%目的:建立注射用丹参多酚酸盐的细菌内毒素检查方法.方法:按2010年版(二部)附录ⅪE细菌内毒素检查法进行操作.结果:注射用丹参多酚酸盐稀释至0.8 mg·ml-1及以下浓度对鲎试剂干扰试验无干扰作用.结论:注射用丹参多酚酸盐可采用细菌内毒素检查法(凝胶法)进行细菌内毒素的限量检查.

  7. 叔丁醇细菌内毒素检查方法的建立%Establishment of Bacterial Endotoxins Test for Tertiary Butyl Alcohol

    Institute of Scientific and Technical Information of China (English)

    于海洲; 都婧; 于风平

    2015-01-01

    目的:建立叔丁醇细菌内毒素检查方法。方法:采用2个不同厂家的鲎试剂和3批样品,进行预干扰试验和干扰试验。结果:供试品溶液对鲎试剂与细菌内毒素之间的凝集反应无干扰;样品的细菌内毒素限值确定为0.25 EU·ml-1。结论:所建立的方法可行,可用于叔丁醇的细菌内毒素检查。%Objective:To establish a method for the detection of bacterial endotoxins in tertiary butyl alcohol. Methods:The in-terference experiment and bacterial endotoxins test for 3 batches of the products from 2 companies were carried out with tachypleus ame-bocyte lysate. Results:There was no interference in the bacterial endotoxins test. The limit of bacterial endotoxins was 0. 25 EU/ml. Conclusion:The established method of bacterial endotoxins test is feasible for tertiary butyl alcohol.

  8. 一种简单实用的鲎试剂检测细菌内毒素法%A New Assay for Determination of Bacterial Endotoxins Using Limulus amebocyte Lysate

    Institute of Scientific and Technical Information of China (English)

    黄义德; 叶锦林; 唐良华

    2000-01-01

    在鲎试剂检测内毒素原理的基础上对传统的凝胶法进行改良.经改良后的新方法与传统方法相比具有检测结果一致,鲎试剂用量少,检测时间短,操作误差小等优点.

  9. Bacteria endotoxin test of Vitamin B Co. Injection%复合维生素B注射液细菌内毒素检查法的研究

    Institute of Scientific and Technical Information of China (English)

    刘骅; 武谷

    2011-01-01

    Aim To establish the bacterial endotoxin test for Vitamin B Co. Injection. Methods The experiment was carried out according to the appendix of Chinese Pharmacopeia in 2005 Edition (Vol Ⅱ ). Results The reactions with two Tachypleus Amebocyte Lysate manufacturers were not disturbed in minimus diluted multiple for Vitamin B Co. Injection,which was one ratio sixteen. The endotoxin limit of Vitamin B Co. Injection was 5 EU · ml -1. Conclusion It is feasible to control quality of Vitamin B Co. Injection by bacterial endotoxin test.%目的 建立复合维生素B注射液的细菌内毒素检查方法.方法 根据2005年版二部附录细菌内毒素检查法,采用2个不同生产厂家的鲎试剂对复合维生素B注射液进行研究.结果 复合维生素B注射液最小不干扰稀释倍数为1: 16,细菌内毒素限值为5 EU·ml-1.结论 采用细菌内毒素检查法控制复合维生素B注射液的质量是可行的.

  10. Bacteria Endotoxin Test of Vitamin B6 for Injection%注射用维生素B6细菌内毒素检查法的研究

    Institute of Scientific and Technical Information of China (English)

    刘骅

    2012-01-01

    OBJECTIVE To establish the bacterial endotoxin test of vitamin B6 for injection. METHODS The experiment was carried out according to the appendix of ChP 2010 Vol Ⅱ. tachypleus amabocyte lysate from two manufacturers were used to test the bacterial endotoxin. RESULTS The endotoxin limit of vitamin B6 for injection was 0.30 EU·mg‐1. The reactions were not disturbed in 0.208 mg·mL‐1 for vitamin B6, which was the maximum nondisrurbing concentration. It was effective that endotoxin was tested by amebocyte lysate agent with the sensitivity of 0.06 EU·mL ‐1 or much higher. CONCLUSION ft is feasible to control quality of vitamin B6 for injection by bacterial endotoxin test.%目的 建立注射用维生素B6的细菌内毒素检查方法.方法 根据中国药典2010年版二部附录细菌内毒素检查法,采用2个不同厂家的鲎试剂对注射用维生素B6进行研究.结果 注射用维生素B6细菌内毒素检查限值为0.30 EU·mg-1,其最大不干扰浓度为0.208 mg·mL-1,用灵敏度为0.06 EU·mL-1或更高灵敏度的鲎试剂可进行检验.结论 细菌内毒素检查法可用于注射用维生素B6中内毒素的检查.

  11. Production of eukaryotic cell-free lysate from Leishmania tarentolae.

    Science.gov (United States)

    Johnston, Wayne A; Alexandrov, Kirill

    2014-01-01

    In this chapter, we describe the production and application of a eukaryotic cell-free expression system based on Leishmania tarentolae. This single-celled flagellate allows straightforward and inexpensive cultivation in flasks or bioreactors. Unlike many other Leishmania species, it is nonpathogenic to humans and does not require special laboratory precautions. An additional reason it is a convenient source organism for cell-free lysate production is that all endogenous protein expression can be suppressed by a single antisense oligonucleotide targeting splice leader sequence on the 5'-end of all protein coding RNAs. We describe simple procedures for cell disruption and lysate processing starting from bioreactor culture. We also describe introduction of genetic information via vectors containing species-independent translation initiation sites (SITS). We consider that such an inexpensive eukaryotic cell-free production system has many advantages when expressing multi-subunit proteins or difficult to express proteins. PMID:24395406

  12. Exosomes: novel effectors of human platelet lysate activity

    OpenAIRE

    E Torreggiani; F Perut; Roncuzzi, L; N Zini; SR Baglìo; N Baldini

    2014-01-01

    Despite the popularity of platelet-rich plasma (PRP) and platelet lysate (PL) in orthopaedic practice, the mechanism of action and the effectiveness of these therapeutic tools are still controversial. So far, the activity of PRP and PL has been associated with different growth factors (GF) released during platelet degranulation. This study, for the first time, identifies exosomes, nanosized vesicles released in the extracellular compartment by a number of elements, including platelets, as one...

  13. Induction of protective antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer model

    Institute of Scientific and Technical Information of China (English)

    Xu Danfeng; Liu Yushan; Gao Yi; Cui Xingang; Xing Jizhang; Yin Lei; Yao Yacheng; Min Zhilian

    2009-01-01

    Objective: To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine therapy. Methods: C57BL/6 mice were immunized on the dorsal flank by s.c. Inoculation of Lysate-DC, ova-DC, and non-DC on day -7. On day 0, 2x 106cells of RM-1 tumor cells (H-2b) were injected s.c. In C57BL/6 mice pre-treated by s.c. Inoculation of modified DCs, correspondingly. DTH assay was performed with modified DCs. In partial test, for the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CD8, or natural killer (NK) NKI.1 cells with the corresponding monoclonal antibodies. The survival time of nude mice loaded with tumor cells was calculated and the size of tumor measured. Results: In RM-1 mice prostate cancer model, immunized with lysate-DC, compared with ova-DC and non-DC, the pre-infection vaccine resulted in 100% clearance of primary tumors, whereas on day 0 of injection vaccine cleared 40-60% of primary tumors. On day 0, C57BL/6 mice (H-2b) were immunized with Lysate-DC, compared with ova-DC and non-DC by caudal vein injection, then on day 15, RM-I cells were inoculated. On day 30, average diameters of tumor in different groups of modified DC were 23.7+5.4 mm, 22.1+4.9 mm, 4.3 ~2.6 mm, respectively. Lysate-DC, compared with ova-DC and non-DC, can greatly depressed RM-I tumor cell growth (P<0.01). The mean survival time of C57BL/6 mice in Lysate-DC, ova-DC and non-DC groups were 15.8 ~2.6, 16.6 ~3.2, 39.0 ~5.6, respectively, and there was a significant difference in the mean survival time in lysate-DC group between ova-DC and non-DC group (P<0.01). DTH test showed that lysate-DC could prime T lymphocyte and elicit tumor antigen specific immune response, and over 80% mice in groups of lysate-DC showed obvious swelling in their foot pad. This response was strengthened with repeating

  14. Enhanced target-specific signal detection using an Escherichia coli lysate in multiplex microbead immunoassays with E. coli-derived recombinant antigens.

    Science.gov (United States)

    Crestani, Sandra; Leitolis, Amanda; Lima, Lucianna Freitas Oliveira; Krieger, Marco A; Foti, Leonardo

    2016-08-01

    Diverse techniques have been developed to analyze antibody-mediated responses to infections. However, the most common tests, i.e., enzyme-linked immunosorbent assays, require separate reactions for each antigen and consequently necessitate large sample volumes. Luminex technology allows the detection of multiple antibodies in a single experiment, but nonspecific binding can impair the results. Therefore, we examined the use of Escherichia coli lysates to reduce nonspecific binding and improve the results of liquid microarrays based on Luminex technology. Anti-bacteria antibodies were detected in human serum samples, as evidenced by high median fluorescence intensity (MFI) in assays performed with paramagnetic microspheres coupled with E. coli lysates. Moreover, the addition of an E. coli lysate as a blocker reduced the nonspecific binding of antigens produced by E. coli in a concentration-dependent manner. Tris-HCl reduced MFI values in negative samples, but did not affect MFI for positive samples. For microspheres coupled with different antigens, an E. coli lysate blocker significantly improved the fluorescence signals from positive samples. The addition of Tris-HCl and the E. coli lysate induced antigen-specific differences in MFI. This combination of the E. coli lysate blocker and Tris-HCl yielded a statistically significant improvement in MFI in the assays for Chagas disease and hepatitis C virus samples. However, for the Treponema pallidum p47 antigen improvement in MFI was only observed for the preparation with the E. coli blocker at a concentration of 3%. In conclusion, the addition of an E. coli lysate and Tris-HCl to the microarray assay reduced the nonspecific binding of human anti-bacteria antibodies and, therefore, increased the specific MFI. PMID:27156997

  15. Influence of radiation on endotoxin test using the PTS TM for 18-FDG radiopharmaceutical

    Directory of Open Access Journals (Sweden)

    Ralph Santos-Oliveira

    2010-09-01

    Full Text Available F-18 FDG (2-[18-F] fluoro-2-deoxy-D-glucose is the most frequently used radiopharmaceutical for PET and PET CT imaging exams. The FDA recently approved the use of the PTS TM (Portable Test System as an alternative to the standard test proposed by the United States Pharmacopeia using the LAL (Limulus Amebocyte Lysates, that takes longer to perform (about 1h than the PTS TM (15 min. Recent studies have demonstrated that radiation could interfere with the PTS TM test. In order to study the effects of radiation on the PTS TM test and/or equipment, 27 batches of F-18 FDG produced in the Nuclear Engineering Institute were analyzed. The results showed that no direct correlation with radiation was found in any of the cases.O FDG-18 é o radiofármaco mais utilizado nos exames de PET e PET CT. O FDA recentemente aprovou o uso do PTS TM (Portable Test System como método alternativo ao teste padrão de endotoxina, proposto pela Farmacopéia Americana, considerando que no primeiro há um tempo de espera de 1 hora frente a somente 15 minutos do segundo. Estudo recentes demonstram que a radiação poderia interferir no teste do PTS TM. De modo a avaliar os efeitos da radiação no teste PTS TM foram analisados 27 lotes de F-18 FDG produzidos no Instituto de Engenharia Nuclear. Os resultados demonstraram que em todos os casos nenhuma correlação direta com a radiação foi observada.

  16. The Effect of Autologous Platelet Lysate Eye Drops: An In Vivo Confocal Microscopy Study

    Science.gov (United States)

    Fea, Antonio M.; Testa, Valeria; Machetta, Federica; Parisi, Simone; D'Antico, Sergio; Spinetta, Roberta; Fusaro, Enrico; Grignolo, Federico M.

    2016-01-01

    Purpose. To determine the effectiveness of autologous platelet lysate (APL) eye drops in patients with primary Sjögren syndrome (SS) dry eye, refractory to standard therapy, in comparison with patients treated with artificial tears. We focused on the effect of APL on cornea morphology with the in vivo confocal microscopy (IVCM). Methods. Patients were assigned to two groups: group A used autologous platelet lysate QID, and group B used preservative-free artificial tears QID, for 90 days. Ophthalmological assessments included ocular surface disease index (OSDI), best corrected visual acuity (BCVA), Schirmer test, fluorescein score, and breakup time (BUT). A subgroup of patients in group A underwent IVCM: corneal basal epithelium, subbasal nerves, Langerhans cells, anterior stroma activated keratocytes, and reflectivity were evaluated. Results. 60 eyes of 30 patients were enrolled; in group A (n = 20 patients) mean OSDI, fluorescein score, and BUT showed significant improvement compared with group B (n = 10 patients). The IVCM showed a significant increase in basal epithelium cells density and subbasal nerve plexus density and number and a decrease in Langerhans cells density (p < 0.05). Conclusion. APL was found effective in the treatment of SS dry eye. IVCM seems to be a useful tool to visualize cornea morphologic modifications. PMID:27200376

  17. The Effect of Autologous Platelet Lysate Eye Drops: An In Vivo Confocal Microscopy Study

    Directory of Open Access Journals (Sweden)

    Antonio M. Fea

    2016-01-01

    Full Text Available Purpose. To determine the effectiveness of autologous platelet lysate (APL eye drops in patients with primary Sjögren syndrome (SS dry eye, refractory to standard therapy, in comparison with patients treated with artificial tears. We focused on the effect of APL on cornea morphology with the in vivo confocal microscopy (IVCM. Methods. Patients were assigned to two groups: group A used autologous platelet lysate QID, and group B used preservative-free artificial tears QID, for 90 days. Ophthalmological assessments included ocular surface disease index (OSDI, best corrected visual acuity (BCVA, Schirmer test, fluorescein score, and breakup time (BUT. A subgroup of patients in group A underwent IVCM: corneal basal epithelium, subbasal nerves, Langerhans cells, anterior stroma activated keratocytes, and reflectivity were evaluated. Results. 60 eyes of 30 patients were enrolled; in group A (n=20 patients mean OSDI, fluorescein score, and BUT showed significant improvement compared with group B (n=10 patients. The IVCM showed a significant increase in basal epithelium cells density and subbasal nerve plexus density and number and a decrease in Langerhans cells density (p<0.05. Conclusion. APL was found effective in the treatment of SS dry eye. IVCM seems to be a useful tool to visualize cornea morphologic modifications.

  18. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture

    International Nuclear Information System (INIS)

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  19. dnaC-dependent reconstitution of replication forks in Escherichia coli lysates.

    OpenAIRE

    Nüsslein-Crystalla, V; Niedenhof, I; Rein, R.

    1982-01-01

    Lysates of Escherichia coli exhibit a DNA-synthesizing activity that depends on the presence of replication forks and of replication proteins. Replicative activity was reconstituted in vitro by mixing lysates prepared from temperature-sensitive dnaB mutants with wild-type dnaB protein. Lysates of double mutants deficient in both dnaB and dnaC genes could only be complemented by the addition of both dnaB and dnaC proteins, whereas lysates deficient in dnaC protein did not require the addition ...

  20. Effect of lysate of Sarcocystis gigantea in rats

    Directory of Open Access Journals (Sweden)

    N. S. Al-Hyali

    2011-01-01

    Full Text Available This study was conducted to examine the effect of lysate isolated from macrosarcocysts of Sarcocystis gigantea after inoculation into albino rats to follow up the pathological effect on heart, lung and skeletal muscle fibers. The results showed the presence of pathological changes, represented by hyperatrophy, congestion, associated with hyaline degeneration in myocardium of heart. In lung tissues severe congestion in each right and left lobes concomitant with emphysema, thickening was observed in wall of branchioles and blood vessels at 3-7 days have been seen. In skeletal muscles the result revealed the presence of multinecrotic areas which infiltrated with inflammatory cells accompanied by hyaline degeneration in muscle fibers at 3-7 days after treatment.

  1. Detection of circulating tumor lysate-reactive CD4+ T cells in melanoma patients

    DEFF Research Database (Denmark)

    Ladekarl, Morten; Agger, Ralf; Fleischer, Charlotte C;

    2004-01-01

    donors with high background levels of spontaneous IFN-gamma production, indicating an inhibitory effect of the lysate. CONCLUSIONS: This method for detection of a peripheral T-cell immune response in melanoma patients has several advantages for clinical use. The tumor lysate preparations may contain......PURPOSE: We wanted to study whether an allogeneic melanoma lysate would be a feasible stimulatory antigen source for detection of a peripheral CD4+ T-cell immune response in patients with medically untreated malignant melanoma. The lysate was produced from a melanoma cell line (FM3.29) which...... expresses high amounts of melanoma antigens. METHODS: Fresh peripheral blood was incubated with and without lysate for 6 h in the presence of anti-CD28/anti-CD49d MoAb (for costimulation). After flow cytometric estimation of the frequency of CD69+/IFN-gamma+ cells in the CD4+ population, the response to...

  2. Establishment of Bacterial Endotoxins Test of Xiyanping Injection%喜炎平注射液细菌内毒素检查法的建立

    Institute of Scientific and Technical Information of China (English)

    吴寒寅; 孟德胜

    2012-01-01

    目的 建立喜炎平注射液细菌内毒素检查法.方法 按2010年版《中国药典(二部)》附录细菌内毒素检查法,用不同厂家的鲎试剂对3个批号的喜炎平注射液分别进行干扰试验和细菌内毒素检查.结果 喜炎平注射液稀释到40倍(质量浓度为1.25 g/L)时对细菌内毒素检查法无干扰,细菌内毒素限值L=5 EU/mL,按照拟订标准,3批样品细菌内毒素检查均符合规定.结论 所建立的鲎试剂法可用于喜炎平注射液的细菌内毒素检查.%Objective To establish the method of bacterial endotoxin test of Xiyanping injection. Methods Based on the bacterial endo-toxin test in the appendix of the Chinese Pharmacopeia(edition 2010,part 2),the interference test and the bacterial endotoxin test were conducted on the samples from 3 batches of Xiyanping injection by using tachypleus amebocyte lysate(TAL) from different manufacturers. Results Xiyanping injection solution with dilution to 40 times (mass concentration 1. 25 g/L) had no interference on the bacterial endotoxin test. The limit value of the bacterial endotoxin was 5 EU/mL. According to the established standard, the bacterial endotoxin test in the 3 batches of samples conformed to the stipulation. Conclusion The established TAL method may be applicable for the bacterial endotoxin test of Xiyanping injection.

  3. Exosomes: novel effectors of human platelet lysate activity

    Directory of Open Access Journals (Sweden)

    E Torreggiani

    2014-09-01

    Full Text Available Despite the popularity of platelet-rich plasma (PRP and platelet lysate (PL in orthopaedic practice, the mechanism of action and the effectiveness of these therapeutic tools are still controversial. So far, the activity of PRP and PL has been associated with different growth factors (GF released during platelet degranulation. This study, for the first time, identifies exosomes, nanosized vesicles released in the extracellular compartment by a number of elements, including platelets, as one of the effectors of PL activity. Exosomes were isolated from human PL by differential ultracentrifugation, and analysed by electron microscopy and Western blotting. Bone marrow stromal cells (MSC treated with three different exosome concentrations (0.6 μg, 5 μg and 50 μg showed a significant, dose-dependent increase in cell proliferation and migration compared to the control. In addition, osteogenic differentiation assays demonstrated that exosome concentration differently affected the ability of MSC to deposit mineralised matrix. Finally, the analysis of exosome protein content revealed a higher amount of basic fibroblast growth factor (bFGF, vascular endothelial growth factor (VEGF, platelet-derived growth factor (PDGF-BB and transforming growth factor beta 1 (TGF-β1 as compared to PL. In regards to RNA content, an enrichment of small RNAs in exosomes as compared to donor platelets has been found. These results suggest that exosomes consistently contribute to PL activity and could represent an advantageous nanodelivery system for cell-free regeneration therapies.

  4. Exosomes: novel effectors of human platelet lysate activity.

    Science.gov (United States)

    Torreggiani, E; Perut, F; Roncuzzi, L; Zini, N; Baglìo, S R; Baldini, N

    2014-01-01

    Despite the popularity of platelet-rich plasma (PRP) and platelet lysate (PL) in orthopaedic practice, the mechanism of action and the effectiveness of these therapeutic tools are still controversial. So far, the activity of PRP and PL has been associated with different growth factors (GF) released during platelet degranulation. This study, for the first time, identifies exosomes, nanosized vesicles released in the extracellular compartment by a number of elements, including platelets, as one of the effectors of PL activity. Exosomes were isolated from human PL by differential ultracentrifugation, and analysed by electron microscopy and Western blotting. Bone marrow stromal cells (MSC) treated with three different exosome concentrations (0.6 μg, 5 μg and 50 μg) showed a significant, dose-dependent increase in cell proliferation and migration compared to the control. In addition, osteogenic differentiation assays demonstrated that exosome concentration differently affected the ability of MSC to deposit mineralised matrix. Finally, the analysis of exosome protein content revealed a higher amount of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF-BB) and transforming growth factor beta 1 (TGF-β1) as compared to PL. In regards to RNA content, an enrichment of small RNAs in exosomes as compared to donor platelets has been found. These results suggest that exosomes consistently contribute to PL activity and could represent an advantageous nanodelivery system for cell-free regeneration therapies. PMID:25241964

  5. Black cobra (Naja naja karachiensis) lysates exhibit broad-spectrum antimicrobial activities

    Science.gov (United States)

    Sagheer, Mehwish; Siddiqui, Ruqaiyyah; Iqbal, Junaid; Khan, Naveed Ahmed

    2014-01-01

    It is hypothesized that animals living in polluted environments possess antimicrobials to counter pathogenic microbes. The fact that snakes feed on germ-infested rodents suggests that they encounter pathogenic microbes and likely possess antimicrobials. The venom is used only to paralyze the rodent, but the ability of snakes to counter potential infections in the gut due to disease-ridden rodents requires robust action of the immune system against a broad range of pathogens. To test this hypothesis, crude lysates of different organs of Naja naja karachiensis (black cobra) were tested for antimicrobial properties. The antimicrobial activities of extracts were tested against selected bacterial pathogens (neuropathogenic Escherichia coli K1, methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Streptococcus pneumonia), protist (Acanthamoeba castellanii), and filamentous fungus (Fusarium solani). The findings revealed that plasma and various organ extracts of N. n. karachiensis exhibited antimicrobial activity against E. coli K1, MRSA, P. aeruginosa, S. pneumoniae, A. castellanii, and F. solani in a concentration-dependent manner. The results of this study are promising for the development of new antimicrobials. PMID:24625321

  6. Positive Limulus amoebocyte lysate reactions with polyriboinosinic acid x polyribocytidylic acid.

    OpenAIRE

    Elin, R J; Utter, A E

    1980-01-01

    The annealed copolymer polyriboinosinic acid x polyriboinosinic acid reacted with Limulus amoebocyte lysate to cause gelation at a concentration approximately 2,000-fold or greater than bacterial endotoxins. This copolymer was pyrogenic in rabbits and demonstrated hypochromicity, but no significant correlation was noted among Limulus amoebocyte lysate reactivity, progenicity, and hypochromicity. Like endotoxin, polyriboinosinic acid x polyribocytidylic acid did not react with purified Limulus...

  7. Aflatoxin B1 degradation by liquid cultures and lysates of three bacterial strains.

    Science.gov (United States)

    Adebo, Oluwafemi Ayodeji; Njobeh, Patrick Berka; Sidu, Sibusiso; Tlou, Matsobane Godfrey; Mavumengwana, Vuyo

    2016-09-16

    Aflatoxin contamination remains a daunting issue to address in food safety. In spite of the efforts geared towards prevention and elimination of this toxin, it still persists in agricultural commodities. This has necessitated the search for other measures such as microbial degradation to combat this hazard. In this study, we investigated the biodegradation of aflatoxin B1 (AFB1), using lysates of three bacterial strains (Pseudomonas anguilliseptica VGF1, Pseudomonas fluorescens and Staphylococcus sp. VGF2) isolated from a gold mine aquifer. The bacterial cells were intermittently lysed in the presence and absence of protease inhibitors to obtain protease free lysates, subsequently incubated with AFB1 for 3, 6, 12, 24, and 48h to investigate whether any possible AFB1 degradation occurred using high performance liquid chromatography (HPLC) for detection. Results obtained revealed that after 6h of incubation, protease inhibited lysates of Staphylococcus sp. VGF2 demonstrated the highest degradation capacity of 100%, whereas P. anguilliseptica VGF1 and P. fluorescens lysates degraded AFB1 by 66.5 and 63%, respectively. After further incubation to 12h, no residual AFB1 was detected for all the lysates. Lower degrading ability was however observed for liquid cultures and uninhibited lysates. Data on cytotoxicity studies against human lymphocytes showed that the degraded products were less toxic than the parent AFB1. From this study, it can thus be deduced that the mechanism of degradation by these bacterial lysates is enzymatic. This study shows the efficacy of crude bacterial lysates for detoxifying AFB1 indicating potential for application in the food and feed industry. PMID:27294556

  8. Culture of human cell lines by a pathogen-inactivated human platelet lysate.

    Science.gov (United States)

    Fazzina, R; Iudicone, P; Mariotti, A; Fioravanti, D; Procoli, A; Cicchetti, E; Scambia, G; Bonanno, G; Pierelli, L

    2016-08-01

    Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety. PMID:25944665

  9. Layer-by-layer assembled cell instructive nanocoatings containing platelet lysate.

    Science.gov (United States)

    Oliveira, Sara M; Santo, Vítor E; Gomes, Manuela E; Reis, Rui L; Mano, João F

    2015-04-01

    Great efforts have been made to introduce growth factors (GFs) onto 2D/3D constructs in order to control cell behavior. Platelet lysate (PL) presents itself as a cost-effective source of multiple GFs and other proteins. The instruction given by a construct-PL combination will depend on how its instructive cues are presented to the cells. The content, stability and conformation of the GFs affect their instruction. Strategies for a controlled incorporation of PL are needed. Herein, PL was incorporated into nanocoatings by layer-by-layer assembling with polysaccharides presenting different sulfation degrees (SD) and charges. Heparin and several marine polysaccharides were tested to evaluate their PL and GF incorporation capability. The consequent effects of those multilayers on human adipose derived stem cells (hASCs) were assessed in short-term cultures. Both nature of the polysaccharide and SD were important properties that influenced the adsorption of PL, vascular endothelial growth factor (VEGF), fibroblast growth factor b (FGFb) and platelet derived growth factor (PDGF). The sulfated polysaccharides-PL multilayers showed to be efficient in the promotion of morphological changes, serum-free adhesion and proliferation of high passage hASCs (P > 5). These biomimetic multilayers promise to be versatile platforms to fabricate instructive devices allowing a tunable incorporation of PL. PMID:25701032

  10. GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts.

    Science.gov (United States)

    Carducci, Augusto; Scafetta, Gaia; Siciliano, Camilla; Carnevale, Roberto; Rosa, Paolo; Coccia, Andrea; Mangino, Giorgio; Bordin, Antonella; Vingolo, Enzo Maria; Pierelli, Luca; Lendaro, Eugenio; Ragona, Giuseppe; Frati, Giacomo; De Falco, Elena

    2016-01-01

    Tenon's fibroblasts (TFs), widely employed as in vitro model for many ophthalmological studies, are routinely cultured with FBS. Platelet Lysate (PL), a hemoderivate enriched with growth factors and cytokines has been largely tested in several clinical applications and as substitute of FBS in culture. Here, we investigate whether PL can exert biological effects on TF populations similarly to other cell types. Results show that PL significantly enhances cell proliferation and migration vs. FBS, without influencing cell size/granularity. Upregulation of EGF, VEGF, KDR, MMP2-9, FAK mRNA levels also occurs and phosphorylation of AKT but not of ERK1/2 is significantly enhanced. The inhibition of the PI3kinase/AKT pathway with the specific inhibitor wortmannin, decreases PL-induced cell migration but not proliferation. Condition supernatants containing PL show increased bioavailability of Nitric Oxide and reduced levels of 8-Iso-PGF2-alpha, correlating with cell proliferation and migration. Pro-angiogenic/inflammatory soluble factors (GRO, Angiogenin, EGF, I-309, PARC) are exclusively or greater expressed in media containing PL than FBS. GMP-grade PL preparations positively influence in vitro biological effects of TFs representing a suitable and safer alternative to FBS. PMID:26709648

  11. エンドトキシン血症の実験的・臨床的研究

    OpenAIRE

    川山, 照雄

    1980-01-01

    The Limulus test is a useful and highly sensitive method for detecting endotoxin. Lysate which was extracted from the amebocytes of Tachypleus tridentatus showed almost the same reactivity for endotoxin, as described for Limulus polyphemus. The results obtained in the present experimental and clinical application of the Limulus test are follows : 1. The lysate from Tachypleus tridentatus was capable of detecting endotoxin of as little as nanogram per ml. 2. The gel-formation activity of the l...

  12. Aplicabilidade do Teste de Ativação de Monócitos (MAT no Brasil: importância da sua utilização como teste para detecção de pirogênios no controle da qualidade de produtos injetáveis | Applicability of the Monocyte Activation Test (MAT in Brazil: the importance of its use as a test for the detection of pyrogens in the quality control of injectable products

    Directory of Open Access Journals (Sweden)

    Cristiane Caldeira da Silva

    2015-08-01

    Full Text Available O MAT (sigla do inglês Monocyte Activation Test é considerado um potencial substituto do Teste de Pirogênios, entretanto: i. não foi avaliado para um número suficiente de produtos; ii. faltam dados que possam garantir sua capacidade em detectar pirogênios não endotoxinas; e iii. deve ser realizada a validação do método para cada classe de produtos. O objetivo foi identificar as monografias que requerem testes de pirogenicidade e propor os produtos que têm por base somente o teste de pirogênios como um ponto de partida para futuros estudos. As monografias específicas nas Farmacopeias Americana, Europeia e Brasileira que recomendam o Teste de Pirogênios ou Teste de Endotoxina Bacteriana ou LAL (sigla do inglês Limulus Amebocyte Lysate foram: Teste de Pirogênios: 20 monografias na Americana, 37 na Europeia e 28 na Brasileira. LAL: 619 monografias na Americana, 157 na Europeia e 41 na Brasileira. Somente quatro produtos requerem testes de pirogenicidade nas três farmacopeias analisadas. O Teste de Pirogênios e LAL são recomendados em seis monografias na Brasileira e 15 na Europeia. Na Brasileira, a maior parte dessas monografias é referente a produtos biológicos, sugerindo, assim, que estes devam ser os primeiros a ser testados, uma vez que são ensaiados em animais. ----------------------------------------------------------------------------------------------- Monocyte Activation Test (MAT is thought to be a good replacement for rabbit pyrogen test (RPT; however, MAT remains controversial. MAT was not adequately evaluated in a sufficient number of products, and there is no sufficient data that support the ability of MAT to detect non-endotoxin pyrogens. Furthermore, MAT was used subject to validation for each specific product. The aim of this study was to identify in main pharmacopoeias, whose monographs require pyrogenicity tests, and propose those products for which only the rabbit pyrogen test is required to be used as a

  13. Regulation of protein synthesis by phosphorylation of eukaryotic initiation factor 2 alpha in intact reticulocytes and reticulocyte lysates.

    OpenAIRE

    Leroux, A; London, I M

    1982-01-01

    Studies in intact rabbit reticulocytes and reticulocyte lysates provide further evidence of a functional role for the phosphorylation of eukaryotic initiation factor 2 alpha (eIF-2 alpha) in the regulation of initiation of protein synthesis in eukaryotic cells. In intact reticulocytes treated with isonicotinic acid hydrazide to inhibit heme synthesis, the phosphorylation of eIF-2 alpha was significantly greater than in control cells. In heme-deficient reticulocyte lysates and in lysates treat...

  14. Quantitative assessment of the p53-Mdm2 feedback loop using protein lysate microarrays.

    Science.gov (United States)

    Ramalingam, Sundhar; Honkanen, Peter; Young, Lynn; Shimura, Tsutomu; Austin, John; Steeg, Patricia S; Nishizuka, Satoshi

    2007-07-01

    Mathematical simulations of the p53-Mdm2 feedback loop suggest that both proteins will exhibit impulsive expression characteristics in response to high cellular stress levels. However, little quantitative experimental evaluation has been done, particularly of the phosphorylated forms. To evaluate the mathematical models experimentally, we used lysate microarrays from an isogenic pair of gamma-ray-irradiated cell lysates from HCT116 (p53(+/+) and p53(-/-)). Both p53 and Mdm2 proteins showed expected pulses in the wild type, whereas no pulses were seen in the knockout. Based on experimental observations, we determined model parameters and generated an in silico "knockout," reflecting the experimental data, including phosphorylated proteins.

  15. Quantitative analysis of HIV-1 protease inhibitors in cell lysates using MALDI-FTICR mass spectrometry.

    NARCIS (Netherlands)

    Kampen, JJ van; Burgers, P.C.; Groot, R. de; Osterhaus, A.D.; Reedijk, M.L.; Verschuren, E.J.; Gruters, R.A.; Luider, T.M.

    2008-01-01

    In this report we explore the use of MALDI-FTICR mass spectrometry for the quantitative analysis of five HIV-1 protease inhibitors in cell lysates. 2,5-Dihydroxybenzoic acid (DHB) was used as the matrix. From a quantitative perspective, DHB is usually a poor matrix due to its poor shot-to-shot and p

  16. Optimizing parameters for clinical-scale production of high IL-12 secreting dendritic cells pulsed with oxidized whole tumor cell lysate

    Directory of Open Access Journals (Sweden)

    Chiang Cheryl L-L

    2011-11-01

    replated in fresh media. Conclusions This study examined criteria including DC phenotype, viability, IL-12p70 production and the ability to stimulate MLR as metrics of whole oxidized tumor lysate-pulsed DC immunogenicity and functionality. Development and optimization of this unique method is now being tested in a clinical trial of autologous oxidized tumor lysate-pulsed DC in clinical-scale in recurrent ovarian, primary peritoneal or fallopian tube cancer (NCT01132014.

  17. MICROFLUIDIC MODULES FOR ISOLATION OF RECOMBINANT CYTOKINE FROM BACTERIAL LYSATES

    Energy Technology Data Exchange (ETDEWEB)

    Retterer, Scott T [ORNL; Doktycz, Mitchel John [ORNL

    2014-01-01

    The portability and personalization of health-care diagnostics and treatments benefits from advancements and applications of micro and nanotechnology. Modularization and miniaturization of standardized biochemical processes and tests facilitates the advancement and customization of analyte detection and diagnosis on-chip. The goal of our work here is to develop modular platforms for on-chip biochemical processing of synthesized biologics for a range of on-demand applications. Our report focuses on the initial development, characterization and application of microfluidic size exclusion/gel filtration and ion exchange protein concentration modules for cytokine isolation from spiked cell extracts.

  18. Antitumor immunity by a dendritic cell vaccine encoding secondary lymphoid chemokine and tumor lysate on murine prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Jun Lu; Qi Zhang; Chun-Min Liang; Shu-Jie Xia; Cui-Ping Zhong; Da-Wei Wang

    2008-01-01

    Aim: To investigate the antitumor immunity by a dendritic cell (DC) vaccine encoding secondary lymphoid chemokine gene and tumor lysate on murine prostate cancer. Methods: DC from bone marrow of C57BL/6 were transfected with a plasmid vector expressing secondary lymphoid chemokine (SLC) cDNA by Lipofectamine2000 liposome and tumor lysate. Total RNA extracted from SLC+lysate-DC was used to verify the expression of SLC by reverse transcriptase-polymerase chain reaction (RT-PCR). The immunotherapeutic effect of DC vaccine on murine prostate cancer was assessed. Results: We found that in the prostate tumor model of C57BL/6 mice, the adminstration of SLC+lysate-DC inhibited tumor growth most significantly when compared with SLC-DC, lysate-DC, DC or phos-phate buffer solution (PBS) counterparts (P<0.01). Immunohistochemical fluorescent staining analysis showed the infiltration of more CD4+, CD8+ T cell and CD11c+ DC within established tumor treated by SLC+lysate-DC vaccine than other DC vaccines (P<0.01). Conclusion: DC vaccine encoding secondary lymphoid chemokine and tumor lysate can elicit significant antitumor immunity by infiltration of CD4+, CD8+ T cell and DC, which might provide a potential immunotherapy method for prostate cancer.

  19. Active, soluble recombinant melittin purified by extracting insoluble lysate of Escherichia coli without denaturation

    OpenAIRE

    Buhrman, Jason S; Cook, Laura C.; Rayahin, Jamie E; Federle, Michael J.; Gemeinhart, Richard A.

    2013-01-01

    Cell lytic peptides are a class of drugs that can be used to selectively kill invading organisms or diseased cells. Several of these peptides have been identified as potential therapeutics. Herein, we report a novel process for purifying recombinant melittin, a cell lytic peptide that inserts into the membranes of cells causing cell lysis, from Escherichia coli. The process involves surfactant and low pH to solubilize melittin fusion proteins from the insoluble fraction of bacterial lysates. ...

  20. Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate

    OpenAIRE

    Sokolova, Ekaterina; Spruijt, Evan; Hansen, Maike M. K.; Dubuc, Emilien; Groen, Joost; Chokkalingam, Venkatachalam; Piruska, Aigars; Heus, Hans A.; Huck, Wilhelm T. S.

    2013-01-01

    Liquid–liquid phase transitions in complex mixtures of proteins and other molecules produce crowded compartments supporting in vitro transcription and translation. We developed a method based on picoliter water-in-oil droplets to induce coacervation in Escherichia coli cell lysate and follow gene expression under crowded and noncrowded conditions. Coacervation creates an artificial cell-like environment in which the rate of mRNA production is increased significantly. Fits to the measured tran...

  1. Mechanical fibrinogen-depletion supports heparin-free mesenchymal stem cell propagation in human platelet lysate

    OpenAIRE

    Laner-Plamberger, Sandra; Lener, Thomas; Schmid, Doris; Streif, Doris A.; Salzer, Tina; Öller, Michaela; Hauser-Kronberger, Cornelia; Fischer, Thorsten; Jacobs, Volker R.; Schallmoser, Katharina; Gimona, Mario; Rohde, Eva

    2015-01-01

    Background Pooled human platelet lysate (pHPL) is an efficient alternative to xenogenic supplements for ex vivo expansion of mesenchymal stem cells (MSCs) in clinical studies. Currently, porcine heparin is used in pHPL-supplemented medium to prevent clotting due to plasmatic coagulation factors. We therefore searched for an efficient and reproducible medium preparation method that avoids clot formation while omitting animal-derived heparin. Methods We established a protocol to deplete fibrino...

  2. Effective biotic elicitation of Ruta graveolens L. shoot cultures by lysates from Pectobacterium atrosepticum and Bacillus sp.

    Science.gov (United States)

    Orlita, A; Sidwa-Gorycka, M; Malinski, E; Czerwicka, M; Kumirska, J; Golebiowski, M; Lojkowska, E; Stepnowski, P

    2008-03-01

    Growth of Ruta graveolens shoots was induced when Bacillus sp. cell lysates were added to the culture medium. Elicitation of coumarin by this lysate was also very effective; the concentrations of isopimpinelin, xanthotoxin and bergapten increased to 610, 2120 and 1460 microg g(-1) dry wt, respectively. It also had a significant effect on the production of psoralen and rutamarin (680 and 380 microg g(-1) dry wt) and induced the biosynthesis of chalepin, which was not detected in the control sample, up to 47 microg g(-1) dry wt With lysates of the Pectobacterium atrosepticum, their effect on growth was not so significant and had no effect on the induction of coumarin accumulation. But elicitation with this lysate was much more effective for inducing the production of furoquinolone alkaloids; the concentrations of gamma-fagarine, skimmianine, dictamnine and kokusaginine rose to 99, 680, 172 and 480 microg g(-1) dry wt, respectively.

  3. Effect of Conditioned Medium and Bone Marrow Stem Cell Lysate on the Course of Acetaminophen-Induced Liver Failure.

    Science.gov (United States)

    Khubutiya, M Sh; Temnov, A A; Vagabov, V A; Sklifas, A N; Rogov, K A; Zhgutov, Yu A

    2015-05-01

    A composition containing culture medium conditioned by mesenchymal stem cells and mesenchymal stem cell lysate improves biochemical parameters, reduces inflammation, and stimulates regenerative processes in the liver. PMID:26033600

  4. Treatment of established colon carcinoma-bearing mice by dendritic cells pulsed with lysates of heat-treated tumor cells

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    To investigate the therapeutic effect of dendritic cells pulsed with lysates of heat-treated CT26 colon carcinoma cells. Bone marrow-derived DCs were pulsed with lysates of heat-treated tumor cells and were used to immunize BALB/c mice with established colon carcinoma. Cytotoxic T lymphocyte (CTL) response was detected. The therapeutic effect induced by DCs was observed by tumor weight and survival time. DCs pulsed with lysates of heat-treated tumor cells markedly induced specific cytotoxic activity of CTLs. Tumor growth in the immunized BALB/c mice was significantly inhibited and the survival time of the tumor-bearing mice was prolonged. DCs pulsed with lysates of heat-treated tumor cells have an observable therapeutic effect on established colon carcinoma-bearing mice.

  5. Treatment of established colon carcinoma-bearing mice by dendritic cells pulsed with lysates of heat-treated tumor cells

    Institute of Scientific and Technical Information of China (English)

    YING MinGang; ZHEN QiuHong; LIU Sheng; GONG FuSheng; XIE YunQing

    2009-01-01

    To investigate the therapeutic effect of dendritic cells pulsed with lysates of heat-treated CT26 colon carcinoma cells. Bone marrow-derived DCs were pulsed with lysates of heat-treated tumor cells and were used to immunize BALB/c mice with established colon carcinoma. Cytotoxic T lymphocyte (CTL) response was detected. The therapeutic effect induced by DCs was observed by tumor weight and survival time. DCs pulsed with lysates of heat-treated tumor cells markedly induced specific cytotoxic activity of CTLs. Tumor growth in the immunized BALB/c mice was significantly inhibited and the survival time of the tumor-bearing mice was prolonged, DCs pulsed with lysates of heat-treated tumor cells have an observable therapeutic effect on established colon carcinoma-bearing mice.

  6. Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products.

    Directory of Open Access Journals (Sweden)

    Günter Klein

    Full Text Available Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS has been used as a multidimensional and high throughput method for the identification and validation of biomarkers of disease in complex biological samples such as biofluids. In this study we evaluate the suitability of CE/MS to measure the consistency of different lots of the probiotic formulation Pro-Symbioflor which is a bacterial lysate of heat-inactivated Escherichia coli and Enterococcus faecalis. Over 5000 peptides were detected by CE/MS in 5 different lots of the bacterial lysate and in a sample of culture medium. 71 to 75% of the total peptide content was identical in all lots. This percentage increased to 87-89% when allowing the absence of a peptide in one of the 5 samples. These results, based on over 2000 peptides, suggest high similarity of the 5 different lots. Sequence analysis identified peptides of both E. coli and E. faecalis and peptides originating from the culture medium, thus confirming the presence of the strains in the formulation. Ontology analysis suggested that the majority of the peptides identified for E. coli originated from the cell membrane or the fimbrium, while peptides identified for E. faecalis were enriched for peptides originating from the cytoplasm. The bacterial lysate peptides as a whole are recognised as highly conserved molecular patterns by the innate immune system as microbe associated molecular pattern (MAMP. Sequence analysis also identified the presence of soybean, yeast and casein protein fragments that are part of the formulation of the culture medium. In conclusion CE/MS seems an appropriate QC tool to analyze complex biological products such as inactivated probiotic formulations and allows determining the similarity between

  7. Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products.

    Science.gov (United States)

    Klein, Günter; Schanstra, Joost P; Hoffmann, Janosch; Mischak, Harald; Siwy, Justyna; Zimmermann, Kurt

    2013-01-01

    Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC) should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS) has been used as a multidimensional and high throughput method for the identification and validation of biomarkers of disease in complex biological samples such as biofluids. In this study we evaluate the suitability of CE/MS to measure the consistency of different lots of the probiotic formulation Pro-Symbioflor which is a bacterial lysate of heat-inactivated Escherichia coli and Enterococcus faecalis. Over 5000 peptides were detected by CE/MS in 5 different lots of the bacterial lysate and in a sample of culture medium. 71 to 75% of the total peptide content was identical in all lots. This percentage increased to 87-89% when allowing the absence of a peptide in one of the 5 samples. These results, based on over 2000 peptides, suggest high similarity of the 5 different lots. Sequence analysis identified peptides of both E. coli and E. faecalis and peptides originating from the culture medium, thus confirming the presence of the strains in the formulation. Ontology analysis suggested that the majority of the peptides identified for E. coli originated from the cell membrane or the fimbrium, while peptides identified for E. faecalis were enriched for peptides originating from the cytoplasm. The bacterial lysate peptides as a whole are recognised as highly conserved molecular patterns by the innate immune system as microbe associated molecular pattern (MAMP). Sequence analysis also identified the presence of soybean, yeast and casein protein fragments that are part of the formulation of the culture medium. In conclusion CE/MS seems an appropriate QC tool to analyze complex biological products such as inactivated probiotic formulations and allows determining the similarity between lots. PMID

  8. Comparative Analysis of Click Chemistry Mediated Activity-Based Protein Profiling in Cell Lysates

    Directory of Open Access Journals (Sweden)

    Yinliang Yang

    2013-10-01

    Full Text Available Activity-based protein profiling uses chemical probes that covalently attach to active enzyme targets. Probes with conventional tags have disadvantages, such as limited cell permeability or steric hindrance around the reactive group. A tandem labeling strategy with click chemistry is now widely used to study enzyme targets in situ and in vivo. Herein, the probes are reacted in live cells, whereas the ensuing detection by click chemistry takes place in cell lysates. We here make a comparison of the efficiency of the activity-based tandem labeling strategy by using Cu(I-catalyzed and strain-promoted click chemistry, different ligands and different lysis conditions.

  9. [Use of reactions with Limulus amoebocyte lysate (LAL) to determine biological activity of lipopolysaccharides from reference and clinical strains of the Bacteroides fragilis group].

    Science.gov (United States)

    Rokosz, Alicja; Fiejka, Maria; Górska, Paulina; Aleksandrowicz, Janina; Meisel-Mikołajczyk, Felicja; Łuczak, MirosŁaw

    2002-01-01

    The aim of this study was to determine and compare a biological activity of lipopolysaccharides (LPS) from reference and clinical strains of strictly anaerobic bacteria belonging to the Bacteroides fragilis group (BFG) by means of quantitative, photometric BET (LAL) method with Limulus polyphemus amoebocyte lysate and chromogenic substrate S-2423. Lipopolysaccharides of five BFG species were extracted by Westphal and Jann method (1965) from eight reference and two clinical strains of B. fragilis group. Crude LPS preparations were purified according to the procedure described by Gmeiner (1975) with ultracentrifugation and nuclease treatment. Biological activities of bacterial endotoxins were determined by quantitative BET method with chromogenic substrate S-2423 (ENDOCHROME kit, Charles River Endosafe Ltd., USA). Tests were performed according to the producer's recommendations. E. coli O55:B5 LPS was applied to compare its activity in reaction with LAL reagent with activities of LPS preparations from rods of the Bacteroides genus. Among examined bacterial compounds the most active in BET method was E. coli O55:B5 LPS. Activities of lipopolysaccharides from five species of BFG rods in reaction with Limulus amoebocyte lysate were differentiated. Greater ability to activate LAL proenzyme revealed lipopolysaccharides of these species of the Bacteroides genus, which are important from the clinical point of view--B. fragilis and B. thetaiotaomicron.

  10. Is buffer a good proxy for a crowded cell-like environment? A comparative NMR study of calmodulin side-chain dynamics in buffer and E. coli lysate.

