Sample records for amplification reagent kit

  1. 21 CFR 864.9650 - Quality control kit for blood banking reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Quality control kit for blood banking reagents... Manufacture Blood and Blood Products § 864.9650 Quality control kit for blood banking reagents. (a) Identification. A quality control kit for blood banking reagents is a device that consists of sera,...

  2. Evaluation of commercial enzyme reagent kits by use of a semiautomated chemistry analyzer. (United States)

    Beckala, H R; Agrell, J; Forsman, R W; Homburger, H A


    The overall performances of several enzyme reagent kits for alkaline phosphatase, creatine kinase, lactic dehydrogenase, and aspartate aminotransferase were evaluated using an ABA-100 Bichromatic Analyzer. Interassay precision using this instrument with commercial reagents compared well with published data for similar analyses performed at university hospitals and referral laboratories. Significantly poorer precision with lower limits of linearity was observed when reagents recommended for use at 30 C were used at 37 C. Significant differences in measured levels of creatine kinase, lactic dehydrogenase, and aspartate aminotransferase due to different lots of expendable cuvettes were found for elevated levels of these enzymes. All kit reagents met manufacturers' claims for stability; however, different absolute levels of lactic dehydrogenase were observed with one kit reagent on successive days. Slight hemolysis affected creatine kinase levels measured with some reagent kits significantly more than others.

  3. Liposomes as signal amplification reagents for bioassays in microfluidic channels. (United States)

    Locascio, Laurie E; Hong, Jennifer S; Gaitan, Michael


    Liposomes with encapsulated carboxyfluorescein were used in an affinity-based assay to provide signal amplification for small-volume fluorescence measurements. Microfluidic channels were fabricated by imprinting in a plastic substrate material, poly(ethylene terephthalate glycol) (PETG), using a silicon template imprinting tool. Streptavidin was linked to the surface through biotinylated-protein for effective immobilization with minimal nonspecific adsorption of the liposome reagent. Lipids derivatized with biotin were incorporated into the liposome membrane to make the liposomes reactive for affinity assays. Specific binding of the liposomes to microchannel walls, dependence of binding on incubation time, and nonspecific adsorption of the liposome reagent were evaluated. The results of a competitive assay employing liposomes in the microchannels are presented.

  4. Validation of the AmpFlSTR« SEfiler Plus(TM) PCR Amplification kit for forensic STR analysis

    DEFF Research Database (Denmark)

    Fredslund, Stine Frisk; Mogensen, Helle Smidt; Morling, Niels


    Validation of the AmpFlSTR« SEfiler Plus(TM) PCR Amplification kit with 29 and 30 PCR cycles for forensic STR analysis demonstrated that the kit had fewer artefacts than the AmpFlSTR« SGM Plus(TM) kit (28 PCR cycles). The SEfiler Plus kit was more sensitive and devoid of colour artefacts, but sho......, but showed more stutters, drop-ins, drop-outs and allelic imbalances...

  5. Validation of the AmpliFLP D1S80 PCR Amplification Kit for forensic casework analysis according to TWGDAM guidelines. (United States)

    Cosso, S; Reynolds, R


    The validation of the AmpliFLP D1S80 PCR Amplification Kit for use in forensic casework was accomplished by performing all the relevant experiments outlined in the TWGDAM guidelines. Standard specimen and reproducibility studies were performed using organic and rapid DNA extraction techniques on both stain and liquid samples (blood, semen and saliva). Over 300 samples from three different populations (US Caucasians, African Americans and US Hispanics) were analyzed to determine allele and genotype frequencies. Purified DNA was mixed in defined ratios (ranging from unmixed DNA samples to 1:9 mixtures of 2 different DNA samples) prior to amplification to demonstrate that samples containing DNA from more than one individual can be detected and, in many cases, that the genotypes contributing to the mixture can be identified. Since casework samples frequently are exposed to environmental insults that can result in DNA degradation, purified DNA was degraded in the laboratory to analyze the effect of DNA fragment length on D1S80 amplification. It is crucial in the validation process to examine actual casework evidentiary material. This D1S80 kit can be used successfully by forensic scientists to amplify and type nonprobative evidentiary material, including bloodstains collected from crime scenes and rape kit materials collected for sexual assault cases. The D1S80 kit is specific to human DNA, and the D1S80 alleles are inherited according to the laws of Mendel. The sensitivity of the novel gel electrophoresis gel matrix allowed the PCR cycle number to be reduced to 29 cycles and the D1S80 kit sensitivity to be increased to 2.5 ng from the previous D1S80 Reagent Set specifications of 30 cycles and 5 ng, respectively.

  6. Development of PCR internal controls for DNA profiling with the AmpFℓSTR® SGM Plus® amplification kit. (United States)

    Nathalie, Zahra; Hadi, Sibte; Goodwin, William


    Forensic DNA profiling uses a series of commercial kits that co-amplify several loci in one reaction; the products of the PCR are fluorescently labelled and analysed using CE. Before CE, an aliquot of the PCR is mixed with formamide and an internal lane size standard. Using the SGM Plus amplification kit, we have developed two internal non-amplified controls of 80 bp and 380 bp that are labelled with ROX fluorescent dye and added to the PCR. Combined with two internal amplification controls of 90 bp and 410 bp, they provide additional controls for the PCR, electrokinetic injection, and CE and also function as an internal size standard.

  7. Developmental validation of the GlobalFiler(®) Express PCR Amplification Kit: A 6-dye multiplex assay for the direct amplification of reference samples. (United States)

    Wang, Dennis Y; Gopinath, Siddhita; Lagacé, Robert E; Norona, Wilma; Hennessy, Lori K; Short, Marc L; Mulero, Julio J


    In order to increase the power of discrimination, reduce the possibility of adventitious matches, and expand global data sharing, the CODIS Core Loci Working Group made a recommendation to expand the CODIS core loci from the "required" 13 loci to 20 plus three additional "highly recommended" loci. The GlobalFiler(®) Express Kit was designed to incorporate all 20 required and 3 highly recommended loci along with a novel male-specific Y insertion/deletion marker. The GlobalFiler(®) Express Kit allows simultaneous amplification of the following loci: D3S1358, vWA, D16S539, CSF1PO, TPOX, Yindel, AMEL, D8S1179, D21S11, D18S51, DYS391, D2S441, D19S433, TH01, FGA, D22S1045, D5S818, D13S317, D7S820, SE33, D10S1248, D1S1656, D12S391, and D2S1338. The kit enables direct amplification from blood and buccal samples stored on paper or swab and the chemistry features an optimized PCR protocol that yields time to results in less than an hour. Developmental validation testing followed SWGDAM guidelines and demonstrated the quality and robustness of the GlobalFiler(®) Express Kit over a number of variables. The validation results demonstrate that the 24-locus multiplex kit is a robust and reliable identification assay as required for forensic DNA typing and databasing.

  8. Using multiplex PCR amplification and typing kits for the analysis of DNA evidence in a serial killer case. (United States)

    Hochmeister, M N; Budowle, B; Eisenberg, A; Borer, U V; Dirnhofer, R


    Analysis of DNA evidence in a serial killer case was performed using the AmpliType HLA-DQ alpha-, AmpliType PM-, and the GenePrint STR Multiplex System PCR Amplification Kits. In addition, a sex typing procedure using the X-Y homologous gene amelogenin was carried out. DNA profiles from a single hair with attached sheath material, recovered from underneath the seat cover of the suspect's car seat were compared with DNA profiles derived from reference head hairs from a homicide victim. From the evidentiary sample only 9 ng of human DNA could be recovered. In a sample, where the quantity of DNA becomes a critical issue a powerful route is the simultaneous amplification of several loci (multiplex PCR). This is the first report where commercially available multiplex PCR amplification and typing kits have been introduced for the analysis of DNA evidence in a serial killer case and the analysis has been admitted in court.


    Directory of Open Access Journals (Sweden)

    M. A. Loginova


    Full Text Available During the screening typing of recruited volunteers with Volga Federal District for unrelated hematopoietic stem cell registry on the loci (HLA-A, B, DRB1, DRB345 in sample No 1758 identified a new allele at locus A. The use of basic kit AlleleSEQR HLA-A Sequencing in combination with HARP – A2F98A allowed to determine the genotype of this sample – А*30:01:01, a new allele А*25, В*13, 44, DRB1*03, 09, DRB3*02, DRB4*01. 

  10. The Development and Use of Internal Amplification Controls (IACs) with DNA Profiling Kits for Forensic DNA Analysis. (United States)

    Zahra, Nathalie; Goodwin, William


    Biological samples recovered for forensic investigations are often degraded and/or have low amounts of DNA; in addition, in some instances the samples may be contaminated with chemicals that can act as PCR inhibitors. As a consequence this can make interpretation of the results challenging with the possibility of having partial profiles and false negative results. Because of the impact of DNA analysis on forensic investigations, it is important to monitor the process of DNA profiling, in particular the amplification reaction. In this chapter we describe a method for the in-house generation and use of internal amplification controls (IACs) with DNA profiling kits to monitor the success of the PCR proces. In the example we show the use of the SGM Plus® kit. These controls can also be used to aid the interpretation of the DNA profile.

  11. Development of internal amplification controls for DNA profiling with the AmpFℓSTR(®) SGM Plus(®) kit. (United States)

    Zahra, Nathalie; Hadi, Sibte; Smith, Judith A; Iyengar, Arati; Goodwin, William


    DNA extracted from forensic samples can be degraded and also contain co-extracted contaminants that inhibit PCR. The effects of DNA degradation and PCR inhibition are often indistinguishable when examining a DNA profile. Two internal amplification controls (IACs) were developed to improve quality control of PCR using the AmpFℓSTR® SGM Plus® kit. The co-amplification of these controls with DNA samples was used to monitor amplification efficiency and detect PCR inhibitors. IAC fragments of 90 and 410 bp (IAC₉₀ and IAC₄₁₀) were generated from the plasmid pBR322 using tailed primers and then amplified with ROX-labelled primers. Co-amplification of IAC₉₀ and IAC₄₁₀ was performed with varying amounts of template DNA, degraded DNA and DNA contaminated with humic acid, heme and indigo dye. Both IAC₉₀ and IAC₄₁₀ were successfully amplified with human DNA without significantly affecting the quality of the DNA profile, even with DNA amounts lower than 0.5 ng. In the presence of inhibitors, the IAC₉₀ signal was still present after all human DNA loci fail to amplify; in contrast, the IAC₄₁₀ signal was reduced or absent at low levels of inhibition. Amplification of the two IACs provided an internal PCR control and allowed partial profiles caused by inhibition to be distinguished from degraded DNA profiles.

  12. Forensic Application of Expressmarker 22 STR Loci Direct PCR Amplification Kit%Expressmarker 22 STR荧光检测试剂盒的法医学应用

    Institute of Scientific and Technical Information of China (English)

    邹凯南; 曹禹; 夏子芳; 郑卫国; 周怀谷


    Objective To explore the application value of Expressmarker 22 STR loci direct PCR amplification kit. Methods One thousand nine hundred and forty-eight samples (including samples spotted on FTA cards, filter papers and case samples) were tested using Expressmarker 22 STR loci direct PCR amplification kit. At the same time, all were tested using Sinofiler? kit, Identifiler(R) kit and PowerPlex(R) 16 kit respectively for comparison. The genotypes were compared at the same STR loci among these four kits to test the sensitivity and accuracy of Expressmarker 22 STR loci direct PCR amplification kit. Results 97.79% samples were successfully typed using Expressmarker 22 STR loci direct PCR amplification kit. The genotype profiles of the same samples using Expressmarker 22 STR loci direct PCR amplification kit were consistent with Sinofiler? kit, Identifiler(R) kit and PowerPlex(R) 16 kit at the same STR loci. Conclusion Expressmarker 22 STR loci direct PCR amplification kit can provide huge information and accurate results and be applied to forensic DNA genotyping.%目的 验证Expressmarker 22 STR荧光检测试剂盒的法医学应用价值.方法 取FTA卡、滤纸上保存的建库血样和各类涉案生物性检材1948份,用Expressmarker 22 STR荧光检测试剂盒进行STR分型检测,同时采用SinofilerTM、Identifiler(R)、PowerPlex(R) 16试剂盒进行平行试验.对4个试剂盒相同基因座的分型进行比对,以确认Expressmarker 22 STR荧光检测试剂盒的灵敏度和准确性.结果 Expressmarker 22 STR荧光检测试剂盒的检测成功率为97.79%,同一样本在相同基因座与SinofilerTM、Identifiler(R)、PowerPlex(R) 16试剂盒的STR分型结果相同.结论 利用Expressmarker 22 STR荧光检测试剂盒进行STR分型,信息量大、结果准确可靠.可应用于法庭科学.

  13. KIT amplification and gene mutations in acral/mucosal melanoma in Korea. (United States)

    Yun, Jina; Lee, Jeeyun; Jang, Jiryeon; Lee, Eui Jin; Jang, Kee Taek; Kim, Jung Han; Kim, Kyoung-Mee


    Mucosal and acral melanomas have demonstrated different genetic alterations and biological behavior compared with more common cutaneous melanomas. It was recently reported that gain-of-function KIT mutations and/or copy number increases are more common in mucosal and acral melanomas. Thus, we studied the frequency and pattern of KIT aberrations in mucosal and acral melanomas in Korea. We analyzed 97 patients who were pathologically confirmed with mucosal or acral melanoma between 1997 and 2010 at Samsung Medical Center. Of the 97 melanoma patients, 92 were screened for mutations in KIT exons 11, 13, 17, and 18, BRAF and NRAS genes. KIT copy number was assessed by quantitative, real-time PCR. Of the 97 patients, 55 (56.7%) were mucosal, 40 (41.2%) were acral melanoma, and two were of unknown primary origin. Among seven cases with KIT mutation, five (60.0%) occurred in exon 11, one (20.0%) in exon 17, and one (20.0%) in exon 13. Point mutations were the most common, resulting in substitutions in exon 11 (K558R, T574A, L576P, and V559A), exon 13 (N655K), and exon 17 (N822K). A novel Thr574Ala (c.1720A>G) KIT mutation, which has not been reported in melanoma or other tumor types, was identified in one genital melanoma case. Of the 97 mucosal or acral melanoma specimens, 49 were tested for KIT gene copy number changes using quantitative PCR. Increased KIT copy number was identified in 15 patients: seven (40%) of 20 acral melanomas and eight (31%) of 26 mucosal melanomas. Our study implicates that a significant proportion of acral and mucosal melanomas have KIT mutations in Asian population.

  14. [The validation of kit of reagents for quantitative detection of DNA of human cytomegalovirus in biological material using polymerase chain reaction technique in real time operation mode]. (United States)

    Sil'veĭstrova, O Iu; Domonova, É A; Shipulina, O Iu


    The validation of kit of reagents destined to detection and quantitative evaluation of DNA of human cytomegalovirus in biological material using polymerase chain reaction technique in real time operation mode was implemented. The comparison was made against international WHO standard--The first WHO international standard for human cytomegalovirus to implement measures the kit of reagents "AmpliSens CMV-screen/monitor-FL" and standard sample of enterprise DNA HCMV (The central research institute of epidemiology of Rospotrebnadzor) was applied. The fivefold dilution of international WHO standard and standard sample of enterprise were carried out in concentrations of DNA HCMV from 106 to 102. The arrangement of polymerase chain reaction and analysis of results were implemented using programed amplifier with system of detection of fluorescent signal in real-time mode "Rotor-Gene Q" ("Qiagen", Germany). In the total of three series of experiments, all stages of polymerase chain reaction study included, the coefficient of translation of quantitative evaluation of DNA HCMV from copy/ml to ME/ml equal to 0.6 was introduced for this kit of reagents.

  15. Forensic and population genetic analyses of Danes, Greenlanders and Somalis typed with the Yfiler® Plus PCR amplification kit. (United States)

    Olofsson, Jill Katharina; Mogensen, Helle Smidt; Buchard, Anders; Børsting, Claus; Morling, Niels


    Recently, the Yfiler® Plus PCR Amplification Kit (Yfiler® Plus, Thermo Fisher Scientific, Waltham, MA, USA) was introduced. Yfiler® Plus amplifies 27 Y-chromosomal short tandem repeat loci (Y-STRs) and adds ten new Y-STRs to those analysed with the commonly used AmpFlSTR® Yfiler® PCR Amplification Kit (Yfiler®, Thermo Fisher Scientific, Waltham, MA, USA). Seven of the new Y-STRs are rapidly mutating Y-STRs (RM Y-STRs). In this study, 551 male individuals from Denmark, Greenland and Somalia were typed with Yfiler® Plus. The results were compared to those obtained with Yfiler® in the same individuals. Forensic and population genetic parameters were estimated for Yfiler® Plus. Yfiler® Plus had a higher power of discrimination than Yfiler® in all three populations. Compared to Yfiler®, Yfiler® Plus offers increased power of discrimination, which is obviously an advantage in crime case investigations. However, the inclusion of seven RM Y-STRs in Yfiler® Plus makes it less attractive for relationship testing because of the relatively high combined mutation rate, approximately 15%.

  16. Forensic and population genetic analyses of Danes, Greenlanders and Somalis typed with the Yfiler® Plus PCR amplification kit

    DEFF Research Database (Denmark)

    Olofsson, Jill Katharina; Mogensen, Helle Smidt; Buchard, Anders


    Recently, the Yfiler(®) Plus PCR Amplification Kit (Yfiler(®) Plus, Thermo Fisher Scientific, Waltham, MA, USA) was introduced. Yfiler(®) Plus amplifies 27 Y-chromosomal short tandem repeat loci (Y-STRs) and adds ten new Y-STRs to those analysed with the commonly used AmpFlSTR(®) Yfiler(®) PCR...... Amplification Kit (Yfiler(®), Thermo Fisher Scientific, Waltham, MA, USA). Seven of the new Y-STRs are rapidly mutating Y-STRs (RM Y-STRs). In this study, 551 male individuals from Denmark, Greenland and Somalia were typed with Yfiler(®) Plus. The results were compared to those obtained with Yfiler......(®) in the same individuals. Forensic and population genetic parameters were estimated for Yfiler(®) Plus. Yfiler(®) Plus had a higher power of discrimination than Yfiler(®) in all three populations. Compared to Yfiler(®), Yfiler(®) Plus offers increased power of discrimination, which is obviously an advantage...

  17. Casework testing of the multiplex kits AmpFlSTR SEfiler Plus PCR amplification kit (AB), PowerPlex S5 System (Promega) and AmpFlSTR MiniFiler PCR amplification kit (AB). (United States)

    Müller, Kathrin; Sommerer, Thomas; Miltner, Erich; Schneider, Harald; Wiegand, Peter


    The short tandem repeat (STR) kits SEfiler Plus (D3S1358, FGA, D8S1179, D18S51, D21S11, TH01, VWA, SE33, D2S1338, D16S539, D19S433 and Amelogenin), PowerPlex S5 System (D18S51, D8S1179, TH01, FGA and Amelogenin) and MiniFiler (D13S317, D7S820, Amelogenin, D2S1338, D21S11, D16S539, D18S51, CSF1PO and FGA) were comparatively tested for their robustness and sensitivity. About 50 stains with highly degraded DNA and little DNA quantity served as examination material (e.g., hair with a telogen root, bones, degraded saliva stains on drinking vessels and skin cell mixtures). The PowerPlex S5 with five German DNA database (DAD) systems and the MiniFiler kit with four topical DAD systems and further STR markers show reduced amplicon lengths. The SEfiler Plus kit represents no MiniSTR multiplex, but contains the nine current DAD systems and further three systems D2S1338, D16S539 and D19S433, which are the potential expansion markers for the German DNA database. We have found on the basis of our comparative stain investigations, that the SEfiler Plus kit was less sensitive than the PowerPlex S5 and the MiniFiler kits. The MiniFiler and the PowerPlex S5 kit showed comparatively high sensitivity. Especially in analysing skin cell mixtures, the MiniFiler kit showed larger differences with regard to the performance of the fluorescent dyes/primer concentration co-ordination than the PowerPlex S5. The SEfiler Plus kit generated - just as both MiniSTR kits - relative robust typing results, but there appeared an increased sensitivity for 'allelic drop-outs' and 'imbalances'. Since the SEfiler Plus kit was not planned as MiniSTR concept, 'allelic drop-outs' were observed, as expected, more frequent in typing stains with degraded DNA and little DNA quantity, especially in the long polymerase chain reaction (PCR) products (e.g., D18S51).

  18. Comprehensive mutation analysis of 17 Y-chromosomal short tandem repeat polymorphisms included in the AmpF lSTR® Yfiler® PCR amplification kit

    NARCIS (Netherlands)

    M.A. Goedbloed (Miriam); M. Vermeulen (Mark); R.N. Fang (Rixun); M. Lembring (Maria); A. Wollstein (Andreas); K. Ballantyne (Kaye); O. Lao Grueso (Oscar); S. Brauer (Silke); C. Krüger (Carmen); L. Roewer (Lutz); R. Lessig (Rüdiger); R. Ploski (Rafal); T. Dobosz (Tadeusz); J. Henke (Jürgen); M.R. Furtado (Manohar); M.H. Kayser (Manfred)


    textabstractThe Y-chromosomal short tandem repeat (Y-STR) polymorphisms included in the AmpF lSTR® Yfiler® polymerase chain reaction amplification kit have become widely used for forensic and evolutionary applications where a reliable knowledge on mutation properties is necessary for correct data in

  19. Oncogene abnormalities in a series of primary melanomas of the sinonasal tract: NRAS mutations and cyclin D1 amplification are more frequent than KIT or BRAF mutations. (United States)

    Chraybi, Meriem; Abd Alsamad, Issam; Copie-Bergman, Christiane; Baia, Maryse; André, Jocelyne; Dumaz, Nicolas; Ortonne, Nicolas


    Primary malignant melanoma of sinonasal tract is a rare but severe form of melanoma. We retrospectively analyzed 17 cases and focused on the histologic presentation and the expression of c-Kit, epidermal growth factor receptor (EGFR), cyclin D1/Bcl-1, PS100, and HMB45 and searched for BRAF, NRAS, and KIT mutations that are known to be associated with melanoma subtypes, together with amplifications of KIT, cyclin D1, cyclin-dependent kinase 4, MDM2, and microphthalmia-associated transcription factor using quantitative polymerase chain reaction. In most cases (78%), an in situ component was evidenced. Invasive components were composed of diffuse areas of rhabdoid, epithelioid, or spindle cells and, in most cases, lacked inflammatory reaction, suggesting that an immune escape phenomenon probably develops when the disease progresses. EGFR was rarely and weakly expressed in the in situ component of 2 cases. None of the investigated case showed BRAF V600E, but 1 had a D594G mutation. NRAS mutations in exon 2 (G12D or G12A) were found in 3 cases (18%), and a KIT mutation in exon 11 (L576P), in 1, whereas c-Kit was expressed at the protein level in half of the cases. Amplifications of cyclin D1 were evidenced in 5 cases, confirmed in 3 by fluorescence in situ hybridization, but this was not always correlated with protein expression, found in 8 patients (62.5%), 3 having no significant amplification. In conclusion, primary malignant melanoma of sinonasal tract is not associated with BRAF V600E mutations. Instead, NRAS or KIT mutations and cyclin D1 amplification can be found in a proportion of cases, suggesting that primary malignant melanoma of sinonasal tract is heterogeneous at the molecular level and should not be sensitive to therapeutic approaches aiming at BRAF.

  20. Developmental validation of the Yfiler(®) Plus PCR Amplification Kit: An enhanced Y-STR multiplex for casework and database applications. (United States)

    Gopinath, Siddhita; Zhong, Chang; Nguyen, Vivian; Ge, Jianye; Lagacé, Robert E; Short, Marc L; Mulero, Julio J


    Y-chromosomal loci have proven useful in solving investigations where low levels of male DNA are present in a high female DNA background. An intrinsic limitation of Y-STRs compared with autosomal STRs is a reduced power of discrimination due to a lack of recombination throughout most of the Y-chromosome. Thus, in an effort to increase the power of discrimination we have developed a new 6-dye, 27-plex Y-STR system that includes the 17 loci from the Yfiler(®) and Yfiler(®) Direct kits (DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635 (Y GATA C4), and Y GATA H4) plus three highly polymorphic Y-STR loci (DYS460, DYS481, and DYS533), and seven rapidly mutating Y-STR loci (DYF387S1a/b, DYS449, DYS518, DYS570, DYS576, DYS627) which allow for improved discrimination of related individuals. The Yfiler(®) Plus PCR Amplification Kit is a dual application assay designed to amplify DNA from extracted casework and database samples from storage cards and swab lysates via direct amplification. Compared to the Yfiler PCR Amplification Kit, the new multiplex shows increased discrimination of male lineages and also improved performance in inhibited samples, improved balance in male DNA samples mixed with female DNA at ratios >1:1000, and faster time to results. The Yfiler Plus Kit shows very high concordance to the Yfiler Kit but discordance with the PowerPlex(®) Y23 Kit at the DYS481 locus was observed in 2 out of 30 samples tested. This developmental validation work follows the SWGDAM guidelines and demonstrates that the assay is robust and suitable for use on forensic casework and database samples.

  1. Loop-mediated isothermal amplification with the Procedure for Ultra Rapid Extraction kit for the diagnosis of pneumocystis pneumonia. (United States)

    Kawano, Shuichi; Maeda, Takuya; Suzuki, Takefumi; Abe, Tatsuhiro; Mikita, Kei; Hamakawa, Yusuke; Ono, Takeshi; Sonehara, Wataru; Miyahira, Yasushi; Kawana, Akihiko


    Loop-mediated isothermal amplification (LAMP) is an innovative molecular technique requiring only a heating device and isothermal conditions to amplify a specific target gene. The results of current microscopic diagnostic tools for pneumocystis pneumonia are not sufficiently consistent for detecting infection with a low-density of Pneumocystis jirovecii. Although polymerase chain reaction (PCR) is highly sensitive, it is not suitable for resource-limited facilities. LAMP is a potential diagnostic replacement for PCR in such settings but a critical disadvantage of DNA extraction was still remained. Therefore, we employed the Procedure for Ultra Rapid Extraction (PURE) kit, which uses a porous material, to isolate the DNA from clinical samples in a simple way in combination with previously reported LAMP procedure for diagnosing PCP. The detection limit of the PURE-LAMP method applied to artificial bronchoalveolar lavage fluid samples was 100 copies/tube, even with the use of massive blood-contaminated solutions. In addition, we concluded the diagnostic procedure within 1 h without the need for additional equipment. PURE-LAMP coupled with suitable primers for specific pathogens has good potential for diagnosing various infectious diseases.

  2. Comparative performance of AmpFLSTR® Identifiler® Plus PCR amplification kit and QIAGEN® Investigator® IDplex Plus kit. (United States)

    Mattayat, Dalad; Kitpipit, Thitika; Phetpeng, Sukanya; Asawutmangkul, Watee; Thanakiatkrai, Phuvadol


    Many forensic STR kits are currently available in the market. The AmpFLSTR® Identifiler® Plus kit, which targets 15 STRs, is commonly used worldwide. The Thai forensic DNA community is built around it in terms of instrument, databases, and interpretation. QIAGEN's IDplex Plus kit targets the same loci, but the PCR cycling time is shorter by about 90min. A direct comparison that assesses forensic parameters and applicability to casework between the two kits has never been carried out. In this study, we performed a direct comparison between the two kits using serial dilutions of two control DNA samples and 60 randomly selected casework samples, including samples taken from improvised explosive devices and terrorist raids. We statistically compared the performance of the two kits in terms of peak height, number of allele detected (allelic drop-out), intra-locus balance, inter-locus balance, inhibitor tolerance, stutter ratio, concordance, and allelic drop-in. The results demonstrate that both kits are statistically similar in performance. IDplex Plus gave higher peak heights in sensitivity test and tolerated inhibitors better, but had slightly worse inter-locus balances and stutter ratios. However, these differences were not practically significant, as seen by the resulting profiles of the casework samples (p=0.601). The performance on low-template samples also was not different. In conclusion, laboratories looking to replace the aging Identifiler® Plus might consider the IDplex Plus as a faster, more robust alternative that fits right into their existing structure without further investment in instrument and DNA database. Having more kits available worldwide by different companies could help bring the technology to different forensic laboratories and the justice system as a whole.

  3. Developmental validation of a single-tube amplification of the 13 CODIS STR loci, D2S1338, D19S433, and amelogenin: the AmpFlSTR Identifiler PCR Amplification Kit. (United States)

    Collins, Patrick J; Hennessy, Lori K; Leibelt, Craig S; Roby, Rhonda K; Reeder, Dennis J; Foxall, Paul A


    Analysis of length polymorphism at short tandem repeat (STR) loci utilizing the polymerase chain reaction (PCR) process has proven to be an ideal assay for human identification purposes. The short length of STR loci coupled with the amplification of target sequence through PCR allows for a robust, sensitive, and specific assay for highly polymorphic markers. A multiplex containing fifteen STR loci plus the gender-determining locus Amelogenin was developed to provide a single amplification/detection of all CODIS (Combined DNA Index System) STR loci (CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, FGA, TH01, TPOX, and vWA) as well as two internationally-accepted STRs (D2S1338 and D19S433). By incorporating five-dye fragment analysis technology and non-nucleotide linkers, previously optimized AmpFlSTR kit primer sequences have been maintained. This kit has been developed in accordance with the standards of the forensic community as defined by the DNA Advisory Board. Validation studies were performed to include developmental validation, and the results support the use of the AmpFlSTR Identifiler PCR Amplification Kit for human identity and parentage testing.

  4. Evaluation of the loop mediated isothermal DNA amplification (LAMP kit for malaria diagnosis in P. vivax endemic settings of Colombia.

    Directory of Open Access Journals (Sweden)

    Andrés F Vallejo


    Full Text Available Most commonly used malaria diagnostic tests, including microscopy and antigen-detecting rapid tests, cannot reliably detect low-density infections which are frequent in low transmission settings. Molecular methods such as polymerase chain reaction (PCR are highly sensitive but remain too laborious for field deployment. In this study, the applicability of a malaria diagnosis kit based on loop-mediated isothermal amplification (mLAMP was assessed in malaria endemic areas of Colombia with Plasmodium vivax predominance.First, a passive case detection (PCD study on 278 febrile patients recruited in Tierralta (department of Cordoba was conducted to assess the diagnostic performance of the mLAMP method. Second, an active case detection (ACD study on 980 volunteers was conducted in 10 sentinel sites with different epidemiological profiles. Whole blood samples were processed for microscopic and mLAMP diagnosis. Additionally RT-PCR and nested RT-PCR were used as reference tests. In the PCD study, P. falciparum accounted for 23.9% and P. vivax for 76.1% of the infections and no cases of mixed-infections were identified. Microscopy sensitivity for P. falciparum and P. vivax were 100% and 86.1%, respectively. mLAMP sensitivity for P. falciparum and P. vivax was 100% and 91.4%, respectively. In the ACD study, mLAMP detected 65 times more cases than microscopy. A high proportion (98.0% of the infections detected by mLAMP was from volunteers without symptoms.mLAMP sensitivity and specificity were comparable to RT-PCR. LAMP was significantly superior to microscopy and in P. vivax low-endemicity settings and under minimum infrastructure conditions, it displayed sensitivity and specificity similar to that of single-well RT-PCR for detection of both P. falciparum and P. vivax infections. Here, the dramatically increased detection of asymptomatic malaria infections by mLAMP demonstrates the usefulness of this new tool for diagnosis, surveillance, and screening in

  5. Electrochemical aptasensor based on the dual-amplification of G-quadruplex horseradish peroxidase-mimicking DNAzyme and blocking reagent-horseradish peroxidase. (United States)

    Yuan, Yali; Gou, Xuxu; Yuan, Ruo; Chai, Yaqin; Zhuo, Ying; Mao, Li; Gan, Xianxue


    A simple electrochemical aptasensor for sensitive detection of thrombin was fabricated with G-quadruplex horseradish peroxidase-mimicking DNAzyme (hemin/G-quadruplex system) and blocking reagent-horseradish peroxidase as dual signal-amplification scheme. Gold nanoparticles (nano-Au) were firstly electrodeposited onto single wall nanotube (SWNT)-graphene modified electrode surface for the immobilization of electrochemical probe of nickel hexacyanoferrates nanoparticles (NiHCFNPs). Subsequently, another nano-Au layer was electrodeposited for further immobilization of thrombin aptamer (TBA), which later formed hemin/G-quadruplex system with hemin. Horseradish peroxidases (HRP) then served as blocking reagent to block possible remaining active sites and avoided the non-specific adsorption. In the presence of thrombin, the TBA binded to thrombin and the hemin released from the hemin/G-quadruplex electrocatalytic structure, increasing steric hindrance of the aptasensor and decomposing hemin/G-quadruplex electrocatalytic structure, which finally decreased the electrocatalytic efficiency of aptasensor toward H(2)O(2) in the presence of NiHCFNPs with a decreased electrochemical signal. On the basis of the synergistic amplifying action, a detection limit as low as 2 pM for thrombin was obtained.

  6. 不同PCR扩增试剂盒检验血样DNA的检验结果对比研究%Comparative Study between Test Results of Different PCR Amplification Kits in Testing Blood Sample DNA

    Institute of Scientific and Technical Information of China (English)



    目的:讨论研究Profiler PlusTM、IdentifilerTM以及Powerplex16扩增试剂盒用于检验血样DNA检验结果的差异,并研究其扩张不平衡和基因丢失现象的发生几率。方法:选取150例完全无血缘关系的个体作为研究对象并采集其血样,分别使用两种扩增试剂盒进行检验。对所得不同结果的同一对象再使用3种扩增试剂盒进行检验。将检验所得结果进行比较。结果:3种试剂盒检测的等位基因缺失率比较差异无统计学意义(P>0.05)。Profiler PlusTM检测扩张不平衡率显著高于其余两种试剂盒(P<0.05)。结论:使用PCR扩增试剂盒对血样DNA进行检测时,会出现不同位置的异常基因,可表现为基因缺失及扩增不平衡。但Profiler PlusTM检验扩增不平衡发生率显著高于其余两种试剂盒。在实际生活对血样DNA进行检测时,除需准备主要检测扩增试剂盒外,还需准备其他不同类备用试剂盒用于互相验证及对比,尽量降低基因等位缺失及扩张不平衡发生率。%To discuss and study the difference between testing results of Profiler PlusTM,IdentifilerTM and Powerplex16 amplification kits in testing blood sample DNA,and study the occurrence rate of amplification imbalance and gene deletion phenomenon.Method:150 individuals who had no genetic connection with each other were chosen as research objects,and their blood samples were collected and tested by 2 different amplification kits. Objects with 2 different results were tested again by the third kit. All testing results were compared.Result:There were no significant differences between the 3 kits on allelic gene deletion(P<0.05). Amplification imbalance rate of Profiler PlusTM was obviously higher than the other 2 kits(P<0.05). Conclusion:When testing blood sample DNA with different PCR amplification kits, there might be abnormal genes of different positions which might manifested as gene deletion and

  7. 血清总蛋白参考方法复现与常用试剂盒比对研究%Reproduction of reference method for serum total protein and evaluation of commercial reagent kits

    Institute of Scientific and Technical Information of China (English)

    何美琳; 翟静; 梁国威; 张捷


    目的:复现血清总蛋白测定的双缩脲参考方法(紫外分光光度法),并验证其性能.同时评价4种常用商品试剂盒.方法:参考方法的建立参照美国AACC的要求.通过参加国际临床化学与检验医学联合会(IFCC)主办的参考实验室间的国际环形比对实验(RELA)以验证其准确性.按照美国临床和实验室标准委员会(CLSI)EP-9A2方案,对4种常用商品试剂盒进行评价.结果:参考方法的CVs在0.47% ~ 0.85%间.线性方程为y=1.0022x -0.2121(r=0.9999).回收率为100.2%~102.4%.RELA比对结果在等效限内.与参考方法相比,4种商品试剂盒在医学决定点(Xc=45 g/L,60 g/L和80 g/L)处的95%可信区间位于可接受偏差范围内.结论:本实验室已复现并验证了血清总蛋白双缩脲法参考方法;4种商品试剂盒的检测结果与参考方法有差异,但仍在可接受范围内,可用于临床实验室检测.%Objective:To reproduce a reference measurement procedure and evaluate commercial reagent kits by such established procedure. Methods:The reference measurement procedure of serum total protein was established according to the American Association for Clinical Chemistry ( AACC) , and the accuracy of this procedure was verified by External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine ( RELA) held by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) . Subsequently, four commercial reagent kits were evaluated by the reproduced reference procedure following CLSI guideline EP -9A2. Results:The performance of the reproduced reference procedure was as follows; CVs ranged from 0.47% toO.85%. Linearity was Y = 1.0022X -0.2121 (r=0.9999) , and the recovery ranged from 100.2% tol02.4%. The External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine ( RELA) was applied, and the result was within the limit of equivalence. Compared with the reference method, the 95

  8. Enzymatic electrochemical detection of epidemic-causing Vibrio cholerae with a disposable oligonucleotide-modified screen-printed bisensor coupled to a dry-reagent-based nucleic acid amplification assay. (United States)

    Yu, Choo Yee; Ang, Geik Yong; Chan, Kok Gan; Banga Singh, Kirnpal Kaur; Chan, Yean Yean


    In this study, we developed a nucleic acid-sensing platform in which a simple, dry-reagent-based nucleic acid amplification assay is combined with a portable multiplex electrochemical genosensor. Preparation of an amplification reaction mix targeting multiple DNA regions of interest is greatly simplified because the lyophilized reagents need only be reconstituted with ultrapure water before the DNA sample is added. The presence of single or multiple target DNAs causes the corresponding single-stranded DNA (ssDNA) amplicons to be generated and tagged with a fluorescein label. The fluorescein-labeled ssDNA amplicons are then analyzed using capture probe-modified screen-printed gold electrode bisensors. Enzymatic amplification of the hybridization event is achieved through the catalytic production of electroactive α-naphthol by anti-fluorescein-conjugated alkaline phosphatase. The applicability of this platform as a diagnostic tool is demonstrated with the detection of toxigenic Vibrio cholerae serogroups O1 and O139, which are associated with cholera epidemics and pandemics. The platform showed excellent diagnostic sensitivity and specificity (100%) when challenged with 168 spiked stool samples. The limit of detection was low (10 colony-forming units/ml) for both toxigenic V. cholerae serogroups. A heat stability assay revealed that the dry-reagent amplification reaction mix was stable at temperatures of 4-56 °C, with an estimated shelf life of seven months. The findings of this study highlight the potential of combining a dry-reagent-based nucleic acid amplification assay with an electrochemical genosensor in a more convenient, sensitive, and sequence-specific detection strategy for multiple target nucleic acids.

  9. 三种方法检测尿中肌酐的结果比对%The results comparison of three methods to assay creatinine in urine with HPLC, spectrophotometry and creatinine diagnostic reagent kit

    Institute of Scientific and Technical Information of China (English)

    李凤贞; 区文凯; 何凤群; 程允杰


    目的:验证反相高效液相色谱法(WS/T 98-1996)、分光光度法(WS/T 97-1996)和肌酐测定试剂盒(苦味酸法)三种方法检测尿中肌酐的结果可比性.方法:以同一尿样,在采集后第1天、第3天、第5天以上述三种检测方法进行尿中肌酐的检测,卫生部行标方法按标准检验进行操作,临床生化非标方法按说明书要求进行操作.结果:同一尿样在采集后第1天、第3天、第5天按同一方法检测,尿中肌酐检测值无显著性差异(P>0.05);同一尿样按不同的方法进行检测,尿中肌酐的结果有显著性差异(P 0.05 ) ; And there were significant differences of the creatinine in the collected same urine with HPLC, spectrophotometry and creatinine diagnostic reagent kit (P < 0.01). Conclusion: The results were incompared because there were significant differences of the creatinine in the collected same urine with HPLC,spectrophotometry and creatinine diagnostic reagent kit.

  10. Protein crystallography prescreen kit (United States)

    Segelke, Brent W.; Krupka, Heike I.; Rupp, Bernhard


    A kit for prescreening protein concentration for crystallization includes a multiplicity of vials, a multiplicity of pre-selected reagents, and a multiplicity of sample plates. The reagents and a corresponding multiplicity of samples of the protein in solutions of varying concentrations are placed on sample plates. The sample plates containing the reagents and samples are incubated. After incubation the sample plates are examined to determine which of the sample concentrations are too low and which the sample concentrations are too high. The sample concentrations that are optimal for protein crystallization are selected and used.

  11. [The reagents kit to detect Metyorchis biis, Opisthorchis viverrini and Clonorchis sinensis, Opisthorchis felineus--agents of opisthorchiasis using technique of polymerase chain reaction in real-time]. (United States)

    Seredina, T A; Petrenko, V A; Tronin, A V; Sazonov, A Iu; Sapugol'tseva, O B; Katokhin, A V; Odintsova, E S


    The helminths Opisthorchis felineus, Opisthorchis viverrini, Clonorchis sinensis, Metorchis bilis are the agents of opisthorchiasis. The actual diagnostic of parasitic diseases based on microscope analysis of samples of human feces to detect presence of ova of parasites suffers of many shortcomings, in particular low sensitivity especially at earlier stages. The purpose of this study was to compare results of detection of parasites using both classical technique and technique of specific differentiation based on extraction of nucleic acids from samples of human feces and implementation of reaction of amplification of the chosen fragment of DNA with detection of products of polymerase chain reaction in the real time. The study detected 150 out of 165 positive samples and also 6 out of 37 negative samples both validated by coproovoscopy.

  12. Direct Y-STR amplification of body fluids deposited on commonly found crime scene substrates. (United States)

    Dargay, Amanda; Roy, Reena


    Body fluids detected on commonly found crime scene substrates require extraction, purification and quantitation of DNA prior to amplification and generation of short tandem repeat (STR) DNA profiles. In this research Y-STR profiles were generated via direct amplification of blood and saliva deposited on 12 different substrates. These included cigarette butts, straws, grass, leaves, woodchips and seven different types of fabric. After depositing either 0.1 μL of blood or 0.5 μL of saliva, each substrate containing the dry body fluid stain was punched using a Harris 1.2 mm micro-punch. Each of these punched substrates, a total of 720 samples, containing minute amount of blood or saliva was either amplified directly without any pre-treatment, or was treated with one of the four washing reagents or buffer. In each of these five experimental groups the substrates containing the body fluid remained in the amplification reagent during the thermal cycling process. Each sample was amplified with the three direct Y-STR amplification kits; AmpFℓSTR(®) Yfiler(®) Direct, Yfiler(®) Plus Amplification Kits and the PowerPlex(®) Y23 System. Complete and concordant Y-STR profiles were successfully obtained from most of these 12 challenging crime scene objects when the stains were analyzed by at least one of the five experimental groups. The reagents and buffer were interchangeable among the three amplification kits, however, pre-treatment with these solutions did not appear to enhance the quality or the number of the full profiles generated with direct amplification. This study demonstrates that blood and saliva deposited on these simulated crime scene objects can be amplified directly.