    Directory of Open Access Journals (Sweden)

    Michael P Latham

    Full Text Available Biophysical studies of protein structure and dynamics are typically performed in a highly controlled manner involving only the protein(s of interest. Comparatively fewer such studies have been carried out in the context of a cellular environment that typically involves many biomolecules, ions and metabolites. Recently, solution NMR spectroscopy, focusing primarily on backbone amide groups as reporters, has emerged as a powerful technique for investigating protein structure and dynamics in vivo and in crowded "cell-like" environments. Here we extend these studies through a comparative analysis of Ile, Leu, Val and Met methyl side-chain motions in apo, Ca(2+-bound and Ca(2+, peptide-bound calmodulin dissolved in aqueous buffer or in E. coli lysate. Deuterium spin relaxation experiments, sensitive to pico- to nano-second time-scale processes and Carr-Purcell-Meiboom-Gill relaxation dispersion experiments, reporting on millisecond dynamics, have been recorded. Both similarities and differences in motional properties are noted for calmodulin dissolved in buffer or in lysate. These results emphasize that while significant insights can be obtained through detailed "test-tube" studies, experiments performed under conditions that are "cell-like" are critical for obtaining a comprehensive understanding of protein motion in vivo and therefore for elucidating the relation between motion and function.

  11. Fast and mild strategy, using superhydrophobic surfaces, to produce collagen/platelet lysate gel beads for skin regeneration.

    Science.gov (United States)

    Lima, Ana Catarina; Mano, João F; Concheiro, Angel; Alvarez-Lorenzo, Carmen

    2015-02-01

    Platelet lysate (PL) was encapsulated in collagen (Coll) millimetric gel beads, on biomimetic superhydrophobic surfaces, under mild conditions, with the aim of obtaining easy-to-handle formulations able to provide sustained release of multiple growth factors for skin ulcers treatment. The gel particles were prepared with various concentrations of PL incorporating or not stem cells, and tested as freshly prepared or after being freeze-dried or cryopreserved. Coll + PL particles were evaluated regarding degradation in collagenase-rich environment (simulating the aggressive environment of the chronic ulcers), sustained release of total protein, PDGF-BB and VEGF, cell proliferation (using particles as the only source of growth factors), scratch wound recovery and angiogenic capability. Compared to Coll solely particles, incorporation of PL notably enhanced cell proliferation (inside and outside gels) and favored scratch wound recovery and angiogenesis. Moreover, cell-laden gel particles containing PL notably improved cell proliferation and even migration of cells from one particle towards a neighbor one, which led to cell-cell contacts and the spontaneous formation of tissue layers in which the spherical gels were interconnected by the stem cells. PMID:25120225

  12. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture; Cultivo e irradiacao de fibroblastos humanos em meio enriquecido com lisado de plaquetas para obtencao de camada de sustentacao em culturas de celulas da epiderme

    Energy Technology Data Exchange (ETDEWEB)

    Yoshito, Daniele

    2011-07-01

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  13. The effect of chitosan nanospheres on the immunogenicity of Toxoplasma lysate vaccine in mice.

    Science.gov (United States)

    El Temsahy, Mona M; El Kerdany, Eman D H; Eissa, Maha M; Shalaby, Thanaa I; Talaat, Iman M; Mogahed, Nermine M F H

    2016-09-01

    Toxoplasmosis, a zoonotic parasitic disease, is a huge challenge for which there is no effective vaccine up till now. In this study, chitosan nanospheres encapsulated with Toxoplasma lysate vaccine was evaluated for its ability to protect mice against both acute and chronic toxoplasmosis models of infection. Results showed that chitosan nanospheres were equally effective to Freund's incomplete adjuvant (FIA) in enhancing the efficacy of Toxoplasma lysate vaccine. The effectiveness was demonstrated by the delayed death of vaccinated mice following challenge either with virulent RH or avirulent Me49 strains, the significant decrease in parasite density in different organs, significant increase in the humoral and cellular immune response (IgG and IFN γ) with a marked reduction of pathological changes in the different organs. However chitosan nanospheres were superior to FIA due to their cost effective preparation and much less necrotic changes induced in the studied organs. The success of chitosan polymer as an alternative to commonly used adjuvants paves the way for the use of other newly developed polymers to be used in the field of vaccine development. PMID:27605755

  14. Naegleria fowleri lysate induces strong cytopathic effects and pro-inflammatory cytokine release in rat microglial cells.

    Science.gov (United States)

    Lee, Yang-Jin; Park, Chang-Eun; Kim, Jong-Hyun; Sohn, Hae-Jin; Lee, Jinyoung; Jung, Suk-Yul; Shin, Ho-Joon

    2011-09-01

    Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A (51)Cr release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-α, IL-6, and IL-1β, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response.

  15. Naegleria fowleri Lysate Induces Strong Cytopathic Effects and Pro-inflammatory Cytokine Release in Rat Microglial Cells

    Science.gov (United States)

    Lee, Yang-Jin; Park, Chang-Eun; Kim, Jong-Hyun; Sohn, Hae-Jin; Lee, Jinyoung; Jung, Suk-Yul

    2011-01-01

    Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A 51Cr release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-α, IL-6, and IL-1β, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response. PMID:22072830

  16. Protective immune-response of aluminium hydroxide gel adjuvanted phage lysate of Brucella abortus S19 in mice against direct virulent challenge with B. abortus 544.

    Science.gov (United States)

    Jain, Lata; Rawat, Mayank; Prajapati, Awadhesh; Tiwari, Ashok Kumar; Kumar, Bablu; Chaturvedi, V K; Saxena, H M; Ramakrishnan, Sarvanan; Kumar, Jatin; Kerketta, Priscilla

    2015-09-01

    The prophylactic efficacies of plain and alum adsorbed lysate were evaluated by direct virulent challenge in mice model. A recently isolated brucellaphage 'ϕLd' was used for generation of lysates. Twenty four h incubated Brucella abortus S19 broth cultures standardized to contain approximately 10(8) CFU/ml were found suitable for generation of lysates. Three lysate batches produced through separate cycles did not show any significant variation with respect to protein and polysaccharide contents, endotoxin level and phage counts, indicating that compositionally stable lysate preparations can be generated through an optimized production process. Three polypeptides of ∼16, 19 and 23 kDa could be identified as immuno-dominant antigens of the lysate which induced both humoral and cell-mediated immune responses in a dose dependent manner. Results of efficacy evaluation trial confirmed dose-dependent protective potencies of lysate preparation. The lysate with an antigenic dose of 0.52 μg protein and 60 μg CHO adsorbed on aluminium gel (0.1 percent aluminium concentration) exhibited the highest protective potency which was greater than that induced by standard S19 vaccine. Phage lysate methodology provides a very viable option through which an improved immunizing preparation with all desirable traits can be developed against brucellosis, and integrated with immunization programmes in a more efficient manner. PMID:26156404

  17. Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate.

    Science.gov (United States)

    Sokolova, Ekaterina; Spruijt, Evan; Hansen, Maike M K; Dubuc, Emilien; Groen, Joost; Chokkalingam, Venkatachalam; Piruska, Aigars; Heus, Hans A; Huck, Wilhelm T S

    2013-07-16

    Liquid-liquid phase transitions in complex mixtures of proteins and other molecules produce crowded compartments supporting in vitro transcription and translation. We developed a method based on picoliter water-in-oil droplets to induce coacervation in Escherichia coli cell lysate and follow gene expression under crowded and noncrowded conditions. Coacervation creates an artificial cell-like environment in which the rate of mRNA production is increased significantly. Fits to the measured transcription rates show a two orders of magnitude larger binding constant between DNA and T7 RNA polymerase, and five to six times larger rate constant for transcription in crowded environments, strikingly similar to in vivo rates. The effect of crowding on interactions and kinetics of the fundamental machinery of gene expression has a direct impact on our understanding of biochemical networks in vivo. Moreover, our results show the intrinsic potential of cellular components to facilitate macromolecular organization into membrane-free compartments by phase separation. PMID:23818642

  18. Electrical protein detection in cell lysates using high-density peptide-aptamer microarrays

    Directory of Open Access Journals (Sweden)

    Evans David

    2008-01-01

    Full Text Available Abstract Background The dissection of biological pathways and of the molecular basis of disease requires devices to analyze simultaneously a staggering number of protein isoforms in a given cell under given conditions. Such devices face significant challenges, including the identification of probe molecules specific for each protein isoform, protein immobilization techniques with micrometer or submicrometer resolution, and the development of a sensing mechanism capable of very high-density, highly multiplexed detection. Results We present a novel strategy that offers practical solutions to these challenges, featuring peptide aptamers as artificial protein detectors arrayed on gold electrodes with feature sizes one order of magnitude smaller than existing formats. We describe a method to immobilize specific peptide aptamers on individual electrodes at the micrometer scale, together with a robust and label-free electronic sensing system. As a proving proof of principle experiment, we demonstrate the specific recognition of cyclin-dependent protein kinases in whole-cell lysates using arrays of ten electrodes functionalized with individual peptide aptamers, with no measurable cross-talk between electrodes. The sensitivity is within the clinically relevant range and can detect proteins against the high, whole-cell lysate background. Conclusion The use of peptide aptamers selected in vivo to recognize specific protein isoforms, the ability to functionalize each microelectrode individually, the electronic nature and scalability of the label-free detection and the scalability of the array fabrication combine to yield the potential for highly multiplexed devices with increasingly small detection areas and higher sensitivities that may ultimately allow the simultaneous monitoring of tens or hundreds of thousands of protein isoforms.

  19. Vaccination with melanoma lysate-pulsed dendritic cells, of patients with advanced colorectal carcinoma: report from a phase I study

    DEFF Research Database (Denmark)

    Burgdorf, S K; Fischer, A; Claesson, M H;

    2006-01-01

    Immune therapy have shown new and exciting perspectives for cancer treatment. Aim of our study was to evaluate toxicity and possible adverse effects from vaccination of patients with advanced colorectal cancer with autologous dendritic cells (DC) pulsed with lysate from a newly developed melanoma...

  20. Intranasal Coadministration of the Cry1Ac Protoxin with Amoebal Lysates Increases Protection against Naegleria fowleri Meningoencephalitis

    Science.gov (United States)

    Rojas-Hernández, Saúl; Rodríguez-Monroy, Marco A.; López-Revilla, Rubén; Reséndiz-Albor, Aldo A.; Moreno-Fierros, Leticia

    2004-01-01

    Cry1Ac protoxin has potent mucosal and systemic adjuvant effects on antibody responses to proteins or polysaccharides. In this work, we examined whether Cry1Ac increased protective immunity against fatal Naegleria fowleri infection in mice, which resembles human primary amoebic meningoencephalitis. Higher immunoglobulin G (IgG) than IgA anti-N. fowleri responses were elicited in the serum and tracheopulmonary fluids of mice immunized by the intranasal or intraperitoneal route with N. fowleri lysates either alone or with Cry1Ac or cholera toxin. Superior protection against a lethal challenge with 5 × 104 live N. fowleri trophozoites was achieved for immunization by the intranasal route. Intranasal immunization of N. fowleri lysates coadministered with Cry1Ac increased survival to 100%; interestingly, immunization with Cry1Ac alone conferred similar protection to that achieved with amoebal lysates alone (60%). When mice intranasally immunized with Cry1Ac plus lysates were challenged with amoebae, both IgG and IgA mucosal responses were rapidly increased, but only the increased IgG response persisted until day 60 in surviving mice. The brief rise in the level of specific mucosal IgA does not exclude the role that this isotype may play in the early defense against this parasite, since higher IgA responses were detected in nasal fluids of mice intranasally immunized with lysates plus either Cry1Ac or cholera toxin, which, indeed, were the treatments that provided the major protection levels. In contrast, serum antibody responses do not seem to be related to the protection level achieved. Both acquired and innate immune systems seem to play a role in host defense against N. fowleri infection, but further studies are required to elucidate the mechanisms involved in protective effects conferred by Cry1Ac, which may be a valuable tool to improve mucosal vaccines. PMID:15271892

  1. THE USE OF PREPARATIONS BASED ON BACTERIAL LYSATES IN THE TREATMENT OF CHRONIC TONSILLITIS IN PATIENTS WITH RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    Kolyada T.І.

    2014-12-01

    Full Text Available In the therapy of decompensated form of chronic tonsillitis (CT were used as immunomodulatory agents IRS and Ismigen. These bacterial lysates differ in the bacterial setting, the method of preparation (chemical, mechanical and the method of application.Rheumatoid arthritis (RA is one of the important factors that could significantly complicate the therapy of chronic tonsillitis. RA is a chronic immune inflammatory disease that progressively affects connective tissue mostly of the peripheral joints and it has a wide range of extra-articular manifestations. The aim of our study was to explore the dynamics of immunologic indicators during the active disease and treatments in patients with decompensate form of chronic tonsillitis, including tonsillitis complicated with RA. Materials and methods. 33 patients with decompensate form of chronic tonsillitis in active period of disease observed during the study. Patients were divided into the following groups: 24 persons with the decompensate form of CT, 9 persons with the rheumatoid arthritis and 9 persons with the decompensate form of CT complicated with RA in remission stage. The control group consisted of 15 apparently healthy persons. Concentrations of serum IgA, IgM, IgG were determined by the method of radial immune diffusion by Manchini. Levels of sIgA, IFN – γ and rheumatoid factor in the blood serum of patients were evaluated using ELISA test systems of "Vector-best". Patients of group CTD (with decompensate form of chronic tonsillitis were divided into subgroups CTD1 and CTD2, depending on the applied treatment. Both subgroups treated with standard therapy for two weeks and received Derynat during 1 month by 2 drops in each nostril twice a day. After 30 days of the standard therapy beginning the subgroup CTD1 patients received IRS 19 during two weeks, one intranasal inhalation in each nostril 3 times a day. Patients subgroup CTD2 and CTD+RA instead IRS 19 received Ismigen after 30 days of

  2. Influence of various dust sampling and extraction methods on the measurement of airborne endotoxin.

    OpenAIRE

    Douwes, J; Versloot, P.; Hollander, A; Heederik, D; Doekes, G.

    1995-01-01

    The influence of various filter types and extraction conditions on the quantitation of airborne endotoxin with the Limulus amebocyte lysate test was studied by using airborne dusts sampled in a potato processing plant. Samples were collected with an apparatus designed to provide parallel samples. Data from the parallel-sampling experiment were statistically evaluated by using analysis of variance. In addition, the influence of storage conditions on the detectable endotoxin concentration was i...

  3. Grain and bean lysates improve function of endothelial progenitor cells from human peripheral blood: involvement of the endogenous antioxidant defenses.

    Directory of Open Access Journals (Sweden)

    Daniela Lucchesi

    Full Text Available Increased oxidative stress contributes to the functional impairment of endothelial progenitor cells (EPCs, the pivotal players in the servicing of the endothelial cell lining. Several evidences suggest that decreasing oxidative stress by natural compounds with antioxidant properties may improve EPCs bioactivity. Here, we investigated the effects of Lisosan G (LG, a Triticum Sativum grain powder, and Lady Joy (LJ, a bean lysate, on function of EPCs exposed to oxidative stress. Peripheral blood mononuclear cells were isolated and plated on fibronectin-coated culture dishes; adherent cells, identified as early EPCs, were pre-treated with different concentrations of LG and LJ and incubated with hydrogen peroxide (H2O2. Viability, senescence, adhesion, ROS production and antioxidant enzymes gene expression were evaluated. Lysate-mediated Nrf-2 (nuclear factor (erythroid-derived 2-like 2/ARE (antioxidant response element activation, a modulator of oxidative stress, was assessed by immunocytochemistry. Lady Joy 0.35-0.7 mg/ml increases EPCs viability; pre-treatment with either LG 0.7 mg/ml and LJ 0.35-0.7 mg/ml protect EPCs viability against H2O2-induced injury. LG 0.7 and LJ 0.35-0.7 mg/ml improve EPCs adhesion; pre-treatment with either LG 0.35 and 0.7 mg/ml or LJ 0.35, 0.7 and 1.4 mg/ml preserve adhesiveness of EPCs exposed to H2O2. Senescence is attenuated in EPCs incubated with lysates 0.35 mg/ml. After exposure to H2O2, LG pre-treated cells show a lower senescence than untreated EPCs. Lysates significantly decrease H2O2-induced ROS generation. Both lysates increase glutathione peroxidase-1 and superoxide dismutase-2 (SOD-2 expression; upon H2O2 exposure, pre-treatment with LJ allows higher SOD-2 expression. Heme oxigenase-1 increases in EPCs pre-treated with LG even upon H2O2 exposure. Finally, incubation with LG 0.7 mg/ml results in Nrf-2 translocation into the nucleus both at baseline and after the oxidative challenge. Our data suggest a

  4. Topical gel formulation and stability assessment of platelet lysate based on turbidimetric method

    Directory of Open Access Journals (Sweden)

    Soliman Mohammadi Samani

    2015-06-01

    Full Text Available Platelet-rich growth factors have attracted attentions of scientists and clinical practitioners who are involved in wound healing and regenerative medicine extensively, according to their unprecedented potential of promoting and catalyzing healing process. Platelet-rich growth factors are cost-benefit, available and more stable than recombinant human growth factors. These appealing characteristics have converted PRGF to one of the popular candidates for treatment of variety of wounds. According to these valuable properties, we decided to formulate and assess the effect of different excipients on the stability of such valuable protein based formulations. Different excipients have been chosen according to their effective ness on the stability of proteins and their application in other similar formulations. The stabilizing effect of excipients was evaluated by measuring heat-induced aggregation of growth factors by turbidimetric assay. Glycerol, glycine and dextrose were chosen as stabilizing excipients for these formulations. The results show that dextrose has more stabilizing effect on prevention of heat induced aggregation of the platelet lysate growth factors than glycerol and glycine. All of the formulations also contained antioxidant, chelating agents, preservative and carbopol934 in order to form appropriate gel.

  5. An efficient method for refolding the extracellular portion of CD147 from the total bacterial lysate

    Institute of Scientific and Technical Information of China (English)

    Fei Song; Xin Zhang; Yifei Li; Qiang Ru; Xiaobai Ren; Bin Xia; Zhi-Nan Chen

    2011-01-01

    CD147 is a widely expressed transmembrane protein that mediates signal transduction,and it plays important roles in many physiological and pathological processes,such as tumor invasion and metastasis.The extracellular portion of CD147 (CD147EC) is responsible for its functional interactions with different signaling molecules.Due to the existence of two disulfide bonds,CD147EC is mainly expressed as an inclusion body in Escherichia coli.Here,we report a convenient rapid-dilution refolding protocol that enables the refolding of CD147EC efficiently from total bacterial lysate instead of pure inclusion bodies.Using this method,over 25 mg of CD147EC can be purified from 1 1 of bacterial culture in M9 medium.The refolded CD147EC is well folded as characterized by nuclear magnetic resonance (NMR),and it can induce the expression of matrix metalloproteinase-9 in fibroblast cells.The described protocol is also applicable to the refolding of two immunoglobulin domains of CD147EC individually.Interestingly,we noticed that little protein was produced for the C-terminal immunoglobulin (Ig) domain of CD147EC by bacteria in M9 medium,even though it was overexpressed in LuriaBertani (LB) medium.However,when the pH of the bacterial culture in M9 medium was adjusted in accordance with that in LB medium during growth,comparable expression level could be achieved.

  6. Clinical Benefit of Allogeneic Melanoma Cell Lysate-Pulsed Autologous Dendritic Cell Vaccine in MAGE-Positive Colorectal Cancer Patients

    DEFF Research Database (Denmark)

    Toh, Han Chong; Wang, Who-Whong; Chia, Whay Kuang;

    2009-01-01

    patients received a total of 161 vaccinations. Treatment was well tolerated and quality of life measurements did not vary much across time. One patient experienced partial response [5%; 95% confidence interval (CI), 1-24%] and seven achieved stable disease (35%; 95% CI, 18-57%), one of whom also achieved......PURPOSE: We evaluated the clinical benefit of an allogeneic melanoma cell lysate (MCL)-pulsed autologous dendritic cell (DC) vaccine in advanced colorectal cancer patients expressing at least one of six MAGE-A antigens overexpressed by the cell line source of the lysate. EXPERIMENTAL DESIGN: DCs...... were cultured from peripheral blood mononuclear cells (PBMC), pulsed with the allogeneic MCL, and matured using cytokines that achieved high CD83- and CCR7-expressing DCs. Each patient received up to 10 intradermal vaccinations (3-5 x 10(6) cells per dose) at biweekly intervals. RESULTS: Twenty...

  7. Estrogen receptor α determination in serum, cell lysates and breast cancer cells using an amperometric magnetoimmunosensing platform

    Directory of Open Access Journals (Sweden)

    U. Eletxigerra

    2016-03-01

    Full Text Available An electrochemical magnetoimmunosensor for the determination of estrogen receptor α (ERα protein in complex samples (serum and cell lysates able to discriminate between ERα positive and negative breast cancer cells is reported. Specifically functionalized magnetic microbeads with sandwich immunocomplexes and amperometric detection at disposable screen-printed carbon electrodes (SPCEs resulted in highly selective and sensitive ERα detection with a detection limit of 19 pg mL−1. This magnetoimmunosensing platform was successfully applied to the quantitation of ERα in spiked human serum and cell lysates samples without any matrix effect with an advantageous performance in terms of simplicity and assay times over commercial ELISA assays. The biosensor capability for assessing ERα in intact breast cancer cells makes it competitive with conventional strategies providing rapidly quantitative and reliable results on this relevant biomarker currently used in the clinical practice for diagnosis, follow-up and monitoring of metastatic breast cancer.

  8. Autologous Tumor Lysate-pulsed Dendritic Cell Immunotherapy for Pediatric Patients with Newly Diagnosed or Recurrent High-grade Gliomas

    OpenAIRE

    Lasky, Joseph L.; Panosyan, Eduard H.; Plant, Ashley; Davidson, Tom; Yong, William H.; Robert M Prins; Liau, Linda M.; Moore, Theodore B.

    2013-01-01

    Immunotherapy has the potential to improve clinical outcomes with little toxicity for pediatric patients with brain tumors. We conducted a pilot feasibility study of tumor lysate-pulsed dendritic cell (DC) vaccination in pediatric patients (1 to 18 years old) with newly diagnosed or recurrent high-grade glioma (HGG). A total of nine DC vaccine doses, each containing 1×106 cells per dose were administered to three out of the seven originally enrolled patients. Toxicities were limited to mild s...

  9. LPS-Activated Monocytes Are Unresponsive to T4 Phage and T4-Generated Escherichia coli Lysate.

    Science.gov (United States)

    Bocian, Katarzyna; Borysowski, Jan; Zarzycki, Michał; Wierzbicki, Piotr; Kłosowska, Danuta; Weber-Dąbrowska, Beata; Korczak-Kowalska, Grażyna; Górski, Andrzej

    2016-01-01

    A growing body of data shows that bacteriophages can interact with different kinds of immune cells. The objective of this study was to investigate whether T4 bacteriophage and T4-generated Escherichia coli lysate affect functions of monocytes, the key population of immune cells involved in antibacterial immunity. To that end, we evaluated how T4 and E. coli lysate influence the expression of main costimulatory molecules including CD40, CD80 and CD86, TLR2, TLR4 on monocytes, as well as the production of IL-6 and IL-12 in cultures of peripheral blood mononuclear cells (PBMCs). Separate experiments were performed on unactivated and LPS-activated PBMCs cultures. Both studied preparations significantly increased the percentage of CD14(+)CD16(-)CD40(+) and CD14(+)CD16(-)CD80(+) monocytes in unactivated PBMCs cultures, as well as the concentration of IL-6 and IL-12 in culture supernates. However, neither purified T4 nor E. coli lysate had any significant effect on monocytes in LPS-activated PBMCs cultures. We conclude that LPS-activated monocytes are unresponsive to phages and products of phage-induced lysis of bacteria. This study is highly relevant to phage therapy because it suggests that in patients with infections caused by Gram-negative bacteria the administration of phage preparations to patients and lysis of bacteria by phages are not likely to overly stimulate monocytes. PMID:27630621

  10. Engineering Enriched Microenvironments with Gradients of Platelet Lysate in Hydrogel Fibers.

    Science.gov (United States)

    Santo, Vítor E; Babo, Pedro; Amador, Miguel; Correia, Cláudia; Cunha, Bárbara; Coutinho, Daniela F; Neves, Nuno M; Mano, João F; Reis, Rui L; Gomes, Manuela E

    2016-06-13

    Gradients of physical and chemical cues are characteristic of specific tissue microenvironments and contribute toward morphogenesis and tissue regeneration upon injury. Recent advances on microfluidics and hydrogel manipulation raised the possibility of generating biomimetic biomaterials enriched with bioactive factors and encapsulating cells following designs specifically tailored for a target application. The novelty of this work relies on the combination of methacrylated gellan gum (MeGG) with platelet lysate (PL), aiming to generate novel advanced 3D PL-enriched photo-cross-linkable hydrogels and overcoming the lack of adhesion sites provided by the native MeGG hydrogels. This combination takes advantage of the availability, enriched growth factor composition, and potential autologous application of PL while simultaneously preserving the ability provided by MeGG to tailor mechanical properties, protein release kinetics, and shape of the construct according to the desired goal. Incorporation of PL in the hydrogels significantly improved cellular adhesion and viability in the constructs. The use of microfluidic tools allowed the design of a fiber-like hydrogel incorporating a gradient of PL along the length of the fiber. These spatial protein gradients led to the viability and cell number gradients caused by maintenance of human umbilical vein endothelial cells (HUVECs) survival in the fibers toward the PL-enriched sections in comparison with the nonloaded MeGG sections of the fibers. Altogether, we propose a proof of concept strategy to design a PL gradient biomaterial with potential in tissue engineering approaches and analysis of cell-microenvironment interactions. PMID:27203709

  11. Heat-shocked tumor cell lysate-pulsed dendritic cells induce effective anti-tumor immune response in vivo

    Institute of Scientific and Technical Information of China (English)

    Jian Qiu; Guo-Wei Li; Yan-Fang Sui; Hong-Ping Song; Shao-Yan Si; Wei Ge

    2006-01-01

    AIM: To study whether heat-shocked tumor cells could enhance the effect of tumor cell lysate-pulsed dendritic cells (DCs) in evoking anti-tumor immune response in vivo.METHODS: Mouse undifferentiated colon cancer cells(CT-26) were heated at 42℃ for 1 h and then frozenthawed. The bone marrow-derived DCs pulsed with heatshocked CT-26 cell lysate (HSCT-26 DCs) were recruited to immunize syngeneic naive BALB/c mice. The cytotoxic activity of tumor specific cytotoxic T lymphocytes (CTLs)in mouse spleen was evaluated by IFN-enzyme-linked immunospot (ELISpot) and LDH release assay. The immunoprophylactic effects induced by HSCT-26 DCs in mouse colon cancer model were compared to those induced by single CT-26 cell lysate-pulsed DCs (CT-26DCs) on tumor volume, peritoneal metastasis and survival time of the mice.RESULTS: Heat-treated CT-26 cells showed a higher hsp70 protein expression. Heat-shocked CT-26 cell lysate pulsing elevated the co-stimulatory and MHC-Ⅱ molecule expression of bone marrow-derived DCs as well as interleukin-12 p70 secretion. The IFN-γ secreting CTLs induced by HSCT-26 DCs were significantly more than those induced by CT-26 DCs (P= 0.002). The former CTLs' specific cytotoxic activity was higher than the latter CTLs' at a serial E/T ratio of 10:1, 20:1, and 40:1. Mouse colon cancer model showed that the tumor volume of HSCT-26 DC vaccination group was smaller than that of CT-26 DC vaccination group on tumor volume though there was no statistical difference between them(24 mm3 vs 8 mm3, P= 0.480). The median survival time of mice immunized with HSCT-26 DCs was longer than that of those immunized with CT-26 DCs (57 d vs 43 d,P= 0.0384).CONCLUSION: Heat-shocked tumor cell lysate-pulsed DCs can evoke anti-tumor immune response in vivo effectively and serve as a novel DC-based tumor vaccine.

  12. Immune response and functional role of antibodies raised in heifers against a Staphylococcus aureus CP5 lysate and recombinant antigens vaccine formulated with Iscom Matrix adjuvant.

    Science.gov (United States)

    Camussone, C M; Pujato, N; Renna, M S; Veaute, C M; Morein, B; Marcipar, I S; Calvinho, L F

    2014-12-15

    Staphylococcus aureus is the most frequently isolated pathogen from bovine intramammary infections worldwide. Commercially available vaccines for mastitis control are composed either of S. aureus lysates or inactivated whole-cells formulated with traditional adjuvants. We recently showed the ability of a S. aureus CP5 lysate vaccine adjuvanted with Iscom Matrix to generate a longer lasting specific antibody response in blood and milk, with improved opsonic capacity, compared with a S. aureus CP5 whole-cell formulation. The aim of the present study was to obtain an experimental immunogen composed of lysed cells of a CP5 S. aureus strain supplemented with recombinant clumping factor A, fibronectin binding protein A and β-toxin formulated with Iscom Matrix, characterize the immune response generated when immunizing pregnant heifers and assess the functional role of antibodies raised against this immunogen in experimental models. Both a lysate vaccine and a lysate+recombinant antigens vaccine elicited antibodies that promoted neutrophil phagocytosis and inhibited internalization into mammary epithelial cells, in vitro. Incorporation of defined antigenic molecules to the lysate formulation elicited a strong specific humoral immune response against both lysate and recombinant antigens and was associated with higher expression of regulatory and pro-inflammatory cytokines. In addition, antibodies were efficient for blocking S. aureus binding to bovine fibrinogen and fibronectin, and neutralizing β-toxin effect in vitro, placing these antigens as candidates to be included in a formulation directed to prevent staphylococcal bovine mastitis. PMID:25454469

  13. Direct quantification of mRNA and miRNA from cell lysates using reverse transcription real time PCR: a multidimensional analysis of the performance of reagents and workflows.

    Directory of Open Access Journals (Sweden)

    Yoon Khei Ho

    Full Text Available Substantial efforts have been devoted to in vitro testing of candidate chemotherapeutics by profiling transcriptional changes across the collection of NCI-60 cell-lines. A work-flow with reagents that enable the direct quantification of RNA of different molecular sizes simultaneously in the same sample without laborious total RNA isolation will invariably increase the throughput and accuracy of the study. MicroRNAs (miRNAs are known to regulate most cellular functions, acting post-transcriptionally by repressing numerous eukaryotic mRNAs. Recent findings on the remarkable stability of miRNA prompted us to investigate the feasibility of quantifying the expression levels of both mRNA and miRNA directly from cell lysates (cell-to-Ct. Multidimensional analyses of the expressions of mRNA and miRNA across seven NCI-60 cell lines and multiple reagents were conducted to assess the performances of these reagents and workflows for cell-to-Ct measurements using reverse transcription-quantitative polymerase chain reaction (RT-qPCR. Quantification of RNA species using lysates prepared from an in-house and one of the commercial reagents demonstrated comparable performance to those prepared by the more laborious and conventional method of using guanidinium-phenol-chloroform. Additionally, miRNA was found to be highly stable in the cell lysates when incubated at room temperature for prolonged period of time and subjected to multiple freeze-thaw cycles. In summary, this study demonstrated significant differences in pre-analytical performance of a variety of commercially available reagents and described a cost-effective reagent useful for rapid, scalable, and high-throughput workflow for the detection of mRNA and miRNA from the same biological sample.

  14. Dendritic cells loaded with pancreatic Cancer Stem Cells (CSCs lysates induce antitumor immune killing effect in vitro.

    Directory of Open Access Journals (Sweden)

    Tao Yin

    Full Text Available According to the cancer stem cells (CSCs theory, malignant tumors may be heterogeneous in which a small population of CSCs drive the progression of cancer. Because of their intrinsic abilities, CSCs may survive a variety of treatments and then lead to therapeutic resistance and cancer recurrence. Pancreatic CSCs have been reported to be responsible for the malignant behaviors of pancreatic cancer, including suppression of immune protection. Thus, development of immune strategies to eradicate pancreatic CSCs may be of great value for the treatment of pancreatic cancer. In this study, we enriched pancreatic CSCs by culturing Panc-1 cells under sphere-forming conditions. Panc-1 CSCs expressed low levels of HLA-ABC and CD86, as measured by flow cytometry analysis. We further found that the Panc-1 CSCs modulate immunity by inhibiting lymphocyte proliferation which is promoted by phytohemagglutinin (PHA and anti-CD3 monoclonal antibodies. The monocyte derived dendritic cells (DCs were charged with total lysates generated from Panc-1 CSCs obtained from tumor sphere culturing. After co-culturing with lymphocytes at different ratios, the Panc-1 CSCs lysates modified DC effectively promoted lymphocyte proliferation. The activating efficiency reached 72.4% and 74.7% at the ratios of 1∶10 and 1∶20 with lymphocytes. The activated lymphocytes secreted high levels of INF-γ and IL-2, which are strong antitumor cytokines. Moreover, Panc-1 CSCs lysates modified DC induced significant cytotoxic effects of lymphocytes on Panc-1 CSCs and parental Panc-1 cells, respectively, as shown by lactate dehydrogenase (LDH assay. Our study demonstrates that the development of CSCs-based vaccine is a promising strategy for treating pancreatic cancer.

  15. Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: an improved approach in cartilage tissue engineering.

    Science.gov (United States)

    Moroz, Andrei; Bittencourt, Renata Aparecida Camargo; Almeida, Renan Padron; Felisbino, Sérgio Luis; Deffune, Elenice

    2013-01-01

    Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n = 5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1 × 10(5)) were than encapsulated inside 60 µl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI.

  16. Induction of interleukin-8 by Naegleria fowleri lysates requires activation of extracellular signal-regulated kinase in human astroglial cells.

    Science.gov (United States)

    Kim, Jong-Hyun; Sohn, Hae-Jin; Lee, Sang-Hee; Kwon, Daeho; Shin, Ho-Joon

    2012-08-01

    Naegleria fowleri is a pathogenic free-living amoeba which causes primary amoebic meningoencephalitis in humans and experimental animals. To investigate the mechanisms of such inflammatory diseases, potential chemokine gene activation in human astroglial cells was investigated following treatment with N. fowleri lysates. We demonstrated that N. fowleri are potent inducers for the expression of interleukin-8 (IL-8) genes in human astroglial cells which was preceded by activation of extracellular signal-regulated kinase (ERK). In addition, N. fowleri lysates induces the DNA binding activity of activator protein-1 (AP-1), an important transcription factor for IL-8 induction. The specific mitogen-activated protein kinase kinase/ERK inhibitor, U0126, blocks N. fowleri-mediated AP-1 activation and subsequent IL-8 induction. N. fowleri-induced IL-8 expression requires activation of ERK in human astroglial cells. These findings indicate that treatment of N. fowleri on human astroglial cells leads to the activation of AP-1 and subsequent expression of IL-8 which are dependent on ERK activation. These results may help understand the N. fowleri-mediated upregulation of chemokine and cytokine expression in the astroglial cells.

  17. Selective Capture of Histidine-tagged Proteins from Cell Lysates Using TEM grids Modified with NTA-Graphene Oxide

    Science.gov (United States)

    Benjamin, Christopher J.; Wright, Kyle J.; Bolton, Scott C.; Hyun, Seok-Hee; Krynski, Kyle; Grover, Mahima; Yu, Guimei; Guo, Fei; Kinzer-Ursem, Tamara L.; Jiang, Wen; Thompson, David H.

    2016-10-01

    We report the fabrication of transmission electron microscopy (TEM) grids bearing graphene oxide (GO) sheets that have been modified with Nα, Nα-dicarboxymethyllysine (NTA) and deactivating agents to block non-selective binding between GO-NTA sheets and non-target proteins. The resulting GO-NTA-coated grids with these improved antifouling properties were then used to isolate His6-T7 bacteriophage and His6-GroEL directly from cell lysates. To demonstrate the utility and simplified workflow enabled by these grids, we performed cryo-electron microscopy (cryo-EM) of His6-GroEL obtained from clarified E. coli lysates. Single particle analysis produced a 3D map with a gold standard resolution of 8.1 Å. We infer from these findings that TEM grids modified with GO-NTA are a useful tool that reduces background and improves both the speed and simplicity of biological sample preparation for high-resolution structure elucidation by cryo-EM.

  18. 鲎试验的临床应用进展%Progress in Clinical Application of Limulus Lysate Test

    Institute of Scientific and Technical Information of China (English)

    刘冰

    2002-01-01

    主要综述了鲎试验在临床检验领域应用的三方面内容与进展:(1)内毒素血症与器官损伤、感染以及其他疾病的相关性;(2)鲎试验方法与临床检验的应用,包括试验方法、血浆/全血的处理方法及试验的专一性;(3)鲎试验进入临床检验领域所面临的问题,主要有内毒素标准的不统一、检测结果的有效性和污染的预防等问题.

  19. Evaluation of ToxA and Vibrio parahaemolyticus lysate on humoral immune response and immune-related genes in Pacific red snapper.

    Science.gov (United States)

    Reyes-Becerril, Martha; Maldonado-García, Minerva; Guluarte, Crystal; León-Gallo, Amalia; Rosales-Mendoza, Sergio; Ascencio, Felipe; Hirono, Ikuo; Angulo, Carlos

    2016-09-01

    Immunogenicity of ToxA and Vibrio parahaemolyticus lysate was evaluated in a double immunostimulation scheme in Pacific red snapper after V. parahaemolyticus infection. Three groups of Pacific red snapper were intraperitonealy (i.p.) injected with phosphate-buffered saline (PBS group), ToxA of V. parahaemolyticus (ToxA-Vp group) or V. parahaemolyticus lysate (lysate-Vp group) (first injection, day 1; second injection, day 7). Fish were subsequently infected with live V. parahaemolyticus. Humoral immune parameters in skin mucus and serum were evaluated on days 1, 7, 8 and 14 days post-immunostimulation and 7 days post-infection. Moreover expression of immune-related genes was quantified by real time PCR in head-kidney leukocytes, spleen, liver, and intestine. The ToxA-Vp-treated group showed a higher anti-protease and catalase activity in skin mucus when compared with the PBS group. Measurements of SOD and CAT activities showed an increment in both activities a day after the second boost with ToxA-Vp or lysate-Vp. Interestingly, IgM levels in mucus and transcripts were enhanced followed the ToxA-Vp treatment even after challenge. Furthermore, IL-1β was strongly expressed in all analyzed cell or tissues followed ToxA-Vp or Vp-lysate treatments. Finally, SOD and CAT gene expression was up-regulated in fish immunostimulated with either treatment ToxA-Vp or lysate-Vp, mainly after infection in head-kidney leukocytes and intestine. This is the first study where the effects of ToxA from V. parahaemolyticus in the immune system of Pacific red snapper was evaluated. These results suggest that ToxA-Vp would positively affect humoral immune response and up-regulate expression of genes involved in the immune system function; and could help in the control of V. parahaemolyticus infection in Pacific red snapper Lutjanus peru, an economic important fish in Mexico.

  20. Detection of endotoxins in radiopharmaceutical preparations. II. Comparison of the sensitivity of methods using the rabbit and the Limulus amoebocyte lysate for the detection of endotoxins

    Energy Technology Data Exchange (ETDEWEB)

    Bruneau, J.; Cohen, Y.; Merlin, L.; Peysson, S.

    1986-01-01

    The rise of the rabbit internal temperature after i.v. injection of an endotoxin solution is proportional to concentration. Gelation of Limulus amoebocyte, when in presence of an endotoxin solution, is also related to concentration. We compared the sensitivity of these two methods. With our experimental procedure, the rabbit is sensitive to a 0.40 ng/mL solution and the Limulus amoebocyte lysate to a 0.14 ng/mL solution. The rabbit sensitivity increase is related to the per kilogramme injected volume, whereas sensitivity is not related to the volume to check in the case of the lysate.

  1. Data from SILAC-based quantitative analysis of lysates from mouse microglial cells treated with Withaferin A (WA

    Directory of Open Access Journals (Sweden)

    Malathi Narayan

    2016-06-01

    Full Text Available Mass spectrometry data collected in a study analyzing the effect of withaferin A (WA on a mouse microglial (N9 cell line is presented in this article. Data was collected from SILAC-based quantitative analysis of lysates from mouse microglial cells treated with either WA or DMSO vehicle control. This article reports all the proteins that were identified in this analysis. The data presented here is related to the published research article on the effect of WA on the differential regulation of proteins in mouse microglial cells [1]. Mass spectrometry data has also been deposited in the ProteomeXchange with the identifier http://www.ebi.ac.uk/pride/archive/projects/PXD003032.

  2. A new MIC1-MAG1 recombinant chimeric antigen can be used instead of the Toxoplasma gondii lysate antigen in serodiagnosis of human toxoplasmosis.

    Science.gov (United States)

    Holec-Gąsior, Lucyna; Ferra, Bartłomiej; Drapała, Dorota; Lautenbach, Dariusz; Kur, Józef

    2012-01-01

    This study presents an evaluation of the MIC1 (microneme protein 1)-MAG1 (matrix antigen 1) Toxoplasma gondii recombinant chimeric antigen for the serodiagnosis of human toxoplasmosis for the first time. The recombinant MIC1-MAG1 antigen was obtained as a fusion protein containing His tags at the N- and C-terminal ends using an Escherichia coli expression system. After purification by metal affinity chromatography, the chimeric protein was tested for usefulness in an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-T. gondii immunoglobulin G (IgG). One hundred ten sera from patients at different stages of infection and 40 sera from seronegative patients were examined. The results obtained for the MIC1-MAG1 chimeric antigen were compared with those of IgG ELISAs using a Toxoplasma lysate antigen (TLA), a combination of recombinant antigens (rMIC1ex2-rMAG1) and single recombinant proteins (rMIC1ex2 and rMAG1). The sensitivity of the IgG ELISA calculated from all of the positive serum samples was similar for the MIC1-MAG1 chimeric antigen (90.8%) and the TLA (91.8%), whereas the sensitivities of the other antigenic samples used were definitely lower, at 69.1% for the mixture of antigens, 75.5% for the rMIC1ex2, and 60% for rMAG1. This study demonstrates that the MIC1-MAG1 recombinant chimeric antigen can be used instead of the TLA in the serodiagnosis of human toxoplasmosis.

  3. 血小板裂解液对人间充质干细胞生物学特性的影响%Effect of platelet lysate on the biological characteristics of human mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    吴迪; 武晓云; 吴岩

    2015-01-01

    BACKGROUND:Platelet lysate has been known as a kind of lysate of autologous or alogeneic platelet-rich products. It not only removes the residual cel structure, reduces immunogenicity, but also retains many growth factors. Platelet lysate has been suggested as a substitute for fetal bovine serum to expand mesenchymal stem cels in vitro. OBJECTIVE:To observe the effects of platelet lysate on biological characteristics of human bone marrow mesenchymal stem cels and adipose mesenchymal stem cels, and provide some experimental data for clinical cel therapy and regenerative medicine. METHODS:Platelet lysate was prepared by repeated freezing and thawing from fresh blood. Healthy adult bone marrow and adipose tissue were colected. Human bone marrow mesenchymal stem cels and adipose mesenchymal stem cels were obtained by density gradient centrifugation and type I colagenase digestion. We tested the morphology, cel phenotype, differentiation characteristics, proliferation capacity, colony forming ability and the level of cytokine secretion of bone marrow mesenchymal stem cels and adipose mesenchymal stem cels after cultured with platelet lysis. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cels and adipose mesenchymal stem cels were successfuly cultured in vitro using platelet lysate. There were no significant differences in morphology, cel phenotype, colony forming ability and the level of cytokine secretion, and chondrogenic, osteogenic and adipogenic capacities between bone marrow mesenchymal stem cels and adipose mesenchymal stem cels. Adipose mesenchymal stem cels had a high cumulative population doublings than bone marrow mesenchymal stem cels (P < 0.05). These findings suggest adipose mesenchymal stem cels had a stronger proliferative ability, and are more suitable for large-scale expansionin vitro cultivation system of platelet lysate compared with bone marrow mesenchymal stem cels.%背景:血小板裂解液是自体或异体血小板富集物经裂解后

  4. Black cobra (Naja naja karachiensis) lysates exhibit broad-spectrum antimicrobial activities

    OpenAIRE

    Sagheer, Mehwish; Siddiqui, Ruqaiyyah; Iqbal, Junaid; Khan, Naveed Ahmed

    2014-01-01

    It is hypothesized that animals living in polluted environments possess antimicrobials to counter pathogenic microbes. The fact that snakes feed on germ-infested rodents suggests that they encounter pathogenic microbes and likely possess antimicrobials. The venom is used only to paralyze the rodent, but the ability of snakes to counter potential infections in the gut due to disease-ridden rodents requires robust action of the immune system against a broad range of pathogens. To test this hypo...

  5. Autologous tumor lysate-pulsed dendritic cell immunotherapy for pediatric patients with newly diagnosed or recurrent high-grade gliomas.

    Science.gov (United States)

    Lasky, Joseph L; Panosyan, Eduard H; Plant, Ashley; Davidson, Tom; Yong, William H; Prins, Robert M; Liau, Linda M; Moore, Theodore B

    2013-05-01

    Immunotherapy has the potential to improve clinical outcomes with little toxicity for pediatric patients with brain tumors. We conducted a pilot feasibility study of tumor lysate-pulsed dendritic cell (DC) vaccination in pediatric patients (1 to 18 years old) with newly diagnosed or recurrent high-grade glioma (HGG). A total of nine DC vaccine doses, each containing 1 × 10(6) cells per dose were administered to three out of the seven originally enrolled patients. Toxicities were limited to mild side-effects, except in one case of elevated alkaline phosphatase, which resolved without clinical consequences. Two patients with primary lesions amongst the three vaccinated were alive at the time of writing, both without evidence of disease. Pre- and post-vaccination tumor samples from a patient with an anaplastic oligoastrocytoma that recurred failed to demonstrate immune cell infiltration by immunohistochemistry. Peripheral cytokine levels were evaluated in one patient following DC vaccination and demonstrated some changes in relation to vaccination. DC vaccine is tolerable and feasible with some limitations for pediatric patients with HGG. Dendritic cell based immunotherapy may provide some clinical benefit in pediatric patients with glioma, especially for patients with minimal residual disease, but further investigation of this modality is required. PMID:23645755

  6. Dynamics of heat shock protein 70 concentrations in peripheral blood lymphocyte lysates during pregnancy in lactating Holstein-Friesian cows.

    Science.gov (United States)

    Yániz, J L; López-Gatius, F; Almería, S; Carretero, T; García-Ispierto, I; Serrano, B; Smith, R F; Dobson, H; Santolaria, P

    2009-11-01

    The aim of this study was to characterize the dynamics of the concentrations of heat shock protein 70 kDa (HSP70) in peripheral blood lymphocytes of lactating Holstein-Friesian dairy cows (Bos taurus) during pregnancy. The detection of pregnancy was carried out and blood samples collected on Days 40, 90, 120, 150, 180, and 210 of gestation from 46 cows (11 primiparous and 35 pluriparous, 34 seropositive and 12 seronegative to Neospora caninum). Peripheral blood lymphocytes were isolated by density gradient centrifugation. Serologic analysis of Neospora infection and determinations of HSP70 concentrations in lymphocyte lysates were carried out using commercial enzyme-linked immunosorbent assay kits. Climate variables were monitored using on-farm data loggers. Heat shock protein 70 concentrations increased in lymphocytes as gestation progressed, particularly in primiparous cows, with no effect from Neospora infection, climate variables, milk production, semen-providing bull, or outcome of gestation (singletons or twins). Our results show that HSP70 concentrations increased in lymphocytes as gestation progressed and were not affected by stressful factors, such as milk production, heat stress, chronic infection (neosporosis), or twin pregnancies.

  7. Protective antitumor immunity induced by tumor cell lysates conjugated with diphtheria toxin and adjuvant epitope in mouse breast tumor models

    Institute of Scientific and Technical Information of China (English)

    Ze-Yu Wang; Rong-Yue Cao; Jie Wu; Tai-Ming LI; Jing-Jing Liu; Yun Xing; Bin Liu; Lei Lu; Xiao Huang; Chi-Yu Ge; Wen-Jun Yao; Mao-Lei Xu; Zhen-Qiu Gao

    2012-01-01

    Cancer cell vaccine-based immunotherapy has received increasing interest in many clinical trials involving patients with breast cancer.Combining with appropriate adjuvants can enhance the weak immunogenic properties of tumor cell lysates (TCL).In this study,diphtheria toxin (DT) and two tandem repeats of mycobacterial heat shock protein 70 (mHSP70) fragment 407-426 (M2) were conjugated to TCL with glutaraldehyde,and the constructed cancer cell vaccine was named DT-TCL-M2.Subcutaneous injection of DT-TCL-M2 in mice effectively elicited tumor-specific polyclonal immune responses,including humoral and cellular immune responses.High levels of antibodies against TCL were detected in the serum of immunized mice with ELISA and verified with Western blot analyses.The splenocytes from immunized mice showed potent cytotoxicity on Ehrlich ascites carcinoma cells.Moreover,the protective antitumor immunity induced by DT-TCL-M2 inhibited tumor growth in a mouse breast tumor model.DTTCL-M2 also attenuated tumor-induced angiogenesis and slowed tumor growth in a mouse intradermal tumor model.These findings demonstrate that TCL conjugated with appropriate adjuvants induced effective antitumor immunity in vivo.Improvements in potency could further make cancer cell vaccines a useful and safe method for preventing cancer recurrence after resection.