  13. Development of lyophilized kit of Tin-Glucoheptonate for in vitro labeling of leucocytes with {sup 99m}Tc; Desenvolvimento de reagente liofilizado de glucoheptonato-estanho para marcacao de leucocitos com Tecnecio-99m in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Nascimento, Rosemeire Fagundes


    The study and localization of inflammatory and infection process in Nuclear Medicine represents a relevant tool in diagnostic procedures. In same cases, the diagnostic is easy and based on anamnesis and clinical observation; in other cases, the patients are asymptomatic or present non specific symptoms that difficult the diagnostic. The early diagnostic of inflammatory or infectious process allow the early introduction of therapy and prevents complications. Farther, the differentiation between inflammation and infection is of extreme importance as well as the localization of the focus. The use of labeled leucocytes, studied and applied in much pathologies, is the method of choice for the visualization of inflammation and infection. The scintigraphy using labeled leucocytes was introduced at 1976 by McAffe and Thakur and since of this is used in the diagnostic of different pathologies related to leucocyte infiltration like intestinal inflammatory disease, bone or prosthetic-vascular infections. The in vitro labeling of leucocytes with {sup 111}In was performed using oxime or tropolone as ligand and with {sup 99m}Tc using hexamethylpropylene amine oxime (HMPAO) as ligand, resulting in a lipophilic complex. The {sup 99m}Tc-HMPAG complex was preferably employed in many indications and countries do to the ideal physical properties of {sup 99m}Tc that results in low dose to the patient. However, the labeling employing the HMPAO complex results in some disadvantages like the low stability of the complex, and some requirements related to the {sup 99m}Tc elution (like the time pos elution), beyond the high cost of the compound that is imported. The aim of this work was the development of a tin-glucoheptonate lyophilized kit for in vitro leucocytes labeling with {sup 99m}Tc using the pre-stannization method. The optimization of the labeling technique was developed using leucocytes isolated from total blood and employing different volumes of the tinglucoheptonate reagent and

  14. Evaluation of self-established assay system in Beckman DXC800 chemistry analyzer measured by prealbumin reagent kits%应用国产前白蛋白在Beckman上自建检测系统及其评价

    Institute of Scientific and Technical Information of China (English)

    张家云; 邓芳; 孙永梅; 李明


    目的:应用EP9-A对某国产前白蛋白在Beckman上的自建检测系统,并进行评价和偏差评估。方法:依据美国临床实验室标准化委员会(NCCLS)EP9-A应用北京利德曼前白蛋白(PA)试剂盒及其配套校准品、质控品在Beckman DXC800全自动生化分析仪的检测系统对40份患者新鲜血清的PA进行检测,并与Beckman公司原装配套的前白蛋白试剂盒(PAB)比对,对检测浓度进行系统评价和偏差分析。结果:北京利德曼前白蛋白(PA)试剂在Beckman DXC800全自动生化分析仪上的准确度、批内精密度、线性均符合要求,两种试剂检测结果无显著差异(P>0.05),具有较好的可比性,其相关系数(r2)>0.99。结论:自建检测系统血清PA检测结果与Beckman可溯源的参考检测系统相比,可被临床接受。%Objective:To evaluation the comparability and clinical acceptability of national prealbumin reagent kits measured by the self-established assay system in Beckman DXC800 chemistry analyzer.Methods:The levels of prealbumin in 40 patients' fresh serum, Calibrators and Controls were analyzed by self-established and Beckman ref-erence assay system. To compared with each other, the bias were estimated according to National Committee for Clini-cal Laboratory Standards (NCCLS) document EP9A2 files. Results:The trueness, inter-precision and linear of Beijing Leadman prealbumin reagent kits measured by the self-established assay system in Beckman DXC800 chemistry an-alyzer were acceptable. There was no statistically significant difference, P>0.05. The correlation coefficient (r2) was more than 0.99. Conclusion:The comparability of national prealbumin measured by the self-established assay system in Beckman DXC800 chemistry analyzer compared with Beckman reference assay system was acceptable.

  15. Molecular Alterations of KIT Oncogene in Gliomas

    Directory of Open Access Journals (Sweden)

    Ana L. Gomes


    Full Text Available Gliomas are the most common and devastating primary brain tumours. Despite therapeutic advances, the majority of gliomas do not respond either to chemo or radiotherapy. KIT, a class III receptor tyrosine kinase (RTK, is frequently involved in tumourigenic processes. Currently, KIT constitutes an attractive therapeutic target. In the present study we assessed the frequency of KIT overexpression in gliomas and investigated the genetic mechanisms underlying KIT overexpression. KIT (CD117 immunohistochemistry was performed in a series of 179 gliomas of various grades. KIT activating gene mutations (exons 9, 11, 13 and 17 and gene amplification analysis, as defined by chromogenic in situ hybridization (CISH and quantitative real-time PCR (qRT-PCR were performed in CD117 positive cases. Tumour cell immunopositivity was detected in 15.6% (28/179 of cases, namely in 25% (1/4 of pilocytic astrocytomas, 25% (5/20 of diffuse astrocytomas, 20% (1/5 of anaplastic astrocytomas, 19.5% (15/77 of glioblastomas and one third (3/9 of anaplastic oligoastrocytomas. Only 5.7% (2/35 of anaplastic oligodendrogliomas showed CD117 immunoreactivity. No association was found between tumour CD117 overexpression and patient survival. In addition, we also observed CD117 overexpression in endothelial cells, which varied from 0–22.2% of cases, being more frequent in high-grade lesions. No KIT activating mutations were identified. Interestingly, CISH and/or qRT-PCR analysis revealed the presence of KIT gene amplification in 6 glioblastomas and 2 anaplastic oligoastrocytomas, corresponding to 33% (8/24 of CD117 positive cases. In conclusion, our results demonstrate that KIT gene amplification rather than gene mutation is a common genetic mechanism underlying KIT expression in subset of malignant gliomas. Further studies are warranted to determine whether glioma patients exhibiting KIT overexpression and KIT gene amplification may benefit from therapy with anti-KIT RTK

  16. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and

  17. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of interna-tional and intercultural cooperation,operating at the interface between theory and practice

  18. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.

  19. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation. The

  20. 新型均相酶法检测sd LDL-C试剂盒的性能评价%Performance Evaluation of New Homogeneous Enzymatic Reagent Kit for Measurement of Small Dense LDL CholesterolLIN Wen-tao1,LI Jiang2,SUN Fei3,ZHAO Xing-bo2,Yasuki Ito4,Motoko Ohta4,YAN

    Institute of Scientific and Technical Information of China (English)

    林文涛; 李江; 孙菲; 赵兴波; Yasuki Ito; Motoko Ohta; 鄢盛恺


    Objective To evaluate the analytical performance of new homogeneous enzymatic method reagent kit for small dense LDL cholesterol (sd LDL-C) assay hy using automatic chemistry analyzers capahle of accommodating two reagent assays. Methods Based on CSLI EP documents and other literatures,the sensitivity,precision,linear range,and cross-contami-native rate of reagent kit were analyzed. The accuracy of new homogeneous method was evaluated hy comparing with the density gradient ultracentrifugation method (DGUC). Results All kind of precisions and specificity of new homogeneous method were demonstrated to he good. The sensitivity was 0. 048 mmol/L. The total precisions of low value sample and high value sample were 8. 69% and 4. 64%. There was a good relativity hetween the results of homogeneous method and DGUC (Y=l. 065 4X-1. 8354). And the upper range of linear was 2. 634 mmol/L. All these parameters met the requirements of the manual of reagent kit and clinical applications. Conclusion All performances of new homogenous reagent kit were good for sd LDL-C assay. This new homogenous test is a direct method for the measurement of sd LDL-C and could he used in clinical routine laboratories.%目的 评价小而密低密度脂蛋白胆固醇(sd LDL-C)均相酶法检测试剂盒的性能.方法参照CLSI EP文件及其它文献,评估新型sd LDL-C均相酶法液态双试剂检测试剂盒的灵敏度、精密度、线性范围、抗干扰能力和携带污染率,并以密度梯度超速离心法(DUGC)为参比方法,评价新型sd LDL-C均相酶法液态试剂盒的准确度.结果均相法各项精密度及特异度良好,灵敏度可达0.048 mmol/L,低值样本和高值样本的总精密度分别为8.69%和4.64%,测定结果与DUGC法相关性良好(Y=1.065 4X-1.8354),线性范围上限可达2.634 mmol/L,可以抵抗临床中常见的干扰现象,符合试剂说明书的参数和临床应用的要求.结论新型sd LDL-C均相酶法检测试剂盒各项性能良好,是

  1. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  2. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of internationa and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Insti-

  3. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation, operating at the interface between theory and practice and between policy and implementation.The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  4. About KIT

    Institute of Scientific and Technical Information of China (English)


    The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  5. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation. The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  6. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of internationaland intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Institute contributes

  7. About KIT

    Institute of Scientific and Technical Information of China (English)


    The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  8. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute ( KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation. The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  9. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute (KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of internationaland intercultural cooperation,operating at the interface between theory and practice and between policy and implementation. The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  10. About KIT

    Institute of Scientific and Technical Information of China (English)


    The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of internationaland intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Insti-tutecontributestosustainabledevelopment,poverty alleviationandculturalpreservationand exchange.

  11. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The

  12. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute(KIT)in Amsterdam is an independent centre of knowledge and expertise in the areas of international and intercultural cooperation,operating at the interface between theory and practice and between policy and implementation.The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  13. About KIT

    Institute of Scientific and Technical Information of China (English)


    <正>The Royal Tropical Institute (KIT) in Amsterdam is an independent centre of knowledge and expertise in the areas of interna-tional and intercultural cooperation,operating at the interface between theory and practice and between policy and implementa-tion. The Institute contributes to sustainable development,poverty alleviation and cultural preservation and exchange.

  14. Analysis of KIT expression and KIT exon 11 mutations in canine oral malignant melanomas. (United States)

    Murakami, A; Mori, T; Sakai, H; Murakami, M; Yanai, T; Hoshino, Y; Maruo, K


    KIT, a transmembrane receptor tyrosine kinase, is one of the specific targets for anti-cancer therapy. In humans, its expression and mutations have been identified in malignant melanomas and therapies using molecular-targeted agents have been promising in these tumours. As human malignant melanoma, canine malignant melanoma is a fatal disease with metastases and the poor response has been observed with all standard protocols. In our study, KIT expression and exon 11 mutations in dogs with histologically confirmed malignant oral melanomas were evaluated. Although 20 of 39 cases were positive for KIT protein, there was no significant difference between KIT expression and overall survival. Moreover, polymerase chain reaction amplification and sequencing of KIT exon 11 in 17 samples did not detect any mutations and proved disappointing. For several reasons, however, KIT expression and mutations of various exons including exon 11 should be investigated in more cases.

  15. Preparation of lyophilized kit of HYNIC-[Tyr3]-Octreotate and labelling studies with 99m-Technetium; Preparo do reagente liofilizado HYNIC-[Ty3{sup 3}]-Octreotato e estudo de marcacao com Tecnecio-99m

    Energy Technology Data Exchange (ETDEWEB)

    Melo, Ivani Bortoleti


    The development of radiolabeled molecules with high specificity for an organ ar tumor has been contributed to the precise diagnostic in nuclear medicine. Somatostatin labeled derivatives constitutes a particular example of labeled peptide applied in the localization of neuroendocrine tumors. Nowadays, the {sup 111}In DTPA-octreotide is the radiopharmaceutical applied in diagnostic procedures for the visualization of tumors with high expression of somatostatin receptors. However, the 111-indium is a radionuclide that presents some limitations related to availability (cyclotron production), half-life (67 hours) and the emission of medium energy photons (171 keV e 245 keV), not favorable to the acquisition of images in SPECT (Single Photon Emission Computed Tomography). The favorable physical properties of the {sup 99m}-technetium ({sup 99m}Tc) make this radionuclide the more favorable to substitute the 111-indium on peptide labeling procedures. This work studied the preparation and labeling of a lyophilized kit of HYNIC-Tyr{sup 3}-octreotate (HYNIC-octreotate) with {sup 99m}Tc, base on previously described procedures and using tricine and EDDA (ethylendiaminediacetic acid) as coligands. It was studied the labeling parameters (incubation time, temperature, volume and perthecnetate activity) and the stability of the lyophilized preparation. Additionally, it was studied the influence of the pre-freezing using liquid nitrogen in the stability of the lyophilized preparation, as well as the influence of manitol in the labeling yield and biological distribution of the complex. The stability studies showed that the lyophilization using liquid nitrogen pre-freezing resulted in a lyophilized preparation with stability over 4 month when stored under refrigeration. The stability of the lyophilized preparation obtained without liquid nitrogen pre-freezing was similar.The labeling studies determined the best labeling conditions, resulting in a radiochemical yield superior than 90

  16. Comparing viral metagenomics methods using a highly multiplexed human viral pathogens reagent. (United States)

    Li, Linlin; Deng, Xutao; Mee, Edward T; Collot-Teixeira, Sophie; Anderson, Rob; Schepelmann, Silke; Minor, Philip D; Delwart, Eric


    Unbiased metagenomic sequencing holds significant potential as a diagnostic tool for the simultaneous detection of any previously genetically described viral nucleic acids in clinical samples. Viral genome sequences can also inform on likely phenotypes including drug susceptibility or neutralization serotypes. In this study, different variables of the laboratory methods often used to generate viral metagenomics libraries were compared for their abilities to detect multiple viruses and generate full genome coverage. A biological reagent consisting of 25 different human RNA and DNA viral pathogens was used to estimate the effect of filtration and nuclease digestion, DNA/RNA extraction methods, pre-amplification and the use of different library preparation kits on the detection of viral nucleic acids. Filtration and nuclease treatment led to slight decreases in the percentage of viral sequence reads and number of viruses detected. For nucleic acid extractions silica spin columns improved viral sequence recovery relative to magnetic beads and Trizol extraction. Pre-amplification using random RT-PCR while generating more viral sequence reads resulted in detection of fewer viruses, more overlapping sequences, and lower genome coverage. The ScriptSeq library preparation method retrieved more viruses and a greater fraction of their genomes than the TruSeq and Nextera methods. Viral metagenomics sequencing was able to simultaneously detect up to 22 different viruses in the biological reagent analyzed including all those detected by qPCR. Further optimization will be required for the detection of viruses in biologically more complex samples such as tissues, blood, or feces.

  17. [Automated RNA amplification for the rapid identification of Mycobacterium tuberculosis complex in respiratory specimens]. (United States)

    Drouillon, V; Houriez, F; Buze, M; Lagrange, P; Herrmann, J-L


    Rapid and sensitive detection of Mycobacterium tuberculosis complex (MTB) directly on clinical respiratory specimens is essential for a correct management of patients suspected of tuberculosis. For this purpose PCR-based kits are available to detect MTB in respiratory specimen but most of them need at least 4 hours to be completed. New methods, based on TRC method (TRC: Transcription Reverse transcription Concerted--TRCRapid M. Tuberculosis--Tosoh Bioscience, Tokyo, Japon) and dedicated monitor have been developed. A new kit (TRC Rapid M. tuberculosis and Real-time monitor TRCRapid-160, Tosoh Corporation, Japan) enabling one step amplification and real-time detection of MTB 16S rRNA by a combination of intercalative dye oxazole yellow-linked DNA probe and isothermal RNA amplification directly on respiratory specimens has been tested in our laboratory. 319 respiratory specimens were tested in this preliminary study and results were compared to smear and culture. Fourteen had a positive culture for MTB. Among theses samples, smear was positive in 11 cases (78.6%) and TRC process was positive in 8 cases (57.1%). Overall sensitivity of TRC compared to smear positive samples is 73%. Theses first results demonstrated that a rapid identification of MTB was possible (less than 2 processing hours for 14 specimens and about 1 hour for 1 specimen) in most cases of smear positive samples using ready to use reagents for real time detection of MTB rRNA in clinical samples. New pretreatment and extraction reagents kits to increase the stability of the sputum RNA and the extraction efficiency are now tested in our laboratory.

  18. Handling Pyrophoric Reagents

    Energy Technology Data Exchange (ETDEWEB)

    Alnajjar, Mikhail S.; Haynie, Todd O.


    Pyrophoric reagents are extremely hazardous. Special handling techniques are required to prevent contact with air and the resulting fire. This document provides several methods for working with pyrophoric reagents outside of an inert atmosphere.

  19. Current Developments in Prokaryotic Single Cell Whole Genome Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Goudeau, Danielle; Nath, Nandita; Ciobanu, Doina; Cheng, Jan-Fang; Malmstrom, Rex


    Our approach to prokaryotic single-cell Whole Genome Amplification at the JGI continues to evolve. To increase both the quality and number of single-cell genomes produced, we explore all aspects of the process from cell sorting to sequencing. For example, we now utilize specialized reagents, acoustic liquid handling, and reduced reaction volumes eliminate non-target DNA contamination in WGA reactions. More specifically, we use a cleaner commercial WGA kit from Qiagen that employs a UV decontamination procedure initially developed at the JGI, and we use the Labcyte Echo for tip-less liquid transfer to set up 2uL reactions. Acoustic liquid handling also dramatically reduces reagent costs. In addition, we are exploring new cell lysis methods including treatment with Proteinase K, lysozyme, and other detergents, in order to complement standard alkaline lysis and allow for more efficient disruption of a wider range of cells. Incomplete lysis represents a major hurdle for WGA on some environmental samples, especially rhizosphere, peatland, and other soils. Finding effective lysis strategies that are also compatible with WGA is challenging, and we are currently assessing the impact of various strategies on genome recovery.

  20. Levitation Kits Demonstrate Superconductivity. (United States)

    Worthy, Ward


    Describes the "Project 1-2-3" levitation kit used to demonstrate superconductivity. Summarizes the materials included in the kit. Discusses the effect demonstrated and gives details on how to obtain kits. Gives an overview of the documentation that is included. (CW)

  1. Use of the Protease Fluorescent Detection Kit to Determine Protease Activity


    Cupp-Enyard, Carrie


    The Protease Fluorescent Detection Kit provides ready-to-use reagents for detecting the presence of protease activity. This simple assay to detect protease activity uses casein labeled with fluorescein isothiocyanate (FITC) as the substrate.

  2. 21 CFR 864.1860 - Immunohistochemistry reagents and kits. (United States)


    ..., by immunological techniques, antigens in tissues or cytologic specimens. Similar devices intended for... hormone receptors in breast cancer. (3) Class III (premarket approval). IHC's intended for any use...

  3. Miniaturized isothermal nucleic acid amplification, a review. (United States)

    Asiello, Peter J; Baeumner, Antje J


    Micro-Total Analysis Systems (µTAS) for use in on-site rapid detection of DNA or RNA are increasingly being developed. Here, amplification of the target sequence is key to increasing sensitivity, enabling single-cell and few-copy nucleic acid detection. The several advantages to miniaturizing amplification reactions and coupling them with sample preparation and detection on the same chip are well known and include fewer manual steps, preventing contamination, and significantly reducing the volume of expensive reagents. To-date, the majority of miniaturized systems for nucleic acid analysis have used the polymerase chain reaction (PCR) for amplification and those systems are covered in previous reviews. This review provides a thorough overview of miniaturized analysis systems using alternatives to PCR, specifically isothermal amplification reactions. With no need for thermal cycling, isothermal microsystems can be designed to be simple and low-energy consuming and therefore may outperform PCR in portable, battery-operated detection systems in the future. The main isothermal methods as miniaturized systems reviewed here include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), and strand displacement amplification (SDA). Also, important design criteria for the miniaturized devices are discussed. Finally, the potential of miniaturization of some new isothermal methods such as the exponential amplification reaction (EXPAR), isothermal and chimeric primer-initiated amplification of nucleic acids (ICANs), signal-mediated amplification of RNA technology (SMART) and others is presented.

  4. Evaluation of a PCR kit for the detection of Erwinia carotovora subsp. atroseptica on potato tubers

    NARCIS (Netherlands)

    Frechon, D.; Exbrayat, P.; Helias, V.; Hyman, L.J.; Jouan, B.; Llop, P.; Lopez, M.M.; Payet, N.; Perombelon, M.C.M.; Toth, I.K.; Beckhoven, van J.R.C.M.; Wolf, van der J.M.; Bertheau, Y.


    A PCR-based kit, Probelia(TM), for the detection of Erwinia carotovora subsp. atroseptica (Eca) on potatoes was evaluated at five laboratories in four countries. The kit is based on DNA-specific PCR amplification followed by detection of amplicons by hybridization to a peroxidase-labelled DNA probe

  5. Evaluation and comparative analysis of direct amplification of STRs using PowerPlex® 18D and Identifiler® Direct systems. (United States)

    Myers, Blake A; King, Jonathan L; Budowle, Bruce


    height ratios for both kits were well balanced with peaks ranging in height >2000 RFUs to those with one or more peaks with heights Direct was an indication of a poor injection. Re-injection effectively overcame the effect manifested by a sloping ILS phenomenon. A subset of samples were subjected to direct amplification using the reagents in Identifiler(®) Plus kit and successful typing results were obtained for the majority of samples. However, the profiles displayed increased amounts of non-adenylated products. The results of this study demonstrate that PP18D and ID Direct are both robust kits for direct amplification. The interpretation guidelines used for this study can form a basis for internal validation studies by databasing laboratories.

  6. Recommendations for adaptation and validation of commercial kits for biomarker quantification in drug development. (United States)

    Khan, Masood U; Bowsher, Ronald R; Cameron, Mark; Devanarayan, Viswanath; Keller, Steve; King, Lindsay; Lee, Jean; Morimoto, Alyssa; Rhyne, Paul; Stephen, Laurie; Wu, Yuling; Wyant, Timothy; Lachno, D Richard


    Increasingly, commercial immunoassay kits are used to support drug discovery and development. Longitudinally consistent kit performance is crucial, but the degree to which kits and reagents are characterized by manufacturers is not standardized, nor are the approaches by users to adapt them and evaluate their performance through validation prior to use. These factors can negatively impact data quality. This paper offers a systematic approach to assessment, method adaptation and validation of commercial immunoassay kits for quantification of biomarkers in drug development, expanding upon previous publications and guidance. These recommendations aim to standardize and harmonize user practices, contributing to reliable biomarker data from commercial immunoassays, thus, enabling properly informed decisions during drug development.

  7. World Disarmament Kit. (United States)

    Woito, Robert, Ed.

    This kit presents a comprehensive introduction for students to arms control and disarmament issues. Included are copies of published and unpublished articles for each topic. Section I provides a self-survey to enable students to assess their own attitudes, values, and knowledge. The survey poses questions for which students select one of several…

  8. ProjectorKit

    DEFF Research Database (Denmark)

    Weigel, Martin; Boring, Sebastian; Steimle, Jürgen


    Researchers have developed interaction concepts based on mobile projectors. Yet pursuing work in this area - particularly in building projector-based interactions techniques within an application - is cumbersome and time-consuming. To mitigate this problem, we contribute ProjectorKit, a flexible ...... open-source toolkit that eases rapid prototyping mobile projector interaction techniques....

  9. Vietnamese Culture Kit. (United States)

    Nguyen, Liem Thanh

    This booklet provides a brief description of the cultural background of the Vietnamese, the geography of the country of Vietnam, the history of the Vietnamese people, their language, beliefs, systems of values, religions, customs, feasts, and holidays. The kit is designed to provide American sponsors and teachers with meaningful information about…

  10. 49 CFR 173.161 - Chemical kits and first aid kits. (United States)


    ... 49 Transportation 2 2010-10-01 2010-10-01 false Chemical kits and first aid kits. 173.161 Section... Class 7 § 173.161 Chemical kits and first aid kits. (a) Chemical kits and First aid kits must conform to... 10 kg. (b) Chemical kits and First aid kits are excepted from the specification...

  11. ISS Expedition 28 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 28 from 04/2011-11/2011. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  12. ISS Expedition 40 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 40 from 03/2014-11/2014. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  13. ISS Expedition 31 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 31 from 12/2011-07/2012. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  14. ISS Expedition 17 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 17 from 04/2008-10/2008. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  15. ISS Expedition 41 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 41 from 05/2014-11/2014. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  16. ISS Expedition 09 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 09 from 04/2004-10/2004. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  17. ISS Expedition 05 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 05 from 06/2002-12/2002. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  18. ISS Expedition 10 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 10 from 10/2004-04/2005. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  19. ISS Expedition 02 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 02 from 03/2001-08/2001. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  20. ISS Expedition 37 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 37 from 05/2013-11/2013. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  1. ISS Expedition 23 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 23 from 12/2009-09/2010. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  2. ISS Expedition 24 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 24 from 04/2010-11/2010. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  3. ISS Expedition 42 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 42 from 09/2014-03/2015. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  4. ISS Expedition 34 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 34 from 12/2012-03/2013. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  5. ISS Expedition 26 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 26 from 10/2010-05/2011. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  6. ISS Expedition 16 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 16 from 10/2007-04/2008. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  7. ISS Expedition 03 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 03 from 08/2001-12/2001. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  8. ISS Expedition 32 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 32 from 05/2012-09/2012. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  9. ISS Expedition 06 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 06 from 11/2002-05/2003. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  10. ISS Expedition 11 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 11 from 04/2005-10/2005. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  11. ISS Expedition 35 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 35 from 03/2013-09/2013. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  12. ISS Expedition 38 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 38 from 09/2013-03/2014. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  13. ISS Expedition 20 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 20 from 05/2009-10/2009. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  14. ISS Expedition 08 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 08 from 10/2003-04/2004. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  15. ISS Expedition 30 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 30 from 11/2011-07/2012. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  16. ISS Expedition 39 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 39 from 11/2013-05/2014. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  17. ISS Expedition 18 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 18 from 10/2008-04/2009. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  18. ISS Expedition 01 Press Kit (United States)

    National Aeronautics and Space Administration — Press kit for ISS mission Expedition 01 from 10/2000-03/2001. Press kits contain information about each mission overview, crew, mission timeline, benefits, and media...

  19. Comparison of three DNA extraction kits to establish maximum yield and quality of coral-associated microbial DNA (United States)

    Baker, Erin J.; Kellogg, Christina A.


    Coral microbiology is an expanding field, yet there is no standard DNA extraction protocol. Although many researchers depend on commercial extraction kits, no specific kit has been optimized for use with coral samples. Both soil and plant DNA extraction kits from MO BIO Laboratories, Inc., have been used by many research groups for this purpose. MO BIO recently replaced their PowerPlant® kit with an improved PowerPlantPro kit, but it was unclear how these changes would affect the kit’s use with coral samples. In order to determine which kit produced the best results, we conducted a comparison between the original PowerPlant kit, the new PowerPlantPro kit, and an alternative kit, PowerSoil, using samples from several different coral genera. The PowerPlantPro kit had the highest DNA yields, but the lack of 16S rRNA gene amplification in many samples suggests that much of the yield may be coral DNA rather than microbial DNA. The most consistent positive amplifications came from the PowerSoil kit.


    Energy Technology Data Exchange (ETDEWEB)

    Susan S. Sorini; John F. Schabron; Joseph F. Rovani, Jr.


    Western Research Institute (WRI) has developed a new commercial product ready for technology transfer, the Diesel Dog{reg_sign} Portable Soil Test Kit, for performing analysis of fuel-contaminated soils in the field. The technology consists of a method developed by WRI (U.S. Patents 5,561,065 and 5,976,883) and hardware developed by WRI that allows the method to be performed in the field (patent pending). The method is very simple and does not require the use of highly toxic reagents. The aromatic components in a soil extract are measured by absorption at 254 nm with a field-portable photometer. WRI added significant value to the technology by taking the method through the American Society for Testing and Materials (ASTM) approval and validation processes. The method is designated as ASTM Method D 5831-96, Standard Test Method for Screening Fuels in Soils. This ASTM designation allows the method to be used for federal compliance activities. In June 2001, the Diesel Dog technology won an American Chemical Society Regional Industrial Innovations Award. To gain field experience with the new technology, Diesel Dog kits have been used for a variety of site evaluation and cleanup activities. Information gained from these activities has led to improvements in hardware configurations and additional insight into correlating Diesel Dog results with results from laboratory methods. The Wyoming Department of Environmental Quality (DEQ) used Diesel Dog Soil Test Kits to guide cleanups at a variety of sites throughout the state. ENSR, of Acton, Massachusetts, used a Diesel Dog Portable Soil Test Kit to evaluate sites in the Virgin Islands and Georgia. ChemTrack and the U.S. Army Corps of Engineers successfully used a test kit to guide excavation at an abandoned FAA fuel-contaminated site near Fairbanks, Alaska. Barenco, Inc. is using a Diesel Dog Portable Soil Test Kit for site evaluations in Canada. A small spill of diesel fuel was cleaned up in Laramie, Wyoming using a Diesel

  1. An evaluation of multiple annealing and looping based genome amplification using a synthetic bacterial community

    Institute of Scientific and Technical Information of China (English)

    WANG Yong; GAO Zhaoming; XU Ying; LI Guangyu; HE Lisheng; QIAN Peiyuan


    The low biomass in environmental samples is a major challenge for microbial metagenomic studies. The amplification of a genomic DNA was frequently applied to meeting the minimum requirement of the DNA for a high-throughput next-generation-sequencing technology. Using a synthetic bacterial community, the amplification efficiency of the Multiple Annealing and Looping Based Amplification Cycles (MALBAC) kit that is originally developed to amplify the single-cell genomic DNA of mammalian organisms is examined. The DNA template of 10 pg in each reaction of the MALBAC amplification may generate enough DNA for Illumina sequencing. Using 10 pg and 100 pg templates for each reaction set, the MALBAC kit shows a stable and homogeneous amplification as indicated by the highly consistent coverage of the reads from the two amplified samples on the contigs assembled by the original unamplified sample. Although GenomePlex whole genome amplification kit allows one to generate enough DNA using 100 pg of template in each reaction, the minority of the mixed bacterial species is not linearly amplified. For both of the kits, the GC-rich regions of the genomic DNA are not efficiently amplified as suggested by the low coverage of the contigs with the high GC content. The high efficiency of the MALBAC kit is supported for the amplification of environmental microbial DNA samples, and the concerns on its application are also raised to bacterial species with the high GC content.

  2. Development of an efficient targeted cell-SELEX procedure for DNA aptamer reagents.

    Directory of Open Access Journals (Sweden)

    Susanne Meyer

    Full Text Available BACKGROUND: DNA aptamers generated by cell-SELEX offer an attractive alternative to antibodies, but generating aptamers to specific, known membrane protein targets has proven challenging, and has severely limited the use of aptamers as affinity reagents for cell identification and purification. METHODOLOGY: We modified the BJAB lymphoblastoma cell line to over-express the murine c-kit cell surface receptor. After six rounds of cell-SELEX, high-throughput sequencing and bioinformatics analysis, we identified aptamers that bound BJAB cells expressing c-kit but not wild-type BJAB controls. One of these aptamers also recognizes c-kit endogenously expressed by a mast cell line or hematopoietic progenitor cells, and specifically blocks binding of the c-kit ligand stem cell factor (SCF. This aptamer enables better separation by fluorescence-activated cell sorting (FACS of c-kit(+ hematopoietic progenitor cells from mixed bone marrow populations than a commercially available antibody, suggesting that this approach may be broadly useful for rapid isolation of affinity reagents suitable for purification of other specific cell types. CONCLUSIONS/SIGNIFICANCE: Here we describe a novel procedure for the efficient generation of DNA aptamers that bind to specific cell membrane proteins and can be used as high affinity reagents. We have named the procedure STACS (Specific TArget Cell-SELEX.

  3. Windows 7 resource kit

    CERN Document Server

    Northrup, Tony; Honeycutt, Jerry; Wilson, Ed


    In-depth and comprehensive, this RESOURCE KIT delivers the information you need to administer your Windows 7 system. You get authoritative technical guidance from those who know the technology best-Microsoft Most Valuable Professionals (MVPs) and the Windows 7 product team-along with essential scripts and resources. In addition, "Direct from the Source" sidebars offer deep insights and troubleshooting tips from the Windows 7 team. Get expert guidance on how to: Use Microsoft Deployment Toolkit best practices and tools. Plan user-state migration and test application compatibility.

  4. Mobile Probing Kit

    DEFF Research Database (Denmark)

    Larsen, Jakob Eg; Sørensen, Lene Tolstrup; Sørensen, J.K.


    characterized as being highly nomadic and thus potential users of mobile and ubiquitous technologies. The methodology has been applied in the 1ST MAGNET Beyond project in order to obtain user needs and requirements in the process of developing pilot services. We report on the initial findings from applying......Mobile Probing Kit is a low tech and low cost methodology for obtaining inspiration and insights into user needs, requirements and ideas in the early phases of a system's development process. The methodology is developed to identify user needs, requirements and ideas among knowledge workers...

  5. Education Payload Operation - Kit D (United States)

    Keil, Matthew


    Education Payload Operation - Kit D (EPO-Kit D) includes education items that will be used to support the live International Space Station (ISS) education downlinks and Education Payload Operation (EPO) demonstrations onboard the ISS. The main objective of EPO-Kit D supports the National Aeronautics and Space Administration (NASA) goal of attracting students to study and seek careers in science, technology, engineering, and mathematics.

  6. Complex amine-based reagents (United States)

    Suslov, S. Yu.; Kirilina, A. V.; Sergeev, I. A.; Zezyulya, T. V.; Sokolova, E. A.; Eremina, E. V.; Timofeev, N. V.


    Amines for a long time have been applied to maintaining water chemistry conditions (WCC) at power plants. However, making use of complex reagents that are the mixture of neutralizing and the filmforming amines, which may also contain other organic components, causes many disputes. This is mainly due to lack of reliable information about these components. The protective properties of any amine with regard to metal surfaces depend on several factors, which are considered in this article. The results of applying complex reagents to the protection of heating surfaces in industrial conditions and estimated behavior forecasts for various reagents under maintaining WCC on heat-recovery boilers with different thermal circuits are presented. The case of a two-drum heat-recovery boiler with in-line drums was used as an example, for which we present the calculated pH values for various brands of reagents under the same conditions. Work with different reagent brands and its analysis enabled us to derive a composition best suitable for the conditions of their practical applications in heat-recovery boilers at different pressures. Testing the new amine reagent performed at a CCPP power unit shows that this reagent is an adequate base for further development of reagents based on amine compounds. An example of testing a complex reagent is shown created with the participation of the authors within the framework the program of import substitution and its possible use is demonstrated for maintaining WCC of power-generating units of combined-cycle power plants (CCPP) and TPP. The compliance of the employed reagents with the standards of water chemistry conditions and protection of heating surfaces were assessed. The application of amine-containing reagents at power-generating units of TPP makes it possible to solve complex problems aimed at ensuring the sparing cleaning of heating surfaces from deposits and the implementation of conservation and management of water chemistry condition

  7. What's In Your Emergency Kit?

    Centers for Disease Control (CDC) Podcasts


    An emergency kit can help you survive during a disaster. This podcast discusses supplies to include in your kit.  Created: 12/4/2012 by Office of Public Health Preparedness and Response (OPHPR).   Date Released: 12/20/2012.

  8. Cosmic Light EDU kit (United States)

    Doran, Rosa


    In 2015 we celebrate the International Year of Light, a great opportunity to promote awareness about the importance of light coming from the Cosmos and what messages it is bringing to mankind. In parallel a unique moment to attract the attention of stakeholders on the dangers of light pollution and its impact in our lives and our pursuit of more knowledge. In this presentation I want to present one of the conrnerstones of IYL2015, a partnership between the Galileo Teacher Training Program, Universe Awareness and Globe at Night, the Cosmic Light EDU kit. The aim of this project is to assemble a core set of tools and resources representing our basic knowledge pilars about the Universe and simple means to preserve our night sky.

  9. Comparison between NuGEN's WT-Ovation Pico and one-direct amplification systems. (United States)

    Morse, Alison M; Carballo, Valentina; Baldwin, Donald A; Taylor, Christopher G; McIntyre, Lauren M


    Differential gene expression between groups of homogenous cell types is a biological question whose time has come. RNA can be extracted from small numbers of cells, such as those isolated by laser-capture microdissection, but the small amounts obtained often require amplification to enable whole genome transcriptome profiling by technologies such as microarray analysis and RNA-seq. Recently, advances in amplification procedures make amplification directly from whole cell lysates possible. The aim of this study was to compare two amplification systems for variations in observed RNA abundance attributable to the amplification procedure for use with small quantities of cells isolated by laser-capture microdissection. Arabidopsis root cells undergoing giant cell formation as a result of nematode infestation and uninfested control root cells were laser-captured and used to evaluate two amplification systems. One, NuGEN's WT-Ovation Pico (Pico) amplification system, uses total RNA as starting material, and the other, NuGEN's WT-One-Direct (One-Direct) amplification system, uses lysate containing the captured cells. The reproducibility of whole genome transcript profiling and correlations of both systems were investigated after microarray analysis. The One-Direct system was less reproducible and more variable than the Pico system. The Pico amplification kit resulted in the detection of thousands of differentially expressed genes between giant cells and control cells. This is in marked contrast to the relatively few genes detected after amplification with the One-Direct amplification kit.

  10. Next-Generation STR Genotyping Kits for Forensic Applications. (United States)

    Mulero, J J; Hennessy, L K


    Forensic DNA typing has been a constantly evolving field driven by innovations from academic laboratories as well as kit manufacturers. Central to these technological advances has been the transition from multilocus-probe restriction fragment length polymorphism (RFLP) methods to short tandem repeat (STR) PCR-based assays. STRs are now the markers of choice for forensic DNA typing and a wide variety of commercial STR kits have been designed to meet the various needs of a forensic lab. This review provides an overview of the commercial STR kits made available since the year 2000 and explains the rationale for creating these kits. Substantial progress has been made in key areas such as sample throughput, speed, and sensitivity. For example, a significant advancement for databasing labs was the capability of direct amplification from a blood or buccal sample without need for DNA extraction or purification, enabling increased throughput. Other key improvements are greater tolerance for inhibitors (e.g., humic acid, hematin, and tannic acid) present in evidence samples, PCR cycling times decreased by 1-1.5 h, and greater sensitivity with improved buffer components and thermal cycling conditions. These improvements that have been made over the last 11 years have enhanced the ability of forensic laboratories to obtain a DNA profile from more challenging samples. However, with the proliferation of kits from different vendors the primer binding sequences of the loci vary, which could result in discordant events that would need to be resolved either via a database-driven software solution or simply by evaluating discordant samples with multiple kits.

  11. 国内外肝炎病毒核酸定量和定性检测试剂的差异及质量控制%Comparison and quality control between domestic and foreign hepatitis virus nucleic acid amplification technology reagents for quantitative and qualitative tests

    Institute of Scientific and Technical Information of China (English)

    吴星; 周诚; 梁争论


    Hepatitis virus NAT reagents are now widely used clinically. However, the qulity of domestic and foreign NAT reagents varies dramatically. The main reasons for these differences including the manufacture technique, test principle and assay procedure were discussed in this paper and current status of the quality control of the NAT reagents were also described. Finally, it was pointed out that strengthening public supervision and laboratory internal control are very important for the quality improvement of the domestic reagents.%肝炎病毒核酸检测试剂在临床广泛应用,国内外相关试剂质量存在较大差异.本文从试剂的生产工艺、检测原理、操作过程等方面分析了国内外试剂质量存在差异的主要原因,同时阐述了国内外相关试剂质量控制的现状.提出为提高国产试剂质量,应加强上市后监管和实验室内部质量控制.

  12. Energy Management Curriculum Starter Kit

    Energy Technology Data Exchange (ETDEWEB)

    Turner, W.C.


    The Energy Management Curriculum Starter Kit was designed to help engineering educators develop and teach energy management courses. Montana State University and Oklahoma State University courses are embodied in the model curriculum given. The curricula offered at many other universities throughout the United States are also presented. The kit was designed specifically to train engineering students to be good energy managers. Courses at both the undergraduate and postgraduate level are presented.

  13. Evaluation of immobilized metal affinity chromatography kits for the purification of histidine-tagged recombinant CagA protein. (United States)

    Karakus, Cebrail; Uslu, Merve; Yazici, Duygu; Salih, Barik A


    Immobilized metal affinity chromatography (IMAC) technique is used for fast and reliable purification of histidine(His)-tagged recombinant proteins. The technique provides purification under native and denaturing conditions. The aim of this study is to evaluate three commercially available IMAC kits (Thermo Scientific, GE Healthcare and Qiagen) for the purification of a 6xHis-tagged recombinant CagA (cytotoxin-associated gene A) protein from IPTG-induced Escherichia coli BL21(DE3) culture. The kits were tested according to the manufacturer instructions and the protein was purified with only GE Healthcare and Qiagen kits under denaturing conditions. 1% (w/v) SDS was used as denaturing agent in PBS instead of extraction reagent of Thermo Scientific kit to lyse bacterial cells from 100ml culture. The 6xHis-tagged recombinant protein was purified by the three kits equally.

  14. An integrated microfluidic sensor for real-time detection of RNA in seawater using preserved reagents (United States)

    Tsaloglou, M.-N.; Loukas, C. M.; Ruano-López, J. M.; Morgan, H.; Mowlem, M. C.


    Quantitation of RNA sequences coding either for key metabolic proteins or highly conserved ribosomal subunits can provide insight on cell abundance, speciation and viability. Nucleic sequence-based amplification (NASBA) is an isothermal alternative to traditional nucleic acid amplification methods, such as quantitative PCR. We present here an integrated microfluidic sensor for cell concentration and lysis, RNA extraction/purification and quantitative RNA detection for environmental applications. The portable system uses pre-loaded reagents, stored as a gel on a disposable microfluidic cartridge, which is manufactured using low-cost injection moulding. The NASBA reaction is monitored real-time using a bespoke control unit which includes: an external fluorescence detector, three peristaltic micro-pumps, two heaters and temperature sensors, a battery, seven pin actuated micro-motors (or valve actuators), and an automatic cartridge insertion mechanism. The system has USB connectivity and none of the expensive components require replacing between reactions. Long-term storage of reagents is critically important for any diagnostic tool that will be used in the field, whether for medical or environmental analysis and has not been previously demonstrated for NASBA reagents on-chip. We have shown effective amplification, for as little as 500 cells of the toxic microalga Karenia brevis using reagents which had been preserved as a gel for 45 days. This is the first reported real-time isothermal RNA amplification using with on-chip preservation. Annealing of primers, amplification at 41 °C and real-time fluorescence detection using, also for the first time, an internal control and sequence-specific molecular beacons was all performed on our microfluidic sensor. Our results show excellent promise as a future quantitative tool of in situ phytoplankton analysis and other environmental applications, where long-term reagent storage and low power consumption is essential.