  8. Autologous tumor lysate-pulsed dendritic cell immunotherapy for pediatric patients with newly diagnosed or recurrent high-grade gliomas.

    Science.gov (United States)

    Lasky, Joseph L; Panosyan, Eduard H; Plant, Ashley; Davidson, Tom; Yong, William H; Prins, Robert M; Liau, Linda M; Moore, Theodore B

    2013-05-01

    Immunotherapy has the potential to improve clinical outcomes with little toxicity for pediatric patients with brain tumors. We conducted a pilot feasibility study of tumor lysate-pulsed dendritic cell (DC) vaccination in pediatric patients (1 to 18 years old) with newly diagnosed or recurrent high-grade glioma (HGG). A total of nine DC vaccine doses, each containing 1 × 10(6) cells per dose were administered to three out of the seven originally enrolled patients. Toxicities were limited to mild side-effects, except in one case of elevated alkaline phosphatase, which resolved without clinical consequences. Two patients with primary lesions amongst the three vaccinated were alive at the time of writing, both without evidence of disease. Pre- and post-vaccination tumor samples from a patient with an anaplastic oligoastrocytoma that recurred failed to demonstrate immune cell infiltration by immunohistochemistry. Peripheral cytokine levels were evaluated in one patient following DC vaccination and demonstrated some changes in relation to vaccination. DC vaccine is tolerable and feasible with some limitations for pediatric patients with HGG. Dendritic cell based immunotherapy may provide some clinical benefit in pediatric patients with glioma, especially for patients with minimal residual disease, but further investigation of this modality is required.

  9. A sensitive fluorescence-based assay for monitoring GM2 ganglioside hydrolysis in live patient cells and their lysates.

    Science.gov (United States)

    Tropak, Michael B; Bukovac, Scott W; Rigat, Brigitte A; Yonekawa, Sayuri; Wakarchuk, Warren; Mahuran, Don J

    2010-03-01

    Enzyme enhancement therapy, utilizing small molecules as pharmacological chaperones, is an attractive approach for the treatment of lysosomal storage diseases that are associated with protein misfolding. However, pharmacological chaperones are also inhibitors of their target enzyme. Thus, a major concern with this approach is that, despite enhancing protein folding within, and intracellular transport of the functional mutant enzyme out of the endoplasmic reticulum, the chaperone will continue to inhibit the enzyme in the lysosome, preventing substrate clearance. Here we demonstrate that the in vitro hydrolysis of a fluorescent derivative of lyso-GM2 ganglioside, like natural GM2 ganglioside, is specifically carried out by the beta-hexosaminidase A isozyme, requires the GM2 activator protein as a co-factor, increases when the derivative is incorporated into anionic liposomes and follows similar Michaelis-Menten kinetics. This substrate can also be used to differentiate between lysates from normal and GM2 activator-deficient cells. When added to the growth medium of cells, the substrate is internalized and primarily incorporated into lysosomes. Utilizing adult Tay-Sachs fibroblasts that have been pre-treated with the pharmacological chaperone Pyrimethamine and subsequently loaded with this substrate, we demonstrate an increase in both the levels of mutant beta-hexosaminidase A and substrate-hydrolysis as compared to mock-treated cells. PMID:19917668

  10. Cytokine-induced killer cells interact with tumor lysate-pulsed dendritic cells via CCR5 signaling.

    Science.gov (United States)

    Lee, Hong Kyung; Kim, Yong Guk; Kim, Ji Sung; Park, Eun Jae; Kim, Boyeong; Park, Ki Hwan; Kang, Jong Soon; Hong, Jin Tae; Kim, Youngsoo; Han, Sang-Bae

    2016-08-10

    The antitumor activity of cytokine-induced killer (CIK) cells can be increased by co-culturing them with tumor lysate-pulsed dendritic cells (tDCs); this phenomenon has been studied mainly at the population level. Using time-lapse imaging, we examined how CIK cells gather information from tDCs at the single-cell level. tDCs highly expressed CCL5, which bound CCR5 expressed on CIK cells. tDCs strongly induced migration of Ccr5(+/+) CIK cells, but not that of Ccr5(-/-) CIK cells or Ccr5(+/+) CIK cells treated with the CCR5 antagonist Maraviroc. Individual tDCs contacted Ccr5(+/+) CIK cells more frequently and lengthily than with Ccr5(-/-) CIK cells. Consequently, tDCs increased the antitumor activity of Ccr5(+/+) CIK cells in vitro and in vivo, but did not increase that of Ccr5(-/-) CIK cells. Taken together, our data provide insight into the mechanism of CIK cell activation by tDCs at the single-cell level.

  11. Highly Efficient In Vitro Reparative Behaviour of Dental Pulp Stem Cells Cultured with Standardised Platelet Lysate Supplementation

    Science.gov (United States)

    Palmieri, Francesca; Marrelli, Massimo

    2016-01-01

    Dental pulp is an accessible source of multipotent mesenchymal stromal cells (MSCs). The perspective role of dental pulp stem cells (DPSCs) in regenerative medicine demands an in vitro expansion and in vivo delivery which must deal with the safety issues about animal serum, usually required in cell culture practice. Human platelet lysate (PL) contains autologous growth factors and has been considered as valuable alternative to fetal bovine serum (FBS) in cell cultures. The optimum concentration to be added of such supplement is highly dependent on its preparation whose variability limits comparability of results. By in vitro experiments, we aimed to evaluate a standardised formulation of pooled PL. A low selected concentration of PL (1%) was able to support the growth and maintain the viability of the DPSCs. The use of PL in cell cultures did not impair cell surface signature typically expressed by MSCs and even upregulated the transcription of Sox2. Interestingly, DPSCs cultured in presence of PL exhibited a higher healing rate after injury and are less susceptible to toxicity mediated by exogenous H2O2 than those cultured with FBS. Moreover, PL addition was shown as a suitable option for protocols promoting osteogenic and chondrogenic differentiation of DPSCs. Taken together, our results indicated that PL is a valid substitute of FBS to culture and differentiate DPSCs for clinical-grade use. PMID:27774106

  12. Development and implementation of a novel assay for L-2-hydroxyglutarate dehydrogenase (L-2-HGDH) in cell lysates: L-2-HGDH deficiency in 15 patients with L-2-hydroxyglutaric aciduria

    DEFF Research Database (Denmark)

    Kranendijk, M; Salomons, G S; Gibson, K M;

    2009-01-01

    /or lymphocyte lysates has hitherto been unavailable. We developed an L-2-HGDH enzyme assay in cell lysates based on the conversion of stable-isotope-labelled L-2-hydroxyglutarate to 2-ketoglutarate, which is converted into L-glutamate in situ. The formation of stable isotope labelled L-glutamate is therefore...... a direct measure of L-2-HGDH activity, and this product is detected by liquid chromatography-tandem mass spectrometry. A deficiency of L-2-HGDH activity was detected in cell lysates from 15 out of 15 L-2-HGA patients. Therefore, this specific assay confirmed the diagnosis unambiguously affirming...

  13. Protection against Naegleria fowleri infection in mice immunized with Cry1Ac plus amoebic lysates is dependent on the STAT6 Th2 response.

    Science.gov (United States)

    Carrasco-Yepez, M; Rojas-Hernandez, S; Rodriguez-Monroy, M A; Terrazas, L I; Moreno-Fierros, L

    2010-01-01

    We previously reported that intranasal administration of Cry1Ac protoxin alone or in combination with amoebic lysates increases protection against Naegleria fowleri meningoencephalitis in mice. Those results suggested that both antibody responses and innate immune mechanisms may be participating in the protective effects observed. The present study was aimed to investigate whether the STAT6-induced Th2 immune response is essential for the resistance to N. fowleri infection, conferred by immunization with amoebic lysates plus Cry1Ac. STAT6-deficient (STAT6-/-) and wild-type (STAT6+/+) BALB/c mice were immunized by the intranasal route with a combination of N. fowleri lysates plus Cry1Ac, and subsequently challenged with lethal doses of N. fowleri trophozoites. STAT6+/+ mice displayed 100% protection, while no protection was observed in STAT6-/- mice. Significantly higher titres of Th2-associated IgG1 as well as interleukin-4 (IL-4) were found in STAT6+/+ mice, whereas in STAT6-/- mice significantly more IL-12 and IFN-gamma as well as significantly higher titres of Th1-associated IgG2a were detected. Thus, whereas protected STAT6+/+-immunized mice elicited a Th-2 type inclined immune response that produced predominantly humoral immunity, unprotected STAT6-/- mice exhibited a polarized Th1 type cellular response. These findings suggest that the STAT6-signalling pathway is critical for defence against N. fowleri infection.

  14. Distribution of AAV8 particles in cell lysates and culture media changes with time and is dependent on the recombinant vector.

    Science.gov (United States)

    Piras, Bryan A; Drury, Jason E; Morton, Christopher L; Spence, Yunyu; Lockey, Timothy D; Nathwani, Amit C; Davidoff, Andrew M; Meagher, Michael M

    2016-01-01

    With clinical trials ongoing, efficient clinical production of adeno-associated virus (AAV) to treat large numbers of patients remains a challenge. We compared distribution of AAV8 packaged with Factor VIII (FVIII) in cell culture media and lysates on days 3, 5, 6, and 7 post-transfection and found increasing viral production through day 6, with the proportion of viral particles in the media increasing from 76% at day 3 to 94% by day 7. Compared to FVIII, AAV8 packaged with Factor IX and Protective Protein/Cathepsin A vectors demonstrated a greater shift from lysate towards media from day 3 to 6, implying that particle distribution is dependent on recombinant vector. Larger-scale productions showed that the ratio of full-to-empty AAV particles is similar in media and lysate, and that AAV harvested on day 6 post-transfection provides equivalent function in mice compared to AAV harvested on day 3. This demonstrates that AAV8 production can be optimized by prolonging the duration of culture post-transfection, and simplified by allowing harvest of media only, with disposal of cells that contain 10% or less of total vector yield. Additionally, the difference in particle distribution with different expression cassettes implies a recombinant vector-dependent processing mechanism which should be taken into account during process development. PMID:27069949

  15. LC-MS/MS method for the simultaneous quantification of 11 compounds of Ginkgo biloba extract in lysates of mesangial cell cultured by high glucose.

    Science.gov (United States)

    Qiu, Jing-ying; Chen, Xu; Li, Zheng; Wang, Shi-rui; Wu, Xiao-wen; Li, Yin-jie; Yang, Dong-zhi; Yu, Yan-yan; Yin, Xiao-xing; Tang, Dao-quan

    2015-08-01

    The mesangial cell (MC) cultured with high glucose has been used to observe the protective effect of Ginkgo biloba extract (GBE) against diabetic nephropathy (DN), but the compounds interacting with cell are still unknown, which may be potential bioactive components. Based on this, the determination of GBE in MC lysates was proposed by high performance liquid chromatography tandem mass spectrometry (LC-MS/MS) in this study. The MC was cultured with normal or high glucose with GBE for 4, 8, 12, 16, 24 and 48h. The harvested cell was extracted with 40% acetic acid in water and further analyzed by LC-MS/MS. All the validation data including linearity, intra-day and inter-day precision, limit of detection and quantification, matrix effect, and stability were within the required limits. The validated method was successfully applied to quantify 11 compounds of GBE in cell lysates. The results showed that high glucose prolonged the peak time of all observed 11 compounds and peak concentrations of bilobalide, ginkgolide C, ginkgolide B, quercetin, luteolin, kaempferol, isorhamnetin and genkwanin in cell lysates, which hinted that these compounds may be the potential bioactive components of GBE with preventive effect against DN.

  16. Characteristics of prolinase against various iminodipeptides in erythrocyte lysates from a normal human and a patient with prolidase deficiency.

    Science.gov (United States)

    Wang, Weifang; Liu, Gang; Yamashita, Koichi; Manabe, Masanobu; Kodama, Hiroyuki

    2004-01-01

    The effect of various amino acids and MnCl2 on prolinase activity in erythrocyte lysates from a healthy individual and a patient with prolidase deficiency was investigated. A concentration of 0.1 mM MnCl2 increased prolinase activity in normal erythrocytes against pro-gly, pro-glu, pro-leu, pro-ser and pro-phe, but inhibited that against pro-ala, pro-val, pro-met and pro-asp. However, prolinase activity against these iminodipeptides was enhanced by pre-incubation with glycine, independent of MnCl2. The same studies on erythrocytes from a prolidase-deficient patient showed almost the same results as the normal control, except that prolinase activity against pro-gly and pro-ser was slightly inhibited by adding 0.1 mM MnCl2. Some amino acids, glutamic acid and glutamine, slightly enhanced prolinase activity against pro-gly in erythrocytes from both the normal control and the prolidase-deficient patient, but N-acetyl-L-glutamic acid, gamma-aminobutyric acid (GABA) and beta-alanine showed no effect. Branched amino acids, L-valine, L-leucine and L-isoleucine strongly inhibited the prolinase activity against pro-gly. However, conversely, their isomers, D-valine, D-leucine and D-isoleucine, enhanced it. The kinetics of prolinase activity in the erythrocytes from both the normal individual and the prolidasedeficient patient were also studied. Their Km values were changed by adding glycine or 0.1 mM MnCl2, but Vmax values were almost the same. PMID:15552267

  17. High throughput ranking of recombinant avian scFv antibody fragments from crude lysates using the Biacore A100.

    Science.gov (United States)

    Leonard, Paul; Säfsten, Pär; Hearty, Stephen; McDonnell, Barry; Finlay, William; O'Kennedy, Richard

    2007-06-30

    Advances in molecular evolution strategies have made it possible to identify antibodies with exquisite specificities and also to fine-tune their biophysical properties for practically any specified application. Depending on the desired function, antibody/antigen interactions can be long-lived or short-lived and, therefore, particular attention is needed when seeking to identify antibodies with specific reaction-rate and affinity properties. Surface plasmon resonance (SPR) biosensors routinely generate sensitive and reliable kinetic data from antibody/antigen interactions for both therapeutic and diagnostic applications. However, many kinetic-based screening assays require rigorous sample preparation and purification prior to analysis. To ameliorate this problem, we developed a rapid and reliable assay for characterising recombinant scFv antibody fragments, directly from crude bacterial lysates. Ninety-six scFv antibodies derived from chickens immunised with C-reactive protein (CRP) were selected by phage display and evaluated using the Biacore A100 protein interaction array system. Antibodies were captured from crude bacterial extracts on the sensor chip surface and ranked based on the percentage of the complex left (% left) after dissociation in buffer. Kinetic rate constants (k(a) and k(d)) and affinity (K(D)) data were obtained for six clones that bound monomeric CRP across a broad affinity range (2.54 x 10(-8) to 3.53 x 10(-10) M). Using this assay format the A100 biosensor yielded high quality kinetic data, permitting the screening of nearly 400 antibody clones per day. PMID:17532001

  18. Preparation, quality criteria, and properties of human blood platelet lysate supplements for ex vivo stem cell expansion.

    Science.gov (United States)

    Shih, Daniel Tzu-Bi; Burnouf, Thierry

    2015-01-25

    Most clinical applications of human multipotent mesenchymal stromal cells (MSCs) for cell therapy, tissue engineering, regenerative medicine, and treatment of immune and inflammatory diseases require a phase of isolation and ex vivo expansion allowing a clinically meaningful cell number to be reached. Conditions used for cell isolation and expansion should meet strict quality and safety requirements. This is particularly true for the growth medium used for MSC isolation and expansion. Basal growth media used for MSC expansion are supplemented with multiple nutrients and growth factors. Fetal bovine serum (FBS) has long been the gold standard medium supplement for laboratory-scale MSC culture. However, FBS has a poorly characterized composition and poses risk factors, as it may be a source of xenogenic antigens and zoonotic infections. FBS has therefore become undesirable as a growth medium supplement for isolating and expanding MSCs for human therapy protocols. In recent years, human blood materials, and most particularly lysates and releasates of platelet concentrates have emerged as efficient medium supplements for isolating and expanding MSCs from various origins. This review analyzes the advantages and limits of using human platelet materials as medium supplements for MSC isolation and expansion. We present the modes of production of allogeneic and autologous platelet concentrates, measures taken to ensure optimal pathogen safety profiles, and methods of preparing PLs for MSC expansion. We also discuss the supply of such blood preparations. Produced under optimal conditions of standardization and safety, human platelet materials can become the future 'gold standard' supplement for ex vivo production of MSCs for translational medicine and cell therapy applications.

  19. Differentiation of Rat bone marrow Mesenchymal stem cells into Adipocytes and Cardiomyocytes after treatment with platelet lysate

    Science.gov (United States)

    Homayouni Moghadam, Farshad; Tayebi, Tahereh; Barzegar, Kazem

    2016-01-01

    Background: Mesenchymal stem cells (MSCs) are multipotential cells and their therapeutic potency is under intense investigation. Studying the effect of different induction factors on MSCs could increase our knowledge about the differentiation potency of these cells. One of the most important sources of these factors in mammalian body is platelet. Platelet lysate (PL) contains many growth factors and therefore, it can be used as a differentiation inducer. In the present study, the effect of PL on differentiation of rat bone marrow MSCs into cardiomyocytes was studied. Materials and Methods: To study the differentiation-inducing effect of PL, MSCs were treated with 2.5, 5 and 10% PL. Early results of this study showed that PL in high concentrations (10%) induces adipogenic differentiation of MSCs. Therefore, to evaluate differentiation to cardiomyocytes, MSCs were cultured in media containing lower levels of PL (2.5% and 5%) and then cardiomyogenic differentiation was induced by treatment with 5-azacytidine. Differentiation of MSCs was evaluated using direct observation of beating cells, immunostaining and real-time PCR techniques. Results: The results of qPCR showed that treatment with PL alone increased the expression of cardiac alpha actinin (CAA) being predictable by earlier observation of beating cells in PL-treated groups. The results of staining assays against cardiac alpha actinin also showed that there were stained cells in PL-treated groups. Conclusion: The results of the present study showed that PL is a powerful induction factor for differentiation of MSCs into different cell lines such as cardiomyocytes and adipocytes. PMID:27047647

  20. Test

    DEFF Research Database (Denmark)

    Bendixen, Carsten

    2014-01-01

    Bidrag med en kortfattet, introducerende, perspektiverende og begrebsafklarende fremstilling af begrebet test i det pædagogiske univers.......Bidrag med en kortfattet, introducerende, perspektiverende og begrebsafklarende fremstilling af begrebet test i det pædagogiske univers....

  1. 1-MT Enhances Potency of Tumor Cell Lysate-pulsed Dendritic Cells against Pancreatic Adenocarcinoma by Downregulating the Percentage of Tregs

    Institute of Scientific and Technical Information of China (English)

    李元栋; 徐钧; 邹浩军; 王春友

    2010-01-01

    This study examined whether 1-methyl-tryptophan [1-MT,an indoleamine 2,3-dioxygenase(IDO) inhibitor] could reduce CD4+CD25+ regulatory T cells(Tregs) proliferation and improve the anti-tumor efficacy of dendritic cells(DCs) pulsed with tumor cell lysate in the mice bearing pancreatic adenocarcinoma.The models of pancreatic adenocarcinoma were established in C57BL/6 mice by subcutaneous injection of Pan02 cells.Eight mice which were subcutaneously injected with PBS served as control.The expression of IDO was...

  2. Overcoming inefficient secretion of recombinant VEGF-C in baculovirus expression vector system by simple purification of the protein from cell lysate.

    Science.gov (United States)

    Klaus, Tomasz; Kulesza, Małgorzata; Bzowska, Monika; Wyroba, Barbara; Kilarski, Witold W; Bereta, Joanna

    2015-06-01

    The first reports about successfully expressed recombinant proteins with the use of a baculovirus vector were published over 30years ago. Despite the long time of refining this expression system, early problems with the production of baculovirus-derived secretory proteins are still not satisfactorily solved. The high expression level driven by baculoviral promoters often does not result in the desired yield of secreted recombinant proteins, which frequently accumulate inside insect cells and are only partially processed. During our attempts to produce vascular endothelial growth factor C (VEGF-C) with the use of a baculovirus vector we also faced an inefficient secretion of the recombinant protein to culture medium. We were not able to improve the outcome and obtain an acceptable concentration of VEGF-C in the medium by changing the culture conditions or utilizing different signal peptides. However, as a significant amount of native VEGF-C was detected inside the baculovirus-infected cells, we developed a simple method to purify recombinant, glycosylated VEGF-C from a lysate of the cells. The presented results indicate that the lack of a secretory protein in the insect cell culture medium after baculovirus infection does not necessarily signify failure in the production of the protein. As demonstrated by us and contrary to generally accepted views, the lysate of baculovirus-infected cells may constitute a valuable source of the biologically active, secretory protein.

  3. Effects of Tamarindus indica fruit pulp extract on abundance of HepG2 cell lysate proteins and their possible consequential impact on metabolism and inflammation.

    Science.gov (United States)

    Chong, Ursula R W; Abdul-Rahman, Puteri S; Abdul-Aziz, Azlina; Hashim, Onn H; Mat-Junit, Sarni

    2013-01-01

    The fruit pulp extract of Tamarindus indica has been reported for its antioxidant and hypolipidemic properties. In this study, the methanol extract of T. indica fruit pulp was investigated for its effects on the abundance of HepG2 cell lysate proteins. Cell lysate was extracted from HepG2 cells grown in the absence and presence of the methanol extract of T. indica fruit pulp. Approximately 2500 spots were resolved using two-dimensional gel electrophoresis and the abundance of 20 cellular proteins was found to be significantly reduced. Among the proteins of reduced abundance, fourteen, including six proteins involved in metabolism (including ethanolamine phosphate cytidylyltransferase), four mitochondrial proteins (including prohibitin and respiratory chain proteins), and four proteins involved in translation and splicing, were positively identified by mass spectrometry and database search. The identified HepG2 altered abundance proteins, when taken together and analyzed by Ingenuity Pathways Analysis (IPA) software, are suggestive of the effects of T. indica fruit pulp extract on metabolism and inflammation, which are modulated by LXR/RXR. In conclusion, the methanol fruit pulp extract of T. indica was shown to cause reduced abundance of HepG2 mitochondrial, metabolic, and regulatory proteins involved in oxidative phosphorylation, protein synthesis, and cellular metabolism.

  4. Effects of Tamarindus indica Fruit Pulp Extract on Abundance of HepG2 Cell Lysate Proteins and Their Possible Consequential Impact on Metabolism and Inflammation

    Directory of Open Access Journals (Sweden)

    Ursula R. W. Chong

    2013-01-01

    Full Text Available The fruit pulp extract of Tamarindus indica has been reported for its antioxidant and hypolipidemic properties. In this study, the methanol extract of T. indica fruit pulp was investigated for its effects on the abundance of HepG2 cell lysate proteins. Cell lysate was extracted from HepG2 cells grown in the absence and presence of the methanol extract of T. indica fruit pulp. Approximately 2500 spots were resolved using two-dimensional gel electrophoresis and the abundance of 20 cellular proteins was found to be significantly reduced. Among the proteins of reduced abundance, fourteen, including six proteins involved in metabolism (including ethanolamine phosphate cytidylyltransferase, four mitochondrial proteins (including prohibitin and respiratory chain proteins, and four proteins involved in translation and splicing, were positively identified by mass spectrometry and database search. The identified HepG2 altered abundance proteins, when taken together and analyzed by Ingenuity Pathways Analysis (IPA software, are suggestive of the effects of T. indica fruit pulp extract on metabolism and inflammation, which are modulated by LXR/RXR. In conclusion, the methanol fruit pulp extract of T. indica was shown to cause reduced abundance of HepG2 mitochondrial, metabolic, and regulatory proteins involved in oxidative phosphorylation, protein synthesis, and cellular metabolism.

  5. Clinical and Immunological Effects in Patients with Advanced Non-Small Cell Lung-Cancer after Vaccination with Dendritic Cells Exposed to an Allogeneic Tumor Cell Lysate*

    DEFF Research Database (Denmark)

    Engell-Noerregaard, Lotte; Kvistborg, Pia; Zocca, Mai-Britt;

    2013-01-01

    and celecoxib were used as adjuvants to the vaccines. The objective of the study was to evaluate specific T cell response in vitro by IFN EliSpot. Secondary objec- tives were overall survival, response and quality of life (QoL). Results: Twenty-two patients initiated the vaccination program consisting of ten...... patients showed an un- expectedly prolonged survival. The treatment was well tolerated and only minor adverse events were reported. Quality of life did not change during the study period. In four of the seven patients with SD, vaccine-specific T cells were de- tected by IFNγ EliSpot assays, whereas only......Background: We evaluated the clinical and immunological effects of dendritic cell (DC) vaccination of patients with NSCLC. Autologous DCs were pulsed with a MAGE containing allogeneic melanoma cell lysate (MelCancerVac®, Dandrit Biotech, Copenhagen, Denmark). Imiquimod cream, proleukin...

  6. Thermodynamic Analysis of the Geldanamycin-Hsp90 Interaction in a Whole Cell Lysate Using a Mass Spectrometry-Based Proteomics Approach

    Science.gov (United States)

    Xu, Yingrong; Wallace, M. Ariel Geer; Fitzgerald, Michael C.

    2016-08-01

    Geldanamycin is a natural product with well-established and potent anti-cancer activities. Heat shock protein 90 (Hsp90) is the known target of geldanamycin, which directly binds to Hsp90's N-terminal ATP binding domain and inhibits Hsp90's ATPase activity. The affinity of geldanamycin for Hsp90 has been measured in multiple studies. However, there have been large discrepancies between the reported dissociation constants (i.e., Kd values), which have ranged from low nanomolar to micromolar. Here the stability of proteins from rates of oxidation (SPROX) technique was used in combination with an isobaric mass tagging strategy to measure the binding affinity of geldanamycin to unpurified Hsp90 in an MCF-7 cell lysate. The Kd values determined here were dependent on how long geldanamycin was equilibrated with the lysate prior to SPROX analysis. The Kd values determined using equilibration times of 0.5 and 24 h were 1 and 0.03 μM, respectively. These Kd values, which are similar to those previously reported in a geldanamycin-Hsp90 binding study that involved the use of a fluorescently labeled geldanamycin analogue, establish that the slow-tight binding behavior previously observed for the fluorescently labeled geldanamycin analogue is not an artifact of the fluorescent label, but rather an inherent property of the geldanamycin-Hsp90 binding interaction. The slow-tight binding property of this complex may be related to time-dependent conformational changes in Hsp90 and/or to time-dependent chemical changes in geldanamycin, both of which have been previously proposed to explain the slow-tight binding behavior of the geldanamycin-Hsp90 complex.

  7. In vitro assessment of the susceptibility of planktonic and attached cells of foodborne pathogens to bacteriophage p22-mediated salmonella lysates.

    Science.gov (United States)

    Ahn, Juhee; Kim, Songrae; Jung, Lae-Seung; Biswas, Debabrata

    2013-12-01

    This study was designed to evaluate the lytic activity of bacteriophage P22 against Salmonella Typhimurium ATCC 19585 (Salmonella Typhimurium P22(-)) at various multiplicities of infections (MOIs), the susceptibility of preattached Salmonella cells against bacteriophage P22, and the effect of P22-mediated bacterial lysates (extracellular DNA) on the attachment ability of Listeria monocytogenes ATCC 7644 and enterohemorrhagic Escherichia coli ATCC 700927 to surfaces. The numbers of attached Salmonella Typhimurium P22(-) cells were effectively reduced to below the detection limit (1 log CFU/ml) at the fixed inoculum levels of 3 × 10(-) CFU/ml (MOI = 3.12) and 3 × 10(3) CFU/ml (MOI = 4.12) by bacteriophage P22. The attached Salmonella Typhimurium P22(-) cells remained more than 2 log CFU/ml, with increasing inoculum levels from 3 × 10(4) to 3 × 10(7) CFU/ml infected with 4 × 10(8) PFU/ml of P22. The number of preattached Salmonella Typhimurium P22(-) cells was noticeably reduced by 2.72 log in the presence of P22. The highest specific attachment ability values for Salmonella Typhimurium P22(-), Salmonella Typhimurium ATCC 23555 carrying P22 prophage (Salmonella Typhimurium P22(+)), L. monocytogenes, and enterohemorrhagic E. coli were 2.09, 1.06, 1.86, and 1.08, respectively, in the bacteriophage-mediated cell-free supernatants (CFS) containing high amounts of extracellular DNA. These results suggest that bacteriophages could potentially be used to effectively eliminate planktonic and preattached Salmonella Typhimurium P22(-) cells with increasing MOI. However, further research is needed to understand the role of bacteriophage-induced lysates in bacterial attachment, which can provide useful information for the therapeutic use of bacteriophage in the food system.

  8. Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Rajesh, P S; Ravishankar Rai, V

    2014-01-01

    Quorum sensing mechanism allows the microorganisms to resist the antibiotic treatment by forming biofilms. Quorum quenching is one of the mechanisms to control the development of drug resistance in microbes. Endophyte bacteria are beneficial to plant growth as they support the immune system against the pathogen attack. The endophytic bacteria present in Pterocarpus santalinus were screened for the presence of N-acyl homoserine lactones (AHLs) degrading bacteria using biosensor strains and further confirmed by quantifying the violacein production. Cell-free lysate of endophytic bacteria, Bacillus firmus PT18 and Enterobacter asburiae PT39 exhibited potent AHL degrading ability by inhibiting about 80% violacein production in biosensor strain. Furthermore, when the cell-free lysate was applied to Pseudomonas aeruginosa PAO1 and PAO1-JP2 biofilm it resulted in significant (p<0.01) inhibition of biofilm formation. The biofilm inhibition was confirmed by visualization of biofilm slides under fluorescence microscopy, which showed decrease in total biomass formation in treated slides. Isolation and amplification of the gene (aiiA) indicated that the presence of AHL lactonase in cell-free lysate and sequence alignment indicated that AiiA contains a "HXHXDH" zinc-binding motif that is being conserved in several groups of metallohydrolases. Therefore, the study shows the potential of AHLs degradation by AHL lactonase present in cell-free lysate of isolated endophytic bacteria and inhibition of quorum sensing regulated biofilm formation in P. aeruginosa PAO1. PMID:24268182

  9. Sponge-Like Dressings Based on the Association of Chitosan and Sericin for the Treatment of Chronic Skin Ulcers. II. Loading of the Hemoderivative Platelet Lysate.

    Science.gov (United States)

    Mori, Michela; Rossi, Silvia; Ferrari, Franca; Bonferoni, Maria C; Sandri, Giuseppina; Riva, Federica; Tenci, Marika; Del Fante, Claudia; Nicoletti, Giovanni; Caramella, Carla

    2016-03-01

    Platelet lysate (PL) was loaded into dressings based on chitosan glutamate (CSG) low and high molecular weight, sericin (Ser), and glycine (Gly). A synergic effect of Ser and PL on fibroblast proliferation was proved in vitro. Two different PL loading approaches were considered: the first provided to prepare dressings by freeze-drying a mixture of PL and CSG/Gly/Ser solution, the second approach consisted in the extemporarily loading of PL in the CSG/Gly/Ser freeze-dried dressings. As for the first approach, PL loading did not produce any variation in dressing mechanical properties. Such dressings absorbed a high amount (about 8-fold of dry weight) of phosphate-buffered saline (fluid mimicking wound exudate), forming a gel with pseudoplastic and elastic properties. Platelet-derived growth factor AB assay indicated that neither freeze-drying nor the excipients alter PL growth factor content. As for the second approach, mechanical and rheological properties of the gel formed upon PL absorption enabled to choose a PL loading of about 90 μL/cm(2). Upon contact with fibroblasts, all PL loaded formulations increased the number not only of viable cells but also of those in the proliferative phase. Histological studies effected on human skin strips pointed out the positive effect of PL loaded dressings on dermal matrix reconstruction. PMID:26886335

  10. Using "On/Off" (19)F NMR/Magnetic Resonance Imaging Signals to Sense Tyrosine Kinase/Phosphatase Activity in Vitro and in Cell Lysates.

    Science.gov (United States)

    Zheng, Zhen; Sun, Hongbin; Hu, Chen; Li, Gongyu; Liu, Xiaomei; Chen, Peiyao; Cui, Yusi; Liu, Jing; Wang, Junfeng; Liang, Gaolin

    2016-03-15

    Tyrosine kinase and phosphatase are two important, antagonistic enzymes in organisms. Development of noninvasive approach for sensing their activity with high spatial and temporal resolution remains challenging. Herein, we rationally designed a hydrogelator Nap-Phe-Phe(CF3)-Glu-Tyr-Ile-OH (1a) whose supramolecular hydrogel (i.e., Gel 1a) can be subjected to tyrosine kinase-directed disassembly, and its phosphate precursor Nap-Phe-Phe(CF3)-Glu-Tyr(H2PO3)-Ile-OH (1b), which can be subjected to alkaline phosphatase (ALP)-instructed self-assembly to form supramolecular hydrogel Gel 1b, respectively. Mechanic properties and internal fibrous networks of the hydrogels were characterized with rheology and cryo transmission electron microscopy (cryo-TEM). Disassembly/self-assembly of their corresponding supramolecular hydrogels conferring respective "On/Off" (19)F NMR/MRI signals were employed to sense the activity of these two important enzymes in vitro and in cell lysates for the first time. We anticipate that our new (19)F NMR/magnetic resonance imaging (MRI) method would facilitate pharmaceutical researchers to screen new inhibitors for these two enzymes without steric hindrance. PMID:26901415

  11. The Potential of GMP-Compliant Platelet Lysate to Induce a Permissive State for Cardiovascular Transdifferentiation in Human Mediastinal Adipose Tissue-Derived Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Camilla Siciliano

    2015-01-01

    Full Text Available Human adipose tissue-derived mesenchymal stem cells (ADMSCs are considered eligible candidates for cardiovascular stem cell therapy applications due to their cardiac transdifferentiation potential and immunotolerance. Over the years, the in vitro culture of ADMSCs by platelet lysate (PL, a hemoderivate containing numerous growth factors and cytokines derived from platelet pools, has allowed achieving a safe and reproducible methodology to obtain high cell yield prior to clinical administration. Nevertheless, the biological properties of PL are still to be fully elucidated. In this brief report we show the potential ability of PL to induce a permissive state of cardiac-like transdifferentiation and to cause epigenetic modifications. RTPCR results indicate an upregulation of Cx43, SMA, c-kit, and Thy-1 confirmed by immunofluorescence staining, compared to standard cultures with foetal bovine serum. Moreover, PL-cultured ADMSCs exhibit a remarkable increase of both acetylated histones 3 and 4, with a patient-dependent time trend, and methylation at lysine 9 on histone 3 preceding the acetylation. Expression levels of p300 and SIRT-1, two major regulators of histone 3, are also upregulated after treatment with PL. In conclusion, PL could unravel novel biological properties beyond its routine employment in noncardiac applications, providing new insights into the plasticity of human ADMSCs.

  12. Two-dimensional blue native/SDS-PAGE analysis of whole cell lysate protein complexes of rice in response to salt stress.

    Science.gov (United States)

    Hashemi, Amenehsadat; Gharechahi, Javad; Nematzadeh, Ghorbanali; Shekari, Faezeh; Hosseini, Seyed Abdollah; Salekdeh, Ghasem Hosseini

    2016-08-01

    To understand the biology of a plant in response to stress, insight into protein-protein interactions, which almost define cell behavior, is thought to be crucial. Here, we provide a comparative complexomics analysis of leaf whole cell lysate of two rice genotypes with contrasting responses to salt using two-dimensional blue native/SDS-PAGE (2D-BN/SDS-PAGE). We aimed to identify changes in subunit composition and stoichiometry of protein complexes elicited by salt. Using mild detergent for protein complex solubilization, we were able to identify 9 protein assemblies as hetero-oligomeric and 30 as homo-oligomeric complexes. A total of 20 proteins were identified as monomers in the 2D-BN/SDS-PAGE gels. In addition to identifying known protein complexes that confirm the technical validity of our analysis, we were also able to discover novel protein-protein interactions. Interestingly, an interaction was detected for glycolytic enzymes enolase (ENO1) and triosephosphate isomerase (TPI) and also for a chlorophyll a-b binding protein and RuBisCo small subunit. To show changes in subunit composition and stoichiometry of protein assemblies during salt stress, the differential abundance of interacting proteins was compared between salt-treated and control plants. A detailed exploration of some of the protein complexes provided novel insight into the function, composition, stoichiometry and dynamics of known and previously uncharacterized protein complexes in response to salt stress. PMID:27362847

  13. Optimization of the isolation and expansion method of human mediastinal-adipose tissue derived mesenchymal stem cells with virally inactivated GMP-grade platelet lysate.

    Science.gov (United States)

    Siciliano, Camilla; Ibrahim, Mohsen; Scafetta, Gaia; Napoletano, Chiara; Mangino, Giorgio; Pierelli, Luca; Frati, Giacomo; De Falco, Elena

    2015-01-01

    Mesenchymal stem cells (MSCs) are adult multipotent cells currently employed in several clinical trials due to their immunomodulating, angiogenic and repairing features. The adipose tissue is certainly considered an eligible source of MSCs. Recently, putative adipose tissue derived MSCs (ADMSCs) have been isolated from the mediastinal depots. However, very little is known about the properties, the function and the potential of human mediastinal ADMSCs (hmADMSCs). However, the lack of standardized methodologies to culture ADMSCs prevents comparison across. Herein for the first time, we report a detailed step by step description to optimize the isolation and the expansion methodology of hmADMSCs using a virally inactivated good manufacturing practice (GMP)-grade platelet lysate, highlighting the critical aspects of the procedure and providing useful troubleshooting suggestions. Our approach offers a reproducible system which could provide standardization across laboratories. Moreover, our system is time and cost effective, and it can provide a reproducible source of adipose stem cells to enable future studies to unravel new insights regard this promising stem cell population. PMID:24306273

  14. Quantification of Saquinavir from Lysates of Peripheral Blood Mononuclear Cells Using Microarrays and Standard MALDI-TOF-MS

    Science.gov (United States)

    Pabst, Martin; Fagerer, Stephan Rupert; Köhling, Rudolf; Eyer, Klaus; Krismer, Jasmin; Jefimovs, Konstantins; Ibáñez, Alfredo Jesus; Zenobi, Renato

    2014-06-01

    Drug monitoring is usually performed by liquid chromatography coupled with optical detection or electrospray ionization mass spectrometry. More recently, matrix-assisted laser desorption/ionization (MALDI) in combination with triple quadrupole or Fourier-transform (FT) mass analyzers has also been reported to allow accurate quantification. Here, we present a strategy that employs standard MALDI time-of-flight (TOF) mass spectrometry (MS) for the sensitive and accurate quantification of saquinavir from an extract of blood peripheral mononuclear cells. Unambiguous identification of saquinavir in the mass spectra was possible because of using internal mass calibration and by an overall low chemical noise in the low mass range. Exact mass determination of the constant background peaks of the cell extract, which were used for recalibration, was performed by an initial MALDI-FT-MS analysis. Fast and multiplexed sample analysis was enabled by microarray technology, which provided 10 replicates in the lower nL range for each sample in parallel lanes on a chip. In order to validate the method, we employed various statistical tests, such as confidence intervals for linear regressions, three quality control samples, and inverse confidence limits of the estimated concentration ratios.

  15. Escherichia coli O157:H7 Cells Exposed to Lettuce Leaf Lysate in Refrigerated Conditions Exhibit Differential Expression of Selected Virulence and Adhesion-Related Genes with Altered Mammalian Cell Adherence.

    Science.gov (United States)

    Kennedy, Nicole M; Mukherjee, Nabanita; Banerjee, Pratik

    2016-07-01

    Contamination by and persistence of pathogenic bacteria in ready-to-eat produce have emerged as significant food safety and public health concerns. Viable produceborne pathogens cope with several stresses (e.g., temperature fluctuations and lowtemperature storage) during production and storage of the commodities. In this study, we investigated the impact of transient cold shock on Escherichia coli O157:H7 (EcO157) cells in a produce matrix (romaine lettuce leaf lysate). EcO157 cells were exposed to 25°C for 1 h, 4°C for 1 h, and 4°C for 10 min in lettuce lysate. The expression of selected genes coding for virulence, stress response, and heat and cold shock proteins was quantified by real-time quantitative reverse transcription PCR assay. Treated EcO157 cells adhered to MAC-T mammalian cells were enumerated by in vitro bioassay. Expression of the Shiga toxin 1 gene (stx1a) was upregulated significantly (P lettuce lysate. The adhesion assay revealed a temperature-dependent reduction in the attachment of cold-shocked EcO157 cells. The results of the current study indicate a reduction in the attachment of cold-shocked EcO157 to epithelial cells and higher levels of Shiga toxin gene expression at the molecular level. PMID:27357048

  16. Prime-boost vaccination with toxoplasma lysate antigen, but not with a mixture of recombinant protein antigens, leads to reduction of brain cyst formation in BALB/c mice.

    Directory of Open Access Journals (Sweden)

    Angelika Wagner

    Full Text Available Infection with the ubiquitous parasite Toxoplasma gondii is a threat for immunocompromised patients and pregnant women and effective immune-prophylaxis is still lacking.Here we tested a mixture of recombinant T. gondii antigens expressed in different developmental stages, i.e., SAG1, MAG1 and GRA7 (SMG, and a lysate derived from T. gondii tachyzoites (TLA for prophylactic vaccination against cyst formation. Both vaccine formulations were applied systemically followed by an oral TLA-booster in BALB/c mice.Systemic priming with SMG and oral TLA-booster did not show significant induction of protective immune responses. In contrast, systemic priming and oral booster with TLA induced higher levels of Toxoplasma-specific IgG, IgG1 and IgG2a in sera as well as high levels of Toxoplasma-specific IgG1 in small intestines. Furthermore, high levels of Toxoplasma-specific Th1-, Th17- and Th2-associated cytokines were only detected in restimulated splenocytes of TLA-vaccinated mice. Importantly, in mice orally infected with T. gondii oocysts, only TLA-vaccination and booster reduced brain cysts. Furthermore, sera from these mice reduced tachyzoites invasion of Vero cells in vitro, indicating that antibodies may play a critical role for protection against Toxoplasma infection. Additionally, supernatants from splenocyte cultures of TLA-vaccinated mice containing high levels of IFN-γ lead to substantial production of nitric oxide (NO after incubation with macrophages in vitro. Since NO is involved in the control of parasite growth, the high levels of IFN-γ induced by vaccination with TLA may contribute to the protection against T. gondii.In conclusion, our data indicate that prime-boost approach with TLA, but not with the mixture of recombinant antigens SMG, induces effective humoral and cellular Toxoplasma-specific responses and leads to significant reduction of cerebral cysts, thereby presenting a viable strategy for further vaccine development against T

  17. Platelet lysate as a substitute for animal serum for the ex-vivo expansion of mesenchymal stem/stromal cells: present and future.

    Science.gov (United States)

    Astori, Giuseppe; Amati, Eliana; Bambi, Franco; Bernardi, Martina; Chieregato, Katia; Schäfer, Richard; Sella, Sabrina; Rodeghiero, Francesco

    2016-01-01

    The use of fetal bovine serum (FBS) as a cell culture supplement is discouraged by regulatory authorities to limit the risk of zoonoses and xenogeneic immune reactions in the transplanted host. Additionally, FBS production came under scrutiny due to animal welfare concerns. Platelet derivatives have been proposed as FBS substitutes for the ex-vivo expansion of mesenchymal stem/stromal cells (MSCs) since platelet-derived growth factors can promote MSC ex-vivo expansion. Platelet-derived growth factors are present in platelet lysate (PL) obtained after repeated freezing-thawing cycles of the platelet-rich plasma or by applying physiological stimuli such as thrombin or CaCl2.PL-expanded MSCs have been used already in the clinic, taking advantage of their faster proliferation compared with FBS-expanded preparations. Should PL be applied to other biopharmaceutical products, its demand is likely to increase dramatically. The use of fresh platelet units for the production of PL raises concerns due to limited availability of platelet donors. Expired units might represent an alternative, but further data are needed to define safety, including pathogen reduction, and functionality of the obtained PL. In addition, relevant questions concerning the definition of PL release criteria, including concentration ranges of specific growth factors in PL batches for various clinical indications, also need to be addressed. We are still far from a common definition of PL and standardized PL manufacture due to our limited knowledge of the mechanisms that mediate PL-promoting cell growth. Here, we concisely discuss aspects of PL as MSC culture supplement as a preliminary step towards an agreed definition of the required characteristics of PL for the requirements of manufacturers and users. PMID:27411942

  18. Platelet lysate as a substitute for animal serum for the ex-vivo expansion of mesenchymal stem/stromal cells: present and future.

    Science.gov (United States)

    Astori, Giuseppe; Amati, Eliana; Bambi, Franco; Bernardi, Martina; Chieregato, Katia; Schäfer, Richard; Sella, Sabrina; Rodeghiero, Francesco

    2016-01-01

    The use of fetal bovine serum (FBS) as a cell culture supplement is discouraged by regulatory authorities to limit the risk of zoonoses and xenogeneic immune reactions in the transplanted host. Additionally, FBS production came under scrutiny due to animal welfare concerns. Platelet derivatives have been proposed as FBS substitutes for the ex-vivo expansion of mesenchymal stem/stromal cells (MSCs) since platelet-derived growth factors can promote MSC ex-vivo expansion. Platelet-derived growth factors are present in platelet lysate (PL) obtained after repeated freezing-thawing cycles of the platelet-rich plasma or by applying physiological stimuli such as thrombin or CaCl2.PL-expanded MSCs have been used already in the clinic, taking advantage of their faster proliferation compared with FBS-expanded preparations. Should PL be applied to other biopharmaceutical products, its demand is likely to increase dramatically. The use of fresh platelet units for the production of PL raises concerns due to limited availability of platelet donors. Expired units might represent an alternative, but further data are needed to define safety, including pathogen reduction, and functionality of the obtained PL. In addition, relevant questions concerning the definition of PL release criteria, including concentration ranges of specific growth factors in PL batches for various clinical indications, also need to be addressed. We are still far from a common definition of PL and standardized PL manufacture due to our limited knowledge of the mechanisms that mediate PL-promoting cell growth. Here, we concisely discuss aspects of PL as MSC culture supplement as a preliminary step towards an agreed definition of the required characteristics of PL for the requirements of manufacturers and users.

  19. Limulus test for pyrogens and radiometric sterility tests on radiopharmaceuticals. Part of a coordinated programme

    International Nuclear Information System (INIS)

    Sterility testing of radiopharmaceuticals prepared at BARC were carried out using the radiometric technique (Radiometric detection of the metabolic product 14Co2). Batches of different radiopharmaceuticals were tested for pyrogen using the limulus lysate method and the results were compared with the rabbit method. The results of sterility test on 202 batches of 19 different radiopharmaceuticals show that the radiometric method can be used for sterility testing of radiopharmaceuticals labelled with 35S, 51Cr, 57Co, 59Fe, 82Br, 86Rb, sup(99m)Tc, sup(113m)In, 125I and 169Yb. The radiometric test proves to be more rapid than the conventional one for the sterility testing of such radiopharmaceuticals. Detection time is between 6-21 hours. In the case of 131I-labelled radiopharmaceuticals and in the case of chlormerodrin-Hg-203, it was found an interference due to volatile species (sup(131m)Xe in the case of 131I and some volatile mercury form in the case of chlormerodrin). In these cases it would be possible to carry out the radiometric sterility test after separation of the microorganisms from the radioactive material (by filtration). The limulus lysate method can be employed for control of various pyrogen-prone raw materials and radiopharmaceuticals. Such method is the only method at present available for detecting the low level pyrogen contamination in intrathecal injections. The limulus test is more rapid than the rabbit test

  20. Proteasome Assay in Cell Lysates

    Science.gov (United States)

    Maher, Pamela

    2016-01-01

    The ubiquitin-proteasome system (UPS) mediates the majority of the proteolysis seen in the cytoplasm and nucleus of mammalian cells. As such it plays an important role in the regulation of a variety of physiological and pathophysiological processes including tumorigenesis, inflammation and cell death (Ciechanover, 2005; Kisselev and Goldberg, 2001). A number of recent studies have shown that proteasome activity is decreased in a variety of neurological disorders including Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis and stroke as well as during normal aging (Chung et al., 2001; Ciechanover and Brundin, 2003; Betarbet et al., 2005). This decrease in proteasome activity is thought to play a critical role in the accumulation of abnormal and oxidized proteins. Protein clearance by the UPS involves two sequential reactions. The first is the tagging of protein lysine residues with ubiquitin (Ub) and the second is the subsequent degradation of the tagged proteins by the proteasome. We herein describe an assay for the second of these two reactions (Valera et al., 2013). This assay uses fluorogenic substrates for each of the three activities of the proteasome: chymotrypsin-like activity, trypsin-like activity and caspase-like activity. Cleavage of the fluorophore from the substrate by the proteasome results in fluorescence that can be detected with a fluorescent plate reader.