  15. Roche/BIOTECON Diagnostics LightCycler foodproof L. monocytogenes detection kit in combination with ShortPrep foodproof II Kit. Performance-Tested Method 070401. (United States)

    Junge, Benjamin; Berghof-Jäger, Kornelia


    A method was developed for the detection of L. monocytogenes in food based on real-time polymerase chain reaction (PCR). This advanced PCR method was designed to reduce the time needed to achieve results from PCR reactions and to enable the user to monitor the amplification of the PCR product simultaneously, in real-time. After DNA isolation using the Roche/BIOTECON Diagnostics ShortPrep foodproof II Kit (formerly called Listeria ShortPrep Kit) designed for the rapid preparation of L. monocytogenes DNA for direct use in PCR, the real-time detection of L. monocytogenes DNA is performed by using the Roche/BIOTECON Diagnostics LightCycler foodproof L. monocytogenes Detection Kit. This kit provides primers and hybridization probes for sequence-specific detection, convenient premixed reagents, and different controls for reliable interpretation of results. For repeatability studies, 20 different foods, covering the 15 food groups recommended from the AOAC Research Institute (AOAC RI) for L. monocytogenes detection were analyzed: raw meats, fresh produce/vegetables, processed meats, seafood, egg and egg products, dairy (cultured/noncultured), spices, dry foods, fruit/juices, uncooked pasta, nuts, confectionery, pet food, food dyes and colorings, and miscellaneous. From each food 20, samples were inoculated with a low level (1-10 colony-forming units (CFU)/25 g) and 20 samples with a high level (10-50 CFU/25 g) of L. monocytogenes. Additionally, 5 uninoculated samples were prepared from each food. The food samples were examined with the test kits and in correlation with the cultural methods according to U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) or U.S. Department of Agriculture (USDA)/Food Safety and Inspection Service (FSIS) Microbiology Laboratory Guidebook. After 48 h of incubation, the PCR method in all cases showed equal or better results than the reference cultural FDA/BAM or USDA/FSIS methods. Fifteen out of 20 tested food types

  16. An efficient multistrategy DNA decontamination procedure of PCR reagents for hypersensitive PCR applications.

    Directory of Open Access Journals (Sweden)

    Sophie Champlot

    Full Text Available BACKGROUND: PCR amplification of minute quantities of degraded DNA for ancient DNA research, forensic analyses, wildlife studies and ultrasensitive diagnostics is often hampered by contamination problems. The extent of these problems is inversely related to DNA concentration and target fragment size and concern (i sample contamination, (ii laboratory surface contamination, (iii carry-over contamination, and (iv contamination of reagents. METHODOLOGY/PRINCIPAL FINDINGS: Here we performed a quantitative evaluation of current decontamination methods for these last three sources of contamination, and developed a new procedure to eliminate contaminating DNA contained in PCR reagents. We observed that most current decontamination methods are either not efficient enough to degrade short contaminating DNA molecules, rendered inefficient by the reagents themselves, or interfere with the PCR when used at doses high enough to eliminate these molecules. We also show that efficient reagent decontamination can be achieved by using a combination of treatments adapted to different reagent categories. Our procedure involves γ- and UV-irradiation and treatment with a mutant recombinant heat-labile double-strand specific DNase from the Antarctic shrimp Pandalus borealis. Optimal performance of these treatments is achieved in narrow experimental conditions that have been precisely analyzed and defined herein. CONCLUSIONS/SIGNIFICANCE: There is not a single decontamination method valid for all possible contamination sources occurring in PCR reagents and in the molecular biology laboratory and most common decontamination methods are not efficient enough to decontaminate short DNA fragments of low concentration. We developed a versatile multistrategy decontamination procedure for PCR reagents. We demonstrate that this procedure allows efficient reagent decontamination while preserving the efficiency of PCR amplification of minute quantities of DNA.

  17. Active Parenting Now: Program Kit. (United States)

    Popkin, Michael H.

    Based largely on the theories of Alfred Adler and Rudolf Dreikurs, this parent education curriculum is a video-based interactive learning experience that teaches a comprehensive model of parenting to parents of children ages 5 to 12 years. The kit provides parents with the skills needed to help their children develop courage, responsibility, and…

  18. Natural Gas Energy Educational Kit. (United States)

    American Gas Association, Arlington, VA. Educational Services.

    Prepared by energy experts and educators to introduce middle school and high school students to natural gas and its role in our society, this kit is designed to be incorporated into existing science and social studies curricula. The materials and activities focus on the origin, discovery, production, delivery, and use of natural gas. The role of…

  19. The Free Universal Construction Kit

    DEFF Research Database (Denmark)

    Stephensen, Jan Løhmann; Hansen, Lone Koefoed


    With the increasing economic accessibility of 3D printers, the lessons learned and the logics cultivated on digital Web 2.0 now seems applicable to the world of material things. Released in early 2012 by the artist groups F.A.T. and Sy-lab, the Free Universal Construction Kit is a set of 3D...


    Directory of Open Access Journals (Sweden)

    T. Yu. Trohimchuk


    Full Text Available The problem of early diagnosis of human immunodeficiency virus remains relevant and can be solved by introduction the highly sensitive 4th generation ELISA test kits in the laboratory practice. The advantage of this analysis is simultaneous determination of serum antigen p24 HIV-1 and HIV-specific total antibody (IgG, IgM, IgA, that can detect the infection in the early stages of development. In Private Joint Stock Company «Scientific and Production Company Diaproph-Med» a modified version of the test kit of the 4th generation DIA-HIV-Ag/Ab hypersensitivity was developed, which uses a mixture of synthetic analogs of viral antigens in the immune-enzyme conjugates at two stages of reaction. Significant amplification of specific signal is achieved by introducing a streptavidin which is conjugated to horseradish peroxidase polymeric form. The analytical sensitivity of the test kit to identify the 1st International Standard WHO HIV p24 (NIBSC was 0.78 IU/ml. The ability to test kit to detect early HIV infection has been examined for 9 seroconversion panels of sera (7 production ZeptoMetrix Corp. and 2 – SeraCare Life Sciences, USA and is comparable to its peers, the leading manufacturers of test kits. The diagnostic characteristics of the test kit tested on standard panels, including dilution of serum of HIV- infected people, and not diluted sera with antibodies to HIV-1 (1 869 samples and HIV-2 ( 41 sample. In these studies, the test kit detected HIV in a large ratio OD/cut off compared to similar commercial test kits for the 4th generation. The diagnostic specificity of the test kit DIA-HIV-Ag/Ab according to the results obtained in the study of blood donors from various transfusion services in Ukraine (93 788 researches was 99.95%.

  1. Accuracy of the TRUGENE HIV-1 Genotyping Kit (United States)

    Grant, Robert M.; Kuritzkes, Daniel R.; Johnson, Victoria A.; Mellors, John W.; Sullivan, John L.; Swanstrom, Ronald; D'Aquila, Richard T.; Van Gorder, Mark; Holodniy, Mark; Lloyd, Jr., Robert M.; Reid, Caroline; Morgan, Gillian F.; Winslow, Dean L.


    Drug resistance and poor virological responses are associated with well-characterized mutations in the viral reading frames that encode the proteins that are targeted by currently available antiretroviral drugs. An integrated system was developed that includes target gene amplification, DNA sequencing chemistry (TRUGENE HIV-1 Genotyping Kit), and hardware and interpretative software (the OpenGene DNA Sequencing System) for detection of mutations in the human immunodeficiency virus type 1 (HIV-1) protease and reverse transcriptase sequences. The integrated system incorporates reverse transcription-PCR from extracted HIV-1 RNA, a coupled amplification and sequencing step (CLIP), polyacrylamide gel electrophoresis, semiautomated analysis of data, and generation of an interpretative report. To assess the accuracy and robustness of the assay system, 270 coded plasma specimens derived from nine patients were sent to six laboratories for blinded analysis. All specimens contained HIV-1 subtype B viruses. Results of 270 independent assays were compared to “gold standard” consensus sequences of the virus populations determined by sequence analysis of 16 to 20 clones of viral DNA amplicons derived from two independent PCRs using primers not used in the kit. The accuracy of the integrated system for nucleotide base identification was 98.7%, and the accuracy for codon identification at 54 sites associated with drug resistance was 97.6%. In a separate analysis of plasma spiked with infectious molecular clones, the assay reproducibly detected all 72 different drug resistance mutations that were evaluated. There were no significant differences in accuracy between laboratories, between technologists, between kit lots, or between days. This integrated assay system for the detection of HIV-1 drug resistance mutations has a high degree of accuracy and reproducibility in several laboratories. PMID:12682149

  2. Statistical assessment of DNA extraction reagent lot variability in real-time quantitative PCR (United States)

    Bushon, R.N.; Kephart, C.M.; Koltun, G.F.; Francy, D.S.; Schaefer, F. W.; Lindquist, H.D. Alan


    Aims: The aim of this study was to evaluate the variability in lots of a DNA extraction kit using real-time PCR assays for Bacillus anthracis, Francisella tularensis and Vibrio cholerae. Methods and Results: Replicate aliquots of three bacteria were processed in duplicate with three different lots of a commercial DNA extraction kit. This experiment was repeated in triplicate. Results showed that cycle threshold values were statistically different among the different lots. Conclusions: Differences in DNA extraction reagent lots were found to be a significant source of variability for qPCR results. Steps should be taken to ensure the quality and consistency of reagents. Minimally, we propose that standard curves should be constructed for each new lot of extraction reagents, so that lot-to-lot variation is accounted for in data interpretation. Significance and Impact of the Study: This study highlights the importance of evaluating variability in DNA extraction procedures, especially when different reagent lots are used. Consideration of this variability in data interpretation should be an integral part of studies investigating environmental samples with unknown concentrations of organisms. ?? 2010 The Society for Applied Microbiology.

  3. A simple, inexpensive device for nucleic acid amplification without electricity-toward instrument-free molecular diagnostics in low-resource settings.

    Directory of Open Access Journals (Sweden)

    Paul LaBarre

    Full Text Available BACKGROUND: Molecular assays targeted to nucleic acid (NA markers are becoming increasingly important to medical diagnostics. However, these are typically confined to wealthy, developed countries; or, to the national reference laboratories of developing-world countries. There are many infectious diseases that are endemic in low-resource settings (LRS where the lack of simple, instrument-free, NA diagnostic tests is a critical barrier to timely treatment. One of the primary barriers to the practicality and availability of NA assays in LRS has been the complexity and power requirements of polymerase chain reaction (PCR instrumentation (another is sample preparation. METHODOLOGY/PRINCIPAL FINDINGS: In this article, we investigate the hypothesis that an electricity-free heater based on exothermic chemical reactions and engineered phase change materials can successfully incubate isothermal NA amplification assays. We assess the heater's equivalence to commercially available PCR instruments through the characterization of the temperature profiles produced, and a minimal method comparison. Versions of the prototype for several different isothermal techniques are presented. CONCLUSIONS/SIGNIFICANCE: We demonstrate that an electricity-free heater based on exothermic chemical reactions and engineered phase change materials can successfully incubate isothermal NA amplification assays, and that the results of those assays are not significantly different from ones incubated in parallel in commercially available PCR instruments. These results clearly suggest the potential of the non-instrumented nucleic acid amplification (NINA heater for molecular diagnostics in LRS. When combined with other innovations in development that eliminate power requirements for sample preparation, cold reagent storage, and readout, the NINA heater will comprise part of a kit that should enable electricity-free NA testing for many important analytes.

  4. 21 CFR 866.4100 - Complement reagent. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Complement reagent. 866.4100 Section 866.4100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL... Complement reagent. (a) Identification. A complement reagent is a device that consists of complement,...

  5. 乙型和丙型肝炎病毒感染检测试剂的标准化:问题与对策%Problems and solutions on standardization of reagents for detection of HBV and HCV infection

    Institute of Scientific and Technical Information of China (English)



    国产HBV和HCV感染检测试剂与国外同类试剂相比,存在过度追求操作简便化和定量检测缺乏量值溯源等问题.将HBsAg的双抗体夹心ELISA试剂的"一步法"改为"两步法",并保证足够的温育时间;同时,选择多个单抗作为包被抗体,不但可以避免"钩状效应"或HBsAg变异造成的假阴性结果,而且将改善试剂的测定下限.将HBV和HCV核酸检测试剂的标本处理由简单的煮沸裂解改为核酸纯化,并增加用于核酸提取的标本量和提取后扩增加样量,同时加入内标,不但可以改善测定下限和检测重复性,而且可以有效地监控假阴性结果的出现.定量检测试剂标准品系列与国家或国际标准物质的量值溯源,可使不同试剂得到的检测结果具有可比性.%Compared with commercial reagents manufactured by foreign companies for detection of HBV and HCV infection, domestic reagents have poorer performance because of the over-pursuit of easy operation and lack of metrological traceability in quantitative measurement. If "two-step" sandwich ELISA model and multiple monoclonal coating antibodies were used, false-negative results caused by the hook-effect and HBsAg mutant would be avoided. Moreover, sufficient incubation time in each step would improve the detection-limit of the reagents. By replacing the boiling lysis with nucleic acid purification in sample preparation and increasing the sample volume of nucleic acid purification and amplification detection could improve the detection-limit and reproducibility of HBV and HCV nucleic acid testing. The use of internal control could effectively monitor the of false negative results as well Application of international or national reference materials for metrological traceability of calibrators in reagents also plays an important role in assuring result concordance among different commercial reagent kits, methods and clinical laboratories.

  6. The Seneca Amplification Construction

    Directory of Open Access Journals (Sweden)

    Wallace Chafe


    Full Text Available The polysynthetic morphology of the Northern Iroquoian languages presents a challenge to studies of clause combining. The discussion here focuses on a Seneca construction that may appear within a single clause but may also straddle clause boundaries. It amplifies the information provided by a referent, here called the trigger, that is introduced by the pronominal prefix within a verb or occasionally in some other way. The particle neh signals that further information about that referent will follow. This construction is found at four levels of syntactic complexity. At the first level the trigger and its amplification occur within the same prosodic phrase and the amplification is a noun. At the second level the amplification occurs in a separate prosodic phrase but remains a noun. At the third level the amplification exhibits verb morphology but has been lexicalized with a nominal function. At the fourth level the amplification functions as a full clause and neh serves as a marker of clause combining. Several varieties of amplification are discussed, as are cases in which the speaker judges that no amplification is needed. It is suggested that the typologically similar Caddo language illustrates a situation in which this construction could never arise, simply because Caddo verbs lack the pronominal element that triggers the construction in Seneca.

  7. KIT exon 11 codon 557/558 deletion/insertion mutations define a subset of gastrointestinal stromal tumors with malignant potential

    Institute of Scientific and Technical Information of China (English)

    Katerina Kontogianni-Katsarou; Euthimios Dimitriadis; Constantina Lariou; Evi Kairi-Vassilatou; Nikolaos Pandis; Agatha Kondi-Paphiti


    AIM: To study the association of the frequency and pattern of KIT and PDGFRA mutations and dinicopathological factors in a group of patients with gastrointestinal stromal tumors (GIST).METHODS: Thirty patients with GIST were examined. Exons 9, 11,13, and 17 of the KIT and exons 12 and 18 of the PDGFRA gene were analyzed for the presence of mutations by PCR amplification and direct sequencing.RESULTS: KIT or PDGFRA mutations were detected in 21 of the 30 patients (70%). Sixteen patients had mutations within KIT exon 11, three within KIT exon 9, and two within PDGFRA exon 18. GISTs with KIT exon 9 mutations were predominantly located in the small intestine, showed a spindle cell phenotype, and were assessed as potentially malignant. GISTs with KIT exon 11 mutations were located in the stomach and intestine, showed mainly a spindle cell phenotype, and were scored as potentially malignant (P < 0.05). Tumors with KIT exon 11 codon 557/558 deletion/insertion mutations were found to be associated with a potentially malignant clinical behaviour (P < 0.003). GISTs with PDGFRA mutations located in stomach showed a mixed cell phenotype and were classified as of very low or low moderate malignant potential.CONCLUSION: Determination of KIT and PDGFRA mutations should be additional parameters for the better prediction of GISTs clinical behaviour. Tumors with deletion/insertion mutations affecting codons 557/558 of the KIT gene seem to represent a distinct subset of malignant GISTs.

  8. Development of a PCR Diagnostic Kit for Cryptosporidium andersoni in Dairy Cow

    Institute of Scientific and Technical Information of China (English)

    ZHOU Rong-qiong; LI Guo-qing; XIAO Shu-min; LI Wei-hua


    Cryptosporidiosis is an important zoonosis caused by the Cryptosporidium species. To develop a PCR diagnostic kit for molecular detection and differential diagnosis of Cryptosporidiurn spp., a portion of ITS-1 sequence of Cryptosporidium. Andersoni was chosen as the target DNA for designing the species-specific primers (ZRQF/ZR). The kit components were determined after the PCR amplification conditions were serially optimized. A series of tests were conducted in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, respectively. The results showed that only C. Andersoni were amplified specific band of about 500 bp, while Cryptosporidium. Parvum, Cryptosporidium. Baileyi, Eimeria sp of dairy cow, Toxoplasma gondii, Eimeria sp of pig, Ascaris suum, Cyclospora sp, and E. Coli could not be amplified. 254 oocysts of C. Andersoni was the lowest number that could be detected using the kit. The kit worked well after being stored at room temperature, 4 and -20℃ for nine months. Fecal specimens, which were collected from a total of 243 calves on four different dairy farms in Guangdong Province, China, and one dairy farm in Henan Province, China, were examined using the kit; the positive rate of the kit was 2-13% higher than that of the routine methods. The results indicated that the kit can detect fecal samples faster, more sensitively, and conveniently, and can provide a useful tool for the identification and differentiation of C. Andersoni from the other Cryptosporidium species; it also has implications for further studies on molecular epidemiology and differential diagnostics of cryptosporidiosis in animals.

  9. Amplification of NOON States

    CERN Document Server

    Agarwal, G S; Rai, Amit


    We examine the behavior of a Non Gaussian state like NOON state under phase insensitive amplification. We derive analytical result for the density matrix of the NOON state for arbitrary gain of the amplifier. We consider cases of both symmetric and antisymmetric amplification of the two modes of the NOON state. We quantitatively evaluate the loss of entanglement by the amplifier in terms of the logarithmic negativity parameter. We find that NOON states are more robust than their Gaussian counterparts.

  10. Amplification of NOON States



    We examine the behavior of a Non Gaussian state like NOON state under phase insensitive amplification. We derive analytical result for the density matrix of the NOON state for arbitrary gain of the amplifier. We consider cases of both symmetric and antisymmetric amplification of the two modes of the NOON state. We quantitatively evaluate the loss of entanglement by the amplifier in terms of the logarithmic negativity parameter. We find that NOON states are more robust than their Gaussian coun...

  11. Evaluation of a commercial loop-mediated isothermal amplification assay for diagnosis of Bordetella pertussis infection. (United States)

    Kamachi, Kazunari; Moriuchi, Takumi; Hiramatsu, Yukihiro; Otsuka, Nao; Shibayama, Keigo


    We evaluated a commercial loop-mediated isothermal amplification (LAMP) assay kit for Bordetella pertussis detection. The LAMP primers were designed to target the ptxP1 allele of the pertussis toxin promoter, but the assay could detect B. pertussis ptxP3 and ptxP8 strains in addition to ptxP1 strains, with high analytical sensitivity.

  12. Genetic engineering approach to develop next-generation reagents for endotoxin quantification (United States)

    Ogura, Norihiko; Aketagawa, Jun; Aizawa, Maki; Kobayashi, Yuki; Kawabata, Shun-ichiro; Oda, Toshio


    The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The standard curve of the RCR containing mammalian cell-derived recombinant factor C had a steeper slope than the curves for those containing natural lysate reagents, suggesting a greater sensitivity to endotoxin. The present study supports the future production of recombinant reagents that do not require the use of natural resources. PMID:27913792

  13. Business Plans Kit For Dummies

    CERN Document Server

    Barrow, Colin


    An interactive guide to every element of the business planning processAre you an entrepreneur who wants to make it big? A manager grappling with logistics? A small business owner trying to map out a future in shaky economic times? Then look no further! Business Plans Kit For Dummies is a nuts-and-bolts guide to evaluating and invigorating your commercial strategy, no matter how big or small your business.Lay the foundations - identify your long-term goals and brainstorm your business options Develop the skills - sketch out a working model, size up competitors and fire up your marketing machine

  14. Gene amplification in carcinogenesis

    Directory of Open Access Journals (Sweden)

    Lucimari Bizari


    Full Text Available Gene amplification increases the number of genes in a genome and can give rise to karyotype abnormalities called double minutes (DM and homogeneously staining regions (HSR, both of which have been widely observed in human tumors but are also known to play a major role during embryonic development due to the fact that they are responsible for the programmed increase of gene expression. The etiology of gene amplification during carcinogenesis is not yet completely understood but can be considered a result of genetic instability. Gene amplification leads to an increase in protein expression and provides a selective advantage during cell growth. Oncogenes such as CCND1, c-MET, c-MYC, ERBB2, EGFR and MDM2 are amplified in human tumors and can be associated with increased expression of their respective proteins or not. In general, gene amplification is associated with more aggressive tumors, metastases, resistance to chemotherapy and a decrease in the period during which the patient stays free of the disease. This review discusses the major role of gene amplification in the progression of carcinomas, formation of genetic markers and as possible therapeutic targets for the development of drugs for the treatment of some types of tumors.

  15. 恒温扩增-防污染核酸试纸条在快速检测结核分枝杆菌中的应用%Application of Mycobacterium tuberculosis isothermal amplification-DNA strip diagnostic kit for rapid detection of the Mycobacterium tuberculosis patients

    Institute of Scientific and Technical Information of China (English)

    武洁; 梅建; 沈鑫; 张阳奕; 桂晓虹; 李静; 江渊; 王莉莉; 方仁东; 高谦


    @@ 结核分枝杆菌生长缓慢的特性一直以来限制了临床对患者的诊断及治疗.为了快速准确的鉴定结核分枝杆菌,为临床诊断提供有效依据,基于分子生物学技术的诊断方法越来越多的应用到日常的检测工作中,核酸扩增试验(nucleic acid amplification tests,NAAT)自应用到结核病的早期诊断以来,就显示出其强大的优越性.它比培养的速度快,特异性高,可以在3~6 h得到结果[1].商业化的检测试剂盒更是让早期快速诊断结核病成为可能[2-3].高通量的自动检测装置虽然提高了工作效率,但是高昂的价格和实验室配套设备的要求限制了方法的使用范围.为更好的控制结核病的蔓延,临床迫切需求一种简便、快速、准确的结核病早期诊断方法.

  16. KIT mutation analysis in mast cell neoplasms

    DEFF Research Database (Denmark)

    Arock, M; Sotlar, K; Akin, C;


    Although acquired mutations in KIT are commonly detected in various categories of mastocytosis, the methodologies applied to detect and quantify the mutant type and allele burden in various cells and tissues are poorly defined. We here propose a consensus on methodologies used to detect KIT...

  17. Culture Kits for the Elementary Classroom. (United States)

    Hickey, M. Gail


    Outlines an instructional unit where students construct culture kits illustrating a specific culture. Culture kits are constructed out of realia and other material including maps, travel brochures, photographs, newspapers, souvenirs, and other items. Discusses collecting these items and possible multicultural applications. (MJP)

  18. Podcasting the Ultimate Starter Kit

    CERN Document Server

    Shipside, Steve


    Podcasting doesn't require an iPod; anyone with a computer, an MP3 player, or in some cases even a phone or a pair of shades can play podcasts. It requires very little technological know-how to set up, listen to, or even make your own programmes. Podcasting: The ultimate starter kit takes a light-hearted, friendly and refreshingly jargon-free look at eveything you need to know to get started, and with its free start-up CD it couldn't be easier. With the help of Podcasting, you can find out how to set up your software and record podcasts, where to go to find programmes on anything from religion

  19. Improved rapid amplification of cDNA ends (RACE) for mapping both the 5' and 3' terminal sequences of paramyxovirus genomes. (United States)

    Li, Zhuo; Yu, Meng; Zhang, Hong; Wang, Hai-Yan; Wang, Lin-Fa


    Rapid amplification of cDNA ends (RACE) is a powerful PCR-based technique for determination of RNA terminal sequences. However, most of the RACE methods reported in the literature are developed specifically for the mapping of eukaryotic transcripts with 3' poly-A tail and 5' cap structure. In this study, an improved RACE strategy was developed which allows both 5' and 3' RACE of paramyxovirus genomic RNA using the same set of common molecular biology reagents without having to rely on expensive RACE kits. Mapping of RNA genome terminal sequences is an essential part of characterizing novel paramyxoviruses since these sequences contain important signals for genome replication and transcription, and are important molecular markers for studying virus evolution. The usefulness of this strategy was demonstrated by rapid characterization of both genome ends for a novel paramyxovirus recently isolated from human kidney primary cells. The RACE strategy described in this paper is simple, cost-effective and can be used to map genome ends of any RNA viruses.

  20. Biomaterials in light amplification (United States)

    Mysliwiec, Jaroslaw; Cyprych, Konrad; Sznitko, Lech; Miniewicz, Andrzej


    Biologically produced or inspired materials can serve as optical gain media, i.e. they can exhibit the phenomenon of light amplification. Some of these materials, under suitable dye-doping and optical pumping conditions, show lasing phenomena. The emerging branch of research focused on obtaining lasing action in highly disordered and highly light scattering materials, i.e. research on random lasing, is perfectly suited for biological materials. The use of biomaterials in light amplification has been extensively reported in the literature. In this review we attempt to report on progress in the development of biologically derived systems able to show the phenomena of light amplification and random lasing together with the contribution of our group to this field. The rich world of biopolymers modified with molecular aggregates and nanocrystals, and self-organized at the nanoscale, offers a multitude of possibilities for tailoring luminescent and light scattering properties that are not easily replicated in conventional organic or inorganic materials. Of particular importance and interest are light amplification and lasing, or random lasing studies in biological cells and tissues. In this review we will describe nucleic acids and their complexes employed as gain media due to their favorable optical properties and ease of manipulation. We will report on research conducted on various biomaterials showing structural analogy to nucleic acids such as fluorescent proteins, gelatins in which the first distributed feedback laser was realized, and also amyloids or silks, which, due to their dye-doped fiber-like structure, allow for light amplification. Other materials that were investigated in that respect include polysaccharides, like starch exhibiting favorable photostability in comparison to other biomaterials, and chitosan, which forms photonic crystals or cellulose. Light amplification and random lasing was not only observed in processed biomaterials but also in living

  1. False negative results from using common PCR reagents

    Directory of Open Access Journals (Sweden)

    Atwood Allison A


    Full Text Available Abstract Background The sensitivity of the PCR reaction makes it ideal for use when identifying potentially novel viral infections in human disease. Unfortunately, this same sensitivity also leaves this popular technique open to potential contamination with previously amplified PCR products, or "carry-over" contamination. PCR product carry-over contamination can be prevented with uracil-DNA-glycosylase (UNG, and it is for this reason that it is commonly included in many commercial PCR master-mixes. While testing the sensitivity of PCR assays to detect murine DNA contamination in human tissue samples, we inadvertently discovered that the use of this common PCR reagent may lead to the production of false-negative PCR results. Findings We show here that contamination with minute quantities of UNG-digested PCR product or any negative control PCR reactions containing primer-dimers regardless of UNG presence can completely block amplification from as much as 60 ng of legitimate target DNA. Conclusions These findings could potentially explain discrepant results from laboratories attempting to amplify MLV-related viruses including XMRV from human samples, as none of the published reports used internal-tube controls for amplification. The potential for false negative results needs to be considered and carefully controlled in PCR experiments, especially when the target copy number may be low - just as the potential for false positive results already is.

  2. Microsampling homogeneous immunoassay with Cedia digoxin reagents on the Technicon CHEM 1 chemistry analyzer. (United States)

    Lua, A C; Chu, D K; Vlastelica, D


    We report the determination of digoxin concentration in serum with Microgenics Cedia digoxin reagents on the Technicon CHEM 1. The Technicon CHEM 1 clinical chemistry analyzer has a throughput of 720 tests per hour and uses only 7 microliters each of two reagents. A 100 test kit can perform 2,640 tests. The within-run coefficient of variation (CV) range is 2.3-0.9% and the total CV is 6.3-2.9% at concentrations tested ranging from 1.10 to 2.94 ng/ml. The results of the Technicon CHEM 1 (y) assay correlated well with those by the Technicon RA 1000 system (x) with 31 clinical serum samples (y = -0.03 + 1.11x, r = 0.96). We concluded that the Cedia digoxin assay on the Technicon CHEM 1 provides a very cost-effective, precise, rapid, and accurate means to determine digoxin concentration in serum.

  3. Determination of designer drug cross-reactivity on five commercial immunoassay screening kits. (United States)

    Regester, Laura E; Chmiel, Jeffrey D; Holler, Justin M; Vorce, Shawn P; Levine, Barry; Bosy, Thomas Z


    The detection of new designer drugs is often a difficult issue in forensic urine drug testing as immunoassays are the primary screening methodology for drugs of abuse in many of these laboratories. Cross-reactivity of compounds with immunoassay kits can either aid or complicate the detection of a variety of drug and drug metabolites. For instance, emerging designer drugs that share structural similarities to amphetamines and phencyclidine (PCP) have the potential to cross-react with assays designed to detect these compounds. This study evaluates the cross-reactivity of five commercially available immunoassay reagent kits for 94 designer drugs on a Roche/Hitachi Modular P automated screening instrument. The compounds used in this study are grouped by structural class as follows: 2,5-dimethoxyamphetamines, 2C (2,5-dimethoxyphenethylamines), β-keto amphetamines, substituted amphetamines, piperazines, α-pyrrolidinopropiophenones, tryptamines and PCP analogs. A drug concentration of 100 µg/mL was used to determine cross-reactivity for each assay and resulted in the following positive rates: Microgenics DRI(®) Ecstasy enzyme assay (19%), Microgenics DRI(®) Phencyclidine enzyme assay (20%), Lin-Zhi Methamphetamine enzyme immunoassay (39%), Siemens/Syva(®) EMIT(®)II Plus Amphetamines assay (43%) and CEDIA(®) DAU Amphetamine/Ecstasy assay (57%). Of the 94 designer drugs tested, 14% produced a negative response for all five kits. No designer drug used in this study generated a positive result for all five immunoassay kits.

  4. Improving DNA data exchange: validation studies on a single 6 dye STR kit with 24 loci. (United States)

    Martín, Pablo; de Simón, Lourdes Fernández; Luque, Gracia; Farfán, María José; Alonso, Antonio


    The idea of developing a new multiplex STR amplification system was conceived in 2011 as an effective way to implement the new European standard set (ESS) of 12 STR markers adopted by The Council of the European Union in 2009 while maintaining an effective compatibility and information exchange with the historical DNA profiles contained in the Spanish national DNA database (around 200,000 DNA profiles) mainly based on the 13 CODIS core STR loci plus D19S433 and D2S1338 markers. With this goal in mind we proposed to test and validate a single STR amplification system for simultaneous analysis of 21 STR markers covering both CODIS and ESS core STR loci plus three additional markers (D19S433, D2S1338, and SE33) also contained in commonly used STR kits and national DNA databases. In 2012, we started the first beta-testing with a 6-dye STR kit prototype containing 24 loci (now known as the GlobalFiler™ PCR Amplification Kit) developed by Life Technologies in response to the CODIS Core Loci Working Group's recommendation to expand the CODIS Core Loci. This prototype included our proposal of 21 autosomal STR markers and two Y-chromosome markers (DYS391 and Y-indel) and maximizes concordance with established databases and previously analyzed samples by maintaining primer sequences of previous Identifiler(®)/NGM SElect™ kits for the 21 STR markers except for TPOX. This paper describes the validation studies conducted with the first commercial available 6-dye STR kit for casework using a 3500 genetic analyzer for fragment detection that included the analysis of the following parameters and aspects: analytical threshold, sensitivity & stochastic threshold, heterozygous balance, stutter threshold, precision and accuracy, repeatability and reproducibility, genotype concordance, DNA mixtures, species specificity, and stability studies with case type samples. The studies demonstrated that the GlobalFiler™ system provided equivalent overall performance to previous forensic

  5. Degradable conjugates from oxanorbornadiene reagents. (United States)

    Kislukhin, Alexander A; Higginson, Cody J; Hong, Vu P; Finn, M G


    Oxanorbornadienedicarboxylate (OND) reagents were explored for purposes of binding and releasing drugs from serum albumins as representative macromolecular carriers. Being highly reactive Michael acceptors, ONDs form adducts with thiols and amines, which then undergo retro-Diels-Alder fragmentation. A study of more than 30 model adducts revealed a number of modifications that can be used to influence adduct stability. For the most reactive OND linkers, the labeling of the single available bovine serum albumin (BSA) cysteine residue was complete within minutes at a mid-micromolar concentration of reactants. While a selectivity of greater than 1000-fold for thiol over amine was observed with model amino acids, the labeling of protein amines with ONDs is fast enough to be practical, as demonstrated by the reaction with thiol-depleted BSA. The OND-amine adducts were found to be up to 15 times more stable than OND-thiol adducts, and to be sensitive to acid by virtue of a stereochemically dependent acceleration of cycloreversion. The release rate of fluorescent cargo from serum albumins was tuned by selecting the coupling partners: the available half-lives ranged from 40 min to 7 days at 37 °C. Such versatility of release profiles from protein carriers, controlled by the nature of the OND linkage, is a useful addition to the drug delivery toolbox.

  6. 100G Deployment@(DE-KIT) (United States)

    Hoeft, Bruno; Petzold, Andreas


    The Steinbuch Centre for Computing (SCC) at Karlsruhe Institute of Technology (KIT) has been involved fairly early in 100GE network technology. Initiated by DFN1 (the German NREN), a first 100GE wide area network testbed over a distance of approx. 450 km was deployed between the national research organizations KIT and FZ-Jülich in 2010. Three years later in 2013. KIT joined the Caltech SuperComputing 2013 (SC132) 100GE "show floor" initiative using the transatlantic ANA-100GE link to transfer LHC data from a storage at DE-KIT (GridKa) in Europe to hard disks at the show floor of SC13 in Denver (USA). The network infrastructure of KIT as well as of the German Tier-1 installation DE-KIT (GridKa). however. is still based on 10Gbps. As highlighted in the contribution "Status and Trends in Networking at LHC Tier1 Facilities" to CHEP 2012. proactive investment is required at the Tier-1 sites. Bandwidth requirements will grow beyond current capacity and the required upgrades are expected in 2015. In close cooperation with DFN. KIT drives the upgrade from 10GE to 100GE. The process is divided into several phases. due to upgrade costs and differing requirements in different parts of the network infrastructure. The requirements of the different phases as well as the planned topology will be described. Some of the obstacles we discovered during the deployment will be discussed and solutions or workarounds presented.


    Directory of Open Access Journals (Sweden)

    Ivona Djurkin Kušec


    Full Text Available In the present study four commercially available DNA extraction kits (Wizard® Genomic DNA Purification Kit, High Pure PCR Template Kit, DNeasy mericon Food and GeneJET PCR Purification Kit, as well as standard phenol/chloroform isolation technique have been evaluated regarding their concentration, purity and suitability for amplification of porcine DNA in dry/fermented sausages. The isolates were assessed for quantity and quality using spectrophotometer (IMPLEN GmbH, Germany. To verify template usability and quality of isolated DNA, the polymerase chain reaction (PCR targeting at porcine cytochrome b by species specific primers was used. The comparison of extraction methods revealed satisfactory efficiency and purity of all extraction kits, while with standard phenol/chloroform isolation method high concentrations of DNA with low A260/280 were obtained. However, all the investigated techniques proved to be suitable for identification of porcine DNA in dry/fermented sausage. Thus, the standard phenol/chloroform DNA extraction method, as the cost-effective one, can be recommended as a good alternative to more expensive isolation kits when investigating the presence of pork DNA in dry/ fermented meat products.

  8. Multicentre evaluation of commercial kit methods

    DEFF Research Database (Denmark)

    Gram, J; Declerck, P J; Sidelmann, Johannes Jakobsen;


    in the production of data from each of the commercial kits tested. A considerable variation of PAI activity results reported from the laboratories testing the same commercial kits was observed. The range of reported results could in individual samples exceed the median value indicating an interlaboratory variation...... activity levels and a CV of about 16% for high PAI activity levels. The high imprecision of the kits in the low concentration range of PAI activity indicates that unspecific factors in plasma may interfere with determination of active PAI. This was confirmed by the evaluation of the results from one...

  9. 21 CFR 866.3010 - Acinetobacter calcoaceticus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Acinetobacter calcoaceticus serological reagents... Acinetobacter calcoaceticus serological reagents. (a) Identification. Acinetobacter calcoaceticus serological reagents are devices that consist of Acinetobacter calcoaceticus antigens and antisera used to...

  10. Clinical Evaluation on Several anti- HIV Diagnostic Reagents

    Institute of Scientific and Technical Information of China (English)


    Objective We Joined clinical evaluation on 6 anti - HIV diagnostic reagents which was organized by National Reference Laboratory of National Center for AIDS Prevention and Control. Method 100 sera of known result and 100 sera of unknown result were detected with 6 reagents according to test procedure of the reagents. Result The crude agreement (99.5 % ) of Organon Teknika and Determine reagents were higher than that of the other reagents. No anti - HIV positive serum was detected negative with Organon Teknika and Determine reagents. The sensitivity and specificity of Organon Teknika and Determine reagents were higher than those of the other reagents. The capacity of Organon Teknika reagent to detect the mild positive serum was greater than that of the other reagent. Conclusion Organon Teknika and Determine antiHIV diagnostic reagents were qualified for anti - HIV screening test while the other 4 reagents should be improved on sensitivity and specificity.

  11. The SCRAM tool-kit (United States)

    Tamir, David; Flanigan, Lee A.; Weeks, Jack L.; Siewert, Thomas A.; Kimbrough, Andrew G.; McClure, Sidney R.


    This paper proposes a new series of on-orbit capabilities to support the near-term Hubble Space Telescope, Extended Duration Orbiter, Long Duration Orbiter, Space Station Freedom, other orbital platforms, and even the future manned Lunar/Mars missions. These proposed capabilities form a toolkit termed Space Construction, Repair, and Maintenance (SCRAM). SCRAM addresses both intra-Vehicular Activity (IVA) and Extra-Vehicular Activity (EVA) needs. SCRAM provides a variety of tools which enable welding, brazing, cutting, coating, heating, and cleaning, as well as corresponding nondestructive examination. Near-term IVA-SCRAM applications include repair and modification to fluid lines, structure, and laboratory equipment inside a shirt-sleeve environment (i.e. inside Spacelab or Space Station). Near-term EVA-SCRAM applications include construction of fluid lines and structural members, repair of punctures by orbital debris, refurbishment of surfaces eroded by contaminants. The SCRAM tool-kit also promises future EVA applications involving mass production tasks automated by robotics and artificial intelligence, for construction of large truss, aerobrake, and nuclear reactor shadow shields structures. The leading candidate tool processes for SCRAM, currently undergoing research and development, include Electron Beam, Gas Tungsten Arc, Plasma Arc, and Laser Beam. A series of strategic space flight experiments would make SCRAM available to help conquer the space frontier.

  12. 21 CFR 864.4020 - Analyte specific reagents. (United States)


    ... reagents. (a) Identification. Analyte specific reagents (ASR's) are antibodies, both polyclonal and... Detect Antibodies to the Human Immunodeficiency Virus, Type I” (54 FR 48943, November 28, 1989)....

  13. Village Green Project and Air Sensor Kits (United States)

    This is a presentation for the OAQPS Teachers Workshop. Will provide a background overview on the Village Green Project and our air sensor kit for outreach, then have the teachers try putting it together.

  14. Comparison of commercial DNA extraction kits for isolation and purification of bacterial and eukaryotic DNA from PAH-contaminated soils

    Energy Technology Data Exchange (ETDEWEB)

    Mahmoudi, Nagissa; Slater, Greg F.; Fulthorpe, Roberta R.


    Polycyclic aromatic hydrocarbons (PAHs) are in the environment and are carcinogenic, teratogenic and mutagenic. Their hydrophobic structure gives them low water solubility and makes them readily absorbed onto soils and sediments, where they persists until they are degraded. Microbial degradation of PAHs has been well documented and is thought to be an important process in remediating contaminated sediments and soils. Obtaining high quality purified DNA is an essential requirement for the successful DNA amplifications that underlie all subsequent procedures. Several commercial DNA extraction kits exist that provide consistent solutions for the central problems - cell lysis and humic acid removal. This study compared four commercial DNA extraction kits to extract pure, high quality bacterial and eukaryotic DNA from PAH contaminated soils and concluded that they can be used on a wide variety of soils, including heavily contaminated soils. The PowerSoil kit was the most effective and reliable.

  15. Directional and Attitude Stability Control Kit (United States)


    and Attitude Stability Control Kit Final Progress Report This report outlines progress on the DARPA M3 Program, project “Directional and Attitude ...2 ABSTRACT Number of Papers published in peer-reviewed journals: Number of Papers published in non peer-reviewed journals: Directional and Attitude ...Stability Control Kit Final Progress Report Report Title This report outlines progress on the DARPA M3 Program, project “Directional and Attitude

  16. 46 CFR 184.710 - First-aid kits. (United States)


    ... 46 Shipping 7 2010-10-01 2010-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent...