  1. The development and assessment of high-throughput mass spectrometry-based methods for the quantification of a nanoparticle drug delivery agent in cellular lysate.

    Science.gov (United States)

    Buse, Joshua; Purves, Randy W; Verrall, Ronald E; Badea, Ildiko; Zhang, Haixia; Mulligan, Christopher C; Peru, Kerry M; Bailey, Jonathan; Headley, John V; El-Aneed, Anas

    2014-11-01

    The safe use of lipid-based drug delivery agents requires fast and sensitive qualitative and quantitative assessment of their cellular interactions. Many mass spectrometry (MS) based analytical platforms can achieve such task with varying capabilities. Therefore, four novel high-throughput MS-based quantitative methods were evaluated for the analysis of a small organic gene delivery agent: N,N-bis(dimethylhexadecyl)-1,3-propane-diammonium dibromide (G16-3). Analysis utilized MS instruments that detect analytes using low-resolution tandem MS (MS/MS) analysis (i.e. QTRAP or linear ion trap in this work) or high-resolution MS analysis (i.e. time of flight (ToF) or Orbitrap). Our results indicate that the validated fast chromatography (FC)-QTRAP-MS/MS, FC- LTQ-Orbitrap-MS, desorption electrospray ionization-collision-induced dissociation (CID)-MS/MS and matrix assisted laser desorption ionization-ToF/ToF-MS MS methods were superior in the area of method development and sample analysis time to a previously developed liquid chromatography (LC)-CID-MS/MS. To our knowledge, this is the first evaluation of the abilities of five MS-based quantitative methods that target a single pharmaceutical analyte. Our findings indicate that, in comparison to conventional LC-CID-MS/MS, the new MS-based methods resulted in a (1) substantial reduction in the analysis time, (2) reduction in the time required for method development and (3) production of either superior or comparable quantitative data. The four new high-throughput MS methods, therefore, were faster, more efficient and less expensive than a conventional LC-CID-MS/MS for the quantification of the G16-3 analyte within tissue culture. When applied to cellular lysate, no significant change in the concentration of G16-3 gemini surfactant within PAM212 cells was observed between 5 and 53 h, suggesting the absence of any metabolism/excretion from PAM212 cells. PMID:25395133

  2. The development and assessment of high-throughput mass spectrometry-based methods for the quantification of a nanoparticle drug delivery agent in cellular lysate.

    Science.gov (United States)

    Buse, Joshua; Purves, Randy W; Verrall, Ronald E; Badea, Ildiko; Zhang, Haixia; Mulligan, Christopher C; Peru, Kerry M; Bailey, Jonathan; Headley, John V; El-Aneed, Anas

    2014-11-01

    The safe use of lipid-based drug delivery agents requires fast and sensitive qualitative and quantitative assessment of their cellular interactions. Many mass spectrometry (MS) based analytical platforms can achieve such task with varying capabilities. Therefore, four novel high-throughput MS-based quantitative methods were evaluated for the analysis of a small organic gene delivery agent: N,N-bis(dimethylhexadecyl)-1,3-propane-diammonium dibromide (G16-3). Analysis utilized MS instruments that detect analytes using low-resolution tandem MS (MS/MS) analysis (i.e. QTRAP or linear ion trap in this work) or high-resolution MS analysis (i.e. time of flight (ToF) or Orbitrap). Our results indicate that the validated fast chromatography (FC)-QTRAP-MS/MS, FC- LTQ-Orbitrap-MS, desorption electrospray ionization-collision-induced dissociation (CID)-MS/MS and matrix assisted laser desorption ionization-ToF/ToF-MS MS methods were superior in the area of method development and sample analysis time to a previously developed liquid chromatography (LC)-CID-MS/MS. To our knowledge, this is the first evaluation of the abilities of five MS-based quantitative methods that target a single pharmaceutical analyte. Our findings indicate that, in comparison to conventional LC-CID-MS/MS, the new MS-based methods resulted in a (1) substantial reduction in the analysis time, (2) reduction in the time required for method development and (3) production of either superior or comparable quantitative data. The four new high-throughput MS methods, therefore, were faster, more efficient and less expensive than a conventional LC-CID-MS/MS for the quantification of the G16-3 analyte within tissue culture. When applied to cellular lysate, no significant change in the concentration of G16-3 gemini surfactant within PAM212 cells was observed between 5 and 53 h, suggesting the absence of any metabolism/excretion from PAM212 cells.

  3. Endotoxins in surgical instruments of hip arthroplasty

    Directory of Open Access Journals (Sweden)

    Vania Regina Goveia

    2016-06-01

    Full Text Available Abstract OBJECTIVE To investigate endotoxins in sterilized surgical instruments used in hip arthroplasties. METHOD A descriptive exploratory study conducted in a public teaching hospital. Six types of surgical instruments were selected, namely: acetabulum rasp, femoral rasp, femoral head remover, chisel box, flexible bone reamer and femoral head test. The selection was based on the analysis of the difficulty in removing bone and blood residues during cleaning. The sample was made up of 60 surgical instruments, which were tested for endotoxins in three different stages. The EndosafeTM Gel-Clot LAL (Limulus Amebocyte Lysate method was used. RESULT There was consistent gel formation with positive analysis in eight instruments, corresponding to 13.3%, being four femoral rasps and four bone reamers. CONCLUSION Endotoxins in quantity ≥0.125 UE/mL were detected in 13.3% of the instruments tested.

  4. Escherichia coli O157:H7 Cells Exposed to Lettuce Leaf Lysate in Refrigerated Conditions Exhibit Differential Expression of Selected Virulence and Adhesion-Related Genes with Altered Mammalian Cell Adherence.

    Science.gov (United States)

    Kennedy, Nicole M; Mukherjee, Nabanita; Banerjee, Pratik

    2016-07-01

    Contamination by and persistence of pathogenic bacteria in ready-to-eat produce have emerged as significant food safety and public health concerns. Viable produceborne pathogens cope with several stresses (e.g., temperature fluctuations and lowtemperature storage) during production and storage of the commodities. In this study, we investigated the impact of transient cold shock on Escherichia coli O157:H7 (EcO157) cells in a produce matrix (romaine lettuce leaf lysate). EcO157 cells were exposed to 25°C for 1 h, 4°C for 1 h, and 4°C for 10 min in lettuce lysate. The expression of selected genes coding for virulence, stress response, and heat and cold shock proteins was quantified by real-time quantitative reverse transcription PCR assay. Treated EcO157 cells adhered to MAC-T mammalian cells were enumerated by in vitro bioassay. Expression of the Shiga toxin 1 gene (stx1a) was upregulated significantly (P < 0.05) upon cold shock treatments, but virulence genes related to EcO157 attachment (eaeA, lpfA, and hcpA) were down-regulated. Two key members of the cold shock regulon, cold shock protein (cspA) and gyrA, were significantly induced (P < 0.05) at the refrigeration temperature (4°C). Significant upregulation of an SOS response gene, recA, was also observed. E. coli heat shock regulon member grpE was induced, but a universal stress protein (uspA) was downregulated at the refrigeration temperatures in lettuce lysate. The adhesion assay revealed a temperature-dependent reduction in the attachment of cold-shocked EcO157 cells. The results of the current study indicate a reduction in the attachment of cold-shocked EcO157 to epithelial cells and higher levels of Shiga toxin gene expression at the molecular level.

  5. Lipidated alpha-Peptide/beta-Peptoid Hybrids with Potent Antiinflammatory Activity

    DEFF Research Database (Denmark)

    Skovbakke, Sarah L.; Larsen, Camilla J.; Heegaard, Peter M. H.;

    2015-01-01

    , in contrast to polymyxin B, displayed only minor activity in the Limulus amebocyte lysate assay. Flow cytometry data showed specific interaction of a fluorophore-labeled lipidated a-peptide/beta-peptoid hybrid with monocytes and granulocytes indicating a cellular target expressed by these leukocyte subsets....

  6. Increased Tryptophan Catabolism is Associated with Increased Frequency of CD161+Tc17/MAIT Cells, and Lower CD4+ T cell Count in HIV-1 infected Patients on cART after Two Years of Follow-up

    DEFF Research Database (Denmark)

    Gaardbo, Julie Christine; Trøseid, Marius; Stiksrud, Birgitte;

    2015-01-01

    were measured using flow cytometry. Soluble CD14, LPS and tryptophan metabolites in plasma were measured retrospectively prior to cART and at inclusion initiation using Limulus Amebocyte Lysate colometric assay, ELISA, and tandem mass spectrometry, respectively. KTR was calculated, and patients were...

  7. Effect of extraction and assay media on analysis of airborne endotoxin

    NARCIS (Netherlands)

    Spaan, S.; Doekes, G.; Heederik, D.; Thorne, P.S.; Wouters, I.M.

    2008-01-01

    The measurement of airborne endotoxins is thus far not standardized. Earlier studies reported higher endotoxin yields when Tween 20 was added to the media used for filter extraction and in the Limulus amebocyte lysate (LAL) assay. This study compared four common media and assessed the effects of Twe

  8. Removal of endotoxins from bacteriophage preparations by extraction with organic solvents.

    Directory of Open Access Journals (Sweden)

    Bożena Szermer-Olearnik

    Full Text Available Lipopolysaccharide (LPS, endotoxin, pyrogen constitutes a very troubling contaminant of crude phage lysates produced in Gram-negative bacteria. Toxicity of LPS depends on the strong innate immunity response including the cytokines. Therefore, its removal is important for bacteriophage applications. In this paper, we present a procedure for extractive removal of endotoxin from bacteriophage preparations with water immiscible solvents (1-octanol or 1-butanol. During extraction most of the phage lytic activity is retained in the aqueous phase, while endotoxin accumulates in the organic solvent. The levels of endotoxin (expressed as endotoxin units, EU in the aqueous bacteriophage-containing fraction determined by limulus amebocyte lysate or EndoLISA assay were exceptionally low. While the initial endotoxin levels in the crude phage lysates ranged between 10(3 and 10(5 EU/ml the average level after organic extraction remaining in the aqueous fraction was 5.3 EU/ml. These values when related to phage titers decreased from 10(3-10(5 EU/10(9 PFU (plaque forming units down to an average of 2.8 EU/10(9 PFU. The purification procedure is scalable, efficient and applicable to all the bacteriophages tested: T4, HAP1 (E. coli and F8 (P. aeruginosa.

  9. Endotoxins in the prostatic secretions of chronic prostatitis patients

    Institute of Scientific and Technical Information of China (English)

    Yu-Ping Dai; Xiang-Zhou Sun; Ke-Li Zheng

    2005-01-01

    Aim: To evaluate the clinical significance of the quantitative determinations of endotoxins in the expressed prostatic secretions (EPS) of chronic prostatitis (CP) patients. Methods: The EPS of 45 patients with CP and 15 normal volunteers were obtained for microscopic examination, bacterial culture and endotoxin determination. The level of endotoxins was determined by the Limulus-amebocyte-lysate test with chromogenic substrate. Results: Patients (P>0.05), type Ⅱ/type Ⅲa vs. Normal controls P < 0.05)]. Conclusion: CP patients have elevated levels of endotoxins in the EPS, which suggests that inflammation is a feature of this disease. EPS endotoxin determination is not only helpful in diagnostic confirmation, but also in evaluating the response to treatment in CP patients.

  10. Repair Following Glutamate-induced Excitotoxic Neuronal Damage Mediated by Intracerebroventricular Injection of Cell-free Filtrate of Neural Stem Cell Lysates in Adult Mice%小鼠脑室内注射神经干细胞裂解液促进谷氨酸盐诱导的兴奋性神经元损伤的修复

    Institute of Scientific and Technical Information of China (English)

    于立坚; 马娟; 马润娣; 张永平; 房娟芝; 张霄瑜; 于廷曦

    2011-01-01

    Our previous study demonstrated that cell-free filtrate of sodium ferulate-induced and differen-tioned PC 12 cell lysates significantly attenuated chronic mild stress-induced depression-like behavioural disorders, up-regulated hippocampal and cerebral cortex expressions of nerve growth factor (NGF) and brain-derived neu-rotrophic factor (BDNF), and increased hippocampal number of neural stem cells (NSC)/neural progenitor cells in mice. The present study was undertaken to investigate the possibility of the repair following glutamate (monosodium glutamate, MSG)-induced excitotoxic brain injury mediated by intracerebroventricular injection of cell-free filtrate of neural stem cell lysates (FNSCL) in adult mice. Mouse NSCs were isolated from the brains of embryos at 15 day postcoitum (dpc). The expression of nestin, a special antigen for NSC, was detected by immunocytochemistry. Cell-free filtrate of NSCs was prepared from the NSC lysates. The animals in the MSG group received intragastric (ig) administration of MSG (2.0 g/(kgd) for 10 days), the animals in the MSG+NSCs group received intracerebroventricular transplantation of NSCs (approximately l.OxlO5 cells), and the animals in the MSG+FNSCL group received intracerebroventricular injection of 10 \\iL of FNSCL (approximately 1.0* 105 cells) separately on day 1 and day 10 after 10-d MSG exposure. The mice in control and MSG groups received intracerebroventricular injection of Dul-becco's modified Eagle's medium (DMEM) instead of NSCs or FNSCL. On 12 day after the last intracerebroventricular injection of FNSCL or transplantation of NSCs, Y-maze test was performed, and then the histopathology of animal brains was studied to analyze MSG-induced functional and morphological changes and the effects of intracerebroventricular injection of FNSCL and transplantation of NSCs on the repair of MSG-induced excitotoxic brain injury. The results showed that both intracerebroventricular injection of FNSCL and

  11. Immunosensor for the ultrasensitive and quantitative detection of bladder cancer in point of care testing.

    Science.gov (United States)

    Chuang, Cheng-Hsin; Du, Yi-Chun; Wu, Ting-Feng; Chen, Cheng-Ho; Lee, Da-Huei; Chen, Shih-Min; Huang, Ting-Chi; Wu, Hsun-Pei; Shaikh, Muhammad Omar

    2016-10-15

    An ultrasensitive and real-time impedance based immunosensor has been fabricated for the quantitative detection of Galectin-1 (Gal-1) protein, a biomarker for the onset of multiple oncological conditions, especially bladder cancer. The chip consists of a gold annular interdigitated microelectrode array (3×3 format with a sensing area of 200µm) patterned using standard microfabrication processes, with the ability to electrically address each electrode individually. To improve sensitivity and immobilization efficiency, we have utilized nanoprobes (Gal-1 antibodies conjugated to alumina nanoparticles through silane modification) that are trapped on the microelectrode surface using programmable dielectrophoretic manipulations. The limit of detection of the immunosensor for Gal-1 protein is 0.0078mg/ml of T24 (Grade III) cell lysate in phosphate buffered saline, artificial urine and human urine samples. The normalized impedance variations show a linear dependence on the concentration of cell lysate present while specificity is demonstrated by comparing the immunosensor response for two different grades of bladder cancer cell lysates. We have also designed a portable impedance analyzing device to connect the immunosensor for regular checkup in point of care testing with the ability to transfer data over the internet using a personal computer. We believe that this diagnostic system would allow for improved public health monitoring and aid in early cancer diagnosis. PMID:26777732

  12. Specific immune responses against hepatocellular carcinoma induced by exosomes derived from dendritic cells loaded with hepatocellular carcinoma cancer cell lysates%肝癌细胞裂解物负载的树突状细胞来源的exosomes制备及其抗肝癌免疫活性的研究

    Institute of Scientific and Technical Information of China (English)

    杨静悦; 孙飞; 付蓉; 薛妍; 刘文超

    2013-01-01

    目的:制备一种比单纯源于DC的、更有效的新型治疗肝癌的DC来源的exosomes瘤苗,进而探讨其生物学特性、免疫学功能及抗肿瘤免疫活性.方法:用细胞因子诱导培养树突状细胞(dendritic cell,DC),将肝癌细胞HepG2裂解物负载DC后,提取exosomes.透射电镜观察exosomes形态,流式细胞术检测exosomes蛋白分子的表达;其后应用exosomes直接刺激效应淋巴细胞,MTT法检测CTL对靶细胞HepG2的杀伤活性.结果:透射电镜下观察到负载组exosomes为直径50-100nm的膜性微囊,圆形或椭圆形,有完整包膜.FCM检测表明,负载组exosomes含有DC的特征分子(包括CD86、CD83).负载组来源的exosomes活化的T细胞对HepG2的杀伤率显著高于未负载组及单纯淋巴细胞组(P<0.05).结论:肝癌细胞裂解物负载能增强DC分泌的exosomes体外诱导CTL效应.本研究为制备高效的exosomes肝癌瘤苗提供了实验依据.%Objective:To obtain high eifficacy hepatocellular carcinoma exosomes cancer vaccine and to assess the properties of these exosomes.Methods:Dendritic cells (DCs) were induced with cytokines and then loaded with whole hepatocellular carcinoma cell lysates.Exosomes were isolated from supematant of DCs.Transmission electron microscopy was used to observe their structu1es.The expressions of several proteins were investigated by flow cytometry.T lymphocytes were pulsed with exosomes directly in the presence of IL-2.After co-cultured with target cells,the stimulated lymphocytes were tested by cytotoxicity capacity assay.Results:Application of the isolation procedure to loaded DCs revealed exosomes vesicles by transmission electron microscopy.Protein analysis by FCM was performed and revealed that the costimulatory molecule CD86 and CD83 was detectable.The loaded DCs derived exosomes could directly activate T lymphocytes which lysed HepG2 target cells much more effectively than unloaded groups and L groups (P < 0.05).Conclusion:The exosomes

  13. Evaluation of a combined lysate/recombinant antigen anti-HTLV-I/II ELISA in high and low endemic areas of HTLV-I/II infection.

    Science.gov (United States)

    Vrielink, H; Sisay, Y; Reesink, H W; Woerdeman, M; Winkel, C; de Leeuw, S J; Lelie, P N; van der Poel, C L

    1995-06-01

    The Wellcozyme HTLV-I/II ELISA (Murex Diagnostics) was evaluated in 7800 samples of various serum panels. Repeat activity was found by Wellcozyme in (A) 1/2181 (0.05%) Dutch blood donors, (B) 44/3036 (1.4%) Curaçao (Caribbean area) blood donors, (C) 46/2533 (1.8%) individuals of different Ethiopian population subsets, (D) 30/30 (100%) confirmed anti-HTLV-I positive samples and (E) 20/20 (100%) HTLV-II PCR-positive samples. All 91 Wellcozyme-positive samples were tested for confirmation by Western blot (WB, Diagnostic Biotechnology). Among Wellcozyme HTLV-I/II ELISA-positive individuals, HTLV-I/II WB positivity was found in 0/1 Dutch blood donors, 40/44 (88.9%) Curaçao blood donors and 20/46 (43.5%) Ethiopian individuals. HTLV-I positivity was found in 40 (1.3%) WB-positive Curaçao blood donors and in 9 (0.35%) Ethiopian individuals. HTLV-II positivity was found in 11 (0.43%) WB-positive Ethiopian individuals. The Wellcozyme HTLV-I/II ELISA had a specificity of 99.95% in Dutch blood donors and a sensitivity of 100% on confirmed HTLV-I- and HTLV-II-positive samples. In Ethiopia 55% of the HTLV-I/II WB-positive individuals were exclusively HTLV-II positive, whereas in Curaçao no HTLV-II infections were found. PMID:7655577

  14. mRNAs containing the unstructured 5' leader sequence of alfalfa mosaic virus RNA 4 translate inefficiently in lysates from poliovirus-infected HeLa cells.

    Science.gov (United States)

    Hann, L E; Gehrke, L

    1995-01-01

    Poliovirus infection is accompanied by translational control that precludes translation of 5'-capped mRNAs and facilitates translation of the uncapped poliovirus RNA by an internal initiation mechanism. Previous reports have suggested that the capped alfalfa mosaic virus coat protein mRNA (AIMV CP RNA), which contains an unstructured 5' leader sequence, is unusual in being functionally active in extracts prepared from poliovirus-infected HeLa cells (PI-extracts). To identify the cis-acting nucleotide elements permitting selective AIMV CP expression, we tested capped mRNAs containing structured or unstructured 5' leader sequences in addition to an mRNA containing the poliovirus internal ribosome entry site (IRES). Translations were performed with PI-extracts and extracts prepared from mock-infected HeLa cells (MI-extracts). A number of control criteria demonstrated that the HeLa cells were infected by poliovirus and that the extracts were translationally active. The data strongly indicate that translation of RNAs lacking an internal ribosome entry site, including AIMV CP RNA, was severely compromised in PI-extracts, and we find no evidence that the unstructured AIMV CP RNA 5' leader sequence acts in cis to bypass the poliovirus translational control. Nevertheless, cotranslation assays in the MI-extracts demonstrate that mRNAs containing the unstructured AIMV CP RNA 5' untranslated region have a competitive advantage over those containing the rabbit alpha-globin 5' leader. Previous reports of AIMV CP RNA translation in PI-extracts likely describe inefficient expression that can be explained by residual cap-dependent initiation events, where AIMV CP RNA translation is competitive because of a diminished quantitative requirement for initiation factors. PMID:7609069

  15. Detection of endotoxins in radiopharmaceutical preparations. III. Limulus test assessment using radiopharmaceutical preparations; correlation with the rabbit pyrogen test

    Energy Technology Data Exchange (ETDEWEB)

    Cohen, Y.; Bahri, F.; Bruneau, J.; Dubuis, M.; Dubuis, N.; Merlin, L.; Michaud, T.; Peysson, S.

    1986-01-01

    Experiments using 17 radiopharmaceuticals containing known amounts of added endotoxin show that none of them inhibits the pyrogenic reaction of the rabbit. Gelation of the Limulus amoebocyte lysate (LAL) is inhibited by 4 of them: colloidal erbium 169Er citrate, colloidal rhenium 186Re sulfide, colloidal technetium /sup 99m/Tc (Re) sulfide for liver scintigraphy and the colloidal technetium /sup 99m/Tc (Re) sulfide for lymphography. This inhibition is cancelled, either by dilution or after neutral pH adjustment. Both controls were performed on 313 batches of various radiopharmaceuticals, 95% of results were identical (93% negative, 2% positive). The remaining 5% correspond to positive LAL tests vs negative rabbit tests on the same batches. No negative LAL test vs positive rabbit test was observed.

  16. Validated LC-MS/MS method for simultaneous determination of SIM and its acid form in human plasma and cell lysate: Pharmacokinetic application%Validated LC-MS/MS method for simultaneous determination of SIM and its acid form in human plasma and cell lysate: Pharmacokinetic application

    Institute of Scientific and Technical Information of China (English)

    Tamer A. Ahmed; Jamie Horn; John Hayslip; Markos Leggas

    2012-01-01

    Simvastatin (SIM) is a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor widely used in hyperlipidemia therapy. SIM has recently been studied for its anticancer activity at doses higher than those used for the hyperlipidemia therapy. This prompted us to study the pharmacokinetics of high-dose SIM in cancer patients. For this purpose, an LC-MS/MS method was developed to measure SIM and its acid form (SIMA) in plasma and peripheral blood mononuclear cells (PBMCs) obtained from patients. Chromatographic analyte separation was carried out on a reverse-phase column using 75:25 (% v/v) acetonitrile:ammonium acetate (0.1 M, pH 5.0) mobile phase. Detection was performed on a triple quadrupole mass spectrometer, equipped with a turbo ion spray source and operated in positive ionization mode. The assay was linear over a range 2.5-500 ng/mL for SIM and 5-500 ng/mL for SIMA in plasma and 2.5-250 ng/mL for SIM and 5-250 ng/mL for SIMA in cell lysate. Recovery was 〉 58% for SIM and 〉 75% for SIMA in both plasma and cell lysate. SIM and SIMA were stable in plasma, cell lysate and the reconstitution solution. This method was successfully applied for the determination of SIM and SIMA in plasma and PBMCs samples collected in the pharmacokinetic study of high-dose SIM in cancer patients.

  17. Polycystic kidney disease: an unrecognized emerging infectious disease?

    OpenAIRE

    Miller-Hjelle, M. A.; Hjelle, J T; Jones, M.; Mayberry, W. R.; Dombrink-Kurtzman, M. A.; Peterson, S.W.; Nowak, D. M.; Darras, F.S.

    1997-01-01

    Polycystic kidney disease (PKD) is one of the most common genetic diseases in humans. We contend that it may be an emerging infectious disease and/or microbial toxicosis in a vulnerable human subpopulation. Use of a differential activation protocol for the Limulus amebocyte lysate (LAL) assay showed bacterial endotoxin and fungal (1-->3)-beta-D-glucans in cyst fluids from human kidneys with PKD. Fatty acid analysis of cyst fluid confirmed the presence of 3-hydroxy fatty acids characteristic o...

  18. 人血小板裂解液替代胎牛血清促进骨髓间质干细胞的增殖%Human platelet lysates promotes the proliferation of mesenchymal stem cells in vitro

    Institute of Scientific and Technical Information of China (English)

    许茹; 夏文杰; 戎霞; 叶欣; 邵媛; 王敏; 罗广平; 付涌水

    2011-01-01

    Objective To investigate the effect of human platelet lysates (HPL) obtained from platelet-rich plasma on the proliferation and biological characteristics of human mesenchymal stem cells (MSCs) in vitro. Methods HPL was obtained by repeated freeze-thawing of human plateletes, and the MSCs separated by density gradient centrifugation from 6 donors were expanded in medium supplemented with 10% fetal bovine serum (FCS) or HPL at different concentrations. The optimal concentration of HPL for cells culture was determined according to the cell proliferation kinetics. The cultured MSCs were characterized for their proliferation, cell phenotype, and cell cycle distribution. Results The HPL-supplemented medium contained 4 essential growth factors for the growth of MSCs, namely platelet-derived growth factors (0.53±0.06 ng/ml), basic fibroblast growth factor (37.5±4.31 pg/ml), insulin-like growth factor-1 (0.15+0.06 mg/ml) and transforming growth factor (5150±463 pg/ml). Cultured in the presence of HPL at the optimal concentration of 7.5%, the MSCs displayed a spindle-shaped fibroblast-like morphology without obvious changes in the proliferation activity till passage 8 (P>0.05), similar to those of cells in FCS-supplemented culture medium. Flow cytometry and cell cycle analysis revealed no differences in the phenotypes or cell cycle distribution between the cells cultured in the presence of 7.5% HPL and 10% FCS. Conclusion The culture medium supplemented by 7.5% HPL can promote the expansion of human MSCs and maintain the basic biological characteristics of the cells.%目的 探讨人血小板裂解液(human platelet lysates,HPL)对骨髓间质干细胞(mesenchymal stem cells,MSCs)增殖以及生物学特性的影响.方法 通过对机采血小板反复冻融获得HPL,采用密度梯度离心法从骨髓中分离MSCs,分别用10%胎牛血清(feotal calf serum,FCS)和LD-DMEM添加不同浓度的HPL进行培养,以确定最佳HPL浓度.分离6株人源MSCs,从原代开

  19. Experimental study of mycobacterium tuberculosis lysates Inducing osteoclast and RANKL/OPG expression changes%结核分枝杆菌裂解物诱导破骨细胞及OPG/RANKL表达变化实验研究

    Institute of Scientific and Technical Information of China (English)

    张元豫; 张浩沙强; 李坤

    2013-01-01

    目的探讨结核分枝杆菌超声裂解产物(Mt sonicate)体外诱导小鼠破骨细胞的形成及 OPG/RANKL表达变化与裂解物的浓度、时间的依赖性,研究结核杆菌引起骨破坏的相关机制。方法原代培养的 BALB/c小鼠骨髓基质细胞以高、中、低浓度的 Mt sonicate进行干预,在1、3、7、14 d进行 TRAP染色,观察破骨细胞的形成,并采用 Realtime RT-PCR检测各浓度、时间的骨保护素(OPG)/核因子Kappa B受体活化因子配基(RANKL)mR-NA表达。结果 Mt sonicate在体外可诱导小鼠骨髓基质细胞向破骨细胞的转化;破骨细胞数量与 Mt sonicate浓度在3、7 d时呈显著的正相关(3 d:r=0.417,P <0.05;7 d:r=0.463,P <0.05);3个浓度干预下,破骨细胞数量在1~7 d逐渐增多,7~14 d则逐渐减少。7 d时,高、中、低浓度 Mt sonicate干预下 OPG/RANKL mRNA表达比分别为(1.35±0.11)、(7.38±1.15)、(9.85±1.26)。结论小鼠骨髓基质细胞向破骨细胞转化的实验中,存在结核分枝杆菌裂解物的浓度和时间依赖性,且与 OPG/RANKL mRNA的表达比呈负相关,破骨细胞形成时的数量与活性受到结核分枝杆菌的调控,其分子机制与 OPG/RANKL系统的变化密切相关。%Objective This paper aims to explore the formation of broken bone cell in vitro mouse bone marrow stromal cells by Mycobacterium tuberculosis ultrasound pyrolysis products (Mt sonicate ) induction,the lysate concentration dependence of the time,and the mechanisms of bone destruction caused by TB bacilli.Methods Primary cultures of BALB/c mice bone marrow stromal cells were intervened with high,medium and low concentrations of Mt sonicate on 1,3,7,14 d;osteoclast formation was observed with TRAP staining and Realtime RT-PCR to detect the concentration and time of the OPG/RANKL mR-NA expression.Results Mt sonicate can induce in vitro transformation of mouse bone

  20. Chlamydia Testing

    Science.gov (United States)

    ... Amplification Test (NAAT); Chlamydia trachomatis Culture; Chlamydia trachomatis DNA Probe Related tests: Gonorrhea Testing , HIV Antibody and HIV Antigen , Syphilis Tests , Herpes Testing , HPV Test , Trichomonas Testing All content on Lab Tests Online has ...

  1. Syphilis Test

    Science.gov (United States)

    ... Gonorrhea Testing , Chlamydia Testing , Herpes Testing , HPV Test , Trichomonas Testing , CSF Analysis All content on Lab Tests ... testing ( polymerase chain reaction, PCR )--this test detects genetic material from the bacteria in the sample from ...

  2. H22 tumor cell lysate plus adjuvants can effectively induce anti-tumor immunity%佐剂增强的肿瘤细胞裂解物疫苗抗小鼠H22肝癌作用

    Institute of Scientific and Technical Information of China (English)

    刘彬; 邢芸; 张秀华; 王泽宇; 路蕾; 葛驰宇; 张龙; 刘景晶; 曹荣月

    2012-01-01

    Whole cell lysate of mouse hepatoma ( H22 ) was prepared and processed to the whole tumor cell vaccine H22-DT-M2-OK432 ( abbreviated as HDTMOK ) by chemical coupling a mixture of DT and microbial HSP70 peptide epitope 407-426 and addition of OK432 as antigen. According to the results of therapeutic immune, the immune response stimulated by HDTMOK had effectively inhibited the growth of H22. Compared with that of the PBS group, the average weight and size of the tumor had both significantly reduced ( P < 0. 05 ). In order to enhance the anti-tumor effect of the vaccines, immunostimulating complex ( ISCOM ) were prepared on the basis of HDTMOK. Therapeutic immune results showed that the mean weight and size of excised tumors significantly reduced compared with that of the PBS group ( P < 0. 01 ). ISCOM also effectively retarded the angiogenesis of intradermal tumor model ( P < 0. 01 ). Meanwhile, the anti-tumor effect was also strengthened in comparison with HDTMOK ( P <0. 05 ).%以鼠源肝癌H22全细胞裂解物为抗原,通过化学偶联白喉毒素(DT)和串联重复的T 辅助表位mHSP70407-426肽段,混合OK432(链球菌A群)制成肿瘤全细胞疫苗H22-DT-M2-OK432(HDTMOK).治疗性免疫结果显示,疫苗激发的免疫应答对H22肿瘤起到了有效的抑制作用.为进一步提高该疫苗的抗肿瘤效果,用偶联的疫苗制备免疫刺激复合物(ISCOM),并验证其抑瘤效果.治疗性免疫结果显示,与PBS组相比,ISCOM疫苗组平均瘤重和瘤体积显著降低(P<0.01),同时有效地抑制了小鼠皮内肿瘤模型中的血管新生(P<0.01); ELISA法从血清中检测到高滴度的抗体.且与HDTMOK疫苗相比,ISCOM疫苗抑瘤作用提升显著(P<0.05).HDTMOK能有效抑制小鼠肝癌实体瘤生长,佐剂配伍后的疫苗对H22的抑制更为显著,能够有效提升肿瘤全细胞疫苗的抗肿瘤能力.

  3. Gonorrhea Test

    Science.gov (United States)

    ... gonorrhoeae Culture; Neisseria gonorrhoeae Gram Stain; Neisseria gonorrhoeae DNA Probe Related tests: Chlamydia Testing , HIV Antibody and HIV Antigen , Syphilis Tests , Herpes Testing , HPV Test , Trichomonas Testing All content on Lab Tests Online has ...

  4. Large-scale expansion of human umbilical cord mesenchymal stem cells in human platelet lysate as a substitute of fetal bovine serum%人血小板裂解液替代胎牛血清大规模扩增人脐带间充质干细胞

    Institute of Scientific and Technical Information of China (English)

    李炳尧; 武晓云; 吴岩

    2014-01-01

    背景:目前国内外大多数实验仍采用经典的培养基和胎牛血清进行脐带间充质干细胞培养,血清培养潜在的不安全因素限制了未来临床应用的可行性。  目的:以人血小板裂解液替代胎牛血清培养、鉴定人间充质干细胞,以期进一步应用于临床低密度扩增人间充质干细胞。  方法:人血小板裂解液通过反复冻融、离心、过滤、浓缩等方法制备。以 IMDM 为基础培养基,添加5%浓缩血小板裂解液作为实验组培养基;以添加体积分数10%胎牛血清为对照组培养基。采用酶消化法分离培养人脐带间充质干细胞,以3000/cm2的浓度进行传代培养,待细胞扩增至P5代后,进行细胞形态与直径、免疫表型、成骨成脂分化、克隆形成率等细胞生物学特性检测,并比较两者之间的差别。  结果与结论:人血小板裂解液扩增的脐带间充质干细胞形态细长,有更高的细胞累积群倍数;克隆形成检测结果显示两者形成的克隆效率差异无显著性意义,流式细胞术检测结果显示两者有相似的细胞表型,体外诱导分化显示两者都具有成骨、成脂分化能力,但人血小板裂解液扩增的间充质干细胞成骨分化能力更强。提示人血小板裂解液可以取代胎牛血清用于临床低密度扩增人间充质干细胞。%BACKGROUND:Classic media and fetal bovine serum are commonly used in the culture of umbilical cord mesenchymal stem cells, but the potential risk of serum culture limits its clinical application. OBJECTIVE:To use human platelet lysate alternative to fetal bovine serum for large-scale production of mesenchymal stem cells for therapeutic applications. METHODS:Human platelet lysate was prepared by repeated freezing and thawing, centrifugation, filtration, and concentration. Human umbilical cord mesenchymal stem cells were cultured in Iscove’s Modified Dulbecco’s Medium

  5. Susceptibility Testing

    Science.gov (United States)

    ... page helpful? Also known as: Sensitivity Testing; Drug Resistance Testing; Culture and Sensitivity; C & S; Antimicrobial Susceptibility Formal name: Bacterial and Fungal Susceptibility Testing Related tests: Urine Culture ; ...

  6. Anti-tumor effect induced by exosomes derived from dendritic cells loaded with lung cancer cell lysates%肺癌细胞裂解物负载对树突状细胞分泌的exosome诱导抗肿瘤作用的影响

    Institute of Scientific and Technical Information of China (English)

    张在云; 李希德; 刘叶; 王志仑; 潘祥林

    2011-01-01

    目的 为制备高效的胞外体(exosome)肿瘤疫苗提供理论依据.方法 用细胞因子诱导培养树突状细胞(DC),将肺癌细胞裂解物负载DC,提取exosome;用exosome活化T细胞(负载组),以未负载DC的exosome(未负载组)及肺癌细胞裂解物负载DC(DC组)活化的T细胞为对照,MTT法检测三组肺癌细胞的杀伤率.结果 exosome中有HSP70、HLA及CEA表达.活化T细胞/肺癌细胞为25∶1、10∶1 、5∶1时负载组杀伤率均明显高于未负载组及DC组(P均<0.05).结论 肺癌细胞裂解物负载能增强DC分泌的exosome诱导的抗肿瘤作用;本研究为制备高效的exosome肿瘤疫苗提供了理论依据.%Objective To obtain theoretical bases for making high efficacy exosome cancer vaccine. Methods Dendritic cells (DC) were induced with cytokines and then loaded with whole lung cancer cell lysates. Exosomes were isolated from supernatant of DC, and T cells activated by the exosomes (group loaded) , T cells activated by exosomes from nonloaded DC (group non-loaded) or activated by lysate-loaded DC(group DC) were taken as control. 11k activity of T cells for killing lung cancer cells were detected by MTT method. Results HSP70, HLA and CEA protein were found in exosomes. The kill rates of activated T cells in group loaded at E: T ratio 25:1, 10= 1, 5:1 were much higher than those in group non-loaded and group DC( all P <0.05). Condnsiong Lung cancer cell lysates loading can promote the anti-tumor activity induced by DC-derived exosomes; this study can provide theoretical bases for making high efficacy exosome cancer vaccine.

  7. A rapid kinetic chromogenic method for quantification of bacterial endotoxins in lyophilized reagents for labeling with {sup 99m}Tc radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Fukumori, Neuza T.O.; Campos, Domingos G.; Silva, Laercio; Fernandes, Adriana V.; Mengatti, Jair; Silva, Constancia P.G.; Matsuda, Margareth M.N. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2009-07-01

    A rapid quantitative kinetic chromogenic test in an automated Portable Test System (PTS) has been developed for determination of bacterial endotoxins in water, in-process and end-products using the Limulus amebocyte lysate (LAL). The aim of this work was to validate the method for lyophilized reagents for labeling with {sup 99m}Tc radiopharmaceuticals with no interfering factors. Experiments were performed in three consecutive batches of the lyophilized reagents Methylenediphosphonic Acid (MDP) and Pyrophosphate (PYRO) produced at IPEN-CNEN/ SP using the PTS from Endosafe, Inc.{sup TM}, Charleston, SC. The Maximum Valid Dilution (MVD) was calculated to establish the extent of dilution to avoid interfering test conditions (MVD=500). Better results were obtained above 1:20 dilution factor for MDP and 1:100 for PYRO. The parameters of coefficient correlation (R) -0.980, RPPC between 50 - 200% and coefficient variation (CV) of the samples less than 25% were satisfied and the endotoxin concentration was lower than the lowest concentration of the standard curve (0.05 EU mL{sup -1}), therefore less than the established limit in pharmacopoeias. The PTS is a rapid, simple and accurate technique using the quantitative kinetic chromogenic method for bacterial endotoxin determination. For this reason, it is very practical in the radiopharmaceutical area and it trends to be the method of choice for the pyrogen test. For MDP and PYRO, the validation was successfully performed. (author)

  8. Test plan :

    Energy Technology Data Exchange (ETDEWEB)

    Dwyer, Stephen F.

    2013-05-01

    This test plan is a document that provides a systematic approach to the planned testing of rooftop structures to determine their actual load carrying capacity. This document identifies typical tests to be performed, the responsible parties for testing, the general feature of the tests, the testing approach, test deliverables, testing schedule, monitoring requirements, and environmental and safety compliance.

  9. Transdifferentiation of Bone Narrow Mesenchymal Stem Cells into Salivary Gland Cells under the Induction with Submandubular Gland Cell Lysate.%下颌下腺细胞裂解液诱导骨髓间充质干细胞向涎腺细胞转分化的实验研究

    Institute of Scientific and Technical Information of China (English)

    吕汉孝; 姜金玲; 杨兆安; 崔丽娟; 张林

    2011-01-01

    目的:用下颌下腺细胞裂解液体外诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BM-MSCs),观察BM-MSCs能否表达涎腺细胞的特异性分泌蛋白-α-淀粉酶.方法:分离并培养新生SD大鼠的下颌下腺细胞(submandubular gland cells,SMGCs),测定各代腺泡细胞分泌α-淀粉酶的量,取分泌量显著一代制成下颌下腺细胞裂解液;自新生SD大鼠股骨、胫骨中分离出BM-MSCs,培养至第3代时用含下颌下腺细胞裂解液的培养基培养1周,细胞免疫组织化学法鉴定诱导后的BM-MSCSα-淀粉酶的表达.结果:细胞免疫组化可检测到所培养的下颌下腺细胞中含α-淀粉酶,淀粉酶试剂盒测定结果显示,P2、P3代SMGCs分泌的α-淀粉酶较其它各代明显(P<0.01).经下颌下腺细胞裂解液诱导BM-MSCs后,可见下颌下腺腺泡样细胞,免疫组化显示其表达淀粉酶,而对照组未表达淀粉酶.结论:下颌下腺细胞裂解液能体外模拟下颌下腺细胞微环境诱导BM-MSCs转分化为具有分泌α-淀粉酶蛋白能力的下颌下腺腺泡样细胞,为解决涎腺组织工程中种子细胞的来源提供一条新途径.%Objective: To understand the effects of submandubular gland cell lysate on the differentiation of bone marrow mesenchymal stem cells into salivary gland-like cells and investigate whether the BM-MSCs express the speaifically expressed salivary gland cell secretory protein-α-amylase. Methods: Submandibular gland cells (SMGCs)were isolated from neonatal SD rats and cultured in vitro. SMGCs were lysed by repeated freezing. BM-MSCs were isolated from neonatal SD rat femur and tibia and cultured for 3 passages, with medium containing submandibular gland cells lysate and control medium for 1 week. Immunocytochemical analysis was used to measure α-amylase ex- pression in induced BM-MSCs. Results: There was expression of α-amylase in normal cultured SMGCs and α-amylase secretion in SMGCs of P2 and P3 was signifycantly

  10. Prenatal Tests

    Science.gov (United States)

    ... Close X Home > Pregnancy > Prenatal care > Prenatal tests Prenatal tests E-mail to a friend Please fill ... even if you’re feeling fine. What are prenatal tests? Prenatal tests are medical tests you get ...

  11. Pinworm test

    Science.gov (United States)

    Oxyuriasis test; Enterobiasis test; Tape test ... diagnose this infection is to do a tape test. The best time to do this is in ... to determine if there are eggs. The tape test may need to be done on 3 separate ...

  12. Testing? Testing? In Literature?

    Science.gov (United States)

    Purves, Alan C.

    The assumptions behind secondary school literature course tests--whether asking students to recall aspects of literary works, to relate literary works to each other, or to analyze unfamiliar literary works--are open to question. They fail to acknowledge some of the most important aspects of literature which, if properly taught, should provide a…

  13. Test Under Test

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    China’s national college entrance examination, regarded as a make-or-break test by many students, leaves much to be desired “We’ve bribed the exam supervisors, paying each one 20,000 yuan. They will make everything go smooth during the exams,” Li Feng, a teacher from No.2 High School in

  14. 益生元和益生菌及细菌溶解物干预对婴幼儿免疫功能的影响研究进展%The Effect of Prebiotics/Probiotics and Bacterial Lysates Intervention on Infants' Immune Function

    Institute of Scientific and Technical Information of China (English)

    刘志刚; 韩波; 王金荣

    2014-01-01

    Epidemiologicalstudieshaveshownthatmicroorganismsandmicrobialcomponentshaveprotectiveeffecta-gainst allergic diseases and chronic inflammatory diseases. Most results show that using prebiotics,probiotics in early life can de-velop a stable internal environment and immune tolerance with gut microbiological colonization,and reduce the incidence of aller-gic and inflammatory diseases by primary prevention as well;while using bacterial lysates in early life can prevent the occurrence of asthma and infectious diseases,and reduce the chances of recurrent infection. This article aims to review the effects of probiot-ics,prebiotics and bacterial lysate on infants'immunity,and discuss the existing problems in current,so as to put forward impro-ving suggestions and directions for future clinical research.%流行病学研究表明,微生物和微生物组分对过敏性疾病及慢性炎性疾病有保护作用。在生命早期使用益生元、益生菌,可以通过早期的肠道菌群定植形成稳定内环境和免疫耐受,以及一级预防减少过敏性疾病及慢性炎性疾病的发生;应用细菌溶解物可以预防哮喘和感染性疾病,减少反复感染的几率。本文对益生元、益生菌和细菌溶解物对婴幼儿免疫功能的作用进行综述,探讨目前研究存在的问题,并对未来的临床研究提出改进设想和方向。

  15. The Effect of Prebiotics/Probiotics and Bacterial Lysates Intervention on Infants' Immune Function%益生元和益生菌及细菌溶解物干预对婴幼儿免疫功能的影响研究进展

    Institute of Scientific and Technical Information of China (English)

    刘志刚; 韩波; 王金荣

    2014-01-01

    流行病学研究表明,微生物和微生物组分对过敏性疾病及慢性炎性疾病有保护作用。在生命早期使用益生元、益生菌,可以通过早期的肠道菌群定植形成稳定内环境和免疫耐受,以及一级预防减少过敏性疾病及慢性炎性疾病的发生;应用细菌溶解物可以预防哮喘和感染性疾病,减少反复感染的几率。本文对益生元、益生菌和细菌溶解物对婴幼儿免疫功能的作用进行综述,探讨目前研究存在的问题,并对未来的临床研究提出改进设想和方向。%Epidemiologicalstudieshaveshownthatmicroorganismsandmicrobialcomponentshaveprotectiveeffecta-gainst allergic diseases and chronic inflammatory diseases. Most results show that using prebiotics,probiotics in early life can de-velop a stable internal environment and immune tolerance with gut microbiological colonization,and reduce the incidence of aller-gic and inflammatory diseases by primary prevention as well;while using bacterial lysates in early life can prevent the occurrence of asthma and infectious diseases,and reduce the chances of recurrent infection. This article aims to review the effects of probiot-ics,prebiotics and bacterial lysate on infants'immunity,and discuss the existing problems in current,so as to put forward impro-ving suggestions and directions for future clinical research.