  17. Powerplex® ES versus Powerplex® S5--casework testing of the new screening kit. (United States)

    Poetsch, Micaela; Kamphausen, Thomas; Bajanowski, Thomas; Schwark, Thorsten; von Wurmb-Schwark, Nicole


    The new Powerplex® S5 Mini STR-System from Promega with the four provided STR loci D18S51, D8S1179, TH01 and FGA as well as the Amelogenin marker (PCR products ranging from 80 to 220 bp not considering the longer FGA fragments) is designed as a screening tool especially in difficult casework samples. To test its suitability we amplified highly degraded DNA from casework samples, which had shown no or only poor results in analyses with the Powerplex® ES kit, as well as artificially degraded DNA or DNA samples containing PCR inhibitors. Despite a tendency for allelic drop-ins in the amplification of highly degraded DNA the Powerplex® S5 kit was a reliable tool for the analysis of casework samples with degraded DNA which gave better results than the Powerplex® ES kit in 64% of analysed swabs. Furthermore, it was especially suitable for the investigation of formalin fixed tissue, tissue samples showing advanced putrefaction or telogen hair samples. However, there was no strict relation between positive Powerplex® S5 results and amplification success with the Powerplex® ES kit.

  18. The effect of whole genome amplification on samples originating from more than one donor

    DEFF Research Database (Denmark)

    Thacker, C.R.; Balogh, M.K.; Børsting, Claus;


    In this study, the GenomiPhi(TM) DNA Amplification Kit (Amersham Biosciences) was used to investigate the potential of whole genome amplification (WGA) when considering samples originating from more than one donor. DNA was extracted from blood samples, quantified and normalised before being mixed...... found to match the expected peak ratios regardless of the starting concentration of DNA. With samples mixed in the ratio of 1:7 and 1:15, and when the concentration of starting material was at the manufacturer's lower limit, too few minor component peaks were found to allow for statistical analysis...

  19. Hardness amplification in nondeterministic logspace


    Gupta, Sushmita


    A hard problem is one which cannot be easily computed by efficient algorithms. Hardness amplification is a procedure which takes as input a problem of mild hardness and returns a problem of higher hardness. This is closely related to the task of decoding certain error-correcting codes. We show amplification from mild average case hardness to higher average case hardness for nondeterministic logspace and worst-to-average amplification for nondeterministic linspace. Finally we explore possible ...

  20. A Homogeneous HLA-B*27 Genotyping Assay Using Dried Reagent Mixtures

    Directory of Open Access Journals (Sweden)

    Minna Kiviniemi


    Full Text Available The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance.

  1. Aging Kit mutant mice develop cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Lei Ye

    Full Text Available Both bone marrow (BM and myocardium contain progenitor cells expressing the c-Kit tyrosine kinase. The aims of this study were to determine the effects of c-Kit mutations on: i. myocardial c-Kit(+ cells counts and ii. the stability of left ventricular (LV contractile function and structure during aging. LV structure and contractile function were evaluated (echocardiography in two groups of Kit mutant (W/Wv and W41/W42 and in wild type (WT mice at 4 and 12 months of age and the effects of the mutations on LV mass, vascular density and the numbers of proliferating cells were also determined. In 4 month old Kit mutant and WT mice, LV ejection fractions (EF and LV fractional shortening rates (FS were comparable. At 12 months of age EF and FS were significantly decreased and LV mass was significantly increased only in W41/W42 mice. Myocardial vascular densities and c-Kit(+ cell numbers were significantly reduced in both mutant groups when compared to WT hearts. Replacement of mutant BM with WT BM at 4 months of age did not prevent these abnormalities in either mutant group although they were somewhat attenuated in the W/Wv group. Notably BM transplantation did not prevent the development of cardiomyopathy in 12 month W41/W42 mice. The data suggest that decreased numbers and functional capacities of c-Kit(+ cardiac resident progenitor cells may be the basis of the cardiomyopathy in W41/W42 mice and although defects in mutant BM progenitor cells may prove to be contributory, they are not causal.

  2. 21 CFR 866.3110 - Campylobacter fetus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Campylobacter fetus serological reagents. 866.3110... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3110 Campylobacter fetus serological reagents. (a) Identification. Campylobacter fetus serological reagents are...

  3. 21 CFR 866.3490 - Rhinovirus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rhinovirus serological reagents. 866.3490 Section... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3490 Rhinovirus serological reagents. (a) Identification. Rhinovirus serological reagents are devices that consist of...

  4. 21 CFR 866.3040 - Aspergillus spp. serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Aspergillus spp. serological reagents. 866.3040... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040 Aspergillus spp. serological reagents. (a) Identification. Aspergillus spp. serological reagents are devices...

  5. 21 CFR 866.3660 - Shigella spp. serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Shigella spp. serological reagents. 866.3660... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella spp. serological reagents. (a) Identification. Shigella spp. serological reagents are devices that consist...

  6. 21 CFR 866.3405 - Poliovirus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Poliovirus serological reagents. 866.3405 Section... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3405 Poliovirus serological reagents. (a) Identification. Poliovirus serological reagents are devices that consist of...

  7. 21 CFR 866.3220 - Entamoeba histolytica serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Entamoeba histolytica serological reagents. 866... Entamoeba histolytica serological reagents. (a) Identification. Entamoeba histolytica serological reagents... Entamoeba histolytica in serum. Additionally, some of these reagents consist of antisera conjugated with...

  8. 21 CFR 866.3500 - Rickettsia serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rickettsia serological reagents. 866.3500 Section... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3500 Rickettsia serological reagents. (a) Identification. Rickettsia serological reagents are devices that consist of...

  9. 21 CFR 866.3510 - Rubella virus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rubella virus serological reagents. 866.3510... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3510 Rubella virus serological reagents. (a) Identification. Rubella virus serological reagents are devices that consist...

  10. 21 CFR 866.3870 - Trypanosoma spp. serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Trypanosoma spp. serological reagents. 866.3870... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3870 Trypanosoma spp. serological reagents. (a) Identification. Trypanosoma spp. serological reagents are devices...

  11. 21 CFR 864.8540 - Red cell lysing reagent. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Red cell lysing reagent. 864.8540 Section 864.8540...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8540 Red cell lysing reagent. (a) Identification. A red cell lysing reagent is a device used to lyse (destroy) red blood cells...

  12. 21 CFR 866.3415 - Pseudomonas spp. serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Pseudomonas spp. serological reagents. 866.3415... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415 Pseudomonas spp. serological reagents. (a) Identification. Pseudomonas spp. serological reagents are devices...

  13. Production of reagents for cleaning fluids

    Energy Technology Data Exchange (ETDEWEB)

    Grunberg, I.V.; Korostyleva, R.N.; Pytel, S.P.; Spasskii, P.I.; Titarenko, N.K.; Trachtenberg, S.I.; Yushkevich, V.I.


    A method for producing reagents for cleaning fluids is proposed using polymerization of acrylonitril, metachrylate or a mixture of the two in water and saponification of the polymers with alakali. To reduce the consumption of monomers and increase the quality of the reagents, 0.4-1.0 parts humic substances, 0.2-1.0 parts hydrolizate from tanning waste products and 1.2-4.0 parts monomers are added to the reaction medium, followed by copolymerization in an acid medium. The proposed method ensures quality reagents which combine lower water yield with a moderate increase in viscosity when acting on clay solutions. Compared with the current method, this method lowers the consumption of an expensive and hard-to-find monomer 1.2-1.4X for one ton of reagent, which lowers the cost of raw material by 1.3-1.7X. This results in a savings of 195-385 rubles per ton of reagent, 600-1200 thousand at 3000 tons/yr.

  14. Chemokine receptor CCR5 Delta 32 genetic analysis using multiple specimen types and the NucliSens Basic Kit. (United States)

    Tetali, S; Lee, E M; Kaplan, M H; Romano, J W; Ginocchio, C C


    Resistance to HIV-1 infection and delayed disease progression have been associated with a 32-bp deletion (Delta32) in the gene encoding the CCR5 chemokine receptor. In the present study we describe the modification of a nucleic acid sequence-based amplification (NASBA)-based CCR5 genotyping assay for a NucliSens Basic Kit (Organon Teknika, Durham, N.C.) format using a new target-specific sandwich oligonucleotide detection methodology. The new method permitted the use of generic electrochemiluminescent probes supplied in the NucliSens Basic Kit, whereas the original NASBA method required expensive target-specific ruthenium detection probes. The Basic Kit CCR5 Delta32 genotypic analysis was in 100% concordance with both the original NASBA assay and DNA PCR results. This study also evaluated the use of multiple specimen types, including peripheral blood mononuclear cells (PBMC), whole blood, dried blood spots, buccal scrapings, and plasma, for CCR5 genotype analysis. The sensitivities of the three assays were comparable when PBMC or whole blood was the specimen source. In contrast, when dried blood spots, buccal scrapings, or plasma was used as the sample source, the sensitivity of DNA PCR was 80.95, 42.8, or 0%, respectively, compared to 100% sensitivity obtained with the original NASBA and Basic Kit NASBA assays. Our study indicates that the NucliSens Basic Kit NASBA assay is very sensitive and specific for CCR5 Delta32 genotyping using multiple sample types.

  15. Verification of multiplex ligation-dependent probe amplification probes in the absence of positive samples. (United States)

    Wooderchak-Donahue, Whitney; Vaughn, Cecily; Chou, Lan-Szu; Lewis, Tracey; Sumner, Kelli; Procter, Melinda; Gedge, Friederike; Bayrak-Toydemir, Pinar; Lyon, Elaine; Pont-Kingdon, Genevieve


    Deletions and duplications of single or multiple exons in specific genes are associated with human diseases. Multiplex ligation-dependant probe amplification (MLPA), a technique recently introduced to clinical laboratories, can detect deletions or duplications at the exon level. MLPA kits have a high multiplexing capability containing mixtures of exon-specific probes that target the gene of interest and control probes that hybridize to other genomic areas before PCR amplification. To verify each probe set, known positive samples with a single-exon deletion or duplication and normal samples are ideally used. Often, positive samples do not exist for each exon and normal samples are not suited to verify the identity of each probe set. We designed a straightforward approach using mixes of exon-specific PCR products as template to unequivocally verify each probe set in MLPA kits. This method can be used to verify the identity of MLPA probes for exons when positive samples are unavailable. Exon-specific probes from 15 MLPA kits were shown to hybridize to the targeted exons of interest. In one kit, this method identified probes that also bind a pseudogene, making them unreliable for clinical analysis. Incorporating this methodology in the analytical validation process will help ensure that MLPA results are interpreted correctly.

  16. [Real-time PCR kits for the detection of the African Swine Fever virus]. (United States)

    Latyshev, O E; Eliseeva, O V; Grebennikova, T V; Verkhovskiĭ, O A; Tsibezov, V V; Chernykh, O Iu; Dzhailidi, G A; Aliper, T I


    The results obtained using the diagnostic kit based on real-time polymerase chain reaction to detect the DNA of the African Swine Fever in the pathological material, as well as in the culture fluid, are presented. A high sensitivity and specificity for detection of the DNA in the organs and tissues of animals was shown to be useful for detection in the European Union referentiality reagent kits for DNA detection by real time PCR of ASFV. More rapid and effective method of DNA extraction using columns mini spin Quick gDNA(TM) MiniPrep was suggested and compared to the method of DNA isolation on the inorganic sorbent. High correlation of the results of the DNA detection of ASFV by real-time PCR and antigen detection results ASFV by competitive ELISA obtained with the ELISA SEROTEST/INGEZIM COMRAC PPA was demonstrated. The kit can be used in the veterinary services for effective monitoring of ASFV to contain, eliminate and prevent further spread of the disease.

  17. Efficient audio power amplification - challenges

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, Michael A.E.


    For more than a decade efficient audio power amplification has evolved and today switch-mode audio power amplification in various forms are the state-of-the-art. The technical steps that lead to this evolution are described and in addition many of the challenges still to be faced and where extensive research and development are needed is covered. (au)

  18. Isothermal Amplification of Nucleic Acids. (United States)

    Zhao, Yongxi; Chen, Feng; Li, Qian; Wang, Lihua; Fan, Chunhai


    Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at constant temperature. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small molecules, and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-molecule analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanalysis, diagnostics, nanotechnology, materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.

  19. Field Test Kit for Gun Residue Detection

    Energy Technology Data Exchange (ETDEWEB)



    One of the major needs of the law enforcement field is a product that quickly, accurately, and inexpensively identifies whether a person has recently fired a gun--even if the suspect has attempted to wash the traces of gunpowder off. The Field Test Kit for Gunshot Residue Identification based on Sandia National Laboratories technology works with a wide variety of handguns and other weaponry using gunpowder. There are several organic chemicals in small arms propellants such as nitrocellulose, nitroglycerine, dinitrotoluene, and nitrites left behind after the firing of a gun that result from the incomplete combustion of the gunpowder. Sandia has developed a colorimetric shooter identification kit for in situ detection of gunshot residue (GSR) from a suspect. The test kit is the first of its kind and is small, inexpensive, and easily transported by individual law enforcement personnel requiring minimal training for effective use. It will provide immediate information identifying gunshot residue.

  20. Shuttle Kit Freezer Refrigeration Unit Conceptual Design (United States)

    Copeland, R. J.


    The refrigerated food/medical sample storage compartment as a kit to the space shuttle orbiter is examined. To maintain the -10 F in the freezer kit, an active refrigeration unit is required, and an air cooled Stirling Cycle refrigerator was selected. The freezer kit contains two subsystems, the refrigeration unit, and the storage volume. The freezer must provide two basic capabilities in one unit. One requirement is to store 215 lbs of food which is consumed in a 30-day period by 7 people. The other requirement is to store 128.3 lbs of medical samples consisting of both urine and feces. The unit can be mounted on the lower deck of the shuttle cabin, and will occupy four standard payload module compartments on the forward bulkhead. The freezer contains four storage compartments.

  1. Low Cost, Advanced, Integrated Microcontroller Training Kit (United States)

    Somantri, Y.; Fushshilat, I.


    This paper describes the design of an AVR microcontroller training kit with a low cost and the additional feature of an integrated downloader. The main components of this device include: Microcontroller, terminal, I/O keypad, push button, LED, seven segment display, LCD, motor stepper, and sensors. The device configuration results in low cost and ease of use; this device is suitable for laboratories with limited funding. The device can also be used as a training kit for the teaching and learning of microcontrollers.

  2. Preparation of a test kit for rapid and on site determination of fluoride in water%水中氟化物现场快速测试盒的研制

    Institute of Scientific and Technical Information of China (English)

    秦惠; 邓金花; 傅妍芳; 陈维


    基于氟试剂分光光度法的显色原理,采用目视比色法和试剂定量包装技术构建氟化物现场快速测试盒.该测试盒具有便于携带、操作简便、快速、测试结果准确等特点,测试范围为0.05 ~1.20 mg,/L.%Based on the principle of the color reaction of fluoride-reagent spectrophotometric method,the visual colorimetric method and reagent quantitative packing technique are used to prepare an on-site flouride test kit. This test kit is easy to carry and use with rapid and accurate determination. The determination range of the kit is 0.05 -1. 2 mg/L.

  3. Comparison of DNA extraction methods for polymerase chain reaction amplification of guanaco (Lama guanicoe) fecal DNA samples. (United States)

    Espinosa, M I; Bertin, A; Squeo, F A; Cortés, A; Gouin, N


    Feces-based population genetic studies have become increasingly popular. However, polymerase chain reaction (PCR) amplification rates from fecal material vary depending on the species, populations, loci, and extraction protocols. Here, we assessed the PCR amplification success of three microsatellite markers and a segment of the mitochondrial control region of DNA extracted from field-collected feces of guanaco (Lama guanicoe) using two protocols - Qiagen DNA Stool Kit and 2 cetyltrimethylammonium bromide/phenol:chloroform:isoamyl alcohol (2CTAB/PCI) method. Chelex resin treatment to remove inhibitors was also tested. Our results show that the mitochondrial locus was the most difficult to amplify. PCR success rates improved for all markers after Chelex treatment of extracted DNA, and 2CTAB/PCI method (95.83%) appeared to perform slightly better than stool kit (91.67%) for the nuclear markers. Amplification success was significantly influenced by the extraction method, Chelex treatment, and locus (P 0.89), but they decreased slightly after treatment for amplification of nuclear markers and markedly after treatment for amplification of the mitochondrial control region. Thus, we showed that Chelex treatment gives high PCR success, especially for nuclear markers, and adequate DNA extraction rates can be achieved from L. guanicoe feces even from non-fresh fecal material. Although not significant, 2CTAB/PCI method tended to provide higher successful amplification rates on a whole set of samples, suggesting that the method could be particularly useful when using small sample sizes.

  4. Development of a qualitative, multiplex real-time PCR kit for screening of genetically modified organisms (GMOs). (United States)

    Dörries, Hans-Henno; Remus, Ivonne; Grönewald, Astrid; Grönewald, Cordt; Berghof-Jäger, Kornelia


    The number of commercially available genetically modified organisms (GMOs) and therefore the diversity of possible target sequences for molecular detection techniques are constantly increasing. As a result, GMO laboratories and the food production industry currently are forced to apply many different methods to reliably test raw material and complex processed food products. Screening methods have become more and more relevant to minimize the analytical effort and to make a preselection for further analysis (e.g., specific identification or quantification of the GMO). A multiplex real-time PCR kit was developed to detect the 35S promoter of the cauliflower mosaic virus, the terminator of the nopaline synthase gene of Agrobacterium tumefaciens, the 35S promoter from the figwort mosaic virus, and the bar gene of the soil bacterium Streptomyces hygroscopicus as the most widely used sequences in GMOs. The kit contains a second assay for the detection of plant-derived DNA to control the quality of the often processed and refined sample material. Additionally, the plant-specific assay comprises a homologous internal amplification control for inhibition control. The determined limits of detection for the five assays were 10 target copies/reaction. No amplification products were observed with DNAs of 26 bacterial species, 25 yeasts, 13 molds, and 41 not genetically modified plants. The specificity of the assays was further demonstrated to be 100% by the specific amplification of DNA derived from reference material from 22 genetically modified crops. The applicability of the kit in routine laboratory use was verified by testing of 50 spiked and unspiked food products. The herein described kit represents a simple and sensitive GMO screening method for the reliable detection of multiple GMO-specific target sequences in a multiplex real-time PCR reaction.

  5. The Kasumi-1 cell line: a t(8;21)-kit mutant model for acute myeloid leukemia. (United States)

    Larizza, Lidia; Magnani, Ivana; Beghini, Alessandro


    The Kasumi-1 cell line is an intensively investigated model system of Acute Myeloid Leukemia with t(8;21) translocation, that represents 1 of the 2 main subtypes of Core Binding Factor Leukemia (CBFL). Since establishment in 1991 the Kasumi-1 cell line has provided the tool to study the peculiar molecular, morphologic, immunophenotypic findings of AML with t(8;21) and the functional consequences of the AML1-ETO fusion oncogene on myeloid differentiation. Leukemogenesis involves multiple genetic changes and, as suggested by murine experiments and other findings in humans, AML1-ETO expression may not be sufficient for full blown leukemia. In agreement with the "two hits" model of leukemogenesis, based on the cooperation between 1 class of mutations that impair hematopoietic differentiation and a second class of mutations that confer a proliferative and/or survival advantage to hematopoietic progenitors an activating mutation in the tyrosine kinase domain of the c-kit gene was identified in the AML1/ETO expressing Kasumi-1 cell line. The dosage of the Asn822Lys mutated allele was shown to be about 5-fold compared to the normal allele and c-kit amplification was found to map to minute 4cen-q11 marker chromosomes, likely derived from the extra chromosome 4 recorded in the newly established cell line. The combination of t(8;21) and trisomy 4 leading to enhanced dosage of a mutated kit allele is a feature of a few CBFL patients reproduced by the Kasumi-1 cell model. The Kasumi-1 cell line, paralleling the commitment stage of CBF leukemia also provides a valuable resource to investigate the effect of tyrosine kinase kit mutant on the main KIT-regulated signal transduction pathways, i.e. MAPK, PI3K/AKT and STAT3 and the diverse inhibitory effect exerted by STI 571 on these KIT mutant activated pathways. PI3K-dependent activation of AKT and STAT activation was observed in Kasumi-1 cells. Contrary to the expectations for an amplified tyrosine kinase kit mutant, we found that

  6. A PCR amplification method without DNA extraction. (United States)

    Li, Hongwei; Xu, Haiyue; Zhao, Chunjiang; Sulaiman, Yiming; Wu, Changxin


    To develop a simple and inexpensive method for direct PCR amplification of animal DNA from tissues, we optimized different components and their concentration in lysis buffer systems. Finally, we acquired the optimized buffer system composed of 10 mmol tris(hydroxymethyl)aminomethane (Tris)-Cl (pH 8.0), 2 mmol ethylene diamine tetraacetic (EDTA) (pH 8.0), 0.2 mol NaCl and 200 μg/mL Proteinase K. Interestingly, the optimized buffer is also very effective when working with common human sample types, including blood, buccal cells and hair. The direct PCR method requires fewer reagents (Tris-Cl, EDTA, Protease K and NaCl) and less incubation time (only 35 min). The cost of treating every sample is less than $0.02, and all steps can be completed on a thermal cycler in a 96-well format. So, the proposed method will significantly improve high-throughput PCR-based molecular assays in animal systems and in common human sample types.