  16. 人骨髓间充质干细胞裂解物对胶原诱导的大鼠关节炎的治疗作用%Therapeutic Effect of Human Bone Marrow Mesenchymal Stem Cell Lysates on Rat Arthritis Induced by Collagen

    Institute of Scientific and Technical Information of China (English)

    曹小芳; 王勇奇; 李志勇; 郭子宽; 王恒湘

    2013-01-01

    Our previous work has shown that mesenchymal stem cells (MSC) have little therapeutic effect on rat atthritts induced by collagen.This study was aimed to further investigate whether the MSC lysates exhibit beneficial effects on rheumatoid arthritis.Aliquots of cell lysates from 1 × 107 human bone marrow MSC were intraperitoneally injected into collagen-induced arthritis(CIA) Wistar rats weekly for 4 consecutive weeks.Methotrexate at a dose of 1 mg/kg or normal saline was served as positive and negative controls respectively.On week 4 the symptom scores were recorded and the hind joints of the rats were pathologically examined and X-ray examination was performed.The results showed that on week 4,the symptom scores of the rats that received MSC lysates (6.87 ± 0.83) and MTX (6.44 ± 1.13) were significantly lower than that of control rats (7.33 ±0.77,P <0.01).Meanwhile,pathological examination on the involved ankle showed that the synovitis and arthritis scores of MSC lysates and control groups were 2.28 ± 0.48 and 2.28 ±0.55 respectively,significantly higher than that of MTX treatment rats (0.71 ±0.48,P <0.05).However,X-ray examination on the ankle joints showed that the injury score of control rats was 4 ± 0.57,greatly higher than those from MSC lysates (2.71 ±0.75) and MTX treatment groups (2.57 ±0.78,P <0.05 for both groups).It is concluded that MSC lysate infusion has beneficial effects on CIA rat,but the effectiveness seems inferior to MTX.%以往的研究表明,间充质干细胞(mesenchymal stem cells,MSC)对Wistar大鼠胶原诱导性关节炎(collagen-induced arthritis,CIA)模型无治疗作用.本研究进一步探讨细胞裂解物是否具有CIA治疗作用.在造模4周后,裂解物治疗组CIA大鼠腹腔内注射1×107人骨髓MSC裂解物,阳性对照组CIA大鼠腹腔注射甲氨蝶呤(methotrexate,MTX)1 mg/kg,阴性对照组CIA大鼠腹腔注射生理盐水,以上各组治疗频率皆为每周1次,连续治疗4周.治疗4周后,记

  17. Enzyme-Cascade Analysis of the Rio Tinto Subsurface Environment: A Biosensor Experiment

    Science.gov (United States)

    McKay, David S.; Lynch, Kennda; Wainwright, Norman; Child, Alice; Williams, Kendra; McKay, David; Amils, Ricardo; Gonzalez, Elena; Stoker, Carol

    2004-01-01

    The Portable Test System (PTS), designed & developed by Charles Rivers Laboratories, Inc. (Charleston, SC) is a portable instrument that was designed to perform analysis of enzymatic assays related to rapid assessment of microbial contamination (Wainwright, 2003). The enzymatic cascade of Limulus Amebocyte Lysate (LAL) is known to be one of the most sensitive techniques available for microbial detection, enabling the PTS to be evaluated as a potential life detection instrument for in situ Astrobiology missions. In the summer of 2003 the system was tested as a part of the Mars Astrobiology Research and Technology Experiment (MARTE) ground truth science campaign in the Rio Tinto Analogue environment near Nerva, Spain. The preliminary results show that the PTS analysis correlates well with the contamination control tests and the more traditional lab-based biological assays performed during the MARTE field mission. Further work will be conducted on this research during a second field campaign in 2004 and a technology demonstration of a prototype instrument that includes autonomous sample preparation will occur in 2005.

  18. Coombs test

    Science.gov (United States)

    Direct antiglobulin test; Indirect antiglobulin test ... No special preparation is necessary for this test. ... There are two types of the Coombs test: Direct Indirect The ... that are stuck to the surface of red blood cells. Many diseases ...

  19. Coombs test

    Science.gov (United States)

    Direct antiglobulin test; Indirect antiglobulin test; Anemia - hemolytic ... No special preparation is necessary for this test. ... There are 2 types of the Coombs test: Direct Indirect The direct ... that are stuck to the surface of red blood cells. Many diseases ...

  20. Ham test

    Science.gov (United States)

    Acid hemolysin test; Paroxysmal nocturnal hemoglobinuria - Ham test; PNH - Ham test ... BJ. In: Chernecky CC, Berger BJ, eds. Laboratory Tests and Diagnostic Procedures . 6th ed. Philadelphia, PA: Elsevier ...

  1. Test Anxiety

    Science.gov (United States)

    ... Can I Help a Friend Who Cuts? Test Anxiety KidsHealth > For Teens > Test Anxiety Print A A ... with their concentration or performance. What Is Test Anxiety? Test anxiety is actually a type of performance ...

  2. 氧氟沙星注射液细菌内毒素检查法的研究%The discussion on the endotoxin inspection method of ofloxacini injection

    Institute of Scientific and Technical Information of China (English)

    隋妍蕾; 张建国; 于秀芬; 李常娥; 郭玉美

    2001-01-01

    OBJECTIVE:To establish the inspection method of endotoxin in ofloxacini injection.METHODS:The experiment is implemented according to the request of endotoxin inspection method collected in the 2nd book,China pharmacopeia,1995.RESULTS:The experiment showed that the interference elements can be excluded by diluting ofloxacini injection 2 times,and it is effective that endotoxin can be tested by amebocyte lysate with accuracy of 0.25Eu/ml.CONCLUSION:For daily pyragen test of ofloxacini injection,enotoxin inspection method can be used instead of dometic rabbit method.%目的:建立氧氟沙星注射液的细菌内毒素检查法。方法:根据中国药典1995年版二部收载的细菌内毒素检查法的要求进行实验。结果:将氧氟沙星注射液经2倍稀释可消除干扰因素,用标示灵敏度为0.25Eu/ml的鲎试剂检测细菌内毒素是有效的。结论:可用鲎试验法取代家兔法对氧氟沙星注射液进行热原检查。

  3. Rapid culture-independent microbial analysis aboard the international space station (ISS) stage two: quantifying three microbial biomarkers.

    Science.gov (United States)

    Morris, Heather C; Damon, Michael; Maule, Jake; Monaco, Lisa A; Wainwright, Norm

    2012-09-01

    Abstract A portable, rapid, microbial detection unit, the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS), was launched to the International Space Station (ISS) as a technology demonstration unit in December 2006. Results from the first series of experiments designed to detect Gram-negative bacteria on ISS surfaces by quantifying a single microbial biomarker lipopolysaccharide (LPS) were reported in a previous article. Herein, we report additional technology demonstration experiments expanding the on-orbit capabilities of the LOCAD-PTS to detecting three different microbial biomarkers on ISS surfaces. Six different astronauts on more than 20 occasions participated in these experiments, which were designed to test the new beta-glucan (fungal cell wall molecule) and lipoteichoic acid (LTA; Gram-positive bacterial cell wall component) cartridges individually and in tandem with the existing Limulus Amebocyte Lysate (LAL; Gram-negative bacterial LPS detection) cartridges. Additionally, we conducted the sampling side by side with the standard culture-based detection method currently used on the ISS. Therefore, we present data on the distribution of three microbial biomarkers collected from various surfaces in every module present on the ISS at the time of sampling. In accordance with our previous experiments, we determined that spacecraft surfaces known to be frequently in contact with crew members demonstrated higher values of all three microbial molecules. Key Words: Planetary protection-Spaceflight-Microbiology-Biosensor. Astrobiology 12, 830-840.

  4. 鲎试剂的灵敏度复核

    Institute of Scientific and Technical Information of China (English)

    邵民象; 于秉新

    2000-01-01

    @@ 细菌内毒素检查法(Bacteridl Endotoxins Test)即鲎试法(Limulus Test, LT)用于大输液热原检查,具有重现性好、操作简便、灵敏度约高于传统兔试法100倍等优点,故先后被美国FDA.USP及我国部颁标准、药典所收载.鲎试剂即鲎变形细胞溶解物(Tachypleus Amebocytes Lysate, TAL),其鲎的个体差异、总蛋白量及各种金属离子浓度、装量均匀度、运输与贮存条件等因素,均可对TAL灵敏度产生一定影响,而TAL灵敏度的准确性直接影响检查结果的可靠性,故药典规定:对每批新的TAL,在用于试验前必须进行灵敏度复核.

  5. Directory of Open Access Journals (Sweden)

    Lilian Eiko MAEKAWA

    2013-03-01

    Full Text Available The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO and ginger (GIN extracts, calcium hydroxide (CH, chlorhexidine (CLX gel and their combinations as ICMs (ICMs against Candida albicans, Enterococcus faecalis, Escherichia coli and endotoxins in root canals.Material and MethodsAfter 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal.Results and ConclusionAfter analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.

  6. Effect of Zingiber officinale and propolis on microorganisms and endotoxins in root canals

    Directory of Open Access Journals (Sweden)

    Lilian Eiko MAEKAWA

    2013-03-01

    Full Text Available The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO and ginger (GIN extracts, calcium hydroxide (CH, chlorhexidine (CLX gel and their combinations as ICMs (ICMs against Candida albicans, Enterococcus faecalis, Escherichia coli and endotoxins in root canals. Material and Methods After 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal. Results and Conclusion After analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.

  7. Antidepressant-like Effect of Cell-free Filtrate of Sodium Ferulate-induced and Differentioned PC12 Cell Lysates%阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液的抗抑郁样效果

    Institute of Scientific and Technical Information of China (English)

    廖铭能; 于立坚; 张永平; 马润娣; 张霄瑜; 于廷曦

    2011-01-01

    阿魏酸(ferulic acid,FA)是一种广泛存在的低毒酚酸,阿魏酸钠(sodium ferulate,SF)则是其钠盐.先前的研究已经证实,阿魏酸钠具有显著的神经保护和神经发生增强作用及抗抑郁效果.该研究的目的在于探讨阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液可能的抗抑郁效果.PC12细胞在含80 μmol/L阿魏酸钠的DMEM培养基中孵育6 d,无菌条件下制备阿魏酸钠诱导分化的PC12细胞液的无细胞滤液,测定PC12细胞裂解液无细胞滤液中残留的阿魏酸钠量.以慢性不可预期的多种刺激制造大鼠抑郁模型,用行为学、形态学、免疫组织化学和BrdU掺入等方法观察并检测阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液对慢性应激大鼠抑郁模型行为学、海马的组织病理学、海马和大脑皮质的神经生长因子(nerve growth factor,NGF)及脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的表达及神经发生的影响.实验证实,阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液能改善抑郁症样模型大鼠的行为学障碍,上调其海马和大脑皮质NGF和BNDF的表达,增加海马神经干细胞/神经前体细胞的增殖.由此可见,阿魏酸钠诱导分化的PC12细胞裂解液的无细胞滤液有明显的抗抑郁效果,而其抗抑郁效果可能源自它的上调NGF和BNDF以及其增强神经发生作用.%Ferulic acid (FA), 3-(4-hydroxy-3-methoxyphenyl)-2-propenoic acid, is one of the most common phenolic acids with low toxicity, and sodium ferulate (SF) is its sodium salt.Our previous work demonstrates that SF has significant neuroprotective and neurogenesis-enhancing actions and antidepressant-like effects.The aim of this study was to investigate a potential antidepressant-like effect of cell-free filtrate of sodium ferulate-induced and differentioned PC 12 cell lysates (SFIDPC 12CL) in the chronic mild stress (CMS)-induced depression-like model rats

  8. MAFF sponsored research: detection tests for irradiated food

    Energy Technology Data Exchange (ETDEWEB)

    Blackburn, C.M. (Ministry of Agriculture Fisheries and Food, Nobel House 17 Smith Square, London (United Kingdom)); Holley, P.A. (Ministry of Agriculture Fisheries and Food, Nobel House 17 Smith Square, London (United Kingdom)); Pryke, D.C. (Ministry of Agriculture Fisheries and Food, Nobel House 17 Smith Square, London (United Kingdom))

    1993-01-01

    In their 1986 report on the safety and wholesomeness of irradiated food the UK Advisory Committee on Irradiated and Novel Foods (ACINF) recognised that a generally applicable test to determine if a food had been irradiated was not available. The committee considered that, although not a pre-requisite, the existence of a detection test would be a useful supplement to a control system and do much to reassure consumers; with this in mind ACINF recommended that detection methods should be kept under review. As a consequence, in 1987 the Ministry initiated a comprehensive R and D detection test programme. Over fifty papers have been published to date as a result of this programme. MAFF (Ministry Of Agriculture Fisheries and Food) has also been involved in other research associated with irradiation and food safety, some of which is described in this paper. This paper aims to give an overview of recent work funded under the food irradiation programme. Twelve projects have been supported over the last two years, ten of which involved the development of detection tests for irradiated food. A summary of these projects is presented: - Thermoluminescence; - Electron Spin Resonance; - 2-alkylcyclobutanones; - Determination Of Hydrogen; - Differential Scanning Calorimetry; - Limulus Amoebocyte Lysate; - DNA; - Pesticide Breakdown; - Neutron Irradiation; - Future Plans. (orig./vhe)

  9. Specific anti-tumor immune responses of dendritic cells pulsed with recombinant human rhHSP70 and freeze-thaw cellular lysates derived from breast cancer%rhHSP70联合冻融抗原修饰树突状细胞诱导的抗乳腺癌作用*

    Institute of Scientific and Technical Information of China (English)

    李斌; 陈鹏; 郑建云

    2013-01-01

      目的:利用rhHSP70联合树突状细胞递呈肿瘤抗原的特性提高细胞毒T淋巴细胞(CTLs)对乳腺癌细胞的杀伤活性。方法:外周血单个核细胞体外经GM-CSF和IL-4诱导产生树突状细胞,负载冻融抗原肽的同时加入新型热休克蛋白(rhHSP70),不同分组分别诱导自体CTLs产生。ELISA测定CTLs杀伤活性和细胞因子的分泌。结果:冻融抗原肽致敏的DCs促进CTLs增殖,上调CTLs中CD3+和CD8+T细胞群及Th1型细胞因子的分泌;体外实验中具有对人乳腺癌细胞MCF-7的杀伤活性,在加入rhHSP70后效果更加明显,并能显著增强CTLs对肿瘤细胞的杀伤率。结论:rhHSP70联合肝癌冻融抗原修饰DCs,能够促进DCs的成熟,增强DCs刺激淋巴细胞增殖的能力,诱导的CTLs在体外对乳腺癌细胞能产生高效杀伤力。rhHSP70增强DCs抗肿瘤能力的机制可能与其促进DCs成熟有关。%Objective:This work aims to use the characteristics of dendritic cells (DCs) pulsed with recombinant human HSP70, which can present and process tumor antigens, to enhance the killing activity of cytotoxic t lymphocytes (CTLs) against breast neoplasms. Methods:Autologous DCs were isolated from peripheral blood mononuclear cells and then stimulated in vitro with granulocyte macrophage-colony stimulating factor and interleukin-4. The DCs were loaded with A549 tumor cell freeze-thaw lysate, and rhHSP70 was added as an immune adjuvant. The specific groups were subjected to tumor-specific cytotoxic assay, enzyme-linked immunosorbent assay, and fluores-cence-activated cell sorting. Results:DCs pulsed with A549 tumor cell lysate enhanced the growth expansion of CTLs, upregulated CD40 and CD80 populations in CTLs, and augmented Th1 cytokines. In addition, the cytotoxicity of specific CTLs against A549 was highly enhanced. The above indications became more obvious after the addition of rhHSP70. Conclusion:DCs pulsed with freeze-thaw cell

  10. Laboratory Tests

    Science.gov (United States)

    Laboratory tests check a sample of your blood, urine, or body tissues. A technician or your doctor ... compare your results to results from previous tests. Laboratory tests are often part of a routine checkup ...

  11. IQ testing

    Science.gov (United States)

    IQ (intelligence quotient) testing is a series of exams used to determine your general intelligence in relation ... Many IQ tests are used today. Whether they measure actual intelligence or simply certain abilities is controversial. IQ tests ...

  12. Laboratory Tests

    Science.gov (United States)

    ... Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & ... or conditions. What are lab tests? Laboratory tests are medical procedures that involve testing samples of blood, urine, or other tissues or ...

  13. Randomization tests

    CERN Document Server

    Edgington, Eugene

    2007-01-01

    Statistical Tests That Do Not Require Random Sampling Randomization Tests Numerical Examples Randomization Tests and Nonrandom Samples The Prevalence of Nonrandom Samples in Experiments The Irrelevance of Random Samples for the Typical Experiment Generalizing from Nonrandom Samples Intelligibility Respect for the Validity of Randomization Tests Versatility Practicality Precursors of Randomization Tests Other Applications of Permutation Tests Questions and Exercises Notes References Randomized Experiments Unique Benefits of Experiments Experimentation without Mani

  14. Biophysical characterization of lipopolysaccharide and lipid A inactivation by lactoferrin.

    Science.gov (United States)

    Brandenburg, K; Jürgens, G; Müller, M; Fukuoka, S; Koch, M H

    2001-08-01

    The interaction of bacterial endotoxins (LPS Re and lipid A, the 'endotoxic principle' of LPS) with the endogenous antibiotic lactoferrin (LF) was investigated using various physical techniques and biological assays. By applying Fourier-transform infrared (FTIR) spectroscopy, we find that LF binds to the phosphate group within the lipid A part and induces a rigidification of the acyl chains of LPS. The secondary structure of the protein - as monitored by the amide I band - is, however, not changed. Concomitant with the IR data, scanning calorimetric data indicate a sharpening of the acyl chain phase transition. From titration calorimetric and zeta potential data, saturation of LF binding to LPS was found to lie at a [LF]:[LPS] ratio of 1:3 to 1:5 M from the former and 1:10 M from the latter technique. X-ray scattering data indicate a change of the lipid A aggregate structure from inverted cubic to multilamellar, and with fluorescence (FRET) spectroscopy, LF is shown to intercalate by itself into phospholipid liposomes and may also block the lipopolysaccharide-binding protein (LBP)-induced intercalation of LPS. The LPS-induced cytokine production of human mononuclear cells exhibits a decrease due to LF binding, whereas the coagulation of amebocyte lysate in the Limulus test exhibited concentration-dependent changes. Based on these results, a model for the mechanisms of endotoxin inactivation by LF is proposed. PMID:11592403

  15. 42批次8种抗菌素药品鲎试验法与兔法检查热原比较

    Institute of Scientific and Technical Information of China (English)

    程玉莲; 柳文今; 于林汇; 祝丽萍

    1992-01-01

    @@ 鲎试验(Limulus test)又称鲎变形细胞溶解物(Limulus amebocyte lysate,LAL)试验,简称LT法,是近年来发展较快的,检测细菌内毒素最敏感的一种生物学检验新方法.LT法在国外被用于部分药品的热原检查法之一.LT法检查热原具有简便,灵敏,快速的特点,与传统的费时费力的兔法(RT法)相比优越很多.但是,由于各种检品本身在鲎试法中的干扰影响很复杂,国家药品标准中仅有4种大输液采用了此法,其它品种正在广泛的研究中,笔者积累日常兔法检查结果,对8个品种40多批次的抗菌素药品进行了LT法试验,与RT法进行比较,以考察LT法的可行性.本试验侧重于检品的稀释及LT法结果与RT法兔温波动数值间的关系.

  16. Masking of endotoxin in surfactant samples: Effects on Limulus-based detection systems.

    Science.gov (United States)

    Reich, Johannes; Lang, Pierre; Grallert, Holger; Motschmann, Hubert

    2016-09-01

    Over the last few decades Limulus Amebocyte Lysate (LAL) has been the most sensitive method for the detection of endotoxins (Lipopolysaccharides) and is well accepted in a broad field of applications. Recently, Low Endotoxin Recovery (LER) in biopharmaceutical drug products has been noticed, whereby the detection of potential endotoxin contaminations is not ensured. Notably, most of these drug products contain surfactants, which can have crucial effects on the detectability of endotoxin. In order to analyze the driving forces of LER, endotoxin detection in samples containing nonionic surfactants in various buffer systems was investigated. The results show that the process of LER is kinetically controlled and temperature-dependent. Furthermore, only the simultaneous presence of nonionic surfactants and components capable of forming metal complexes resulted in LER. In addition, capacity experiments show that even hazardous amounts of endotoxin can remain undetectable within such formulation compositions. In conclusion, the LER phenomenon is caused by endotoxin masking and not by test interference. In this process, the supramolecular structure of endotoxin is altered and exhibits only a limited susceptibility in binding to the Factor C of Limulus-based detection systems. We propose a two-step mechanism of endotoxin masking by complex forming agents and nonionic surfactants. PMID:27464990

  17. Residual endotoxin contaminations in recombinant proteins are sufficient to activate human CD1c+ dendritic cells.

    Directory of Open Access Journals (Sweden)

    Harald Schwarz

    Full Text Available Many commercially available recombinant proteins are produced in Escherichia coli, and most suppliers guarantee contamination levels of less than 1 endotoxin unit (EU. When we analysed commercially available proteins for their endotoxin content, we found contamination levels in the same range as generally stated in the data sheets, but also some that were higher. To analyse whether these low levels of contamination have an effect on immune cells, we stimulated the monocytic cell line THP-1, primary human monocytes, in vitro differentiated human monocyte-derived dendritic cells, and primary human CD1c+ dendritic cells (DCs with very low concentrations of lipopolysaccharide (LPS; ranging from 0.002-2 ng/ml. We show that CD1c+ DCs especially can be activated by minimal amounts of LPS, equivalent to the levels of endotoxin contamination we detected in some commercially available proteins. Notably, the enhanced endotoxin sensitivity of CD1c+ DCs was closely correlated with high CD14 expression levels observed in CD1c+ DCs that had been maintained in cell culture medium for 24 hours. When working with cells that are particularly sensitive to LPS, even low endotoxin contamination may generate erroneous data. We therefore recommend that recombinant proteins be thoroughly screened for endotoxin contamination using the limulus amebocyte lysate test, fluorescence-based assays, or a luciferase based NF-κB reporter assay involving highly LPS-sensitive cells overexpressing TLR4, MD-2 and CD14.

  18. Capture of lipopolysaccharide (endotoxin by the blood clot: a comparative study.

    Directory of Open Access Journals (Sweden)

    Margaret T Armstrong

    Full Text Available In vertebrates and arthropods, blood clotting involves the establishment of a plug of aggregated thrombocytes (the cellular clot and an extracellular fibrillar clot formed by the polymerization of the structural protein of the clot, which is fibrin in mammals, plasma lipoprotein in crustaceans, and coagulin in the horseshoe crab, Limulus polyphemus. Both elements of the clot function to staunch bleeding. Additionally, the extracellular clot functions as an agent of the innate immune system by providing a passive anti-microbial barrier and microbial entrapment device, which functions directly at the site of wounds to the integument. Here we show that, in addition to these passive functions in immunity, the plasma lipoprotein clot of lobster, the coagulin clot of Limulus, and both the platelet thrombus and the fibrin clot of mammals (human, mouse operate to capture lipopolysaccharide (LPS, endotoxin. The lipid A core of LPS is the principal agent of gram-negative septicemia, which is responsible for more than 100,000 human deaths annually in the United States and is similarly toxic to arthropods. Quantification using the Limulus Amebocyte Lysate (LAL test shows that clots capture significant quantities of LPS and fluorescent-labeled LPS can be seen by microscopy to decorate the clot fibrils. Thrombi generated in the living mouse accumulate LPS in vivo. It is suggested that capture of LPS released from gram-negative bacteria entrapped by the blood clot operates to protect against the disease that might be caused by its systemic dispersal.

  19. Intranasal coadministration of Cholera toxin with amoeba lysates modulates the secretion of IgA and IgG antibodies, production of cytokines and expression of pIgR in the nasal cavity of mice in the model of Naegleria fowleri meningoencephalitis.

    Science.gov (United States)

    Carrasco-Yepez, Maricela; Campos-Rodriguez, Rafael; Lopez-Reyes, Israel; Bonilla-Lemus, Patricia; Rodriguez-Cortes, Antonio Yahve; Contis-Montes de Oca, Arturo; Jarillo-Luna, Adriana; Miliar-Garcia, Angel; Rojas-Hernandez, Saul

    2014-11-01

    The nasal mucosa is the first contact with antigens to induce IgA response. The role of this site has rarely been studied. We have shown than intranasal administration with Naegleria fowleri lysates plus Cholera toxin (CT) increased the protection (survival up to 100%) against N. fowleri infection in mice and apparently antibodies IgA and IgG together with polymorphonuclear (PMN) cells avoid the attachment of N. fowleri to apical side of the nasal epithelium. We also observed that nasal immunization resulted in the induction of antigen-specific IgG subclasses (IgG1 and IgG2a) in nasal washes at days 3 and 9 after the challenge and IgA and IgG in the nasal cavity, compared to healthy and infected mice. We found that immunization with both treatments, N. fowleri lysates plus CT or CT alone, increased the expression of the genes for alpha chain, its receptor (pIgR), and it also increased the expression of the corresponding proteins evidenced by the ∼65 and ∼74kDa bands, respectively. Since the production of pIgR, IgA and IgG antibodies, is up-regulated by some factors, we analyzed the expression of genes for IL-10, IL-6, IFN-γ, TNF-α and IL-1β by using RT-PCR of nasal passages. Immunization resulted in an increased expression of IL-10, IL-6, and IFN-γ cytokines. We also aimed to examine the possible influences of immunization and challenge on the production of inflammatory cytokines (TNF-α and IL-1β). We observed that the stimulus of immunization inhibits the production of TNF-α compared to the infected group where the infection without immunization causes an increase in it. Thus, it is possible that the coexistence of selected cytokines produced by our immunization model may provide a highly effective immunological environment for the production of IgA, IgG and pIgR as well as a strong activation of the PMN in mucosal effector tissue such as nasal passages.

  20. Production of Recombinant Echinococcus granulosus Antigen B Subunits, In Order to Using Of Them in Serodiagnostic Tests of Hydatidosis

    Directory of Open Access Journals (Sweden)

    R Pazoki

    2007-06-01

    Full Text Available Background: Hydatidosis is one of the most important helminthiasis, and is a public health problem in many regions of the world. Methods: With the aim of production of recombinant subunits of antigen B, two different sequences of Echinococcus granulosus Antigen B, acquired from Gene Bank and amplified with specific primers via RT-PCR reaction. The amplified fragments (HI, HII cloned into pTZ57R T.vector, and then subcloned into pGEMEX-1 expression vector. Resaults: The SDS-PAGE performed after induction of cloned genes, and production of about 35 K.Da recombinant fusion proteins were confirmed for either two cloned genes. The immunogenicity of the recombinant fusion proteins were tested using double diffusion and immunoblotting. Both recombinant fusion proteins derived from lysate of transformed bacteria, were reactive for antibodies in serum of cystic hydatid patient. Conclusion: The produced recombinant antigen B subunits can be use in seroldiagnostic tests of hydatidosis, after purification.

  1. Enhanced effect of CD8++ T cells activated by tumor lysate -pulsed DCs on killing autologous tumor cells%通过肿瘤致敏的DCs活化的CD8+T细胞可有效地杀死肿瘤细胞

    Institute of Scientific and Technical Information of China (English)

    唐小龙; 江振友; 蔡淑玉

    2008-01-01

    AIM:To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by eonglutination method. The immature dendritic cells were generated in the presence of interleukin -4(IL-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF) from monocytes of healthy individuals.These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes(CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay a gninst autolognns human tumor cells. RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor- pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay a gainst target autologons tumor cells. The CD95 (Fas) expression, IFN-γ, and TNF -α secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autolognns tumor cells was significantly different(P<0. 05). Antigen-specific DCs vaccine can induce T cells activation and proliferation, thus we can obtain higher proportion of tumor specific cytotoxic T cells(CTLs), and enhance the CTLs to secret IFN-γ and TNF-α. CONCLUSION: Our results indicate that monocyte-derived human dendritic cells pulsed with tumor lysate could in duce the specific antitumor effect against autologons tumors. This in vitro model offers a new and simple approach to the development of DC + CTL - based immunotherapy.%目的:探索肿瘤裂解物负载的DCs诱导活化的初始T细胞介导细胞免疫及活化的T细胞杀死肿瘤细胞的能力.方法:应用黏附法分离外周血中的淋巴细胞

  2. The anti-tumor activity of dendritic cell/cytotoxic T lymphocyte induced by cholangiocarcinoma-derived exosome and its ultrafiltered lysate%基于exosome裂解超滤液的树突状细胞/细胞毒性T淋巴细胞诱导及其抗胆管癌研究

    Institute of Scientific and Technical Information of China (English)

    陈炯煌; 丁国平; 陈文超; 曹利平

    2013-01-01

    Objective To observe the impact of cholangiocarcinoma-derived exosome and ultrafiltered exosome lysate on the anti-tumor activity of dendritic celL/cytotoxic T lymphocyte (DC/CTL) cells and discuss the mechanism involved.Methods Exosomes derived from RBE cells (human cholangiocarcinoma line) were collected by ultracentrifugation,ow-osmotic splitting followed by ultrafiltration was performed to remove exosomal microRNAs and purify the ultrafiltered exosome lysate.ImDCs (immature dendritic cells) were induced from peripheral blood and were co-cultured with CTL(cytotoxic lymphocyte),which were impulsed by exosome and ultrafiltered exosome lysate,respectively.The concentrations of tumor necrosis factor (TNF)-αt and perforin in culture medium supernatant were examined by enzyme linked immunosorbent assay (ELISA).Cell counting kit-8 (CCK-8) was used to evaluate the cytotoxic activity of DC/CTL to RBE cells.Results The concentrations of TNF-αand perforin in E-DC/CTL group were 138.61 ng/L and 2.41 μg/L respectively,lower than those of DC/CTL (194.08 ng/L and 3.39 μg/L) and EL-DC/CTL group (210.87 ng/L and 3.79 μg/L).The killing rate of E-DC/CTL was 33.35%,lower than that of DC/CTL (47.35%) and EL-DC/CTL (66.23%) significantly.The killing rate of EL-DC/CTL was significantly higher than that of DC/CTL (P < 0.01).Conclusion RBE cell-derived exosome inhibits the anti-tumor activity of DC/CTL by down-regulating TNF-α and perforin,exosomal miRNA may play important roles in the immune escape of cholangiocarcinoma.We built a new model based on ultrafiltered exosome lysate derived from cholangiocarcinoma to enhance the anti-tumor activity of DC/CTL.%目的 观察人胆管癌细胞来源的exosome及exosome裂解超滤液(UEL)对树突状细胞/细胞毒性T淋巴细胞诱导(DC/CTL)混合细胞抗肿瘤活性的影响及机制.方法 采用超速离心法提取人胆管癌细胞(RBE细胞)释放的exosome,低渗法裂解exosome,超

  3. Tensile testing

    CERN Document Server

    2004-01-01

    A complete guide to the uniaxial tensile test, the cornerstone test for determining the mechanical properties of materials: Learn ways to predict material behavior through tensile testing. Learn how to test metals, alloys, composites, ceramics, and plastics to determine strength, ductility and elastic/plastic deformation. A must for laboratory managers, technicians, materials and design engineers, and students involved with uniaxial tensile testing. Tensile Testing , Second Edition begins with an introduction and overview of the test, with clear explanations of how materials properties are determined from test results. Subsequent sections illustrate how knowledge gained through tensile tests, such as tension properties to predict the behavior (including strength, ductility, elastic or plastic deformation, tensile and yield strengths) have resulted in improvements in materals applications. The Second Edition is completely revised and updated. It includes expanded coverage throughout the volume on a variety of ...

  4. Objective Tests versus Subjective tests

    Institute of Scientific and Technical Information of China (English)

    魏福林

    2007-01-01

    objective test has only one correct answer, while subjective test has a range of possible answers. Because of this feature, reliability will not be difficult to achieve in the marking of the objective item, while the marking of the subjective items is reliable. On the whole, a good test must contain both subjective and objective test items.

  5. Nationale test

    DEFF Research Database (Denmark)

    2009-01-01

    Professor Sven Erik Nordenbo og centerleder Niels Egelund, begge DPU, i samtale om nationale test.......Professor Sven Erik Nordenbo og centerleder Niels Egelund, begge DPU, i samtale om nationale test....

  6. Predictive Testing

    Science.gov (United States)

    ... is used to check for mutations linked to Huntington’s Disease. Predispositional Testing: Positive tests results show that you ... results. Twitter YouTube RSS Genetics and Health How Genes Work Genes, Lifestyle, & Environment Collecting Family Health History ...

  7. Ferritin Test

    Science.gov (United States)

    ... page: Was this page helpful? Also known as: Serum Ferritin Formal name: Ferritin, serum Related tests: Complete Blood Count , Hemoglobin , Hematocrit , Serum Iron , TIBC, UIBC and Transferrin , Iron Tests , Zinc Protoporphyrin , ...

  8. Glucose Tests

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Glucose Tests Share this page: Was this page helpful? ... the meaning of other test results. Fasting Blood Glucose Glucose Level Indication From 70 to 99 mg/ ...

  9. TSH test

    Science.gov (United States)

    ... different samples. Talk to your provider about the meaning of your specific test results. If you are ... iodine in the body (due to receiving iodine contrast used during imaging tests, such as CT scan) ...

  10. Nationale Test

    DEFF Research Database (Denmark)

    2009-01-01

    Hvad er egentlig formålet med de nationale test? Bliver eleverne klogere af at blive testet? Og er der en sammenhæng mellem bandekrig og nationale test? Fysisk medie: dpu.dk/tv......Hvad er egentlig formålet med de nationale test? Bliver eleverne klogere af at blive testet? Og er der en sammenhæng mellem bandekrig og nationale test? Fysisk medie: dpu.dk/tv...

  11. HIV Testing

    Science.gov (United States)

    ... Abroad Treatment Basic Statistics Get Tested Find an HIV testing site near you. Enter ZIP code or city Follow HIV/AIDS CDC HIV CDC HIV/AIDS See RSS | ... All Collapse All Should I get tested for HIV? CDC recommends that everyone between the ages of ...

  12. Schilling test

    Science.gov (United States)

    Vitamin B12 absorption test ... This test may be done in four different stages to find the cause of a low vitamin B12 level. ... can absorb vitamin B12. Stage II of the test can tell whether a low vitamin B12 level ...

  13. Trace test

    OpenAIRE

    Leykin, Anton; Sottile, Frank

    2016-01-01

    We give a brief derivation of the trace test to verify completeness of a partial witness set of an irreducible variety in affine or projective space. We then consider the trace test for subvarieties of products of projective spaces working with multihomogeneous witness sets. We show how a dimension reduction based on Bertini's Theorem leads to a practical trace test in this case involving a curve in a small-dimensional affine space.

  14. Software testing

    Science.gov (United States)

    Price-Whelan, Adrian M.

    2016-01-01

    Now more than ever, scientific results are dependent on sophisticated software and analysis. Why should we trust code written by others? How do you ensure your own code produces sensible results? How do you make sure it continues to do so as you update, modify, and add functionality? Software testing is an integral part of code validation and writing tests should be a requirement for any software project. I will talk about Python-based tools that make managing and running tests much easier and explore some statistics for projects hosted on GitHub that contain tests.

  15. Ultrasonic testing

    Energy Technology Data Exchange (ETDEWEB)

    Song, Sung Jin [Sungkwunkwan Univ., Seoul (Korea, Republic of); Jeong, Hyun Jo [Wonkwang Univ., Iksan (Korea, Republic of)

    2004-02-15

    For the proper performance of ultrasonic testing of steel welded joints, and anisotropic material it is necessary to have sound understanding on the underlying physics. To provide such an understanding, it is beneficial to have simulation tools for ultrasonic testing. In order to address such a need, we develop effective approaches to simulate angle beam ultrasonic testing with a personal computer. The simulation is performed using ultrasonic measurement models based on the computationally efficient multi-Gaussian beams. This reach will describe the developed ultrasonic testing models together with the experimental verification of their accuracy.

  16. Anti-Tumor Effects of B16F10 Tumor Cell Lysate Vaccine in Mouse Melanoma%肿瘤细胞裂解物抗小鼠B16F10黑色素瘤的作用研究

    Institute of Scientific and Technical Information of China (English)

    路蕾; 刘景晶; 陈科; 刘彬; 王泽宇; 葛驰宇; 侯景; 金亮; 邢芸; 曹荣月

    2012-01-01

    反复冻融B16F10肿瘤细胞制备裂解物,以白喉毒素(Diphtheria toxin,DT)为载体,OK432和来源于结核分枝杆菌( Mycobacterium tuberculosis)热休克蛋白70(HSP70)第407-426( mHSP70407-426,M)的两段串联重复序列M2为佐剂,制备了肿瘤细胞疫苗B16F10-DT-M2-OK432(BDTMOK),探讨其能否抑制小鼠B16黑色素瘤,并且对其抗肿瘤的作用机理进行部分探讨.以制备的BDTMOK免疫C57BL/6小鼠,分别检测体液免疫应答和细胞免疫应答.通过ELISA法,从血清中检测到高滴度的抗B16肿瘤细胞裂解物(B16 tumor cell lysate,B16TCL)类抗体.淋巴细胞增殖实验的结果显示,BDTMOK的免疫能够有效的刺激脾淋巴细胞的增殖.预防结合治疗性实验的结果显示,BDTMOK激发的免疫应答对于B16肿瘤攻击起到有效的保护作用,与PBS阴性对照组比较,皮下注射BDTMOK可以延长皮下移植瘤发生的潜伏期(P<0.05),并且平均瘤重显著降低(P<0.05);抑制了小鼠皮内肿瘤模型中的血管新生(P<0.01).疫苗BDTMOK能有效的抑制小鼠B16黑色素瘤的生长.%To make tumor vaccine B16F10-DT-M2-OK432,the freeze-thaw method was adopted to obtain B16F10 tumor cell lysate. Diphtheria toxin( DT) ,OK432 or M2 which was two tandem repeats of sequence 407-426 of microbial HSP70 were selected as carrier and adjuvants respectively. The C57BL/6 mice were immunized with B16F10-DT-M2-OK432 in order to explore whether the tumor cell vaccine can effectively inhibit B16F10 melanoma in tumor-bearing mice and to study the mechanism of B16F10-DT-M2-OK432. After the last immunization, humoral immune and cellular immune response were detected. High tiler of anti-B16F10 tumor cell lysate antibody was detected in immunized mice sera by ELISA. Splenic lymophocyte proliferation assay results showed that the proliferation activity of splenocytes from mice immunized with B16F10-DT-M2-OK432 vaccine was significantly higher. The results of prophylactic/ therapeutic experiment

  17. Nationale test

    DEFF Research Database (Denmark)

    Bundsgaard, Jeppe; Puck, Morten Rasmus

    Nationale test skubber undervisning i en forkert retning. Det er lærerne og skolelederne enige om. Men særligt skolelederne ser også muligheder for at bruge testen til at få viden om elevernes faglige kompetencer og om undervisningen. Det kommer til udtryk i rapporten Nationale test: Danske lærere...

  18. TORCH Test

    Science.gov (United States)

    ... Epstein-Barr Virus Antibodies , Chickenpox and Shingles Tests , Parvovirus B19 All content on Lab Tests Online has been ... enterovirus, Epstein-Barr virus , varicella-zoster virus , and parvovirus B19 . ^ Back to top When is it ordered? The ...

  19. Malnutrition Tests

    Science.gov (United States)

    ... indicate possible nutritional deficits, patients may be provided nutritional support prior to a surgery or procedure and be monitored regularly during recovery. Laboratory tests may include: For general screening and ... protein For nutritional status and deficiencies: Iron tests such as serum ...

  20. Detection of endotoxin using a photonic crystal nanolaser

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Daichi; Hachuda, Shoji; Watanabe, Takumi; Nishijima, Yoshiaki; Baba, Toshihiko, E-mail: baba@ynu.ac.jp [Department of Electrical and Computer Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama 240-8501 (Japan)

    2015-03-30

    Fast and reliable detection of endotoxin (ET) in medical equipment and pharmaceutical products is an essential precursor to clinical treatment. In this study, we demonstrate the use of shifts in wavelength of photonic crystal nanolasers for sensing the Limulus amebocyte lysate reaction, which is a standard method for detecting ET. From working curves of wavelength shift vs ET concentration, whose correlation factors were as high as 98%, we detected a required concentration of 0.001 EU/ml within 33 min and detected a low concentration of 0.0001 EU/ml.

  1. Determination of bacterial endotoxin (pyrogen) in radiopharmaceuticals by the gel clot method. Validation

    International Nuclear Information System (INIS)

    Before the Limulus amebocyte lysate (LAL) test, the only available means of pirogenicity testing for parenteral drugs and medical devices was the United States Pharmacopoeia (USP) rabbit pyrogen test. Especially for radiopharmaceuticals, the LAL assay is the elective way to determine bacterial endotoxin. The aim of this work was to validate the gel clot method for some radiopharmaceuticals without measurable interference. The FDA's LALTest guideline defines interference as a condition that causes a significant difference between the endpoints of a positive water control and positive product control series using a standard endotoxin. Experiments were performed in accordance to the USP bacterial endotoxins test in the 131I- m-iodobenzylguanidine; the radioisotopes Gallium-67 and Thallium-201; the lyophilized reagents DTPA, Phytate, GHA, HSA and Colloidal Tin. The Maximum Valid Dilution (MVD) was calculated for each product based upon the clinical dose of the material and a twofold serial dilution below the MVD was performed in duplicate to detect interferences. The labeled sensitivity of the used LAL reagent was 0.125 EU mL-1 (Endotoxin Units per milliliter). For validation, a dilution series was performed, a twofold dilution of control standard endotoxin (CSE) from 0.5 to 0.03 EU mL-1, to confirm the labeled sensitivity of the LAL reagent being tested in sterile and non pyrogenic water, in quadruplicate. The same dilution series was performed with the CSE and the product in the 1:100 dilution factor, in three consecutive batches of each radiopharmaceutical. The products 131I-m-iodobenzylguanidine, Gallium-67, Thallium-201, DTPA, HSA and Colloidal Tin were found compatible with the LAL test at a 1:100 dilution factor. Phytate and GHA showed some interference in the gel clot test. Other techniques to determine endotoxins as the chromogenic (color development) and the turbidimetric test (turbidity development), were also assessed to get valuable quantitative and

  2. Penetration testing

    OpenAIRE

    Zemaníková, Martina

    2010-01-01

    This work focuses on the practical demonstration of design penetration testing. Testing is carried out based on the order of a particular entity/subject which is at their request anonymous. The work is divided into two parts, theoretical and practical. We will be made in first part familiarize with the process and techniques of testing, as is now used in ethical hacking, then we can find in the end appropriate recommendations 6 how to prevent and fight against it in order to protect the...

  3. Oedometer Tests

    DEFF Research Database (Denmark)

    Thorsen, Grete

    1996-01-01

    The paper describes the results of oedometer tests carried out with samples from Eemian fresh-water deposits and the methods used to determine the preconsolidation pressure from the test results. The influence of creep in the material on the apparent preconsolidation pressure is estimated from a...... model set up by Moust Jacobsen in 1992. The test results do not show any significant difference in the determined values of the overconsolidation ratio (OCR) for the samples from Hollerup and Solsø, east and west of the main stationary line for the last ice sheet in Weichselian, respectively. The...

  4. Hydrolyzing Condition and Immunocompetence of Sheep Bone Protein Enzymatic Lysates

    Institute of Scientific and Technical Information of China (English)

    YANG Hua; LIU Yu-hua; MA Li-zhen; KONG Bao-hua

    2009-01-01

    Utilizing collagen of sheep bone as material to get immunocompetent peptide, enzymatic hydrolysis conditions were optimized using quadratic regression general rotation design. The effect of temperature (T), time (t), enzyme/substrate (E/S) ratio, and substrate concentration (S) on the amount of tricarboxylix acid cycle (TCA) soluble peptides were investigated. The content of soluble peptide in the acquisition was measured by Folin-hydrozybebzebe method, and the correlation between soluble peptide content and immunocompetence was analyzed by SAS software. The best enzymatic hydrolysis condition was gotten from Design Expert 7.1.2 software. The optimal condition under which immunocompetent peptides could be prepared was 1 576 U g-1 (E/S), 64.05℃ (T), 0.271 kg L-1(S), and 7.22 h (t). The correlation coefficient between TCA soluble peptides and the immuneocompetence was 0.045 < r0.05= 0.355, which indicated that they had no significant correlation. The result showed that the soluble peptide contained immunocompetent peptides which content was independent of immunocompetence in the hydrolasates.

  5. Platelets lysate-based membranes for periodontal ligament regeneration

    OpenAIRE

    Babo, P. M.; Santo, Vítor E.; Duarte, Ana Rita C.; Gomes, Manuela E.; Reis, R. L.

    2013-01-01

    The periodontal ligament (PDL) is a group of specialized connective tissue fibers that attach a tooth to the alveolar bone where it is deployed. These fibers help the tooth withstand the substantial compressive forces which occur during chewing and remain embedded in the bone. Periodontitis is a prevalent infection disease that causes the destruction of the tooth supportive tissues including the PDL. Given its low ability of regeneration in adult patients, concerted efforts have been made to ...

  6. Albumin Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Albumin Share this page: Was this page helpful? Also known as: ALB Formal name: Albumin, serum Related tests: Liver Panel , Comprehensive Metabolic Panel , ...

  7. Knowledge Test

    DEFF Research Database (Denmark)

    Sørensen, Ole Henning

    1998-01-01

    The knowledge test is about competing temporal and spatial expressions of the politics of technological development and national prosperity in contemporary society. The discussion is based on literature of national systems of innovation and industrial networks of various sorts. Similarities...

  8. RPR test

    Science.gov (United States)

    ... infections and is routinely used to screen pregnant women for the disease. Several states also require that couples be screened for syphilis before getting a marriage license. The test is also used to see ...

  9. PTT Test

    Science.gov (United States)

    ... when the person has a clinical history of bleeding, such as frequent or excessive nose bleeds and easy bruising, which may indicate the presence of a bleeding disorder ^ Back to top What does the test ...

  10. Fungal Tests

    Science.gov (United States)

    ... or Calcofluor white stain): in general, if fungal elements are seen, then a fungus is the likely the cause of symptoms. These tests, however, do not identify the fungus. Culture: care must be taken when interpreting culture results. ...

  11. Sodium Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Sodium Share this page: Was this page helpful? Also known as: Na Formal name: Sodium Related tests: Chloride , Bicarbonate , Potassium , Electrolytes , Osmolality , Basic ...

  12. Bilirubin Test

    Science.gov (United States)

    ... Bilirubin; Indirect Bilirubin; Unconjugated Bilirubin Formal name: Bilirubin - blood Related tests: Liver Panel ; GGT ; ALP ; AST ; ALT ; Hepatitis A ; Hepatitis B ; Hepatitis C ; Complete Blood Count ; Urinalysis ; Direct Antiglobulin ...

  13. Insulin Test

    Science.gov (United States)

    ... especially as a result of taking non-human (animal or synthetic) insulin, these can interfere with insulin testing. In this case, a C-peptide may be performed as an alternative way to evaluate insulin production. Note also that ...

  14. Tested Demonstrations.

    Science.gov (United States)

    Gilbert, George L.

    1990-01-01

    Included are three demonstrations that include the phase change of ice when under pressure, viscoelasticity and colloid systems, and flame tests for metal ions. The materials, procedures, probable results, and applications to real life situations are included. (KR)

  15. Lipase Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Lipase Share this page: Was this page helpful? Also known as: LPS Formal name: Lipase Related tests: Amylase , Trypsin , Trypsinogen At a Glance ...

  16. Pap test

    Science.gov (United States)

    ... cells - Pap; AGUS - Pap; Atypical squamous cells - Pap; HPV - Pap; Human papilloma virus - Pap cervix - Pap; Colposcopy - Pap ... test to check for the presence of the HPV virus types most likely to cause cancer Cervix cryosurgery- ...

  17. Trypsinogen test

    Science.gov (United States)

    ... is broken) Considerations Other tests used to detect pancreas diseases may include: Serum amylase Serum lipase Alternative Names ... Related MedlinePlus Health Topics Cystic Fibrosis Pancreatic Cancer Pancreatic Diseases Browse the Encyclopedia A.D.A.M., Inc. ...

  18. Amylase Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Amylase Share this page: Was this page helpful? Also known as: Amy Formal name: Amylase Related tests: Lipase , Trypsin , Trypsinogen At a Glance ...

  19. Mono Test

    Science.gov (United States)

    ... white blood cells and reactive lymphocytes on a blood smear in the presence of symptoms associated with mono indicates a likely diagnosis of infectious mononucleosis. A negative mono test requires ...

  20. Lipase test

    Science.gov (United States)

    ... the bowel (bowel obstruction) Celiac disease Duodenal ulcer Cancer of the pancreas Infection or swelling of the pancreas This test may also be done for familial lipoprotein lipase deficiency . Risks ... Update Date 2/4/2015 Updated ...

  1. HPV Test

    Science.gov (United States)

    ... type of HPV infection may benefit from the protection against other types included in the vaccine. ^ Back ... infection. These include having many children, long-term oral contraceptive ... having multiple sex partners, infrequent Pap tests , smoking, a history of ...

  2. Adaptive test

    DEFF Research Database (Denmark)

    Kjeldsen, Lars Peter; Eriksen, Mette Rose

    2010-01-01

    Artikelen er en evaluering af de adaptive tests, som blev indført i folkeskolen. Artiklen sætter særligt fokus på evaluering i folkeskolen, herunder bidrager den med vejledning til evaluering, evalueringsværktøjer og fagspecifkt evalueringsmateriale.......Artikelen er en evaluering af de adaptive tests, som blev indført i folkeskolen. Artiklen sætter særligt fokus på evaluering i folkeskolen, herunder bidrager den med vejledning til evaluering, evalueringsværktøjer og fagspecifkt evalueringsmateriale....

  3. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    Science.gov (United States)

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology.

  4. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    Science.gov (United States)

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology. PMID:25952731

  5. DHEAS Test

    Science.gov (United States)

    ... be ordered, along with other hormone tests, whenever excess (or, more rarely, deficient) androgen production is suspected and/or when a health ... level, in addition to other normal male hormone ( androgen ) levels, likely ... may indicate that excess DHEAS production is causing or contributing to the ...

  6. Rotational testing.

    Science.gov (United States)

    Furman, J M

    2016-01-01

    The natural stimulus for the semicircular canals is rotation of the head, which also might stimulate the otolith organs. Vestibular stimulation usually induces eye movements via the vestibulo-ocular reflex (VOR). The orientation of the subject with respect to the axis of rotation and the orientation of the axis of rotation with respect to gravity together determine which labyrinthine receptors are stimulated for particular motion trajectories. Rotational testing usually includes the measurement of eye movements via a video system but might use a subject's perception of motion. The most common types of rotational testing are whole-body computer-controlled sinusoidal or trapezoidal stimuli during earth-vertical axis rotation (EVAR), which stimulates primarily the horizontal semicircular canals bilaterally. Recently, manual impulsive rotations, known as head impulse testing (HIT), have been developed to assess individual horizontal semicircular canals. Most types of rotational stimuli are not used routinely in the clinical setting but may be used in selected research environments. This chapter will discuss clinically relevant rotational stimuli and several types of rotational testing that are used primarily in research settings. PMID:27638070

  7. Estimativa da incerteza em ensaio de detecção de endotoxina bacteriana pelo método de gelificação Estimation of uncertainty in the detection of bacterial endotoxin by gel-clot method

    Directory of Open Access Journals (Sweden)

    Felipe Rebello Lourenço

    2005-12-01

    Full Text Available Desde a publicação da ISO 17025:1999, o interesse em métodos para estimativa da incerteza em ensaios qualitativos, do tipo "passa/não passa", têm ganho grande importância. Uma forma de estimar e informar a incerteza deste tipo de ensaio é o uso das probabilidades de respostas-falsas, particularmente falsos-positivos e falsos-negativos, determinados a partir do teorema de Bayes. O objetivo deste artigo é estabelecer um método para a estimativa de incerteza em ensaios de detecção de endotoxina bacteriana pelo método in vitro Limulus Amebocyte Lysate (LAL. Considerando a confirmação da sensibilidade do LAL e a validação do teste, a probabilidade de uma resposta falsa corresponde à soma da probabilidade dos resultados falso-negativos e falso-positivos. A partir dos resultados obtidos foi verificado que a etapa da confirmação da sensibilidade do LAL contribui para a incerteza de forma mais significativa (67,6% que a etapa de validação do teste (32,4%. Através de um procedimento simples, descrito neste artigo, e de dados obtidos a partir da confirmação da sensibilidade do LAL e validação do teste para um produto em questão é possível obter uma estimativa de incerteza razoável para o ensaio de detecção de endotoxinas bacterianas pelo método de gelificação.Since the publication of ISO 17025:1999, the interest in methods for estimation of the uncertainty in qualitative analysis, such as 'pass/fail', have became more important. The usual form of estimating and informing the uncertainty in this kind of analysis is the use of false-response rates, particularly false-positive and false-negative, determinated from Bayes theorem. The aim of this paper is establish a method for estimation of the uncertainty in the detection of bacterial endotoxins by in vitro Limulus Amebocyte Lysate (LAL test. Considering the confirmation of LAL sensitivity and the validation of the test, the probability of a false-response corresponds to the

  8. Radiographic Test

    Energy Technology Data Exchange (ETDEWEB)

    Lee, H.J; Yang, S.H. [Korea Electric Power Research Institute, Taejon (Korea)

    2002-07-01

    This report contains theory, procedure technique and interpretation of radiographic examination and written for whom preparing radiographic test Level II. To determine this baseline of technical competence in the examination, the individual must demonstrate a knowledge of radiography physics, radiation safety, technique development, radiation detection and measurement, facility design, and the characteristics of radiation-producing devices and their principles of operation. (author) 98 figs., 23 tabs.

  9. Putting Tests To The Test

    Institute of Scientific and Technical Information of China (English)

    王丽荣

    2005-01-01

    Tests are usually a big part of a teacher's life. They are part of the ritual of the classroom. Students expect them. Administrators, school boards and legislators require them. They are important to us in our work; we think about them often and have a lot to say about them.

  10. Developing a nanoparticle test for prostate cancer scoring

    Directory of Open Access Journals (Sweden)

    Huo Qun

    2012-03-01

    Full Text Available Abstract Background Over-diagnosis and treatment of prostate cancer has been a major problem in prostate cancer care and management. Currently the most relevant prognostic factor to predict a patient's risk of death due to prostate cancer is the Gleason score of the biopsied tissue samples. However, pathological analysis is subjective, and the Gleason score is only a qualitative estimate of the cancer malignancy. Molecular biomarkers and diagnostic tests that can accurately predict prostate tumor aggressiveness are rather limited. Method We report here for the first time the development of a nanoparticle test that not only can distinguish prostate cancer from normal and benign conditions, but also has the potential to predict the aggressiveness of prostate cancer quantitatively. To conduct the test, a prostate tissue lysate sample is spiked into a blood serum or human IgG solution and the spiked sample is incubated with a citrate-protected gold nanoparticle solution. IgG is known to adsorb to citrate-protected gold nanoparticles to form a "protein corona" on the nanoparticle surface. From this study, we discovered that certain tumor-specific molecules can interact with IgG and change the adsorption behavior of IgG to the gold nanoparticles. This change is reflected in the nanoparticle size of the assay solution and detected by a dynamic light scattering technique. Assay data were analyzed by one-way ANOVA for multiple variant analysis, and using the Student t-test or nonparametric Mann-Whitney U-tests for pairwise analyses. Results An inverse, quantitative correlation of the average nanoparticle size of the assay solution with tumor status and histological diagnostic grading was observed from the nanoparticle test. IgG solutions spiked with prostate tumor tissue exhibit significantly smaller nanoparticle size than the solutions spiked with normal and benign tissues. The higher grade the tumor is, the smaller the nanoparticle size is. The test

  11. Systematic review of membrane components of Gram-positive bacteria responsible as pyrogens for inducing human monocyte / macrophage cytokine release

    Directory of Open Access Journals (Sweden)

    Thomas eHartung

    2012-04-01

    Full Text Available Fifty years after the elucidation of lipopolysaccharides (LPS, endotoxin as the principal structure of Gram-negative bacteria activating the human immune system, its Gram-positive counterpart is still under debate. Pyrogen tests based on the human monocyte activation have been validated for LPS detection as an alternative to the rabbit test and, increasingly, the limulus amebocyte lysate test. For full replacement, international validations with non-endotoxin pyrogens are in preparation. Following evidence-based medicine approaches, a systematic review of existing evidence as to the structural nature of the Gram-positive pyrogen was undertaken. For the three major constituents suggested, i.e. peptidoglycan, lipoteichoic acids (LTA, and bacterial lipoproteins, the questions to be answered and a search strategy for relevant literature was developed, starting in MedLine. The evaluation was based on the Koch-Dale criteria for a mediator of an effect. A total of 380 articles for peptidoglycan, 391 for lipoproteins, and 285 for LTA were retrieved of which 12, 8 and 24, respectively, fulfilled inclusion criteria. The compiled data suggest that for peptidoglycan two Koch-Dale criteria are fulfilled, four for LTA, and two for bacterial lipoproteins. In conclusion, based on the best currently available evidence, LTA is the only substance that fulfils all criteria. LTA has been isolated from a large number of bacteria, results in cytokine release patterns inducible also with synthetic LTA. Reduction in bacterial cytokine induction with an inhibitor for LTA was shown. However, this systematic review cannot exclude the possibility that other stimulatory compounds complement or substitute for LTA in being the counterpart to LPS in some Gram-positive bacteria.

  12. Nuclear stress test

    Science.gov (United States)

    ... Persantine stress test; Thallium stress test; Stress test - nuclear; Adenosine stress test; Regadenoson stress test; CAD - nuclear stress; Coronary artery disease - nuclear stress; Angina - nuclear ...

  13. 鲎试剂法测定细菌内毒素影响因素的研究

    Institute of Scientific and Technical Information of China (English)

    张靖贤

    2002-01-01

    @@ 鲎试剂法是利用鲎血细胞溶解物(Limulus Amebocyte Lysate,LAL;Tachypleus Amebocyte Lysate, TAL)与微量(pg*ml-1级)细菌(主要是革兰氏阴性细菌,G-)的内毒素反应,从而检出被检物中内毒素的一种生物体外检测新技术,我国常用的鲎试剂主要为东方鲎的细胞裂解物.因为具有快速、简便、经济、灵敏度高、重现性好、一次可同时测几个样品等优点,正被日益广泛应用于医药卫生、食品卫生、环境卫生、分子生物学以及微生物学中.鲎试剂法大致可分为凝胶法、比浊法、比色法(显色法)等.凝胶法为限度试验,比浊法、比色法(显色法)常作为定量检测.本文综述介绍用此法测定细菌内毒素时发现的一些影响因素及消除方法的研究进展.

  14. APU diaphragm testing. Test plan

    Science.gov (United States)

    Shelley, Richard

    1992-01-01

    Auxiliary Power Unit (APU) fuel (hydrazine) tanks have had to be removed from the Columbia Shuttle (OV-102) because they have been in service for 11 years, which is the limit of their useful life. As part of an effort to determine whether the useful life of the fuel tanks can be extended, examination of the ethylene propylene rubber (EPR) diaphragm and the metal from one of the APU tanks is required. The JSC Propulsion and Power Division has requested White Sands Test Facility (WSTF) to examine the EPR diaphragm thoroughly and the metal casing generally from one tank. The objective is to examine the EPR diaphragm for signs of degradation that may limit the life of its function in the APU propellant tank. The metal casing will also be examined for signs of surface corrosion.

  15. QMix® irrigant reduces lipopolysacharide (LPS) levels in an in vitro model

    Science.gov (United States)

    GRÜNDLING, Grasiela Longhi; de MELO, Tiago André Fontoura; MONTAGNER, Francisco; SCARPARO, Roberta Kochenborger; VIER-PELISSER, Fabiana Vieira

    2015-01-01

    The presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions. Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals. Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C). After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX), and 3% sodium hypochlorite (NaOCl). LPS quantification was determined by Limulus Amebocyte Lysate (LAL) assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls. Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU)/mL (p<0.001). NaOCl (25.50 EU/mL), chlorhexidine (44.10 EU/mL) and positive control group (26.80 EU/mL) samples had similar results. Higher levels were found with EDTA (176.00 EU/mL) when compared to positive control (p<0.001). There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL) had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001). Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001). 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load. PMID:26398517

  16. Lab-on-a-Chip: From Astrobiology to the International Space Station

    Science.gov (United States)

    Maule, Jake; Wainwright, Nor; Steele, Andrew; Gunter, Dan; Monaco, Lisa A.; Wells, Mark E.; Morris, Heather C.; Boudreaux, Mark E.

    2008-01-01

    The continual and long-term habitation of enclosed environments, such as Antarctic stations, nuclear submarines and space stations, raises unique engineering, medical and operational challenges. There is no easy way out and no easy way to get supplies in. This situation elevates the importance of monitoring technology that can rapidly detect events within the habitat that affect crew safety such as fire, release of toxic chemicals and hazardous microorganisms. Traditional methods to monitor microorganisms on the International Space Station (ISS) have consisted of culturing samples for 3-5 days and eventual sample return to Earth. To augment these culture methods with new, rapid molecular techniques, we developed the Lab-on-a-Chip Application Development - Portable Test System (LOCAD-PTS). The system consists of a hand-held spectrophotometer, a series of interchangeable cartridges and a surface sampling/dilution kit that enables crew to collect samples and detect a range of biological molecules, all within 15 minutes. LOCAD-PTS was launched to the ISS aboard Space Shuttle Discovery in December 2006, where it was operated for the first time during March-May 2007. The surfaces of five separate sites in the US Lab and Node 1 of ISS were analyzed for endotoxin, using cartridges that employ the Limulus Amebocyte Lysate (LAL) assay; results of these tests will be presented. LOCAD-PTS will remain permanently onboard ISS with new cartridges scheduled for launch in February and October of 2008 for the detection of fungi (Beta-glucan) and Gram-positive bacteria (lipoteichoic acid), respectively.

  17. QMix® irrigant reduces lipopolysacharide (LPS) levels in an in vitro model.

    Science.gov (United States)

    Gründling, Grasiela Longhi; Melo, Tiago André Fontoura de; Montagner, Francisco; Scarparo, Roberta Kochenborger; Vier-Pelisser, Fabiana Vieira

    2015-01-01

    The presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions.Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals.Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C). After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX), and 3% sodium hypochlorite (NaOCl). LPS quantification was determined by Limulus Amebocyte Lysate (LAL) assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls.Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU)/mL (p<0.001). NaOCl (25.50 EU/mL), chlorhexidine (44.10 EU/mL) and positive control group (26.80 EU/mL) samples had similar results. Higher levels were found with EDTA (176.00 EU/mL) when compared to positive control (p<0.001). There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL) had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001).Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001). 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load. PMID:26398517

  18. In vitro evaluation of the action of irrigating solutions associated with intracanal medications on Escherichia coli and its endotoxin in root canals

    Directory of Open Access Journals (Sweden)

    Lilian Eiko Maekawa

    2011-04-01

    Full Text Available OBJECTIVE: The purpose of this study was to evaluate the effcacy of auxiliary chemical substances and intracanal medications on Escherichia coli and its endotoxin in root canals. MATERIAL AND METHODS: Teeth were contaminated with a suspension of E. coli for 14 days and divided into 3 groups according to the auxiliary chemical substance used: G1 2.5% sodium hypochlorite (NaOCl; G2 2% chlorhexidine gel (CLX; G3 pyrogenfree solution. After, these groups were subdivided according to the intracanal medication (ICM: A Calcium hydroxide paste (Calen®, B polymyxin B, and C Calcium hydroxide paste+2% CLX gel. For the control group (G4, pyrogen-free saline solution was used without application of intracanal medication. Samples of the root canal content were collected immediately after biomechanical preparation (BMP, at 7 days after BMP, after 14 days of intracanal medication activity, and 7 days after removal of intracanal medication. The following aspects were evaluated for all collections: a antimicrobial activity; b quantifcation of endotoxin by the Limulus Amebocyte Lysate test (LAL. Results were analyzed by the Kruskal-Wallis and Dunn's tests at 5% signifcance level. RESULTS: The 2.5% NaOCl and CLX were able to eliminate E. coli from root canal lumen and reduced the amount of endotoxin compared to saline. CONCLUSIONS: It was concluded that 2.5% NaOCl and CLX were effective in eliminating E. coli. Only the studied intracanal medications were to reduce the amount of endotoxin present in the root canals, regardless of the irrigant used.

  19. QMix® irrigant reduces lipopolysacharide (LPS levels in an in vitro model

    Directory of Open Access Journals (Sweden)

    Grasiela Longhi GRÜNDLING

    2015-08-01

    Full Text Available AbstractThe presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions.Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals.Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C. After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX, and 3% sodium hypochlorite (NaOCl. LPS quantification was determined by Limulus Amebocyte Lysate (LAL assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls.Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU/mL (p<0.001. NaOCl (25.50 EU/mL, chlorhexidine (44.10 EU/mL and positive control group (26.80 EU/mL samples had similar results. Higher levels were found with EDTA (176.00 EU/mL when compared to positive control (p<0.001. There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001.Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001. 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load.

  20. Microbiological assessment of house and imported bottled water by comparison of bacterial endotoxin concentration, heterotrophic plate count, and fecal coliform count.

    Science.gov (United States)

    Reyes, Mayra I; Pérez, Cynthia M; Negrón, Edna L

    2008-03-01

    Consumers increasingly use bottled water and home water treatment systems to avoid direct tap water. According to the International Bottled Water Association (IBWA), an industry trade group, 5 billion gallons of bottled water were consumed by North Americans in 2001. The principal aim of this study was to assess the microbial quality of in-house and imported bottled water for human consumption, by measurement and comparison of the concentration of bacterial endotoxin and standard cultivable methods of indicator microorganisms, specifically, heterotrophic and fecal coliform plate counts. A total of 21 brands of commercial bottled water, consisting of 10 imported and 11 in-house brands, selected at random from 96 brands that are consumed in Puerto Rico, were tested at three different time intervals. The Standard Limulus Amebocyte Lysate test, gel clot method, was used to measure the endotoxin concentrations. The minimum endotoxin concentration in 63 water samples was less than 0.0625 EU/mL, while the maximum was 32 EU/mL. The minimum bacterial count showed no growth, while the maximum was 7,500 CFU/mL. Bacterial isolates like P. fluorescens, Corynebacterium sp. J-K, S. paucimobilis, P. versicularis, A. baumannii, P. chlororaphis, F. indologenes, A. faecalis and P. cepacia were identified. Repeated measures analysis of variance demonstrated that endotoxin concentration did not change over time, while there was a statistically significant (p count over time. In addition, multiple linear regression analysis demonstrated that a unit change in the concentration of endotoxin across time was associated with a significant (p count. This analysis evidenced a significant time effect in the average log bacteriological cell count. Although bacterial growth was not detected in some water samples, endotoxin was present. Measurement of Gram-negative bacterial endotoxins is one of the methods that have been suggested as a rapid way of determining bacteriological water quality.

  1. Mononucleosis spot test

    Science.gov (United States)

    Monospot test; Heterophile antibody test; Heterophile agglutination test; Paul-Bunnell test; Forssman antibody test ... The mononucleosis spot test is done when symptoms of mononucleosis are ... Fatigue Fever Large spleen (possibly) Sore throat Tender ...

  2. A1C test

    Science.gov (United States)

    HbA1C test; Glycated hemoglobin test; Glycosylated hemoglobin test; Hemoglobin glycosylated test; Glycohemoglobin test ... have recently eaten does not affect the A1C test, so you do not need to fast to ...

  3. HPV DNA test

    Science.gov (United States)

    ... is generally not recommended for detecting low-risk HPV infections. ... Human papilloma virus - testing; Abnormal Pap smear - HPV testing; LSIL-HPV testing, Low-grade dysplasia - HPV testing; HSIL - HPV testing; High-grade dysplasia - HPV testing; HPV ...

  4. Rapid Monitoring of Bacteria and Fungi aboard the International Space Station (ISS)

    Science.gov (United States)

    Gunter, D.; Flores, G.; Effinger, M.; Maule, J.; Wainwright, N.; Steele, A.; Damon, M.; Wells, M.; Williams, S.; Morris, H.; Monaco, L.

    2009-01-01

    Microorganisms within spacecraft have traditionally been monitored with culture-based techniques. These techniques involve growth of environmental samples (cabin water, air or surfaces) on agar-type media for several days, followed by visualization of resulting colonies or return of samples to Earth for ground-based analysis. Data obtained over the past 4 decades have enhanced our understanding of the microbial ecology within space stations. However, the approach has been limited by the following factors: i) Many microorganisms (estimated > 95%) in the environment cannot grow on conventional growth media; ii) Significant time lags (3-5 days for incubation and up to several months to return samples to ground); iii) Condensation in contact slides hinders colony counting by crew; and iv) Growth of potentially harmful microorganisms, which must then be disposed of safely. This report describes the operation of a new culture-independent technique onboard the ISS for rapid analysis (within minutes) of endotoxin and beta-1, 3-glucan, found in the cell walls of gramnegative bacteria and fungi, respectively. The technique involves analysis of environmental samples with the Limulus Amebocyte Lysate (LAL) assay in a handheld device, known as the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS). LOCADPTS was launched to the ISS in December 2006, and here we present data obtained from Mach 2007 until the present day. These data include a comparative study between LOCADPTS analysis and existing culture-based methods; and an exploratory survey of surface endotoxin and beta-1, 3-glucan throughout the ISS. While a general correlation between LOCAD-PTS and traditional culture-based methods should not be expected, we will suggest new requirements for microbial monitoring based upon culture-independent parameters measured by LOCAD-PTS.

  5. Endotoxin exposure and changes in short-term pulmonary function among sewage workers

    Directory of Open Access Journals (Sweden)

    Marcin Cyprowski

    2015-10-01

    Full Text Available Objectives: The inhaled endotoxin is considered as a causative factor in the process of acute bronchial obstruction, which can be measured by a decrease in forced expiratory volume in 1 s (FEV1. The aim of this study was to assess endotoxin exposure among sewage treatment plant workers (STPW and its effect on across-shift changes in respiratory airflow. Material and Methods: A group of 78 STPW from a large sewage treatment plant was studied. Inhalable dust for endotoxin assessment was collected using personal aerosol samplers. Endotoxin was assayed with the kinetic, chromogenic Limulus amebocyte lysate test. Across-shift spirometric measurements were performed on Mondays, after 2-days absence from work, with the use of portable spirometer. The forced vital capacity (FVC, and FEV1 parameters were analyzed. Multifactor regression modeling was performed to determine parameters significantly associated with endotoxin exposure. Results: The concentration of inhalable dust and endotoxin ranged from 0.01–1.38 mg/m3 and 0.68–214 endotoxin units per cubic meter of air (EU/m3, respectively. Endotoxins were characterized with the skewed distribution (arithmetic mean (AM = 38.8 EU/m3, geometric mean (GM = 15.4 EU/m3, geometric standard deviation (GSD = 4.21. Through the use of multifactor analysis, which excluded the main confounders (inhalable dust and smoking habit it was found that, despite low levels of endotoxin, it had significant impact on the observed across-shift decline in FEV1 (p = 0.044. For this parameter, the regression slope was additionally calculated (r = –0.017, p = 0.071. Conclusions: Relatively low levels of endotoxin among sewage treatment plant workers may cause small, but significant across-shift declines in FEV1. The observed relationship was independent of organic dust concentrations and smoking habit. The respiratory protection should be provided for STPW.

  6. Bioaerosols from a Food Waste Composting Plant Affect Human Airway Epithelial Cell Remodeling Genes

    Directory of Open Access Journals (Sweden)

    Ming-Wei Chang

    2013-12-01

    Full Text Available The composting procedure in food waste plants generates airborne bioaerosols that have the potential to damage human airway epithelial cells. Persistent inflammation and repair responses induce airway remodeling and damage to the respiratory system. This study elucidated the expression changes of airway remodeling genes in human lung mucoepidermoid NCI-H292 cells exposed to bioaerosols from a composting plant. Different types of microorganisms were detectable in the composting plant, using the agar culture method. Real-time polymerase chain reaction was used to quantify the level of Aspergillus fumigatus and the profile of remodeling genes. The real-time PCR results indicated that the amount of A. fumigatus in the composting hall was less than 102 conidia. The endotoxins in the field bioaerosols were determined using a limulus amebocyte lysate test. The endotoxin levels depended on the type of particulate matter (PM, with coarse particles (2.5–10 μm having higher endotoxin levels than did fine particles (0.5–2.5 μm. After exposure to the conditioned medium of field bioaerosol samples, NCI-H292 cells showed increased pro-inflammatory interleukin (IL-6 release and activated epidermal growth factor receptor (EGFR, transforming growth factor (TGF-β1 and cyclin-dependent kinase inhibitor 1 (p21WAF1/CIP1 gene expression, but not of matrix metallopeptidase (MMP-9. Airborne endotoxin levels were higher inside the composting hall than they were in other areas, and they were associated with PM. This suggested that airborne bioaerosols in the composting plant contained endotoxins and microorganisms besides A. fumigatus that cause the inflammatory cytokine secretion and augment the expression of remodeling genes in NCI-H292 cells. It is thus necessary to monitor potentially hazardous materials from bioaerosols in food composting plants, which could affect the health of workers.

  7. Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis

    Directory of Open Access Journals (Sweden)

    Ariane Cassia Salustiano MARINHO

    2014-12-01

    Full Text Available Objective: The aim of this study was to monitor the effectiveness of root canal procedures by using different irrigants and intracanal medication on endotoxin levels found in root canals of teeth with chronic apical periodontitis. Material and Methods: Thirty root canals of teeth with pulpal necrosis associated with periapical lesions were selected and randomly divided into groups according to the irrigants used: GI - 2.5% NaOCl, GII - 2% chlorhexidine (CHX gel, and GIII - saline solution (SS (all, n=10. Samples were collected with sterile/apyrogenic paper points before (S1 and after root canal instrumentation (S2, after use of 17% ethylenediaminetetraacetic acid (EDTA (S3, and after 30 days of intracanal medication (Ca(OH2+SS (S4. A turbidimetric kinetic Limulus Amebocyte Lysate assay was used for endotoxin measurement. Results: Endotoxins were detected in 100% of the root canals investigated (30/30, with a median value of 18.70 EU/mL. After S2, significant median percentage reduction was observed in all groups, irrespective of the irrigant tested: 2.5% NaOCl (99.65% (GI, 2% CHX (94.27% (GII, and SS (96.79% (GIII (all p0.05. Intracanal medication for 30 days was able to significantly reduce residual endotoxins: 2.5% NaOCl (90% (GI, 2% CHX (88.8% (GII, and SS (85.7% (GIII, p<0.05. No differences were found in the endotoxin reduction when comparing s2 and s4 treatment groups. Conclusion: Our findings demonstrated the effectiveness of the mechanical action of the instruments along with the flow and backflow of irrigant enduring root canal instrumentation for the endotoxin removal from root canals of teeth with chronic apical periodontitis. Moreover, the use of intracanal medication for 30 days contributed for an improvement of endotoxin reduction.

  8. Oral Administration of P. gingivalis Induces Dysbiosis of Gut Microbiota and Impaired Barrier Function Leading to Dissemination of Enterobacteria to the Liver.

    Directory of Open Access Journals (Sweden)

    Mayuka Nakajima

    Full Text Available Although periodontitis has been implicated as a risk factor for various systemic diseases, the precise mechanisms by which periodontitis induces systemic disease remain to be elucidated. We have previously revealed that repeated oral administration of Porphyromonas gingivalis elicits endotoxemia via changes in the gut microbiota of the ileum, and thereby induces systemic inflammation and insulin resistance. However, it is not clear to what extent a single administration of P. gingivalis could affect gut microbiota composition, gut barrier function, and subsequent influx of gut microbiota into the liver. Therefore, in the present study, C57BL/6 mice were orally administered P. gingivalis (strain W83 once and compared to sham-inoculated mice. The phylogenetic structure and diversity of microbial communities in the gut and liver were analyzed by pyrosequencing the 16S ribosomal RNA genes. Serum endotoxin activity was determined by a Limulus amebocyte lysate test. Gene expression in the intestine and expression of 16S rRNA genes in the blood and liver were examined by quantitative polymerase chain reaction. Administration of P. gingivalis significantly altered gut microbiota, with an increased proportion of phylum Bacteroidetes, a decreased proportion of phylum Firmicutes, and increased serum endotoxin levels. In the intestinal tissues, gene expression of tjp-1 and occludin, which are involved in intestinal permeability, were downregulated. Higher amounts of bacterial DNA were detected in the liver of infected mice. Importantly, changes in gut microbiota preceded systemic inflammatory changes. These results further support the idea that disturbance of the gut microbiota composition by orally derived periodontopathic bacteria may be a causal mechanism linking periodontitis and systemic disease.

  9. Relationship between plasma )-lactate and intestinal damage after severe injuries in rats

    Institute of Scientific and Technical Information of China (English)

    Xiao-Qing Sun; Xiao-Bing Fu; Rong-Zhang; Yi Lü; Qun Deng; Xiao-Guo Jiang; Zhi-Yong Sheng

    2001-01-01

    AIM To explore the kinetic changes in plasma D (-)lactate and lipopolysaccharide ( LPS ) levels, and investigate whether D (-)-lactate could be used as a marker of intestinal injury in rats following gut ischemia/reperfusion, burn, and acute necrotizing pancreatitis (ANP). METHODS Three models were developed in rats:gut ischemia/reperfusion obtained by one hour of superior severe burn injury created by 30% of total body surface induced by continuous inverse infusion of sodium taurocholate and trypsin into main pancreatic duct.Plasma levels of D( - )-lactate in systemic circulation and LPS in portal circulation were measured by enzymaticspectrophotometric method and limulus amebocyte lysate (LAL) test kit, respectively. Tissue samples of intestine were taken for histological analysis. RESULTS One hour gut ischemia followed by reperfusion injuries resulted in a significant elevation in plasma D( - )lactate and LPS levels, and there was a significant correlation between the plasma D( - )-lactate and LPS (r =0.719, P<0.05). The plasma concentrations of D(-)lactate and LPS increased significantly at 6 h postburn,and there was also a remarkable correlation between them (r = 0.877, P < 0.01). D ( - )-lactate and LPS levels elevated significantly at 2 h after ANP, with a similar significant correlation between the two levels (r-0.798,P < 0.01 ). The desquamation of intestine villi and infiltration of inflammatory cells in the lamina propria were observed in all groups. CONCLUSION The changes of plasma D( - )-lactate levels in systemic blood paralleled with LPS levels in the portal vein blood. The measurement of plasma D (-)-lactate level may be a useful marker to assess the intestinal injury and to monitor an increase of intestinal permeability and endotoxemia following severe injuries in early stage.

  10. The Effect of Residual Endotoxin Contamination on the Neuroinflammatory Response to Sterilized Intracortical Microelectrodes.

    Science.gov (United States)

    Ravikumar, Madhumitha; Hageman, Daniel J; Tomaszewski, William H; Chandra, Gabriella M; Skousen, John L; Capadona, Jeffrey R

    2014-05-01

    A major limitation to the use of microelectrode technologies in both research and clinical applications is our inability to consistently record high quality neural signals. There is increasing evidence that recording instability is linked, in part, to neuroinflammation. A number of factors including extravasated blood products and macrophage released soluble factors are believed to mediate neuroinflammation and the resulting recording instability. However, the roles of other inflammatory stimuli, such as residual endotoxin contamination, are poorly understood. Therefore, to determine the effect of endotoxin contamination we examined the brain tissue response of C57/BL6 mice to non-functional microelectrodes with a range of endotoxin levels. Endotoxin contamination on the sterilized microelectrodes was measured using a limulus amebocyte lysate test following FDA guidelines. Microelectrodes sterilized by autoclave, dry heat, or ethylene oxide gas, resulted in variable levels of residual endotoxins of 0.55 EU/mL, 0.22 EU/mL, and 0.11 EU/mL, respectively. Histological evaluation at two weeks showed a direct correlation between microglia/macrophage activation and endotoxin levels. Interestingly, astrogliosis, neuronal loss, and blood brain barrier dysfunction demonstrated a threshold-dependent response to bacterial endotoxins. However, at sixteen weeks, no histological differences were detected, regardless of initial endotoxin levels. Therefore, our results demonstrate that endotoxin contamination, within the range examined, contributes to initial but not chronic microelectrode associated neuroinflammation. Our results suggest that minimizing residual endotoxins may impact early recording quality. To this end, endotoxins should be considered as a potent stimulant to the neuroinflammatory response to implanted intracortical microelectrodes.

  11. Development and preclinical testing of HNVAC, a cell culture-based H1N1 pandemic influenza vaccine from India.

    Science.gov (United States)

    Hegde, Nagendra R; Kumar, Deepak; Rao, P Panduranga; Kumari, P Krishna; Kaushik, Yashpal; Ravikrishnan, R; Prasad, Sai D; Ella, Krishna M

    2014-06-17

    Several limitations of the use of embryonated eggs and the threat of pandemics have highlighted the need for other platforms for the production of influenza vaccines. We report the indigenous development and pre-clinical testing of an MDCK-based H1N1 pandemic influenza vaccine HNVAC from India. The cell bank and virus seed were characterized extensively. The cells were characterized by PCR, electron microscopy, and karyotyping, and found to be of female canine epithelial origin. The virus was confirmed by neutralization, haemagglutination inhibition, neuraminidase inhibition, and PCR and nucleotide sequencing. Adventitious agent testing was performed by both in vitro and in vivo studies. The in vitro studies included culturing, haemadsorption, haemagglutination, PCR and RT-PCR, whereas in vivo studies included passage in embryonated eggs and in laboratory animals. Both cell bank and virus seed were free of adventitious agents. MDCK cell lysates as well as cellular DNA did not produce tumours in newborn or adult laboratory animals. The bioprocess parameters were standardized to recover antigen with minimal levels of process-related impurities. The vaccine bulk was tested for the presence of specific antigen, and quantified by single radial immunodiffusion. Finally, non-adjuvanted and aluminium hydroxide adjuvanted vaccine formulations were found to be safe in preclinical toxicity studies in mice, rats, guinea pigs and rabbits, and immunogenic in mice and rabbits. This is the first and only cell culture-based influenza vaccine platform developed in any developing country.

  12. Tests in contingency tables as regression tests

    OpenAIRE

    Stanislav Anatolyev; Grigory Kosenok

    2006-01-01

    Applied researchers often use tests based on contingency tables in preliminary data analysis and diagnostic testing. We show that many of such tests may be alternatively implemented by testing for coecient restrictions in linear regression systems (as a rule, employing the Wald test). This uni es the theories of regression analysis and contingency tables, sheds more light on intuitive contents of contingency table tests, and provides a more convenient and familiar tool for practitioners.

  13. Growth hormone suppression test

    Science.gov (United States)

    GH suppression test; Glucose loading test; Acromegaly - blood test; Gigantism - blood test ... during the suppression test, the provider will suspect gigantism or acromegaly. You may need to be retested to confirm ...

  14. Tests Related to Pregnancy

    Science.gov (United States)

    ... to learn. Search form Search Tests related to pregnancy You are here Home Testing & Services Testing for ... to Genetic Counseling . What Are Tests Related to Pregnancy? Pregnancy related testing is done before or during ...

  15. Anthrax - blood test

    Science.gov (United States)

    ... The best test for diagnosing anthrax is a culture of affected tissue or blood. Alternative Names Anthrax serology test; Antibody test for anthrax; Serologic test for B anthracis Images Blood test Bacillus anthracis References Hall GS, Woods GL. Medical bacteriology. ...

  16. Practicable group testing method to evaluate weight/weight GMO content in maize grains.

    Science.gov (United States)

    Mano, Junichi; Yanaka, Yuka; Ikezu, Yoko; Onishi, Mari; Futo, Satoshi; Minegishi, Yasutaka; Ninomiya, Kenji; Yotsuyanagi, Yuichi; Spiegelhalter, Frank; Akiyama, Hiroshi; Teshima, Reiko; Hino, Akihiro; Naito, Shigehiro; Koiwa, Tomohiro; Takabatake, Reona; Furui, Satoshi; Kitta, Kazumi

    2011-07-13

    Because of the increasing use of maize hybrids with genetically modified (GM) stacked events, the established and commonly used bulk sample methods for PCR quantification of GM maize in non-GM maize are prone to overestimate the GM organism (GMO) content, compared to the actual weight/weight percentage of GM maize in the grain sample. As an alternative method, we designed and assessed a group testing strategy in which the GMO content is statistically evaluated based on qualitative analyses of multiple small pools, consisting of 20 maize kernels each. This approach enables the GMO content evaluation on a weight/weight basis, irrespective of the presence of stacked-event kernels. To enhance the method's user-friendliness in routine application, we devised an easy-to-use PCR-based qualitative analytical method comprising a sample preparation step in which 20 maize kernels are ground in a lysis buffer and a subsequent PCR assay in which the lysate is directly used as a DNA template. This method was validated in a multilaboratory collaborative trial.

  17. Spiked proteomic standard dataset for testing label-free quantitative software and statistical methods.

    Science.gov (United States)

    Ramus, Claire; Hovasse, Agnès; Marcellin, Marlène; Hesse, Anne-Marie; Mouton-Barbosa, Emmanuelle; Bouyssié, David; Vaca, Sebastian; Carapito, Christine; Chaoui, Karima; Bruley, Christophe; Garin, Jérôme; Cianférani, Sarah; Ferro, Myriam; Dorssaeler, Alain Van; Burlet-Schiltz, Odile; Schaeffer, Christine; Couté, Yohann; Gonzalez de Peredo, Anne

    2016-03-01

    This data article describes a controlled, spiked proteomic dataset for which the "ground truth" of variant proteins is known. It is based on the LC-MS analysis of samples composed of a fixed background of yeast lysate and different spiked amounts of the UPS1 mixture of 48 recombinant proteins. It can be used to objectively evaluate bioinformatic pipelines for label-free quantitative analysis, and their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. More specifically, it can be useful for tuning software tools parameters, but also testing new algorithms for label-free quantitative analysis, or for evaluation of downstream statistical methods. The raw MS files can be downloaded from ProteomeXchange with identifier PXD001819. Starting from some raw files of this dataset, we also provide here some processed data obtained through various bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold) in different workflows, to exemplify the use of such data in the context of software benchmarking, as discussed in details in the accompanying manuscript [1]. The experimental design used here for data processing takes advantage of the different spike levels introduced in the samples composing the dataset, and processed data are merged in a single file to facilitate the evaluation and illustration of software tools results for the detection of variant proteins with different absolute expression levels and fold change values.

  18. Autologous tumor cell lysate-loaded dendritic cells and cytokine-induced killer cells in combination with autologous hematopoietic stem cell transplatation in the treatment of refractory lymphoma%负载自体肿瘤抗原的DC-CIK细胞联合自体造血干细胞移植治疗难治性淋巴瘤的临床研究

    Institute of Scientific and Technical Information of China (English)

    苏毅; 邵文军; 闵敏; 李莉; 陈健; 范方教; 易海; 付利; 刘阳阳; 邓涛; 孙浩平; 孙薏; 钟国成

    2009-01-01

    目的 观察负载自体肿瘤抗原的DE-CIK细胞联合自体造血干细胞移植治疗难治性淋巴瘤的疗效.方法 选取难治性淋巴瘤35例,采用MAC预处理方案,用自体淋巴瘤抗原致敏DC-CIK细胞,于移植预处理后5-10d,将DC-CIK细胞回输给患者.结果 35例难治性淋巴瘤中,29例完全缓解(82.86%),4例部分缓解(14.43%),移植过程中死亡2例(5.71%)(均死于严重混合性感染).所有完全缓解和部分缓解病例均随访3-49个月:4名部分缓解患者分别于移植后3、6、10、13个月后病情进展死亡;完全缓解患者中有3人于移植后11、17、20个月再次复发死亡;现存活26例.结论 负载自体肿瘤抗原的DC-CIK细胞联合自体造血千细胞移植治疗难治性淋巴瘤高于单纯自体外周血造血干细胞的疗效,且无明显毒副作用.%Objective To study the efficacy and side effect of autologons tumor cell lysate-loaded DC-CIK plus autologous hematopoietic stem cell transplantation in the treatment of refractory lymphoma.Methods Thirty-five cases of refractory lymphoma were recruited,lymphoma antigen-pulsed autologous DE-CIK cells were infused 5-10 days after the MAC conditioning regimen.Results Out of the 35 Cases,complete remission Was achieved in 29(82.8%),partial remission in 4(14.4%).Two patients died during transplantation due to severe mixed infections,with a transplant-related mortality rate of 5.7%.All cases of complete remission and partial remission were followed-up for 3-49 months,4 cases with partial remission died 3,6,10,13 months after transplantation.Three cases with complete remission relapsed 11,17,20 months after transplantation.and the other 26 cases survived until now.Conclusion Autologous tumor cell lysate-loaded DE-CIK combined with autologous hematopoietic stem cell transplantation in the treatment of refractory lymphoma is safe and effective.

  19. Design Driven Testing Test Smarter, Not Harder

    CERN Document Server

    Stephens, M

    2010-01-01

    The groundbreaking book Design Driven Testing brings sanity back to the software development process by flipping around the concept of Test Driven Development (TDD) - restoring the concept of using testing to verify a design instead of pretending that unit tests are a replacement for design. Anyone who feels that TDD is "Too Damn Difficult" will appreciate this book. Design Driven Testing shows that, by combining a forward-thinking development process with cutting-edge automation, testing can be a finely targeted, business-driven, rewarding effort. In other words, you'll learn how to test

  20. Heat pipe testing program test plan

    Energy Technology Data Exchange (ETDEWEB)

    Bienert, W.B.

    1980-03-14

    A test plan is given which describes the tests to be conducted on several typical solar receiver heat pipes. The hardware to be used, test fixtures and rationale of the test program are discussed. The program objective is to perform life testing under simulated receiver conditions, and to conduct performance tests with selected heat pipes to further map their performance, particularly with regard to their transient behavior. Performance requirements are defined. Test fixtures designed for the program are described in detail, and their capabilities for simulating the receiver conditions and their limitations are discussed. The heat pipe design is given. (LEW)

  1. Learning software testing with Test Studio

    CERN Document Server

    Madi, Rawane

    2013-01-01

    Learning Software Testing with Test Studio is a practical, hands-on guide that will help you get started with Test Studio to design your automated solution and tests. All through the book, there are best practices and tips and tricks inside Test Studio which can be employed to improve your solution just like an experienced QA.If you are a beginner or a professional QA who is seeking a fast, clear, and direct to the point start in automated software testing inside Test Studio, this book is for you. You should be familiar with the .NET framework, mainly Visual Studio, C#, and SQL, as the book's

  2. TEST SUITE GENERATION PROCESS FOR AGENT TESTING

    Directory of Open Access Journals (Sweden)

    HOUHAMDI ZINA

    2011-04-01

    Full Text Available Software agents are a promising technology for today's complex, distributed systems. Methodologies and techniques that address testing and reliability of multi agent systems are increasingly demanded, in particular to support automated test case generation and execution. In this paper, we introduce a novel approach for goal-oriented software agent testing. It specifies a testing process that complements the goal oriented methodology Tropos and reinforces the mutual relationship between goal analysis and testing. Furthermore, it defines a structured and comprehensive agent test suite generation process by providing a systematic way of deriving test cases from goal analysis.

  3. Lyme disease blood test

    Science.gov (United States)

    ... antibodies in the blood sample using the ELISA test . If the ELISA test is positive, it must be confirmed with another ... were seen in your blood sample. If the ELISA test is negative, usually no other testing is needed. ...

  4. Ketones urine test

    Science.gov (United States)

    Ketone bodies - urine; Urine ketones; Ketoacidosis - urine ketones test; Diabetic ketoacidosis - urine ketones test ... Urine ketones are usually measured as a "spot test." This is available in a test kit that ...

  5. Coccidioides precipitin test

    Science.gov (United States)

    Coccidioidomycosis antibody test ... There is no special preparation for the test. ... The precipitin test is one of several tests that can be done to determine if you are infected with the fungus ...

  6. Sweat electrolytes test

    Science.gov (United States)

    Sweat test; Sweat chloride; Iontophoretic sweat test ... No special steps are needed before this test. ... The test is not painful. Some people have a tingling feeling at the site of the electrode. This feeling ...

  7. Genetic Testing (For Parents)

    Science.gov (United States)

    ... Story" 5 Things to Know About Zika & Pregnancy Genetic Testing KidsHealth > For Parents > Genetic Testing Print A A ... blood, skin, bone, or other tissue is needed. Genetic Testing During Pregnancy For genetic testing before birth, pregnant ...

  8. Genetic Testing for ALS

    Science.gov (United States)

    ... your area, please visit www.nsgc.org . Genetic Testing Genetic testing can help determine the cause of FALS ... couples planning on having children to pursue prenatal testing. Genetic testing does not: Currently change medical treatment. Diagnose ...

  9. Allergy testing - skin

    Science.gov (United States)

    Patch tests - allergy; Scratch tests - allergy; Skin tests - allergy; RAST test ... There are three common methods of allergy skin testing. The skin prick test involves: Placing a small amount of substances that may be causing your symptoms on the skin, most often ...

  10. To test or not to test

    DEFF Research Database (Denmark)

    Rochon, Justine; Gondan, Matthias; Kieser, Meinhard

    2012-01-01

    Background: Student's two-sample t test is generally used for comparing the means of two independent samples, for example, two treatment arms. Under the null hypothesis, the t test assumes that the two samples arise from the same normally distributed population with unknown variance. Adequate...... control of the Type I error requires that the normality assumption holds, which is often examined by means of a preliminary Shapiro-Wilk test. The following two-stage procedure is widely accepted: If the preliminary test for normality is not significant, the t test is used; if the preliminary test rejects...... the null hypothesis of normality, a nonparametric test is applied in the main analysis. Methods: Equally sized samples were drawn from exponential, uniform, and normal distributions. The two-sample t test was conducted if either both samples (Strategy I) or the collapsed set of residuals from both samples...

  11. Susceptibility Testing by Polymerase Chain Reaction DNA Quantitation: A Method to Measure Drug Resistance of Human Immunodeficiency Virus Type 1 Isolates

    Science.gov (United States)

    Eron, Joseph J.; Gorczyca, Paul; Kaplan, Joan C.; D'Aquila, Richard T.

    1992-04-01

    Polymerase chain reaction (PCR) DNA quantitation (PDQ) susceptibility testing rapidly and directly measures nucleoside sensitivity of human immunodeficiency virus type 1 (HIV-1) isolates. PCR is used to quantitate the amount of HIV-1 DNA synthesized after in vitro infection of peripheral blood mononuclear cells. The relative amounts of HIV-1 DNA in cell lysates from cultures maintained at different drug concentrations reflect drug inhibition of virus replication. The results of PDQ susceptibility testing of 2- or 3-day cultures are supported by assays measuring HIV-1 p24 antigen production in supernatants of 7- or 10-day cultures. DNA sequence analyses to identify mutations in the reverse transcriptase gene that cause resistance to 3'-azido-3'-deoxythymidine also support the PDQ results. With the PDQ method, both infectivity titration and susceptibility testing can be performed on supernatants from primary cultures of peripheral blood mononuclear cells. PDQ susceptibility testing should facilitate epidemiologic studies of the clinical significance of drug-resistant HIV-1 isolates.

  12. To Test or Not to Test?

    Science.gov (United States)

    Circle, David

    2005-01-01

    This paper discusses whether music educators should push for national testing of music students. The National Assessment of Educational Progress (NAEP) did test music students in 1997. Even though the results of that test did not indicate students were very accomplished, there was a general feeling that at least NAEP and the nation recognized…

  13. Impact of bacterial lysates on the expression of TGF-β1 and Foxp3 in mouse%细菌溶解产物对支气管哮喘小鼠TGF-β1及Foxp3表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘静; 周小建; 洪建国

    2014-01-01

    达显著低于正常组,f组较c组表达明显增加,e组、d组次之,差异有统计学意义(P<0.01).结论 口服OM-85BV干预有利于改善哮喘小鼠模型气道炎症,显著提高外周血及外周免疫器官-脾脏的Treg功能相关细胞因子TGF-β1及特异性转录因子Foxp3的表达,二者可能是预防哮喘气道炎症的发生和发展的治疗靶点.%Objective To establish mouse allergic bronchial asthma (asthma) model and observe the effect of bacterial lysates (OM-85BV) on airway inflammation,as well as the expression of TGF-β1 and Foxp3 in serum and spleen.Methods Forty-eight 4 to 6 weeks healthy male BALB/c mice were used as research subjects and randomly divided into six groups,a:contral group,b:OM-85BV contral group,c:allergic asthma model,d:dexamethasone group (Dex group),e:OM-85BV A group,f:OM-85BV B group (the intervention time was prolonged 10 days than group e).BALB/c mice were sensitized and challenged by ovalbumin (OVA).Mice in groups c,d,e and f were intraperitoneally administered by antigen (OVA)-Al(OH)3 on days 1,8 and 15,others were administered by PBS.From the 17th day to the 26th day,mice in the group f were treated with OM-85BV and others were treated with normal saline.In the next days,mice in groups c,d,e and f were intranasal given OVA for 5 consecutive days.Additionally,mice in groups b,e and f were treated with OM-85BV before challenge,while mice in the group d were administered by Dex,others were treated with normal saline at the same dose.Twenty four hours after the last intranasal administration,mice were anesthetized and dissected.The TGF-β1 levels of blood serum were detected.The removed parts of lung tissue were for histological to observe airway inflammation and the thymuses were prepared for immunohistochemistry examination to detect the expression of Foxp3.The expression of TGF-β1 mRNA and Foxp3 mRNA in spleen were detected by Real time-PCR.Results Compared with groups a and b,lung tissue biopsies by HE

  14. Web Security Testing Cookbook

    CERN Document Server

    Hope, Paco

    2008-01-01

    Among the tests you perform on web applications, security testing is perhaps the most important, yet it's often the most neglected. The recipes in the Web Security Testing Cookbook demonstrate how developers and testers can check for the most common web security issues, while conducting unit tests, regression tests, or exploratory tests. Unlike ad hoc security assessments, these recipes are repeatable, concise, and systematic-perfect for integrating into your regular test suite.

  15. Get Tested for HIV

    Science.gov (United States)

    ... Submit Home > HIV/AIDS > Get tested for HIV HIV/AIDS This information in Spanish ( en español ) Get tested for HIV When should I get tested for HIV? Types ... to top More information on Get tested for HIV Explore other publications and websites HIV Anonymous Testing, ...

  16. Test Construction: Automated

    NARCIS (Netherlands)

    Veldkamp, Bernard P.

    2014-01-01

    Optimal test construction deals with automated assembly of tests for educational and psychological measurement. Items are selected from an item bank to meet a predefined set of test specifications. Several models for optimal test construction are presented, and two algorithms for optimal test assemb

  17. Test Construction: Automated

    NARCIS (Netherlands)

    Veldkamp, Bernard P.

    2016-01-01

    Optimal test construction deals with automated assembly of tests for educational and psychological measurement. Items are selected from an item bank to meet a predefined set of test specifications. Several models for optimal test construction are presented, and two algorithms for optimal test assemb

  18. CO2 blood test

    Science.gov (United States)

    Bicarbonate test; HCO3-; Carbon dioxide test; TCO2; Total CO2; CO2 test - serum ... Many medicines can interfere with blood test results. Your health care provider will tell you if you need to stop taking any medicines before you have this test. DO ...

  19. Growth hormone stimulation test

    Science.gov (United States)

    Arginine test; Arginine-GHRH test ... of re-inserting the needle each time. The test takes between 2 to 5 hours. The procedure ... eat for 10 to 12 hours before the test. Eating food can change the test results. Some ...

  20. Vendor System Vulnerability Testing Test Plan

    Energy Technology Data Exchange (ETDEWEB)

    James R. Davidson

    2005-01-01

    The Idaho National Laboratory (INL) prepared this generic test plan to provide clients (vendors, end users, program sponsors, etc.) with a sense of the scope and depth of vulnerability testing performed at the INL’s Supervisory Control and Data Acquisition (SCADA) Test Bed and to serve as an example of such a plan. Although this test plan specifically addresses vulnerability testing of systems applied to the energy sector (electric/power transmission and distribution and oil and gas systems), it is generic enough to be applied to control systems used in other critical infrastructures such as the transportation sector, water/waste water sector, or hazardous chemical production facilities. The SCADA Test Bed is established at the INL as a testing environment to evaluate the security vulnerabilities of SCADA systems, energy management systems (EMS), and distributed control systems. It now supports multiple programs sponsored by the U.S. Department of Energy, the U.S. Department of Homeland Security, other government agencies, and private sector clients. This particular test plan applies to testing conducted on a SCADA/EMS provided by a vendor. Before performing detailed vulnerability testing of a SCADA/EMS, an as delivered baseline examination of the system is conducted, to establish a starting point for all-subsequent testing. The series of baseline tests document factory delivered defaults, system configuration, and potential configuration changes to aid in the development of a security plan for in depth vulnerability testing. The baseline test document is provided to the System Provider,a who evaluates the baseline report and provides recommendations to the system configuration to enhance the security profile of the baseline system. Vulnerability testing is then conducted at the SCADA Test Bed, which provides an in-depth security analysis of the Vendor’s system.b a. The term System Provider replaces the name of the company/organization providing the system

  1. Estradiol blood test

    Science.gov (United States)

    E2 test ... temporarily stop taking certain medicines that may affect test results. Be sure to tell your provider about ... helps prevent sperm from dying too early. This test may be ordered to check: How well your ...

  2. Hepatitis A Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Hepatitis A Testing Share this page: Was this page ... HAV-Ab total; Anti-HAV Formal name: Viral Hepatitis A Antibody Related tests: Hepatitis B Testing ; Hepatitis ...

  3. CSF-VDRL test

    Science.gov (United States)

    Venereal disease research laboratory slide test - CSF ... provider's instructions on how to prepare for this test. ... The CSF-VDRL test is done to diagnose syphilis in the brain or spinal cord. Brain and spinal cord involvement is often a ...

  4. Mark 1 Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Mark I Test Facility is a state-of-the-art space environment simulation test chamber for full-scale space systems testing. A $1.5M dollar upgrade in fiscal year...

  5. Assessment and Testing.

    Science.gov (United States)

    Clapham, Caroline

    2000-01-01

    Explores the term "applied linguistics" and discusses the role of language testing within this discipline, the relationship between testing and teaching, and the relationship between testing and assessment (Author/VWL)

  6. Vitamin A Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Vitamin A Share this page: Was this page helpful? ... Related tests: Complete Blood Count , Comprehensive Metabolic Panel , Vitamin B12 and Folate , Vitamin D Tests , Iron Tests , ...

  7. Teratology testing under REACH.

    Science.gov (United States)

    Barton, Steve

    2013-01-01

    REACH guidelines may require teratology testing for new and existing chemicals. This chapter discusses procedures to assess the need for teratology testing and the conduct and interpretation of teratology tests where required.

  8. Sickle cell test

    Science.gov (United States)

    Sickledex; Hgb S test ... This test is done to tell if a person has abnormal hemoglobin that causes sickle cell disease and sickle ... and no symptoms, or only mild ones. This test does not tell the difference between these two ...

  9. Dexamethasone suppression test

    Science.gov (United States)

    DST; ACTH suppression test; Cortisol suppression test ... During this test, you will receive dexamethasone. This is a strong man-made (synthetic) glucocorticoid medication. Afterward, your blood is drawn ...

  10. Stomach acid test

    Science.gov (United States)

    Gastric acid secretion test ... The test is done after you have not eaten for a while so fluid is all that remains in ... injected into your body. This is done to test the ability of the cells in the stomach ...

  11. Breath alcohol test

    Science.gov (United States)

    Alcohol test - breath ... There are various brands of breath alcohol tests. Each one uses a different method to test the level of alcohol in the breath. The machine may be electronic or manual. One ...

  12. ALP isoenzyme test

    Science.gov (United States)

    Alkaline phosphatase isoenzyme test ... anything for 10 to 12 hours before the test, unless your health care provider tells you to do so. Many medicines can interfere with blood test results. Your health care provider will tell you ...

  13. Bone mineral density test

    Science.gov (United States)

    BMD test; Bone density test; Bone densitometry; DEXA scan; DXA; Dual-energy x-ray absorptiometry; p-DEXA; Osteoporosis-BMD ... need to undress. This scan is the best test to predict your risk of fractures. Peripheral DEXA ( ...

  14. PBG urine test

    Science.gov (United States)

    Porphobilinogen test ... temporarily stop taking medicines that may affect the test results. Be sure to tell your provider about ... This test involves only normal urination, and there is no discomfort.

  15. Creatine phosphokinase test

    Science.gov (United States)

    CPK test ... vein. The procedure is called a venipuncture . This test may be repeated over 2 or 3 days ... helps determine which tissue has been damaged. This test may be used to: Diagnose heart attack Evaluate ...

  16. Home vision tests

    Science.gov (United States)

    ... portion. Do the test with each eye separately, first the right and then the left. Hold the test grid ... them for the test. Check each eye separately, first the right and then the left. Keep both eyes open ...

  17. Structural Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — Provides a wide variety of testing equipment, fixtures and facilities to perform both unique aviation component testing as well as common types of materials testing...

  18. Small test SDHW systems

    DEFF Research Database (Denmark)

    Vejen, Niels Kristian

    1999-01-01

    Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different.......Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different....

  19. Textiles Performance Testing Facilities

    Data.gov (United States)

    Federal Laboratory Consortium — The Textiles Performance Testing Facilities has the capabilities to perform all physical wet and dry performance testing, and visual and instrumental color analysis...

  20. Who fails lantern tests?

    Science.gov (United States)

    Cole, B L; Vingrys, A J

    1983-05-01

    A battery of clinical colour vision tests was given to a group of 100 observers with abnormal colour vision who were also tested on the Farnsworth lantern and the Holmes-Wright lanterns types A and B. It was found that clinical colour vision tests are imperfect predictors of lantern test performance. However, observers classified as having a 'severe' colour vision defect were found to fail the lantern tests but only one half to two-thirds of those who fail the lantern tests can be identified in this way. It is not possible to identify with certainty any of the people likely to pass the lantern tests: about one-third to two-thirds of observers classified as being mildly affected fail the lantern tests. The Farnsworth D-15 and City University tests were found to be the best predictors of lantern test performance but other tests such as the Nagel anomaloscope, the H-16, L'Anthony's desaturated test can also be used. The lack of a strong correlation between clinical tests and the recognition of the small coloured stimuli presented by the lantern tests suggests that clinical tests do not test the same aspect of colour vision that is important to the recognition of signal lights. For this reason lantern tests should be retained for occupational testing of colour vision.

  1. Electromagnetic Interface Testing Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Electromagnetic Interface Testing facilitysupports such testing asEmissions, Field Strength, Mode Stirring, EMP Pulser, 4 Probe Monitoring/Leveling System, and...

  2. AUTOMATED API TESTING APPROACH

    Directory of Open Access Journals (Sweden)

    SUNIL L. BANGARE

    2012-02-01

    Full Text Available Software testing is an investigation conducted to provide stakeholders with information about the quality of the product or service under test. With the help of software testing we can verify or validate the software product. Normally testing will be done after development of software but we can perform the software testing at the time of development process also. This paper will give you a brief introduction about Automated API Testing Tool. This tool of testing will reduce lots of headache after the whole development of software. It saves time as well as money. Such type of testing is helpful in the Industries & Colleges also.

  3. Test Control Center (TCC)

    Data.gov (United States)

    Federal Laboratory Consortium — The Test Control Center (TCC) provides a consolidated facility for planning, coordinating, controlling, monitoring, and analyzing distributed test events. ,The TCC...

  4. Mobile Test Capabilities

    Data.gov (United States)

    Federal Laboratory Consortium — The Electrical Power Mobile Test capabilities are utilized to conduct electrical power quality testing on aircraft and helicopters. This capability allows that the...

  5. FOOD SAFETY TESTING LABORATORY

    Data.gov (United States)

    Federal Laboratory Consortium — This laboratory develops screening assays, tests and modifies biosensor equipment, and optimizes food safety testing protocols for the military and civilian sector...

  6. GPS Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Global Positioning System (GPS) Test Facility Instrumentation Suite (GPSIS) provides great flexibility in testing receivers by providing operational control of...

  7. Testing Library Specifications by Verifying Conformance Tests

    DEFF Research Database (Denmark)

    Kiniry, Joseph Roland; Zimmerman, Daniel; Hyland, Ralph

    2012-01-01

    conformance test suites. Rather than executing the conformance tests, FCTD uses them to measure the correctness and utility of specifications through static verification. FCTD is beginning to see significant use within the JML community and is the cornerstone process of the JML Spec-a-thons, meetings...

  8. Test Wiseness and Analogy Test Performance

    Science.gov (United States)

    Moore, James C.

    1971-01-01

    Subjects received self instruction on how to approach analogy questions. Instruction was directed toward knowledge of the general format of analogy questions in standarized tests and the 15 types of relationships commonly asked for in analogy questions. An analogies post-test showed a significant effect for the group. (Author)

  9. Localizing Test Power Consumption for Scan Testing

    Institute of Scientific and Technical Information of China (English)

    XIANG Dong; LI Kai-wei

    2005-01-01

    A two stage scan architecture is proposed to do low powerand low test application cost scan testing. The first stage includes multiple scan chains, where each scan chain is driven by a primary input. Each scan flip-flop in the multiple scan chains drives a group of scan flip-flops. The scan flip-flop in the multiple scan chain and the scan flipflop driven by it are assigned the same values for all test vectors. Scan flip-flops in the multiple scan chains and those in the second stage use separate clock signals, but the design for testability technqiue needs only one clock. The proposed scan architecture localizes test power consumption to the multiple scan chains during test application. Test signals assigned to scan flip-flops in the multiple scan chains are applied to the scan flip-flops in the second stage after the test vector has been applied to the multiple scan chains. This technique can make test power consumption very small.

  10. Digface characterization test plan (remote testing)

    Energy Technology Data Exchange (ETDEWEB)

    Croft, K.; Hyde, R.; Allen, S.

    1993-08-01

    The objective of the Digface Characterization (DFC) Remote Testing project is to remotely deploy a sensor head (Mini-Lab) across a digface to determine if it can characterize the contents below the surface. The purpose of this project is to provide a robotics technology that allows removal of workers from hazards, increases speed of operations, and reduces life cycle costs compared to alternate methods and technologies. The Buried Waste Integrated Demonstration (BWID) is funding the demonstration, testing, and evaluation of DFC. This document describes the test plan for the DFC remote deployment demonstration for the BWID. The purposes of the test plan are to establish test parameters so that the demonstration results are deemed useful and usable and perform the demonstration in a safe manner and within all regulatory requirements.

  11. Digface characterization test plan (remote testing)

    International Nuclear Information System (INIS)

    The objective of the Digface Characterization (DFC) Remote Testing project is to remotely deploy a sensor head (Mini-Lab) across a digface to determine if it can characterize the contents below the surface. The purpose of this project is to provide a robotics technology that allows removal of workers from hazards, increases speed of operations, and reduces life cycle costs compared to alternate methods and technologies. The Buried Waste Integrated Demonstration (BWID) is funding the demonstration, testing, and evaluation of DFC. This document describes the test plan for the DFC remote deployment demonstration for the BWID. The purposes of the test plan are to establish test parameters so that the demonstration results are deemed useful and usable and perform the demonstration in a safe manner and within all regulatory requirements

  12. Role of test motivation in intelligence testing

    Science.gov (United States)

    Duckworth, Angela Lee; Quinn, Patrick D.; Lynam, Donald R.; Loeber, Rolf; Stouthamer-Loeber, Magda

    2011-01-01

    Intelligence tests are widely assumed to measure maximal intellectual performance, and predictive associations between intelligence quotient (IQ) scores and later-life outcomes are typically interpreted as unbiased estimates of the effect of intellectual ability on academic, professional, and social life outcomes. The current investigation critically examines these assumptions and finds evidence against both. First, we examined whether motivation is less than maximal on intelligence tests administered in the context of low-stakes research situations. Specifically, we completed a meta-analysis of random-assignment experiments testing the effects of material incentives on intelligence-test performance on a collective 2,008 participants. Incentives increased IQ scores by an average of 0.64 SD, with larger effects for individuals with lower baseline IQ scores. Second, we tested whether individual differences in motivation during IQ testing can spuriously inflate the predictive validity of intelligence for life outcomes. Trained observers rated test motivation among 251 adolescent boys completing intelligence tests using a 15-min “thin-slice” video sample. IQ score predicted life outcomes, including academic performance in adolescence and criminal convictions, employment, and years of education in early adulthood. After adjusting for the influence of test motivation, however, the predictive validity of intelligence for life outcomes was significantly diminished, particularly for nonacademic outcomes. Collectively, our findings suggest that, under low-stakes research conditions, some individuals try harder than others, and, in this context, test motivation can act as a third-variable confound that inflates estimates of the predictive validity of intelligence for life outcomes. PMID:21518867

  13. Nationale test i naturfag

    DEFF Research Database (Denmark)

    Andreasen, Karen Egedal; Jensen, Lars Bang

    2015-01-01

    Kapitlet rummer en analyse og diskussion af test inden for naturfagsområdet og de fagforståelser de afspejler med fokus på de nationale test.......Kapitlet rummer en analyse og diskussion af test inden for naturfagsområdet og de fagforståelser de afspejler med fokus på de nationale test....

  14. Pre-Test Assessment

    Science.gov (United States)

    Berry, Thomas

    2008-01-01

    Pre-tests are a non-graded assessment tool used to determine pre-existing subject knowledge. Typically pre-tests are administered prior to a course to determine knowledge baseline, but here they are used to test students prior to topical material coverage throughout the course. While counterintuitive, the pre-tests cover material the student is…

  15. Refactoring test code

    NARCIS (Netherlands)

    Deursen, A. van; Moonen, L.M.F.; Bergh, A. van den; Kok, G.

    2001-01-01

    Two key aspects of extreme programming (XP) are unit testing and merciless refactoring. Given the fact that the ideal test code / production code ratio approaches 1:1, it is not surprising that unit tests are being refactored. We found that refactoring test code is different from refactoring product

  16. ITER test programme

    Science.gov (United States)

    Abdou, M.; Baker, C.; Casini, G.

    1991-07-01

    The International Thermonuclear Experimental Reactor (ITER) was designed to operate in two phases. The first phase, which lasts for 6 years, is devoted to machine checkout and physics testing. The second phase lasts for 8 years and is devoted primarily to technology testing. This report describes the technology test program development for ITER, the ancillary equipment outside the torus necessary to support the test modules, the international collaboration aspects of conducting the test program on ITER, the requirements on the machine major parameters and the R and D program required to develop the test modules for testing in ITER.

  17. ITER test programme

    International Nuclear Information System (INIS)

    ITER has been designed to operate in two phases. The first phase which lasts for 6 years, is devoted to machine checkout and physics testing. The second phase lasts for 8 years and is devoted primarily to technology testing. This report describes the technology test program development for ITER, the ancillary equipment outside the torus necessary to support the test modules, the international collaboration aspects of conducting the test program on ITER, the requirements on the machine major parameters and the R and D program required to develop the test modules for testing in ITER. 15 refs, figs and tabs

  18. Numeracy Tests For Dummies

    CERN Document Server

    Beveridge, Colin

    2012-01-01

    The easy way to get practice and excel at numeracy tests Whether you're looking for a new job, applying to certain university courses, or attempting to join the military, you're increasingly likely to face a numeracy test as part of the screening process. And the only way to prepare for a numeracy test is practise. Numeracy Tests For Dummies is an accessible one-stop guide to pass these test. Featuring expert advice, instruction, review, and plenty of practise, Numeracy Tests For Dummies will help you succeed. Numeracy Tests For Dummies contains instruction and revision on:Basic mathematical k

  19. Materials Testing and Automation

    Science.gov (United States)

    Cooper, Wayne D.; Zweigoron, Ronald B.

    1980-07-01

    The advent of automation in materials testing has been in large part responsible for recent radical changes in the materials testing field: Tests virtually impossible to perform without a computer have become more straightforward to conduct. In addition, standardized tests may be performed with enhanced efficiency and repeatability. A typical automated system is described in terms of its primary subsystems — an analog station, a digital computer, and a processor interface. The processor interface links the analog functions with the digital computer; it includes data acquisition, command function generation, and test control functions. Features of automated testing are described with emphasis on calculated variable control, control of a variable that is computed by the processor and cannot be read directly from a transducer. Three calculated variable tests are described: a yield surface probe test, a thermomechanical fatigue test, and a constant-stress-intensity range crack-growth test. Future developments are discussed.

  20. POWER CONSCIOUS TESTING

    OpenAIRE

    BONHOMME Y.; Girard, P.; LANDRAULT C.; PRAVOSSOUDOVITCH S.

    2003-01-01

    Test power relates to the power consumed during test of integrated circuits or embedded cores. Test power is now a big concern in large System-on-Chip designs. In this paper, we propose to shortly review the state-of-the-art in this domain. We first survey the recent approaches proposed for minimizing test power. Next, we propose some interesting directions for the development of new low power testing techniques by enumerating the relevant criteria that have to be satisfied.

  1. Determination of bacterial endotoxin (pyrogen) in radiopharmaceuticals by the gel clot method. Validation; Determinacao de endotoxina bacteriana (pirogenio) em radiofarmacos pelo metodo de formacao de gel. Validacao

    Energy Technology Data Exchange (ETDEWEB)

    Fukumori, Neuza Taeko Okasaki

    2008-07-01

    Before the Limulus amebocyte lysate (LAL) test, the only available means of pirogenicity testing for parenteral drugs and medical devices was the United States Pharmacopoeia (USP) rabbit pyrogen test. Especially for radiopharmaceuticals, the LAL assay is the elective way to determine bacterial endotoxin. The aim of this work was to validate the gel clot method for some radiopharmaceuticals without measurable interference. The FDA's LALTest guideline defines interference as a condition that causes a significant difference between the endpoints of a positive water control and positive product control series using a standard endotoxin. Experiments were performed in accordance to the USP bacterial endotoxins test in the {sup 131}I- m-iodobenzylguanidine; the radioisotopes Gallium-67 and Thallium-201; the lyophilized reagents DTPA, Phytate, GHA, HSA and Colloidal Tin. The Maximum Valid Dilution (MVD) was calculated for each product based upon the clinical dose of the material and a twofold serial dilution below the MVD was performed in duplicate to detect interferences. The labeled sensitivity of the used LAL reagent was 0.125 EU mL{sup -1} (Endotoxin Units per milliliter). For validation, a dilution series was performed, a twofold dilution of control standard endotoxin (CSE) from 0.5 to 0.03 EU mL{sup -1}, to confirm the labeled sensitivity of the LAL reagent being tested in sterile and non pyrogenic water, in quadruplicate. The same dilution series was performed with the CSE and the product in the 1:100 dilution factor, in three consecutive batches of each radiopharmaceutical. The products {sup 131}I-m-iodobenzylguanidine, Gallium-67, Thallium-201, DTPA, HSA and Colloidal Tin were found compatible with the LAL test at a 1:100 dilution factor. Phytate and GHA showed some interference in the gel clot test. Other techniques to determine endotoxins as the chromogenic (color development) and the turbidimetric test (turbidity development), were also assessed to get

  2. Teste de vasorreatividade pulmonar Testing pulmonary vasoreactivity

    Directory of Open Access Journals (Sweden)

    Edmundo Clarindo Oliveira

    2008-10-01

    Full Text Available A hipertensão arterial pulmonar é classificada como idiopática ou secundária (associada a colagenoses, cardiopatias, hipertensão portal, tromboembolismo pulmonar e doenças da vasculatura pulmonar. O teste de vasorreatividade pulmonar é indicado para definir a melhor opção terapêutica. Muitas drogas têm sido utilizadas para a realização desse teste, sendo o óxido nítrico inalado a melhor opção, por apresentar ação específica pulmonar e meia vida muita curta (5-10 s. O resultado desse teste identifica candidatos à cirurgia cardíaca nas cardiopatias congênitas e candidatos ao uso de antagonista de cálcio nas outras formas de hipertensão pulmonar. A realização e interpretação do teste de vasorreatividade pulmonar exigem grande responsabilidade, e erros podem levar a decisões erradas e à ocorrência de óbitos.Pulmonary arterial hypertension is classified as idiopathic or secondary (associated with collagenoses, heart disease, portal hypertension, pulmonary thromboembolism, and pulmonary vascular diseases. Pulmonary vasoreactivity should be tested in order to define the best treatment option. Of the many drugs that have been used to test pulmonary vasoreactivity, inhaled nitric oxide is the best choice, due its specific pulmonary effect and very short half-life (5-10 s. The results of this test identify candidates for heart surgery among patients with congenital heart disease and candidates for the use of calcium antagonists among patients with other forms of pulmonary hypertension. Performing and interpreting the results of such tests are a great responsibility, since mistakes can lead to incorrect treatment decisions, resulting in the death of patients.

  3. Role of test motivation in intelligence testing

    OpenAIRE

    Duckworth, Angela Lee; Quinn, Patrick D.; Lynam, Donald R.; Loeber, Rolf; Stouthamer-Loeber, Magda

    2011-01-01

    Intelligence tests are widely assumed to measure maximal intellectual performance, and predictive associations between intelligence quotient (IQ) scores and later-life outcomes are typically interpreted as unbiased estimates of the effect of intellectual ability on academic, professional, and social life outcomes. The current investigation critically examines these assumptions and finds evidence against both. First, we examined whether motivation is less than maximal on intelligence tests adm...

  4. Advanced test reactor. Testing capabilities and plans

    International Nuclear Information System (INIS)

    The Advanced Test Reactor (ATR), at the Idaho National Laboratory (INL), is one of the world's premier test reactors for providing the capability for studying the effects of intense neutron and gamma radiation on reactor materials and fuels. The physical configuration of the ATR, a 4-leaf clover shape, allows the reactor to be operated at different power levels in the corner 'lobes' to allow for different testing conditions for multiple simultaneous experiments. The combination of high flux (maximum thermal neutron fluxes of 1E15 neutrons per square centimeter per second and maximum fast [E>1.0 MeV] neutron fluxes of 5E14 neutrons per square centimeter per second) and large test volumes (up to 122 cm long and 12.7 cm diameter) provide unique testing opportunities. For future research, some ATR modifications and enhancements are currently planned. In 2007 the US Department of Energy designated the ATR as a National Scientific User Facility (NSUF) to facilitate greater access to the ATR for material testing research by a broader user community. This paper provides more details on some of the ATR capabilities, key design features, experiments, and plants for the NSUF. (author)

  5. The Danish National Tests

    DEFF Research Database (Denmark)

    Beuchert, Louise Voldby; Nandrup, Anne Brink

    In 2010, the Danish National Tests were implemented in the public compulsory schools as a mean of evaluating the performance of the public school system. The extensive test program consists of ten mandatory tests in six subjects in grades 2 through 8. In this paper, we share our insights from...... working with the first four rounds of the test data. We provide a brief introduction to adaptive testing, the available data and general data issues including missing data, test participation and data transformations. Additionally, we construct a standardized measure of the raw pupil ability estimate...... within each test and argue that this is often a more feasible measure for data analyses compared to the transformed test score presented to pupils and teachers. We provide the reader with preliminary analyses of the relation between pupils' national test results and a wide range of pupil background...

  6. Nondestructive testing of materials

    International Nuclear Information System (INIS)

    NUKEM has transferred know-how from reactor technology to materials testing. The high and to a large extent new quality standards in the nuclear industry necessitate reliable measuring and testing equipment of the highest precision. Many of the tasks presented to us could not be solved with the equipment available on the market, for which reason we have developed our own measuring, testing and control systems. We have therefore acquired considerable experience in dealing with specific measuring, testing and control tasks and can handle even out-of-the-way problems that are submitted to us from a wide variety of fields. Our mechanical systems for the checking of close-tolerance gaps, the automatic determination of pellet dimensions and the measurement of absolute lengths and absolute velocities are in use in many different industrial fields. We have succeeded in solving unusual testing and sorting problems with the aid of automated surface testing equipment working on optical principles. Our main activities in the field of non-destructive testing have been concentrated on ultrasonic and eddy current testing and, of late, acoustic emission analysis. NUKEM ultrasonic systems are notable for their high defect detection rate and testing accuracy, combined with high testing speed. The equipment we supply includes ultrasonic rotary systems for the production testing of quality tubes, ultrasonic immersion systems for the final testing of reactor cladding tubes, weld testing equipment, and test equipment for the bonds in multi-layer plates. (orig./RW)

  7. Testing (HIV). Quick test receives Singapore approval.

    Science.gov (United States)

    1996-04-22

    Hema-Strip HIV 1/2 is a rapid HIV antibody immunoassay developed by Saliva Diagnostic Systems, Inc. (SDS) which can be used by anyone who can read the product insert. The test kit is comprised of a small lancet for a finger stick, a cylindrical tube with a capillary tip and a SDS diagnostic strip inside, and a vial of buffer. Once blood is drawn by the lancet, the capillary tip is placed upon the blood droplet and the blood is automatically drawn into the tube. The tube is then inserted tip first into the vial of buffer. The buffer and blood migrate over the diagnostic strip inside, yielding stable results within 15 minutes. Studies have found Hema-Strip HIV 1/2 to have a sensitivity and specificity greater than 99.4%, as accurate as most conventional HIV tests which require the use of laboratory equipment and trained staff, and possibly hours to produce results. Moreover, the test kit requires neither refrigeration nor special storage. Hema-Strip HIV 1/2 has received a certificate of free sale from the Ministry of Health in Singapore and is now being submitted for regulatory approval in Brazil, China, Russia, India, Malaysia, Thailand, and the UK. SDS products in production include Sero-Strip HIV 1/2, a rapid serum-based HIV antibody test; Omni-SAL, a saliva collector which is the principal sample collection device used by British insurance companies for HIV testing with other confirmatory tests; Omni-Swab, a serrated swab which collects body fluids or cells; Saliva-Sampler, a saliva collection device used for general testing purposes; and Saliva Check, a test which checks the composition of saliva samples. SDS is in the final stages of developing Saliva-Strip HIV-1/2, a rapid saliva-based HIV antibody test. The company also intends to complete development in 1996 of a rapid blood-based antibody test for the Helicobacter pylori bacteria, a pathogen linked to 80% of peptic ulcers and gastric cancers. PMID:12290908

  8. Comparative Test Case Specification

    DEFF Research Database (Denmark)

    Kalyanova, Olena; Heiselberg, Per

     This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....... This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....

  9. NIST Statistical Test Suite

    OpenAIRE

    Paul, Rourab; Dey, Hemanta; Chakrabrti, Amlan; Ghosh, Ranjan

    2016-01-01

    The NIST Statistical Test Suite has 15 tests. The principal strategy of the NIST Statistical Test Suite is to judge statistical randomness property of random bit generating algorithms. Based on 300 to 500 different keys, the algorithm generates a series of even number of different long random sequences of n bits, n varying between 13 and 15 lacs, each of which is tested by the 15 tests. Each test has a specific statistic parameter for bit sequences under the assumption of randomness and calcu...

  10. Materials Test Branch

    Science.gov (United States)

    Gordon, Gail

    2012-01-01

    The Materials Test Branch resides at Marshall Space Flight Center's Materials and Processing laboratory and has a long history of supporting NASA programs from Mercury to the recently retired Space Shuttle. The Materials Test Branch supports its customers by supplying materials testing expertise in a wide range of applications. The Materials Test Branch is divided into three Teams, The Chemistry Team, The Tribology Team and the Mechanical Test Team. Our mission and goal is to provide world-class engineering excellence in materials testing with a special emphasis on customer service.

  11. On Reading Test

    Institute of Scientific and Technical Information of China (English)

    孙健

    2005-01-01

    There has been a long discussion over the construct validity of reading tests. In china's reading tests, multiple choice is the main test method in view of the4 long controversy over the validity of multiple choice, construct validation is called for to empirically test the hypothesized relationships between test scores and abilities. The national CET committee conducted a comprehensive validation study. As part of the project, the specialists studied the reading comprehension test's validity by qualitative means, namely "introspective verbal reports". The analysis revealed that an overwhelming majority of the questions items were handled through "expected reading operations".

  12. Evaluation of a von Willebrand factor three test panel and chemiluminescent-based assay system for identification of, and therapy monitoring in, von Willebrand disease.

    Science.gov (United States)

    Favaloro, Emmanuel J; Mohammed, Soma

    2016-05-01

    von Willebrand disease (VWD) is reportedly the most common bleeding disorder and arises from deficiency and/or defects of von Willebrand factor (VWF). Laboratory diagnosis and typing of VWD has important management implications and requires a wide range of tests, including VWF antigen (VWF:Ag) and various activities, involving differential identification of qualitative vs quantitative VWF defects. We have assessed a new hemostasis instrument, the chemiluminescent assay based ACL AcuStar™, and an associated HemosIL AcuStar three test panel comprising VWF:Ag, VWF ristocetin cofactor (VWF:RCo) and VWF collagen binding (VWF:CB) (Instrumentation Laboratory, Bedford, Ma. USA) for ability to identify VWD, to help provisionally type VWD, and for potential use in therapy monitoring. This test system was compared to previously evaluated and validated test systems including VWF:RCo on CS-5100 and BCS analyzers, the new Siemens INNOVANCE assay (VWF Ac) on CS-5100, and VWF:Ag and VWF:CB assays performed by automated ELISA. We employed a large total sample test set (n=535) comprising plasma and platelet-lysate samples from individuals with and without VWD, some on treatment, normal plasmas, and normal and pathological controls. We also evaluated desmopressin (DDAVP) responsiveness, plus differential sensitivity to reduction in high molecular weight (HMW) VWF. The chemiluminescent test panel (VWF:Ag, VWF:RCo, VWF:CB) showed good comparability to similar assays performed by alternate methods, and broadly similar data for identification of VWD, provisional VWD type identification, DDAVP and VWD therapy, and HMW VWF sensitivity, although some notable differences were evident. The chemiluminescent system showed best low level VWF sensitivity, and lowest inter-assay variability, compared to all other systems. In conclusion, we have validated theACL AcuStar and the chemiluminescent HemosIL AcuStar VWF test panel for use in VWD diagnostics, and have identified some favorable

  13. Gas Test Loop Booster Fuel Hydraulic Testing

    Energy Technology Data Exchange (ETDEWEB)

    Gas Test Loop Hydraulic Testing Staff

    2006-09-01

    The Gas Test Loop (GTL) project is for the design of an adaptation to the Advanced Test Reactor (ATR) to create a fast-flux test space where fuels and materials for advanced reactor concepts can undergo irradiation testing. Incident to that design, it was found necessary to make use of special booster fuel to enhance the neutron flux in the reactor lobe in which the Gas Test Loop will be installed. Because the booster fuel is of a different composition and configuration from standard ATR fuel, it is necessary to qualify the booster fuel for use in the ATR. Part of that qualification is the determination that required thermal hydraulic criteria will be met under routine operation and under selected accident scenarios. The Hydraulic Testing task in the GTL project facilitates that determination by measuring flow coefficients (pressure drops) over various regions of the booster fuel over a range of primary coolant flow rates. A high-fidelity model of the NW lobe of the ATR with associated flow baffle, in-pile-tube, and below-core flow channels was designed, constructed and located in the Idaho State University Thermal Fluids Laboratory. A circulation loop was designed and constructed by the university to provide reactor-relevant water flow rates to the test system. Models of the four booster fuel elements required for GTL operation were fabricated from aluminum (no uranium or means of heating) and placed in the flow channel. One of these was instrumented with Pitot tubes to measure flow velocities in the channels between the three booster fuel plates and between the innermost and outermost plates and the side walls of the flow annulus. Flow coefficients in the range of 4 to 6.5 were determined from the measurements made for the upper and middle parts of the booster fuel elements. The flow coefficient for the lower end of the booster fuel and the sub-core flow channel was lower at 2.3.

  14. USA Hire Testing Platform

    Data.gov (United States)

    Office of Personnel Management — The USA Hire Testing Platform delivers tests used in hiring for positions in the Federal Government. To safeguard the integrity of the hiring processes and ensure...

  15. Quadruple screen test

    Science.gov (United States)

    ... screen; Multiple marker screening; AFP plus; Triple screen test; AFP maternal; MSAFP; 4-marker screen ... This test is most often done between the 15th and 22nd weeks of the pregnancy. It is most accurate ...

  16. Directory of Medical Tests

    Science.gov (United States)

    ... KidsHealth in the Classroom What Other Parents Are Reading Upsetting News Reports? What to Say Vaccines: Which ... variety of illnesses or signs of infection. Blood chemistry test. Basic blood chemistry tests measure the levels ...

  17. PHYSICAL SIMULATION & TEST

    Data.gov (United States)

    Federal Laboratory Consortium — Crew Station/Turret Motion Based Simulator (CS/TMBS) Test station simulates operational scenarios The CS/TMBS is a high-capacity six-degrees-of-freedom test device....

  18. The non destructive testings

    International Nuclear Information System (INIS)

    In this paper the author after having indicated the methods used in nondestructive testing presents 4 of these methods - X-ray and gamma radiography - ultrasonic testing - eddy current - acoustic emission

  19. Test Your Asthma Knowledge

    Science.gov (United States)

    ... Issue Past Issues Special Section Test Your Asthma Knowledge Past Issues / Fall 2007 Table of Contents For ... Asthma: A Chance to Heal / Test Your Asthma Knowledge Fall 2007 Issue: Volume 2 Number 4 Page ...

  20. Get Your Eyes Tested

    Science.gov (United States)

    ... Print This Topic En español Get Your Eyes Tested Browse Sections The Basics Overview Eye Exams Vision ... The Basics The Basics: Overview Have your eyes tested (examined) regularly to help find problems early, when ...

  1. Screening Tests and Vaccines

    Science.gov (United States)

    ... Contact Us Text size | Print | Screening Tests and Vaccines This information in Spanish ( en español ) Getting important screening tests and vaccines can save your life. Check this section of ...

  2. Understanding Laboratory Tests

    Science.gov (United States)

    ... and Drug Administration (FDA) regulates the development and marketing of all laboratory tests that use test kits ... at the National Institutes of Health FOLLOW US Facebook Twitter Instagram YouTube Google+ LinkedIn GovDelivery RSS CONTACT ...

  3. CSD skin test

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003385.htm CSD skin test To use the sharing features on this page, please enable JavaScript. The cat scratch disease (CSD) skin test was once used to help ...

  4. Campylobacter serology test

    Science.gov (United States)

    ... a blood test to look for antibodies to bacteria called campylobacter. ... An abnormal (positive) result means that antibodies against ... with the bacteria. Tests are often repeated during the course of ...

  5. Vitamin A blood test

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003570.htm Vitamin A blood test To use the sharing features on this page, please enable JavaScript. The vitamin A test measures the level of vitamin A ...

  6. HPV and HPV Testing

    Science.gov (United States)

    ... Close + - Text Size HPV and HPV Testing Human Papilloma Virus (HPV) What are viruses? Viruses are very ... does it mean? If you have cervical human papilloma virus (HPV) infection and an abnormal Pap test ...

  7. Variable Attitude Test Stand

    Data.gov (United States)

    Federal Laboratory Consortium — The Variable Attitude Test Stand designed and built for testing of the V-22 tilt rotor aircraft propulsion system, is used to evaluate the effect of aircraft flight...

  8. HIV Antibody Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? HIV Antibody and HIV Antigen (p24) Share this page: Was this page helpful? Also known as: HIV Screening Tests; AIDS Test; AIDS Screen; HIV Serology; ...

  9. Integrated Usability Testing

    Directory of Open Access Journals (Sweden)

    Andrei Ternauciuc

    2015-11-01

    Full Text Available It is essential to regularly test the usability of a learning management system, in order to ensure a fast adoption by new users and rapidly shift the focus from the platform to the content and the learning experience. Quantitative testing yields the most reliable results due to the large number of data points acquired, but lacks the in-depth analysis of the qualitative research from a controlled testing setup. We are proposing in this paper an integrated usability testing tool, which can replace a certain type of laboratory testing, where the users’ actions on the real platform are measured and analyzed. We conducted tests with the tool and compared the results with a small scale laboratory test using the same scenarios. The results seem to confirm the proposed tool as a viable alternative to the laboratory test.

  10. Environmental Test Facility (ETF)

    Data.gov (United States)

    Federal Laboratory Consortium — The Environmental Test Facility (ETF) provides non-isolated shock testing for stand-alone equipment and full size cabinets under MIL-S-901D specifications. The ETF...

  11. Quadruple screen test

    Science.gov (United States)

    ... screen; Multiple marker screening; AFP plus; Triple screen test; AFP maternal; MSAFP; 4-marker screen; Down syndrome - ... This test is most often done between the 15th and 22nd weeks of the pregnancy. It is most accurate ...

  12. Hepatitis B Test

    Science.gov (United States)

    ... limited. Home Visit Global Sites Search Help? Hepatitis B Testing Share this page: Was this page helpful? Also known as: HBV Tests; Hep B; anti-HBs; Hepatitis B Surface Antibody; HBsAg; Hepatitis ...

  13. Lyme Disease Tests

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Lyme Disease Tests Share this page: Was this page helpful? ... else I should know? How is it used? Lyme disease tests are used to determine if a person ...

  14. Atlantic Test Range (ATR)

    Data.gov (United States)

    Federal Laboratory Consortium — ATR controls fully-instrumented and integrated test ranges that provide full-service support for cradle-to-grave testing. Airspace and surface target areas are used...

  15. Rapid Lead Screening Test

    Science.gov (United States)

    ... Medical Procedures In Vitro Diagnostics Lab Tests Rapid Lead Screening Test Share Tweet Linkedin Pin it More ... reducing the need for a follow-up visit. Lead Risk Links Centers for Disease Control and Prevention ( ...

  16. Urine specific gravity test

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003587.htm Urine specific gravity test To use the sharing features on this page, please enable JavaScript. Urine specific gravity is a laboratory test that shows the concentration ...

  17. Testing for HIV

    Science.gov (United States)

    ... Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Vaccines, Blood & Biologics Home Vaccines, Blood & Biologics Safety & Availability (Biologics) HIV Home Test Kits Testing for HIV Share Tweet Linkedin Pin it More ...

  18. Insensitive Munitions Testing

    Data.gov (United States)

    Federal Laboratory Consortium — Insensitive Munitions Testing at RTC is conducted (IAW MILSTD-2105) at Test Area 4. Our engineers and technicians obtain data for hazards classification and safety...

  19. HPV DNA test

    Science.gov (United States)

    The HPV DNA test is used to check for high-risk HPV infection in women. HPV infection around the genitals is ... warts spread when you have sex. The HPV-DNA test is generally not recommended for detecting low- ...

  20. Learning Android application testing

    CERN Document Server

    Blundell, Paul

    2015-01-01

    If you are an Android developer looking to test your applications or optimize your application development process, then this book is for you. No previous experience in application testing is required.

  1. Learning Python testing

    CERN Document Server

    Arbuckle, Daniel

    2014-01-01

    This book is ideal if you want to learn about the testing disciplines and automated testing tools from a hands-on, conversational guide. You should already know Python and be comfortable with Python 3.

  2. Growth hormone test

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003706.htm Growth hormone test To use the sharing features on this page, please enable JavaScript. The growth hormone test measures the amount of growth hormone in ...

  3. Cholesterol testing and results

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/patientinstructions/000386.htm Cholesterol testing and results To use the sharing features ... can tell you what your goal should be. Cholesterol Tests Some cholesterol is considered good and some ...

  4. Lung diffusion testing

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003854.htm Lung diffusion testing To use the sharing features on this page, please enable JavaScript. Lung diffusion testing measures how well the lungs exchange ...

  5. Ballistic Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Ballistic Test Facility is comprised of two outdoor and one indoor test ranges, which are all instrumented for data acquisition and analysis. Full-size aircraft...

  6. Static Loads Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — FUNCTION: Provides the capability to perform large-scale structural loads testing on spacecraft and other structures. Results from these tests can be used to verify...

  7. Tests in Print II: An Index to Tests, Test Reviews, and the Literature on Specific Tests.

    Science.gov (United States)

    Buros, Oscar K., Ed.

    Tests in Print II is a comprehensive, annotated bibliography of all in-print tests published as separates for use with English-speaking subjects. The 1,155 two-column pages list 2,467 tests in print as of early 1974; 16,574 references through 1971 on specific tests; a reprinting of the 1974 APA-AERA-NCME Standards for Educational andPsychological…

  8. Blade Testing Trends (Presentation)

    Energy Technology Data Exchange (ETDEWEB)

    Desmond, M.

    2014-08-01

    As an invited guest speaker, Michael Desmond presented on NREL's NWTC structural testing methods and capabilities at the 2014 Sandia Blade Workshop held on August 26-28, 2014 in Albuquerque, NM. Although dynamometer and field testing capabilities were mentioned, the presentation focused primarily on wind turbine blade testing, including descriptions and capabilities for accredited certification testing, historical methodology and technology deployment, and current research and development activities.

  9. Tubing weld cracking test

    International Nuclear Information System (INIS)

    A tubing weld cracking (TWC) test was developed for applications involving advanced austenitic alloys (such as modified 800H and 310HCbN). Compared to the Finger hot cracking test, the TWC test shows an enhanced ability to evaluate the crack sensitivity of tubing materials. The TWC test can evaluate the cracking tendency of base as well as filter materials. Thus, it is a useful tool for tubing suppliers, filler metal producers and fabricators

  10. Sperm function test

    OpenAIRE

    Pankaj Talwar; Suryakant Hayatnagarkar

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give ins...

  11. Uniform Test Assembly

    Science.gov (United States)

    Belov, Dmitry I.

    2008-01-01

    In educational practice, a test assembly problem is formulated as a system of inequalities induced by test specifications. Each solution to the system is a test, represented by a 0-1 vector, where each element corresponds to an item included (1) or not included (0) into the test. Therefore, the size of a 0-1 vector equals the number of items "n"…

  12. Security Testing: A Survey

    OpenAIRE

    Felderer, M.; Büchlein, M.; Johns, M; Brucker, A.D.; Breu, R.; Pretschner, A.

    2015-01-01

    Identifying vulnerabilities and ensuring security functionality by security testing is a widely applied measure to evaluate and improve the security of software. Due to the openness of modern software-based systems, applying appropriate security testing techniques is of growing importance and essential to perform effective and efficient security testing. Therefore, an overview of actual security testing techniques is of high value both for researchers to evaluate and refine the techniques and...

  13. Diagnosing the Diagnostic Test

    Science.gov (United States)

    Popham, W. James

    2009-01-01

    Scads of pseudodiagnostic tests are peddled by commercial vendors who recognize that desperate educators will do almost anything to dodge an impending accountability cataclysm. And this "almost anything" includes buying tests that promise to help a teacher raise test scores--even if they don't. Accordingly, today's educators need to be aware of…

  14. Blood Test: Glucose

    Science.gov (United States)

    ... Things to Know About Zika & Pregnancy Blood Test: Glucose KidsHealth > For Parents > Blood Test: Glucose Print A A A Text Size What's in ... de sangre: glucosa What It Is A blood glucose test measures the amount of glucose (the main ...

  15. Parvovirus B19 Test

    Science.gov (United States)

    ... be done. Viral detection If a parvovirus B19 DNA test is positive, then the person is currently infected ... The PCR assay is used to detect viral DNA and is the optimal method for detecting chronic ... a reticulocyte test may be performed along with parvovirus B19 testing ...

  16. Automated Web Applications Testing

    Directory of Open Access Journals (Sweden)

    Alexandru Dan CĂPRIŢĂ

    2009-01-01

    Full Text Available Unit tests are a vital part of several software development practicesand processes such as Test-First Programming, Extreme Programming andTest-Driven Development. This article shortly presents the software quality andtesting concepts as well as an introduction to an automated unit testingframework for PHP web based applications.

  17. About Instruction Sequence Testing

    NARCIS (Netherlands)

    J.A. Bergstra

    2012-01-01

    Software testing is presented as a so-called theme within which different authors and groups have defined different subjects each of these subjects having a different focus on testing. A uniform concept of software testing is non-existent and the space of possible coherent perspectives on software t

  18. Test Reviewing in Germany

    Science.gov (United States)

    Hagemeister, Carmen; Kersting, Martin; Stemmler, Gerhard

    2012-01-01

    In 2006, a (new) German standard for test reviewing was passed (Testkuratorium, 2006). There was already a European standard in place (European Federation of Psychologists' Associations, 2008). This article presents the German standard for test reviewing and explains how the German test review system was derived from demands in the German standard…

  19. Testing for central symmetry

    NARCIS (Netherlands)

    Einmahl, John; Gan, Zhuojiong

    2016-01-01

    Omnibus tests for central symmetry of a bivariate probability distribution are proposed. The test statistics compare empirical measures of opposite regions. Under rather weak conditions, we establish the asymptotic distribution of the test statistics under the null hypothesis; it follows that they a

  20. Aldosterone and Renin Test

    Science.gov (United States)

    ... are used to treat high blood pressure. Stress, exercise, and pregnancy can also affect the test results. Coffee, tea or cola can affect the 24-hour urine sample test. Your doctor will tell you if you should change the amount of ... routine before aldosterone testing. Licorice may mimic aldosterone ...

  1. Testing Backbone.js

    CERN Document Server

    Roemer, Ryan

    2013-01-01

    This book is packed with the step by step tutorial and instructions in recipe format helping you setup test infrastructure and gradually advance your skills to plan, develop, and test your backbone applications.If you are a JavaScript developer looking for recipes to create and implement test support for your backbone application, then this book is ideal for you.

  2. Testing Technology, June 1992

    Energy Technology Data Exchange (ETDEWEB)

    Getsch, B; Floyd, H L; Parrott, L; Van Arsdall, A

    1992-01-01

    This report highlights the following topics: Photon Correlation Spectroscopy--a new application in jet fuel analysis, Testing news in brief; Solar test facility supports space station research; Shock isolation technique developed for piezoresistive accelerometer; High-speed photography captures Distant Image measurements; and, Radiation effects test revised for CMOS electronics.

  3. Follow-Up Testing

    Science.gov (United States)

    Follow-up Testing Follow-up testing is conducted to ensure that antibody levels are returning to normal, indicating that the intestine is healing on the ... has entered the diet. How often should follow-up testing occur? New celiacs should receive follow-up ...

  4. The Tyranny of Testing.

    Science.gov (United States)

    Sternberg, Robert J.

    1989-01-01

    Standardized tests which measure a narrow span of intelligence unfairly penalize students whose strengths don't fall within that range. Three kinds of intelligence (analytical, creative, practical) are discussed. Sternberg's Triarchic Abilities Test, currently being test-piloted, assesses all three aspects of intelligence in contrast to current…

  5. Toroid magnet test facility

    CERN Multimedia

    2002-01-01

    Because of its exceptional size, it was not feasible to assemble and test the Barrel Toroid - made of eight coils - as an integrated toroid on the surface, prior to its final installation underground in LHC interaction point 1. It was therefore decided to test these eight coils individually in a dedicated test facility.

  6. 100 statistical tests

    CERN Document Server

    Kanji, Gopal K

    2006-01-01

    This expanded and updated Third Edition of Gopal K. Kanji's best-selling resource on statistical tests covers all the most commonly used tests with information on how to calculate and interpret results with simple datasets. Each entry begins with a short summary statement about the test's purpose, and contains details of the test objective, the limitations (or assumptions) involved, a brief outline of the method, a worked example, and the numerical calculation. 100 Statistical Tests, Third Edition is the one indispensable guide for users of statistical materials and consumers of statistical information at all levels and across all disciplines.

  7. Le test detourne (The Test Turned Around).

    Science.gov (United States)

    Valderrania, Isabelle

    1994-01-01

    It is proposed that the multiple-choice test can be useful in helping language students understand the meaning of text better without overwhelming them. Techniques for drafting effective questions are discussed, and a sample list of questions is presented. (MSE)

  8. Nondestructive testing of weldings

    International Nuclear Information System (INIS)

    Today the nondestructive testing of materials with transverse weldings would be often neglect. Because the production of this parts will get more expensive it is interesting to test the parts eg. cans with ultrasound. Within this research program we developed a testing device for automatically testing of materials with transverse weldings, e.g. tubes. Functional characteristics: after putting in the test tube in the testing device the transducer is adjusted automatically to the best adjustment. This takes place with three step motors. The mechanic of adjustments gets its information from an optimal computer. The optimal computer processes the signals, which it gets from the automatic evaluator during the adjustment and stores the optimal adjustment parameters. With an additional equipment on can identify the exact source of error, if the testing device turns out and the repairing of the part can take place very fast. With an appropiate mechanical science the devide is useable in a production line with high flow rate. (orig.)

  9. Pendulum detector testing device

    International Nuclear Information System (INIS)

    A detector testing device is described which provides consistent, cost-effective, repeatable results. The testing device is primarily constructed of PVC plastic and other non-metallic materials. Sensitivity of a walk-through detector system can be checked by: (1) providing a standard test object simulating the mass, size and material content of a weapon or other contraband, (2) suspending the test object in successive positions, such as head, waist and ankle levels, simulating where the contraband might be concealed on a person walking through the detector system; and (3) swinging the suspended object through each of the positions, while operating the detector system and observing its response. The test object is retained in a holder in which the orientation of the test device or target can be readily changed, to properly complete the testing requirements. 5 figs

  10. Control of dialysis water by microbial contamination and bacterial endotoxin tests

    Directory of Open Access Journals (Sweden)

    Fesem BAŞARI

    2012-01-01

    Full Text Available Objective: The aim of this study was to evaluate the microbial contamination and bacterial endotoxin levels in dialysis water samples sent to the Water and Food Microbiology Laboratory of the Adana Hygiene Institute Refik Saydam Hygiene Center, and to assess factors that might influence the results of these tests.Method: Two hundred fourty five dialysis water samples sent to the laboratory between January 2009 and December 2010 were tested; all of them for microbial contamination and 198 of them for bacterial endotoxin levels. Microbial contamination was evaluated by using the Plate Count Agar (PCA method while the Limulus Amoebocyte Lysate (LAL assay was used for the detection of bacterial endotoxins. The results were evaluated according to the criteria of the “Directives on the Water Purification Systems” and “the European Pharmacopoeia”. To compare the annual differences, the chi-square test was used for statistical evaluation and results with p0.05in 2010. On the other hand, in 26.7% of the dialysiswater samples the levels of bacterial endotoxinswas higher than 0.25 İU/ml in 2009, this percentagedropped to 16.5% in 2010 (p>0.05. During 2009-2010,245 microbial contamination tests were performedand in 20 (8.2% of the samples the TAMC values werehigher than 100 CFU/ml, while in 43 (21.7% out of198 of the samples the bacterial endotoxin levelswere higher than 0.25 İU/ml. Out of 20 in 43 samples,a bacterial endotoxin levels were >0.25 İU/ml werealso found as microbiologically contaminated ( >100CFU/ml, while in remaining 23 samples the level was<100 CFU/ml. There were no statistically significantdifferences in the percentages of positives found in2009 and 2010.Conclusion: The results of our study indicatethat the detection of endotoxins is a complementaryfactor to microbial contamination in the bacteriologicalmonitoring of dialysis waters. We recommend thatlevel of endotoxin should be taken into accountwhen the bacterial contamination

  11. Formulation and characterization of a plasma sterilized, pharmaceutical grade chitosan powder.

    Science.gov (United States)

    Crofton, Andrew R; Hudson, Samuel M; Howard, Kristy; Pender, Tyler; Abdelgawad, Abdelrahman; Wolski, Daniel; Kirsch, Wolff M

    2016-08-01

    Chitosan has great potential as a pharmaceutical excipient. In this study, chitosan flake was micronized using cryo-ball and cryo-jet milling and subsequently sterilized with nitrogen plasma. Micronized chitosan was characterized by laser diffraction, scanning electron microscopy (SEM), conductometric titration, viscometry, loss on drying, FTIR, and limulus amebocyte lysate (LAL) assays. Cryo-jet milling produced mean particle size of 16.05μm, 44% smaller than cryo-ball milling. Cryomilled chitosan demonstrated increased hygroscopicity, but reduced molecular weight and degree of deacetylation (DD). SEM imaging showed highly irregular shapes. FTIR showed changes consistent with reduced DD and an unexplained shift at 1100cm(-1). Plasma treated chitosan was sterile with plasma and plasma treatment. Plasma treatment decreased the reduced viscosity of chitosan flake and powder, with a greater effect on powder. In conclusion, pharmaceutical grade, sterile chitosan powder was produced with cryo-jet milling and plasma sterilization. PMID:27112892

  12. Concentration, physical state, and purity of bacterial endotoxin affect its detoxification by ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Csako, G.; Tsai, C.M.; Hochstein, H.D.; Elin, R.J.

    1986-11-01

    Increasing concentrations of a highly purified bacterial lipopolysaccharide preparation, the U.S. Reference Standard Endotoxin, were exposed to increasing doses of ionizing radiation from a 60Co source. At identical radiation doses both the structural change and Limulus amebocyte lysate (LAL) reactivity were progressively smaller with increasing concentrations of the lipopolysaccharide in an aqueous medium. Under the experimental conditions used, there was a linear relationship between the endotoxin concentration and radiation dose for the structural changes. In contrast to endotoxin in aqueous medium, endotoxin irradiated in its dry state showed no decrease in LAL reactivity and rabbit pyrogenicity. Endotoxin exposed to radiation in water in the presence of albumin showed a much smaller decrease in LAL and pyrogenic activities than expected. The results show that the concentration, physical state, and purity of endotoxin influence its structural and functional alteration by ionizing radiation.

  13. Measurement of endotoxin in Pseudo-epitheliomatous hyperplastic granuloma-like lesions on superficial Ⅱdegree burn wounds and its pathogenic mechanism%烧伤后假上皮瘤肉芽肿样病变内毒素测量及其致病机制

    Institute of Scientific and Technical Information of China (English)

    王波涛; 李新华; 陈璧; 董茂龙; 贾赤宇; 朱雄翔; 胡大海; 徐明达

    2003-01-01

    AIM: To investigate the content of endotoxin in Pseudo-epitheliomatous hyperplastic granuloma-like lesions and its pathogenic mechanism. METHODS: Endotoxin concentrations in the tissue were measured with chromogenic limulus amebocyte lysate ( LAL), which was modified by perchloric acid (PCA) pretreatment for samples. To observe the effect of supernate from peripheral blood mononuclear cells by single or repeated endotoxin stimulation, on proliferation of cultured fibroblasts peripheral blood mononuclear cells. RESULTS: Endotoxin was detected in the injured skin but not detected in the normal skin. Supernate from rom peripheral blood mononuclear cells by single endotoxin stimulation has stronger effect on proliferation of fibroblats than repeated stimulation. CONCLUSION: Pseudo-epitheliomatous hyperplastic granuloma-like lesions were resulted from the bacteria in the injured shin, the bacteria produced endotoxin which stimulated the local inflammatory cells to produce cytokines, and the cytokines led to the overgrowth of repairing cells in the skin.

  14. Nemesis Autonomous Test System

    Science.gov (United States)

    Barltrop, Kevin J.; Lee, Cin-Young; Horvath, Gregory A,; Clement, Bradley J.

    2012-01-01

    A generalized framework has been developed for systems validation that can be applied to both traditional and autonomous systems. The framework consists of an automated test case generation and execution system called Nemesis that rapidly and thoroughly identifies flaws or vulnerabilities within a system. By applying genetic optimization and goal-seeking algorithms on the test equipment side, a "war game" is conducted between a system and its complementary nemesis. The end result of the war games is a collection of scenarios that reveals any undesirable behaviors of the system under test. The software provides a reusable framework to evolve test scenarios using genetic algorithms using an operation model of the system under test. It can automatically generate and execute test cases that reveal flaws in behaviorally complex systems. Genetic algorithms focus the exploration of tests on the set of test cases that most effectively reveals the flaws and vulnerabilities of the system under test. It leverages advances in state- and model-based engineering, which are essential in defining the behavior of autonomous systems. It also uses goal networks to describe test scenarios.

  15. Testing On Computers

    Directory of Open Access Journals (Sweden)

    Michael Russell

    1999-06-01

    Full Text Available Russell and Haney (1997 reported that open-ended test items administered on paper may underestimate the achievement of students accustomed to writing on computers. This study builds on Russell and Haney's work by examining the effect of taking open-ended tests on computers and on paper for students with different levels of computer skill. Using items from the Massachusetts Comprehensive Assessment System (MCAS and the National Assessment of Educational Progress (NAEP, this study focuses on language arts, science and math tests administered to eighth grade students. In addition, information on students' prior computer use and keyboarding speed was collected. Unlike the previous study that found large effects for open-ended writing and science items, this study reports mixed results. For the science test, performance on computers had a positive group effect. For the two language arts tests, an overall group effect was not found. However, for students whose keyboarding speed is at least 0.5 or one-half of a standard deviation above the mean, performing the language arts test on computer had a moderate positive effect. Conversely, for students whose keyboarding speed was 0.5 standard deviations below the mean, performing the tests on computer had a substantial negative effect. For the math test, performing the test on computer had an overall negative effect, but this effect became less pronounced as keyboarding speed increased. Implications are discussed in terms of testing policies and future research.

  16. Sperm function test.

    Science.gov (United States)

    Talwar, Pankaj; Hayatnagarkar, Suryakant

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation). They should be protected from the bad effects of pus cells and reactive oxygen species (ROS) (leukocyte detection test, ROS estimation). Their number should be in sufficient in terms of (count), structure normal to be able to fertilize eggs (semen morphology). Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test), should have good mitochondrial function to be able to provide energy (mitochondrial activity index test). They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test). Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test) to the oocyte during fertilization. PMID:26157295

  17. Sperm function test

    Directory of Open Access Journals (Sweden)

    Pankaj Talwar

    2015-01-01

    Full Text Available With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation. They should be protected from the bad effects of pus cells and reactive oxygen species (ROS (leukocyte detection test, ROS estimation. Their number should be in sufficient in terms of (count, structure normal to be able to fertilize eggs (semen morphology. Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test, should have good mitochondrial function to be able to provide energy (mitochondrial activity index test. They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test. Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test to the oocyte during fertilization.

  18. GIRAFFE test results summary

    Energy Technology Data Exchange (ETDEWEB)

    Yokobori, S.; Arai, K.; Oikawa, H. [Toshiba Corporation, Kawasaki (Japan)

    1996-03-01

    A passive system can provide engineered safety features enhancing safety system reliability and plant simplicity. Toshiba has conducted the test Program to demonstrate the feasibility of the SBWR passive safety system using a full-height, integral system test facility GIRAFFE. The test facility GIRAFFE models the SBWR in full height to correctly present the gravity driving head forces with a 1/400 volume scale. The GIRAFFE test Program includes the certification tests of the passive containment cooling system (PCCS) to remove the post-accident decay heat and the gravity driven cooling system (GDCS) to replenish the reactor coolant inventory during a LOCA. The test results have confirmed the PCCS and GDCS design and in addition, have demonstrated the operation of the pCCS with the presence of a lighter-than-steam noncondensable as well as with the presence of a heavier-than-steam, noncondensable. The GIRAFFE test Program has also provided the database to qualify a best estimate thermal-hydraulic computer code TRAC. The post test analysis results have shown that TRAC can accurately predict the PCCS heat removal Performance and the containment pressure response to a LOCA. This paper summarizes the GIRAFFE test results to investigate post-LOCA PCCS heat removal performance and post-test analysis using TRAC.

  19. Standard Test Method for Sandwich Corrosion Test

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2009-01-01

    1.1 This test method defines the procedure for evaluating the corrosivity of aircraft maintenance chemicals, when present between faying surfaces (sandwich) of aluminum alloys commonly used for aircraft structures. This test method is intended to be used in the qualification and approval of compounds employed in aircraft maintenance operations. 1.2 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information. 1.3 This standard may involve hazardous materials, operations, and equipment. This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements appear in Section 9.

  20. Test Results for CSTR Test 4

    Energy Technology Data Exchange (ETDEWEB)

    Lee, D.D.

    2001-05-31

    One of the 3 technologies currently being developed for the Savannah River Salt Waste Processing Program is the Small-Tank Tetraphenylborate Process (STTP). This process uses sodium tetraphenylborate to precipitate and remove radioactive Cs from the waste and monosodium titanate to sorb and remove radioactive Sr and actinides. ORNL is demonstrating this process at the 1:4000 scale using a 20-liter continuous-flow stirred tank reactor (CSTR) system. The primary goal of Test 4 was to verify that the STTP process could achieve and maintain the necessary Cs decontamination while TPB was actively decomposing. Even with TPB being decomposed by the off-normal conditions of this test, the decontaimination factor for {sup 137}Cs obtained for the filtrate from the Slurry Concentrating Tank ranged from 47,000 to 646,000, exceeding the WAC standard.

  1. Automated Test Stand for HEV Capacitor Testing

    Energy Technology Data Exchange (ETDEWEB)

    Seiber, Larry Eugene [ORNL; Armstrong, Gary [Maverick Systems

    2007-01-01

    As capacitor manufacturers race to meet the needs of the hybrid-electric vehicle (HEV) of the future, many trade-offs at the system level as well as the component level must be considered. Even though the ultra-capacitor has the spot light for recent research and development (R&D) for HEVs, the electrostatic capacitor is also the subject of R&D (for HEVs as well as wireless communications). The Department of Energy has funded the Oak Ridge National Laboratory's Power Electronic and Electric Machinery Research Center to develop an automated test to aid in the independent testing of prototype electrostatic capacitors. This paper describes the design and development of such a stand.

  2. Pilocarpine iontophoresis test

    DEFF Research Database (Denmark)

    Hjortskov, N; Jepsen, Leif; Nielsen, B;

    1995-01-01

    The pilocarpine iontophoresis test (P-test) is used as a predictor of the capacity to produce sweat. Therefore, we studied the reproducibility of this test in 12 normal subjects on 10 consecutive days. Furthermore, we determined whether the P-test reflects whole-body and regional sweat secretion...... during exercise in the heat. Finally, we determined whether the P-test stimulates the eccrine sweat glands to maximal sweat secretion. Six growth hormone-deficient (GHD) patients who are known to have decreased sweating, and 11 healthy control subjects were studied. To induce maximal sweat secretion......, the patients exercised on a bicycle ergometer at a workload corresponding to 40% of their maximal aerobic power (VO2max). The 11 healthy subjects exercised at a workload of 150 W. All subjects exercised for 60 min in ambient air at 35 degrees C, with 50% relative humidity. The P-test showed a mean day...

  3. Dismantling the Mantel tests

    DEFF Research Database (Denmark)

    Guillot, Gilles; Rousset, François

    2012-01-01

    1. The simple and partialMantel tests are routinely used in many areas of evolutionary biology to assess the significance of the association between two ormorematrices of distances relative to the same pairs of individuals or demes. Partial Mantel tests rather than simple Mantel tests are widely...... used to assess the relationship between two variables displaying some form of structure. 2. We show that contrary to a widely shared belief, partialMantel tests are not valid in this case, and their bias remains close to that of the simpleMantel test. 3. We confirm that strong biases are expected under...... a sampling design and spatial correlation parameter drawn from an actual study. 4. TheMantel tests should not be used in case autocorrelation is suspected in both variables compared under the null hypothesis.We outline alternative strategies. The R code used for our computer simulations is...

  4. Standard environmental test methods

    Energy Technology Data Exchange (ETDEWEB)

    Schafer, D R

    1983-12-01

    This guide to uniformity in testing is intended primarily as an aid to persons responsible for designing, developing, and performing environmental tests. It will also be of use to those concerned with production, evaluation, and quality control and assurance. Checklists for preparing the environmental testing portion of product specifications are included, as are copies of Process Standards covering the instrumentation, equipment, and methods for use in environmental testing of Sandia National Laboratories components. Techniques and equipment are constantly improving. This version of SC-4452 reflects current state-of-the-art and practice in environmental testing. Previously existing sections of the document have ben updated and new ones have been added, e.g., Transient Testing on Vibration Machines.

  5. Crowdsourcing for Usability Testing

    CERN Document Server

    Liu, Di; Kuipers, Rebecca; Bias, Randolph

    2012-01-01

    While usability evaluation is critical to designing usable websites, traditional usability testing can be both expensive and time consuming. The advent of crowdsourcing platforms such as Amazon Mechanical Turk and CrowdFlower offer an intriguing new avenue for performing remote usability testing with potentially many users, quick turn-around, and significant cost savings. To investigate the potential of such crowdsourced usability testing, we conducted two similar (though not completely parallel) usability studies which evaluated a graduate school's website: one via a traditional usability lab setting, and the other using crowdsourcing. While we find crowdsourcing exhibits some notable limitations in comparison to the traditional lab environment, its applicability and value for usability testing is clearly evidenced. We discuss both methodological differences for crowdsourced usability testing, as well as empirical contrasts to results from more traditional, face-to-face usability testing.

  6. Tests, testing, and tested - we need to critically evaluate the meaning of tests in psychiatry.

    Science.gov (United States)

    Berger, Douglas M

    2013-04-01

    This article describes clinical pitfalls in our concepts of what it means for an illness, diagnosis, or evaluation and treatment methods to say that they have been "tested". This articles begins with the problems encountered in newborn testing for Krabbe Disease of the nervous system in New York State over the last few years as an example of a test that did not live up to its promise to help the society. Next, the article gives 3 examples of testing in psychiatry, 1. Psychological testing to make treatment decisions in children with depression, 2. Patient's and parents who have been told, or believe, that they have Asperger's disorder, and 3. The conclusions made about the efficacy of cognitive behavioral therapy based on clinical studies. The article's conclusion sums up these examples as reasons why we need to have a more practical and scientific approach to our understanding and implementation of tests used in our field. PMID:23825862

  7. Dismantling the Mantel tests

    DEFF Research Database (Denmark)

    Guillot, Gilles

    The simple and partial Mantel tests are routinely used in many areas of evolutionary biology to assess the significance of the association between two or more matrices of distances relative to the same pairs of individuals or demes. Partial Mantel tests rather than simple Mantel tests are widely...... used to assess the relationship between two variables displaying some form of structure. We show that contrarily to a widely shared belief, partial Mantel tests are not valid in this case, and their bias remains close to that of the simple Mantel test. We confirm that strong biases are expected under a...... sampling design and spatial correlation parameter drawn from an actual study. The Mantel tests should not be used in case auto-correlation is suspected in both variables compared under the null hypothesis. We outline alternative strategies. The R code used for our computer simulations is distributed as...

  8. Team Exploratory Testing Sessions

    OpenAIRE

    Soili Saukkoriipi; Ilkka Tervonen

    2012-01-01

    Exploratory testing (ET) is popular, especially among agile development teams. In this paper, we study the team aspect in the ET context and explore how to use ET in team sessions to complement other testing activities. The goal was to define a team exploratory testing (TET) session approach and to provide evidence that the approach is worth using. A TET session approach is defined by means of parameters, roles, and process. Also, instructions for using the approach are given. The team is the...

  9. Nondestructive testing method

    International Nuclear Information System (INIS)

    Nondestructive testing (NDT) is the use of physical and chemical methods for evaluating material integrity without impairing its intended usefulness or continuing service. Nondestructive tests are used by manufaturer's for the following reasons: 1) to ensure product reliability; 2) to prevent accidents and save human lives; 3) to aid in better product design; 4) to control manufacturing processes; and 5) to maintain a uniform quality level. Nondestructive testing is used extensively on power plants, oil and chemical refineries, offshore oil rigs and pipeline (NDT can even be conducted underwater), welds on tanks, boilers, pressure vessels and heat exchengers. NDT is now being used for testing concrete and composite materials. Because of the criticality of its application, NDT should be performed and the results evaluated by qualified personnel. There are five basic nondestructive examination methods: 1) liquid penetrant testing - method used for detecting surface flaws in materials. This method can be used for metallic and nonmetallic materials, portable and relatively inexpensive. 2) magnetic particle testing - method used to detect surface and subsurface flaws in ferromagnetic materials; 3) radiographic testing - method used to detect internal flaws and significant variation in material composition and thickness; 4) ultrasonic testing - method used to detect internal and external flaws in materials. This method uses ultrasonics to measure thickness of a material or to examine the internal structure for discontinuities. 5) eddy current testing - method used to detect surface and subsurface flaws in conductive materials. Not one nondestructive examination method can find all discontinuities in all of the materials capable of being tested. The most important consideration is for the specifier of the test to be familiar with the test method and its applicability to the type and geometry of the material and the flaws to be detected

  10. Tests of Machine Intelligence

    CERN Document Server

    Legg, Shane

    2007-01-01

    Although the definition and measurement of intelligence is clearly of fundamental importance to the field of artificial intelligence, no general survey of definitions and tests of machine intelligence exists. Indeed few researchers are even aware of alternatives to the Turing test and its many derivatives. In this paper we fill this gap by providing a short survey of the many tests of machine intelligence that have been proposed.

  11. Psychological Test Validity

    Directory of Open Access Journals (Sweden)

    Gessmann H.-W.

    2015-02-01

    Full Text Available This paper presents a theoretical overview of current aspects about the validity problems of psychological tests. The article demonstrates the importance of the development of psychological tests and the scientific studies of their validity, describes the different types of validity and the possible ways of measurement and determination of the validity coefficients. The paper is recommended for researchers, whose work is dedicated to the development, modification or adaptation of the psychological test.

  12. Web Application Security Testing

    OpenAIRE

    Bukovský, Ondřej

    2012-01-01

    The purpose of this bachelor's thesis is to present the topic of web applications security. The purpose of the first, theoretical part of this work is to introduce and describe fundamentals like web security or penetration testing. OWASP (Open Web Application Security Project) and their ten most critical web applications security risks are presented in the rest of the first part. Second, practical part describes tested web application and defines purpose and scope of penetration tests. Then t...

  13. Accelerated cyclic corrosion tests

    OpenAIRE

    Prošek T.

    2016-01-01

    Accelerated corrosion testing is indispensable for material selection, quality control and both initial and residual life time prediction for bare and painted metallic, polymeric, adhesive and other materials in atmospheric exposure conditions. The best known Neutral Salt Spray (NSS) test provides unrealistic conditions and poor correlation to exposures in atmosphere. Modern cyclic accelerated corrosion tests include intermittent salt spray, wet and dry phases and eventually other technical p...

  14. Automatic test suite evolution

    OpenAIRE

    Mirzaaghaei, Mehdi; Pezzè, Mauro

    2013-01-01

    Software testing is one of the most common approaches to verify software systems. Despite of many automated techniques proposed in the literature, test cases are often generated manually. When a software system evolves during development and maintenance to accommodate requirement changes, bug fixes, or functionality extensions, test cases may become obsolete, and software developers need to evolve them to verify the new version of the software system. Due to time pressure and effort requir...

  15. GRE physics test

    CERN Document Server

    Fakhrudin, Hasan

    2010-01-01

    Designed for students and professionals looking to advance their careers, our test prep gives you everything you need to succeed. Focused chapter reviews cover all the information tested on the GRE Physics exam. Each review chapter contains examples to reinforce key concepts. The book includes four full-length practice tests based on the most recent GRE Physics exam. Each test contains every type of question that can be expected on the GRE so you can “practice for real” and boost your confidence before taking the exam.

  16. Reactor Simulator Testing

    Science.gov (United States)

    Schoenfeld, Michael P.; Webster, Kenny L.; Pearson, Boise J.

    2013-01-01

    As part of the Nuclear Systems Office Fission Surface Power Technology Demonstration Unit (TDU) project, a reactor simulator test loop (RxSim) was design & built to perform integrated testing of the TDU components. In particular, the objectives of RxSim testing was to verify the operation of the core simulator, the instrumentation and control system, and the ground support gas and vacuum test equipment. In addition, it was decided to include a thermal test of a cold trap purification design and a pump performance test at pump voltages up to 150 V since the targeted mass flow rate of 1.75 kg/s was not obtained in the RxSim at the originally constrained voltage of 120 V. This paper summarizes RxSim testing. The gas and vacuum ground support test equipment performed effectively in NaK fill, loop pressurization, and NaK drain operations. The instrumentation and control system effectively controlled loop temperature and flow rates or pump voltage to targeted settings. The cold trap design was able to obtain the targeted cold temperature of 480 K. An outlet temperature of 636 K was obtained which was lower than the predicted 750 K but 156 K higher than the cold temperature indicating the design provided some heat regeneration. The annular linear induction pump (ALIP) tested was able to produce a maximum flow rate of 1.53 kg/s at 800 K when operated at 150 V and 53 Hz.

  17. Testing as social practice

    DEFF Research Database (Denmark)

    Kousholt, Kristine

    2016-01-01

    In this article, the author challenges the implicit assumptions of tests as a neutral tool for measuring the individual’s learning achievement. Instead, testing is explored as a social practice which becomes part of children’s conduct of everyday life. The theoretical foundation for the analysis is...... Danish-German Critical Psychology. This approach offers a dialectically developed set of concepts and hereby another basis for understanding school testing than the one implicit in the technology of testing. The analysis is primarily based on a case consisting of an observation conducted in a 2nd grade...

  18. Load testing circuit

    DEFF Research Database (Denmark)

    2009-01-01

    A load testing circuit a circuit tests the load impedance of a load connected to an amplifier. The load impedance includes a first terminal and a second terminal, the load testing circuit comprising a signal generator providing a test signal of a defined bandwidth to the first terminal of the load...... impedance, an energy-storing element being connected to the second terminal of the load impedance and providing an output signal, and a measuring unit that measures the output signal or compares the output signal with a reference....

  19. Tensile testing apparatus

    Science.gov (United States)

    Blackburn, L. B.; Ellingsworth, J. R. (Inventor)

    1985-01-01

    An improved mechanical extensometer is described for use with a constant load creep test machine. The dead weight of the extensometer is counterbalanced by two pairs of weights connected through a pulley system and to rod extension and leading into the furnace where the test sample is undergoing elevated temperature (above 500 F.) tensile testing. Novel gripper surfaces, conical tip and flat surface are provided in each sampling engaging platens to reduce the grip pressure normally required for attachment of the extensometer to the specimen and reduce initial specimen bending normally associated with foil-gage metal testing.

  20. LISA Optical Bench Testing

    Science.gov (United States)

    Tröbs, M.; d'Arcio, L.; Barke, S.; Bogenstahl, J.; Diekmann, C.; Fitzsimons, E. D.; Gerberding, O.; Hennig, J.; Hey, F. G.; Hogenhuis, H.; Killow, C. J.; Lieser, M.; Lucarelli, S.; Nikolov, S.; Perreur-Lloyd, M.; Pijnenburg, J.; Robertson, D. I.; Sohmer, A.; Taylor, A.; Ward, H.; Weise, D.; Heinzel, G.; Danzmann, K.

    2013-01-01

    Each LISA satellite carries optical benches, one for each test mass, that measure the distance to the local test mass and to the remote optical bench on the distant satellite. Currently, an elegant bread board of the optical bench is developed for the European Space Agency (ESA) by EADS Astrium, TNO Science and Technology, University of Glasgow and the Albert Einstein Institute. To test the optical bench the two interferometers mentioned above must be completed by an external simulator, the test mass and telescope simulator. We give an overview of the simulator layout and performance predictions.

  1. Pleural Fluid Analysis Test

    Science.gov (United States)

    ... leukemias , lung cancer , metastatic cancers Other conditions – idiopathic , cardiac bypass surgery, heart or lung transplantation, pancreatitis , or intra-abdominal abscesses Additional testing on ...

  2. Pavement Testing Facility

    Data.gov (United States)

    Federal Laboratory Consortium — Comprehensive Environmental and Structural Analyses The ERDC Pavement Testing Facility, located on the ERDC Vicksburg campus, was originally constructed to provide...

  3. Main Test Floor (MTF)

    Data.gov (United States)

    Federal Laboratory Consortium — Purpose: The MTF is employed to validate advanced structural concepts and verify new analytical methodologies. Test articles range in size from subcomponent to full...

  4. Charcoal filter testing

    Energy Technology Data Exchange (ETDEWEB)

    Lyons, J. [Nuclear Regulatory Commission, Washington, DC (United States)

    1997-08-01

    In this very brief, informal presentation, a representative of the US Nuclear Regulatory Commission outlines some problems with charcoal filter testing procedures and actions being taken to correct the problems. Two primary concerns are addressed: (1) the process to find the test method is confusing, and (2) the requirements of the reference test procedures result in condensation on the charcoal and causes the test to fail. To address these problems, emergency technical specifications were processed for three nuclear plants. A generic or an administrative letter is proposed as a more permanent solution. 1 fig.

  5. Der Progress Test Medizin

    OpenAIRE

    Osterberg, K.; Kölbel, S; Brauns, K

    2006-01-01

    In 1999 a interdiciplinary, formative progress test for medical students has been established at the Charité - Universitätsmedizin Berlin. It contents of 200 MC-questions on a graduate level and is performed at the beginning of each semester since then.The ascertained test data is evaluated and reported back in a detailed written feedback to each participant.After initial problems in recruiting new item authors and an efficient administration of test items the progress test now is a well est...

  6. Automatic Test Builder

    OpenAIRE

    Abbasi, Saad Zeb

    2012-01-01

    In Ericsson, the Automation Team automates test cases that are frequently rerun. This process involves copying data related to a particular Configured Test Case from a database and then pasting it into a java file created to run a test case. In one java file, there can be more than one Configured Test Cases. So information can vary. Then the tester has to add package name, necessary imports, member variables, preamble and post amble methods, help methods and main execution methods. A lot of t...

  7. Abstractions on test design techniques

    OpenAIRE

    Wendland, Marc-Florian

    2014-01-01

    Automated test design is an approach to test design in which automata are utilized for generating test artifacts such as test cases and test data from a formal test basis, most often called test model. A test generator operates on such a test model to meet a certain test coverage goal. In the plethora of the approaches, tools and standards for model-based test design, the test design techniques to be applied and test coverage goals to be met are not part of the test model, which may easily le...

  8. Flushable reagent stool blood test

    Science.gov (United States)

    Stool occult blood test - flushable home test; Fecal occult blood test - flushable home test ... This test is performed at home with disposable pads. You can buy the pads at the drug store without ...

  9. Over the air test

    DEFF Research Database (Denmark)

    2015-01-01

    [1] This invention relates to over-the-air testing of a device in an anechoic chamber. In particular, the invention is suitable for simulating both uplink and downlink over-the-air communication with a device under test even when the anechoic chamber has different numbers of uplink and downlink...

  10. Test dig selv!

    DEFF Research Database (Denmark)

    Hølge-Hazelton, Bibi; Nielsen, Gregers; Langstrup, Henriette;

    2011-01-01

    Teknologirådet omkring rapporten ”Test dig selv! – Vurderinger og anbefalinger vedrørende anvendelse medicinsk udstyr til selvtestning. Offentliggjort d. 25.10.11.......Teknologirådet omkring rapporten ”Test dig selv! – Vurderinger og anbefalinger vedrørende anvendelse medicinsk udstyr til selvtestning. Offentliggjort d. 25.10.11....

  11. Judgments in Language Testing

    Institute of Scientific and Technical Information of China (English)

    J.Charles Alderson

    2009-01-01

    Language testing is an area linguistics that combines the professional judgment and the nature of the of applied exercise of about language, learning, achievement of language learning with empirical data about students' performances and, by inference, their abilities. This paper addresses the relationship between judgments and empirical data in language testing by reporting on three studies.

  12. Cross cultural usability testing

    DEFF Research Database (Denmark)

    Clemmensen, Torkil; Goyal, Shivam

    2005-01-01

    and hereby identify usability problems with the clipart application. Data on the evaluators' and test users' behaviour were recorded and analyzed by coding and summarizing statistics on these behavioural events. The results show that Think Aloud Usability Test of a localized application is most effectively...

  13. Solenoid Testing Facility

    Data.gov (United States)

    Federal Laboratory Consortium — Current Configuration: Accommodate a device under test up to 2.8 m diameter, 0.7 m height and 15,000 lbs. weight. Up to 10 g/s, 4.5 K helium flow. Up to 250 A test...

  14. The Test Validation Summary

    Science.gov (United States)

    Frederick, Richard I.; Bowden, Stephen C.

    2009-01-01

    Common rates employed in classificatory testing are the true positive rate (TPR), false positive rate (FPR), positive predictive power (PPP), and negative predictive power (NPP). FPR and TPR are estimated from research samples representing populations to be distinguished by classificatory testing. PPP and NPP are used by clinicians to classify…

  15. Testing children for allergies

    DEFF Research Database (Denmark)

    Eigenmann, P A; Atanaskovic-Markovic, M; O'B Hourihane, J;

    2013-01-01

    or recurrent or severe symptoms suggestive for allergy should undergo an appropriate diagnostic work-up, irrespective of their age. Adequate allergy testing may also allow defining allergic trigger in common symptoms. We provide here evidence-based guidance on when and how to test for allergy in children based...

  16. Platelet Function Tests

    Science.gov (United States)

    ... of the clotting process in the body ( in vivo ). A person with normal platelet function test results may still experience excessive bleeding or inappropriate clotting during and after a surgery. Most samples for platelet function testing are only stable for a very short period ...

  17. CSF glucose test

    Science.gov (United States)

    Glucose test - CSF; Cerebrospinal fluid glucose test ... The glucose level in the CSF should be 50 to 80 mg/100 mL (or greater than 2/3 of the blood sugar level). Note: Normal value ranges may vary slightly ...

  18. Routine DNA testing

    Science.gov (United States)

    Routine DNA testing. It’s done once you’ve Marker-Assisted Breeding Pipelined promising Qantitative Trait Loci within your own breeding program and thereby established the performance-predictive power of each DNA test for your germplasm under your conditions. By then you are ready to screen your par...

  19. Helicobacter pylori Test

    Science.gov (United States)

    ... or bloody vomit or vomit that looks like coffee grounds. H. pylori testing may also be ordered when a person has completed a regimen of prescribed antibiotics to confirm that the H. pylori bacteria have been eliminated. A follow-up test is ...

  20. Math Sense: Placement Test.

    Science.gov (United States)

    2003

    Math Sense consists of five books that develop from basic to more advanced math skills. This document contains a placement test used with Math Sense to help students and their teachers decide into which Math Sense book to begin working. The placement test is divided into six parts, each consisting of 10 to 22 problems, and is based on exit skill…

  1. Testing object Interactions

    NARCIS (Netherlands)

    Grüner, Andreas

    2010-01-01

    In this thesis we provide a unit testing approach for multi-purposes object-oriented programming languages in the style of Java and C#. Our approach includes the definition of a test specification language which results from extending the programming language with new designated specification constr

  2. Automate functional testing

    Directory of Open Access Journals (Sweden)

    Ramesh Kalindri

    2014-06-01

    Full Text Available Currently, software engineers are increasingly turning to the option of automating functional tests, but not always have successful in this endeavor. Reasons range from low planning until over cost in the process. Some principles that can guide teams in automating these tests are described in this article.

  3. Test Reviewing in Spain

    Science.gov (United States)

    Muniz, Jose; Fernandez-Hermida, Jose R.; Fonseca-Pedrero, Eduardo; Campillo-Alvarez, Angela; Pena-Suarez, Elsa

    2012-01-01

    The proper use of psychological tests requires that the measurement instruments have adequate psychometric properties, such as reliability and validity, and that the professionals who use the instruments have the necessary expertise. In this article, we present the first review of tests published in Spain, carried out with an assessment model…

  4. Dengue Fever Testing

    Science.gov (United States)

    ... the initial fever has passed. Antibody tests for dengue fever can be positive if a person is infected with another arbovirus such as West Nile virus . A health practitioner will consider a person's test results, medical history, and recent travel history in making a diagnosis. ...

  5. Beyond the Test Scores.

    Science.gov (United States)

    Thibodeau, Janice J.

    1985-01-01

    A diagnostic-prescriptive scheme is illustrated using subtests of the Slingerland Screening Tests for Identifying Children with Specific Language Disability and the Detroit Tests of Learning Aptitude. The scheme is intended to focus on the child's learning style by examining the task and the strategies employed. (CL)

  6. Heart failure - tests

    Science.gov (United States)

    ... tests, such as coronary angiography. MRI , CT , or PET scans of the heart may be done to check how much heart muscle damage is present. It can also help detect the reason for a patient's heart failure. Stress tests are done to see whether the heart ...

  7. A monostrain test apparatus

    Science.gov (United States)

    Helf, J. C.; Hill, W. L.; Pontius, H. L.

    1972-01-01

    Test apparatus is designed for determining tensile strength, modulus of elasticity, elongation, and thermal coefficient of contraction or expansion of uniformly shaped plastics, adhesives, and foam materials over temperature range of 700 to 90 K (800 to -300). Tests may be used in design quality control, and in evaluation of new adhesives and plastic materials.

  8. Testing for Subcellular Randomness

    CERN Document Server

    Okunoye, Babatunde O

    2008-01-01

    Statistical tests were conducted on 1,000 numbers generated from the genome of Bacteriophage T4, obtained from GenBank with accession number AF158101.The numbers passed the non-parametric, distribution-free tests.Deoxyribonucleic acid was discovered to be a random number generator, existent in nature.

  9. Preparing for CLIC tests

    CERN Multimedia

    Laurent Guiraud

    1999-01-01

    The Canon 5 undergoes first brazing for preparation in the CLIC study at the CLIC Test Facility 2 (CTF2). This will test injection for a proposed linear collider that will further explore discoveries made at the LHC. Electric fields in the canon will boost electrons into the acceleration fields of the collider.

  10. Neonatal Screening Tests.

    Science.gov (United States)

    Vigue, Charles L.

    1986-01-01

    Describes several laboratory experiments that are adaptations of clinical tests for certain genetic diseases in babies. Information and procedures are provided for tests for phenylketonuria (PKU), galactosemia, tyrosinemia, cystinuria, and mucopolysaccharidosis. Discusses the effects of each disease on the infants' development. (TW)

  11. Poppering the Newell Test

    NARCIS (Netherlands)

    Taatgen, Niels A.

    2003-01-01

    The Newell Test as it is proposed by Anderson & Lebiere (A&L) has the disadvantage of being too positivistic, stressing areas a theory should cover, instead of attempting to exclude false predictions. Nevertheless, Newell's list can be used as the basis for a more stringent test with a stress on the

  12. Identification of the Mycobacterium ulcerans protein MUL_3720 as a promising target for the development of a diagnostic test for Buruli ulcer.

    Directory of Open Access Journals (Sweden)

    Anita Dreyer

    2015-02-01

    Full Text Available Buruli ulcer (BU caused by Mycobacterium ulcerans is a devastating skin disease, occurring mainly in remote West African communities with poor access to health care. Early case detection and subsequent antibiotic treatment are essential to counteract the progression of the characteristic chronic ulcerative lesions. Since the accuracy of clinical BU diagnosis is limited, laboratory reconfirmation is crucial. However, currently available diagnostic techniques with sufficient sensitivity and specificity require infrastructure and resources only accessible at a few reference centres in the African endemic countries. Hence, the development of a simple, rapid, sensitive and specific point-of-care diagnostic tool is one of the major research priorities for BU. In this study, we have identified a previously unknown M. ulcerans protein, MUL_3720, as a promising target for antigen capture-based detection assays. We show that MUL_3720 is highly expressed by M. ulcerans and has no orthologs in other prevalent pathogenic mycobacteria. We generated a panel of anti-MUL_3720 antibodies and used them to confirm a cell wall location for MUL_3720. These antibodies could also specifically detect M. ulcerans in infected human tissue samples as well as in lysates of infected mouse footpads. A bacterial 2-hybrid screen suggested a potential role for MUL_3720 in cell wall biosynthesis pathways. Finally, we demonstrate that a combination of MUL_3720 specific antibody reagents in a sandwich-ELISA format has sufficient sensitivity to make them suitable for the development of antigen capture-based diagnostic tests for BU.

  13. LHC synchronization test successful

    CERN Multimedia

    The synchronization of the LHC's clockwise beam transfer system and the rest of CERN's accelerator chain was successfully achieved last weekend. Tests began on Friday 8 August when a single bunch of a few particles was taken down the transfer line from the SPS accelerator to the LHC. After a period of optimization, one bunch was kicked up from the transfer line into the LHC beam pipe and steered about 3 kilometres around the LHC itself on the first attempt. On Saturday, the test was repeated several times to optimize the transfer before the operations group handed the machine back for hardware commissioning to resume on Sunday. The anti-clockwise synchronization systems will be tested over the weekend of 22 August.Picture:http://lhc-injection-test.web.cern.ch/lhc-injection-test/

  14. Nanomechanical testing system

    Energy Technology Data Exchange (ETDEWEB)

    Vodnick, David James; Dwivedi, Arpit; Keranen, Lucas Paul; Okerlund, Michael David; Schmitz, Roger William; Warren, Oden Lee; Young, Christopher David

    2015-01-27

    An automated testing system includes systems and methods to facilitate inline production testing of samples at a micro (multiple microns) or less scale with a mechanical testing instrument. In an example, the system includes a probe changing assembly for coupling and decoupling a probe of the instrument. The probe changing assembly includes a probe change unit configured to grasp one of a plurality of probes in a probe magazine and couple one of the probes with an instrument probe receptacle. An actuator is coupled with the probe change unit, and the actuator is configured to move and align the probe change unit with the probe magazine and the instrument probe receptacle. In another example, the automated testing system includes a multiple degree of freedom stage for aligning a sample testing location with the instrument. The stage includes a sample stage and a stage actuator assembly including translational and rotational actuators.

  15. Nanomechanical testing system

    Energy Technology Data Exchange (ETDEWEB)

    Vodnick, David James; Dwivedi, Arpit; Keranen, Lucas Paul; Okerlund, Michael David; Schmitz, Roger William; Warren, Oden Lee; Young, Christopher David

    2015-02-24

    An automated testing system includes systems and methods to facilitate inline production testing of samples at a micro (multiple microns) or less scale with a mechanical testing instrument. In an example, the system includes a probe changing assembly for coupling and decoupling a probe of the instrument. The probe changing assembly includes a probe change unit configured to grasp one of a plurality of probes in a probe magazine and couple one of the probes with an instrument probe receptacle. An actuator is coupled with the probe change unit, and the actuator is configured to move and align the probe change unit with the probe magazine and the instrument probe receptacle. In another example, the automated testing system includes a multiple degree of freedom stage for aligning a sample testing location with the instrument. The stage includes a sample stage and a stage actuator assembly including translational and rotational actuators.

  16. Biocide patch tests

    DEFF Research Database (Denmark)

    Andersen, Klaus Ejner; Veien, Niels

    1985-01-01

    Routine patch testing with a series of 6 industrial biocides containing methylene-bis-thiocyanate (Cytox 3522), benzisothiazolin-3-one (BIT), chlorocresol (Preventol CMK), 2-n-octyl-4-isothiazolin-3-one (Kathon 893), polyhydroxymethylene monobenzylether (Preventol D2) or 1,3,5-tris (hydroxy......-ethyl) hexahydrotriazine (Grotan BK) was carried out in 6 Danish out-patient clinics to evaluate guinea pig allergy test results with the same compounds. A total of 1652 consecutive patients with dermatitis were tested. The usefulness of this patch test battery was limited. There were a few positive reactions to Cytox...... male patients and atopics, but significant differences in the frequencies of occupational cases, hand eczemas, and leg ulcers/stasis dermatitis, indicating possible variations in referral patterns, use of patch tests, and/or environmental factors....

  17. Invalid Permutation Tests

    Directory of Open Access Journals (Sweden)

    Mikel Aickin

    2010-01-01

    Full Text Available Permutation tests are often presented in a rather casual manner, in both introductory and advanced statistics textbooks. The appeal of the cleverness of the procedure seems to replace the need for a rigorous argument that it produces valid hypothesis tests. The consequence of this educational failing has been a widespread belief in a “permutation principle”, which is supposed invariably to give tests that are valid by construction, under an absolute minimum of statistical assumptions. Several lines of argument are presented here to show that the permutation principle itself can be invalid, concentrating on the Fisher-Pitman permutation test for two means. A simple counterfactual example illustrates the general problem, and a slightly more elaborate counterfactual argument is used to explain why the main mathematical proof of the validity of permutation tests is mistaken. Two modifications of the permutation test are suggested to be valid in a very modest simulation. In instances where simulation software is readily available, investigating the validity of a specific permutation test can be done easily, requiring only a minimum understanding of statistical technicalities.

  18. LISA Pathfinder ground testing

    Science.gov (United States)

    Guzman, Felipe; LISA Pathfinder Team

    2010-01-01

    The space-based gravitational wave observatory LISA is a joint NASA-ESA mission that requires challenging technology to ensure pure geodetic trajectories of test masses and the interferometric measurement of distance variations between them. The LISA Pathfinder mission is an ESA-launched technology demonstrator of key LISA subsystems such as spacecraft control with micronewton thrusters, test mass drag-free control, and precision laser interferometry between free-flying test masses. Ground testing of pre-flight hardware of the Gravitational Reference Sensor and Optical Metrology subsystems is currently ongoing. Studies have been carried out on very sensitive torsion pendulums that effectively reproduce a free-fall condition for the test mass within a horizontal plane in the lab, down to frequencies loop operation, demonstrating the required optical metrology sensitivity to test mass displacement. This poster presents the current status in the development and implementation of LISA Pathfinder pre-flight systems and latest results of the ongoing ground testing efforts.

  19. [Stroop test and schizophrenia].

    Science.gov (United States)

    Grapperon, J; Delage, M

    1999-01-01

    Utilization of the Stroop test with schizophrenic subjects is reviewed from 32 studies published since 1960. This test, in which subject has to name the color of color-words which are congruent or incongruent, is much used as a selective attention test. Its specific disturbance in schizophrenics would confirm the overload of consciousness with unsignificant non filtered information, probably due to lowering of frontal functions. Stroop effect is not always raised in these patients, especially when general slowing is taken into account. Technical differences modify the test power and can explain contradictory results. The thought disorder symptom seems the most correlated with the results of the test, and particular sensitiveness of paranoïd form has been argued. Neuroleptics do not seem to act on Stroop effect. Factors explaining the great variability of patients in this test are not fully put in light. One hypothesis, already raised, is that the test is linked to a particular attentional constitutional aptitude, and whose relations with the illness are to be explained. PMID:10205734

  20. Instant testing with QUnit

    CERN Document Server

    Sheiko, Dmitry

    2013-01-01

    Filled with practical, step-by-step instructions and clear explanations for the most important and useful tasks. A concise guide full of step-by-step recipes to teach you how to perform automated testing using QUnit.""Instant Testing with QUnit"" is for anyone working with JavaScript who is looking for a powerful and easy-to-use testing framework. The reader does not need to know any particular framework, only the basic principles of JavaScript and HTML.