  7. Evaluation of two new enzyme immunoassay reagents for diagnosis of histoplasmosis in a cohort of clinically characterized patients. (United States)

    Zhang, Chen; Lei, Guang-Sheng; Lee, Chao-Hung; Hage, Chadi A


    The performance characteristics of the recently available analyte-specific reagent based enzyme immunoassay (ASR-EIA) and in vitro diagnostic (IVD) kit for urine Histoplasma antigen detection were evaluated in a cohort of 50 clinically characterized patients with histoplasmosis and 50 control patients. Overall sensitivity and specificity of the ASR-EIA were significantly improved compared with those of the IVD kit (sensitivity 72% vs. 22%, Phistoplasmosis (five with pulmonary histoplasmosis and nine with progressive disseminated histoplasmosis) were falsely negative by ASR-EIA. All 10 specimens from patients with severe symptoms of progressive disseminated histoplasmosis were positive by ASR-EIA, although the average reading value of these 10 specimens was not significantly different from that of others with positive results. Compared to the MiraVista antigen assay, both the IVD kit and the ASR-EIA were significantly less sensitive in detecting Histoplasma antigen in the urine of patients with histoplasmosis. The ASR-EIA and MiraVista assay had comparable specificity. In conclusion, the ASR-EIA has improved performance compared with the IVD kit in the detection of Histoplasma antigen in the urine. However, users should be aware of the potential for false negative results using the currently recommended cutoff value.

  8. Teen PACK: Population Awareness Campaign Kit. (United States)

    Zero Population Growth, Inc., Washington, DC.

    This packet of instructional materials is designed to teach teenagers about the effects of overpopulation on the world and on the individual. Information is presented in three related booklets. The first of the three parts of the "Teen Population Awareness Campaign Kit," illustrates overpopulation through profiles of teens living in…

  9. Countdown to Six Billion Teaching Kit. (United States)

    Zero Population Growth, Inc., Washington, DC.

    This teaching kit features six activities focused on helping students understand the significance of the world population reaching six billion for our society and our environment. Featured activities include: (1) History of the World: Part Six Billion; (2) A Woman's Place; (3) Baby-O-Matic; (4) Earth: The Apple of Our Eye; (5) Needs vs. Wants; and…

  10. Word 2013 elearning kit for dummies

    CERN Document Server

    Wempen, Faithe


    Lois Lowe is the author of several books on Microsoft Office, including Microsoft Word 2010 eLearning Kit For Dummies. She is also an online instructor who develops and teaches courses on Microsoft Office applications, computer purchase and upgrade, home office setup and emerging hardware technologies. Her courses have educated over 250,000 students for clients including Hewlett-Packard and Sony.

  11. Numeric Data Products and Services. SPEC Kit. (United States)

    Cook, Michael N., Comp.; Hernandez, John J., Comp.; Nicholson, Shawn, Comp.


    This SPEC (Systems and Procedures Exchange Center) Kit presents the results of a survey of Association of Research Libraries (ARL) member libraries. The survey addressed the following questions about numeric data (i.e., any information resource, print or non-print, with considerable numeric content) in academic libraries: (1) What relationships…

  12. Synthesis of reagents for fluoride technologies

    Institute of Scientific and Technical Information of China (English)

    Gordienko; P.; S.; Kolzunov; V.; A.; Dostovalov; V.; A.; Kaidalova; T.; A.


    Growing demand for fluorinating reagents to be used in rare-metal industry has stimulated conducting research in the field of production for these reagents. That is why the fluorinating reagents production has recently formed an independent segment of industry. Main industrial fluorinating reagents include hydrofluoric acid, anhydrous hydrogen fluoride, technical ammonium hydrodifluoride, fluorosilicic acid and its salts. To produce technical etching acid, fluor-spar with calcium fluoride content at least 92% is used in most cases. To produce anhydrous hydrogen fluoride, fluor-spar with calcium fluoride content 96 %-97 % is necessary. The fluorine-containing raw materials refinement from silica by means of flotation makes the fluorinating reagents production substantially more expensive. In this work we have attempted to process unconcentrated raw materials by fluorine removal in the form of volatile silicon tetrafluoride. In this process silicon tetrafluoride was recovered by liquid ammonia with subsequent hydrolysis of the formed ammonia hexafluorosilicate. Hydrolysis occurred according to the reaction:(NH4)2 SiF6 + 4NH3 + 2 H2O= 6NH4F+ SiO2 The products of the ammonia hexafluorosilicate hydrolysis included ammonia fluoride and amorphous silica gel ("white soot") as by-product. This "white soot" was of high purity-with main component content 99.95% and total admixture content 0.05%. Silica gel is a superfine material with specific surface of 267.6 m2/g and is recommended as filler in the production of rubber, plastics and for other applications.Ammonia fluoride was transformed into ammonia hydrodifluoride (main processing product) according to the reaction:2NH4F→NH3+NH4 HF2 It was stated that the NH4F: NH4 HF2 ratio depends on boiling point temperature-with its increase the ammonia hydrofluoride concentration in solution increases as well.

  13. Test/QA Plan for Verification of Microcystin Test Kits (United States)

    Microcystin test kits are used to quantitatively measure total microcystin in recreational waters. These test kits are based on enzyme-linked immunosorbent assays (ELISA) with antibodies that bind specifically to microcystins or phosphate activity inhibition where the phosphatas...

  14. L576P KIT mutation in anal melanomas correlates with KIT protein expression and is sensitive to specific kinase inhibition. (United States)

    Antonescu, Cristina R; Busam, Klaus J; Francone, Todd D; Wong, Grace C; Guo, Tianhua; Agaram, Narasimhan P; Besmer, Peter; Jungbluth, Achim; Gimbel, Mark; Chen, Chin-Tung; Veach, Darren; Clarkson, Bayard D; Paty, Philip B; Weiser, Martin R


    Activating mutations in either BRAF or NRAS are seen in a significant number of malignant melanomas, but their incidence appears to be dependent to ultraviolet light exposure. Thus, BRAF mutations have the highest incidence in non-chronic sun damaged (CSD), and are uncommon in acral, mucosal and CSD melanomas. More recently, activating KIT mutations have been described in rare cases of metastatic melanoma, without further reference to their clinical phenotypes. This finding is intriguing since KIT expression is downregulated in most melanomas progressing to more aggressive lesions. In this study, we investigated a group of anal melanomas for the presence of BRAF, NRAS, KIT and PDGFRA mutations. A heterozygous KIT exon 11 L576P substitution was identified in 3 of 20 cases tested. The 3 KIT mutation-carrying tumors were strongly immunopositive for KIT protein. No KIT mutations were identified in tumors with less than 4+ KIT immunostaining. NRAS mutation was identified in one tumor. No BRAF or PDGFRA mutations were identified in either KIT positive or negative anal melanomas. In vitro drug testing of stable transformant Ba/F3 KIT(L576P) mutant cells showed sensitivity for dasatinib (previously known as BMS-354825), a dual SRC/ABL kinase inhibitor, and imatinib. However, compared to an imatinib-sensitive KIT mutant, dasatinib was potent at lower doses than imatinib in the KIT(L576P) mutant. These results suggest that a subset of anal melanomas show activating KIT mutations, which are susceptible for therapy with specific kinase inhibitors.

  15. 46 CFR 121.710 - First-aid kits. (United States)


    ... 46 Shipping 4 2010-10-01 2010-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit... kits, the contents must be stowed in a suitable, watertight container that is marked “First-Aid Kit”....

  16. Next generation Chirped Pulse Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Nees, J.; Biswal, S.; Mourou, G. [Univ. Michigan, Center for Ultrafast Optical Science, Ann Arbor, MI (United States); Nishimura, Akihiko; Takuma, Hiroshi


    The limiting factors of Chirped Pulse Amplification (CPA) are discussed and experimental results of CPA in Yb:glass regenerative amplifier are given. Scaling of Yb:glass to the petawatt level is briefly discussed. (author)

  17. 46 CFR 108.707 - First aid kit. (United States)


    ... 46 Shipping 4 2010-10-01 2010-10-01 false First aid kit. 108.707 Section 108.707 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) A-MOBILE OFFSHORE DRILLING UNITS DESIGN AND EQUIPMENT Miscellaneous Equipment § 108.707 First aid kit. Each unit must have a first-aid kit approved by the Mine...

  18. 33 CFR 144.01-30 - First-aid kit. (United States)


    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall...

  19. 46 CFR 169.725 - First aid kit. (United States)


    ... 46 Shipping 7 2010-10-01 2010-10-01 false First aid kit. 169.725 Section 169.725 Shipping COAST... Control, Miscellaneous Systems, and Equipment § 169.725 First aid kit. Each vessel must carry an approved first aid kit, constructed and fitted in accordance with subpart 160.041 of this chapter....

  20. Escherichia coli and virus isolated from ''sticky kits''

    DEFF Research Database (Denmark)

    Jørgensen, M.; Scheutz, F.; Strandbygaard, Bertel


    A total of 121 Escherichia coli strains isolated from 3-week-old mink kits were serotyped and examined for virulence factors. 56 strains were isolated from healthy kits while 65 were from ''sticky kits''. Among these, 34 different serotypes were detected. No difference in serotypes or the presence...

  1. 21 CFR 864.2260 - Chromosome culture kit. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Chromosome culture kit. 864.2260 Section 864.2260...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture Products § 864.2260 Chromosome culture kit. (a) Identification. A chromosome culture kit is a device containing the necessary...

  2. Home Pregnancy Test Kits: How Readable Are the Instructions? (United States)

    Holcomb, Carol Ann

    At the conclusion of their study on home pregnancy test kits, Valinas and Perlman (1982) suggested that the instructions accompanying the kits be revised to make them easier to read. A study was undertaken to determine the readability of the printed instructions accompanying five home pregnancy test kits (Daisy II, Answer, Acu-Test, Predictor, and…

  3. 21 CFR 870.1350 - Catheter balloon repair kit. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Catheter balloon repair kit. 870.1350 Section 870... repair kit. (a) Identification. A catheter balloon repair kit is a device used to repair or replace the... effect the repair or replacement. (b) Classification. Class III (premarket approval). (c) Date PMA...

  4. 21 CFR 872.3570 - OTC denture repair kit. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false OTC denture repair kit. 872.3570 Section 872.3570...) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3570 OTC denture repair kit. (a) Identification. An OTC denture repair kit is a device consisting of a material, such as a resin monomer system...

  5. 21 CFR 878.3925 - Plastic surgery kit and accessories. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Plastic surgery kit and accessories. 878.3925... (CONTINUED) MEDICAL DEVICES GENERAL AND PLASTIC SURGERY DEVICES Prosthetic Devices § 878.3925 Plastic surgery kit and accessories. (a) Identification. A plastic surgery kit and accessories is a device intended...



  7. Determination of the concentrations of the steroids estradiol, progesterone and testosterone in bovine sera: comparison of commercial dissociation enhanced lanthanide fluorescence immunoassay kits with conventional radio and enzyme immunoassays. (United States)

    Elliott, C T; Francis, K S; Shortt, H D; McCaughey, W J


    The performance of three conventional enzyme and radioimmunoassays routinely used to detect residues of anabolic steroids in cattle sera were compared with dissociation enhanced lanthanide fluorescence immunoassay (DELFIA) kits designed for the hospital market. Slight modifications to the kit reagents were required for the analysis of bovine sera. Owing to the large sample volumes used in conventional assays, detection limits were generally better than those obtained with DELFIA kits, however, assay reproducibility was enhanced using the DELFIA technology. Comparison of sera obtained from cattle implanted with anabolic steroids revealed a good correlation between alternate methods (r2 from 0.91 to 0.97). The DELFIA kits offer a faster method for measuring estradiol, progesterone and testosterone with adequate sensitivity and in a safer environment than that encountered using radioimmunoassays.

  8. Heterogenous high-level HER-2 amplification in a small subset of colorectal cancers. (United States)

    Marx, Andreas H; Burandt, Eike C; Choschzick, Matthias; Simon, Ronald; Yekebas, Emre; Kaifi, Jussuf T; Mirlacher, Martina; Atanackovic, Djordje; Bokemeyer, Carsten; Fiedler, Walter; Terracciano, Luigi; Sauter, Guido; Izbicki, Jakob R


    HER-2 is the molecular target for antibody-based treatment of breast cancer (trastuzumab). The potential benefit of anti-HER-2 therapy is currently investigated in several other HER-2 amplified cancers. For example, trastuzumab was recently shown to be effective in HER-2 positive gastric cancer. To address the potential applicability of anti-HER-2 therapy in colorectal cancer, tissue microarray sections and colorectal resection specimens of 1851 colorectal cancers were analyzed for HER-2 overexpression and amplification using FDA approved reagents for immunohistochemistry and fluorescence in situ hybridization. HER-2 amplification was seen in 2.5% and HER-2 overexpression in 2.7% of 1439 interpretable colorectal cancers. Amplification was often high level with HER-2 copies ranging from 4 to 60 per tumor cell and was strongly related to protein overexpression. HER-2 amplification and overexpression were unrelated to histological tumor type, tumor localization, grading, pT, pN, pM or survival. As heterogeneity of drug target expression could represent a major drawback for targeted cancer therapy we next studied HER-2 heterogeneity in selected cases. Extensive evaluation of all available large sections from patients with HER-2 positive colorectal cancer revealed heterogenous findings in 3 of 4 cases. In summary, high-level HER-2 amplification occurs in a small fraction of colorectal cancers. Heterogeneity of amplification may limit the utility of anti- HER-2 therapy in some of these tumors and therefore, adequate clinical trials are needed to further evaluate this approach.

  9. Rapid diagnosis of meningitis using reagent strips

    Directory of Open Access Journals (Sweden)

    Parmar Ramesh


    Full Text Available OBJECTIVES: Identification of causative agent with estimation of cerebrospinal fluid (CSF glucose, protein, cells is necessary for accurate diagnosis of meningitis. Unfortunately, even these facilities are not available in many areas. Reagent strips that measure glucose and protein in blood and urine can serve this task but have been used with varying results in the past. This study was carried out to evaluate the utility and efficacy of Combur 10 strips in the diagnosis of meningitis. DESIGN, SETTINGS AND METHODS: A prospective clinical single blinded study of 63 children suspected to have meningitis undergoing CSF analysis. Each CSF sample was divided in to two and was utilised for reagent strip analysis in addition to standard laboratory evaluation and a correlation analysis were made. Statistical Method used: Results were analysed using standard statistical tests. Accuracy of the reagent strips as a screening tool was established using Godyn′s test. RESULTS: The sensitivity, specificity of the reagent strips for the diagnosis of meningitis was 97.14%, 96.42%. The sensitivity, specificity for tuberculous meningitis and bacterial meningitis were 100%, and 96.55%. That for the aseptic meningitis was 70% and 96.55%. Accuracy for the diagnosis of meningitis as a whole, bacterial meningitis, tuberculous meningitis, and aseptic meningitis were 96.78%, 98.2%, 98.27% and 83.0% respectively. CONCLUSION: Combur10 strips thus can be used for the rapid CSF analysis and screening with good accuracy. In situations where facilities of routine laboratory testing are not available this can be of an immense help.

  10. 21 CFR 866.3255 - Escherichia coli serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Escherichia coli serological reagents. 866.3255... coli serological reagents. (a) Identification. Escherichia coli serological reagents are devices that consist of antigens and antisera used in serological tests to identify Escherichia coli from...

  11. 21 CFR 866.3930 - Vibrio cholerae serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Vibrio cholerae serological reagents. 866.3930... cholerae serological reagents. (a) Identification. Vibrio cholerae serological reagents are devices that are used in the agglutination (an antigen-antibody clumping reaction) test to identify Vibrio...

  12. 21 CFR 866.3680 - Sporothrix schenckii serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Sporothrix schenckii serological reagents. 866... Sporothrix schenckii serological reagents. (a) Identification. Sporothrix schenckii serological reagents are... Sporothrix schenckii in serum. The identification aids in the diagnosis of sporothrichosis caused by a...

  13. 21 CFR 660.30 - Reagent Red Blood Cells. (United States)


    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Reagent Red Blood Cells. 660.30 Section 660.30...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells § 660.30 Reagent Red Blood Cells. (a) Proper name and definition. The proper name of the product shall...

  14. 21 CFR 866.3550 - Salmonella spp. serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Salmonella spp. serological reagents. 866.3550... spp. serological reagents. (a) Identification. Salmonella spp. serological reagents are devices that consist of antigens and antisera used in serological tests to identify Salmonella spp. from...

  15. 21 CFR 866.3700 - Staphylococcus aureus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Staphylococcus aureus serological reagents. 866... Staphylococcus aureus serological reagents. (a) Identification. Staphylococcus aureus serological reagents are... epidemiological information on these diseases. Certain strains of Staphylococcus aureus produce an...

  16. 21 CFR 866.3330 - Influenza virus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Influenza virus serological reagents. 866.3330... virus serological reagents. (a) Identification. Influenza virus serological reagents are devices that... subpart E of part 807 of this chapter subject to the limitations in § 866.9....

  17. 40 CFR 160.83 - Reagents and solutions. (United States)


    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Reagents and solutions. 160.83 Section... LABORATORY PRACTICE STANDARDS Testing Facilities Operation § 160.83 Reagents and solutions. All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer or concentration,...

  18. 21 CFR 866.3850 - Trichinella spiralis serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Trichinella spiralis serological reagents. 866... Trichinella spiralis serological reagents. (a) Identification. Trichinella spiralis serological reagents are... Trichinella spiralis in serum. The identification aids in the diagnosis of trichinosis caused by...

  19. 21 CFR 606.65 - Supplies and reagents. (United States)


    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Supplies and reagents. 606.65 Section 606.65 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS CURRENT GOOD MANUFACTURING PRACTICE FOR BLOOD AND BLOOD COMPONENTS Equipment § 606.65 Supplies and reagents. All supplies and reagents used in...

  20. Kit preparation of radioiodinated o-iodohippuran

    Energy Technology Data Exchange (ETDEWEB)

    Hinkle, G.H.; Basmadjian, G.P.; Kirschner, A.S.; Ice, R.D.


    The purpose of this study was to evaluate a kit preparation for radioiodinated o-iodohippuran (I). All ingredients, excluding the radionuclide, were packaged in a ready-to-use kit for easy, quick formulation. Electrophoresis was utilized to evaluate the radiochemical purity of the labeled product and indicated that the radiolabeling technique provided a product with greater than 95% radiochemical purity. Biodistribution studies in rats and rabbits provided an indication of the tissue distribution and localization of the radiopharmaceutical. Computer-generated renogram curves plotted from gamma-camera images of rabbits showed the equivalency of the 131I-labeled I and 123I-labeled I to the commercially available radiopharmaceutical.

  1. Development of Mode Conversion Waveguides at KIT

    Directory of Open Access Journals (Sweden)

    Jin Jianbo


    Full Text Available The development of mode conversion waveguides (launchers for high power gyrotrons has gone through three stages at KIT. Formerly, harmonically deformed launchers have been used in the series gyrotrons developed for the stellarator W7-X. In 2009, a numerical method for the analysis and synthesis of mirror-line launchers was developed at KIT. Such a launcher with adapted mode-converting mirrors for a 2 MW TE34,19-mode, 170GHz coaxial-cavity gyrotron has been designed and tested, and also a mirror-line launcher for the 1MW EU ITER gyrotron has been designed. Recently, based on the Helmholtz-Kirchhoff integral theorem, a novel numerical method for the synthesis of hybrid-type gyrotron launchers has been developed. As an example, TE32,9 mode launchers operating at 170GHz that have been designed using the three different methods are being compared.

  2. Monodisperse Picoliter Droplets for Low-Bias and Contamination-Free Reactions in Single-Cell Whole Genome Amplification.

    Directory of Open Access Journals (Sweden)

    Yohei Nishikawa

    Full Text Available Whole genome amplification (WGA is essential for obtaining genome sequences from single bacterial cells because the quantity of template DNA contained in a single cell is very low. Multiple displacement amplification (MDA, using Phi29 DNA polymerase and random primers, is the most widely used method for single-cell WGA. However, single-cell MDA usually results in uneven genome coverage because of amplification bias, background amplification of contaminating DNA, and formation of chimeras by linking of non-contiguous chromosomal regions. Here, we present a novel MDA method, termed droplet MDA, that minimizes amplification bias and amplification of contaminants by using picoliter-sized droplets for compartmentalized WGA reactions. Extracted DNA fragments from a lysed cell in MDA mixture are divided into 105 droplets (67 pL within minutes via flow through simple microfluidic channels. Compartmentalized genome fragments can be individually amplified in these droplets without the risk of encounter with reagent-borne or environmental contaminants. Following quality assessment of WGA products from single Escherichia coli cells, we showed that droplet MDA minimized unexpected amplification and improved the percentage of genome recovery from 59% to 89%. Our results demonstrate that microfluidic-generated droplets show potential as an efficient tool for effective amplification of low-input DNA for single-cell genomics and greatly reduce the cost and labor investment required for determination of nearly complete genome sequences of uncultured bacteria from environmental samples.

  3. Defense Acquisition University Program Managers Tool Kit (United States)


    22 Procurement Appropriations (Account Numbers and Budget Activities) ................ 23–24 Research, Dev ., Test, and Eval...BA 4 Adv. Comp. Dev . and Prototypes amount amount 6.5 BA 5 System Devel. and Demo. appropriated appropriated 6.6 BA 6 RDT&E Management Support...Testing with . . . (To Include the Mod Kits used for Testing) Is DT or IOT &E Required? Procurement $ YES RDT&E $ NO RDT&E Development Costs of PME

  4. Uncertainties in Site Amplification Estimation (United States)

    Cramer, C. H.; Bonilla, F.; Hartzell, S.


    Typically geophysical profiles (layer thickness, velocity, density, Q) and dynamic soil properties (modulus and damping versus strain curves) are used with appropriate input ground motions in a soil response computer code to estimate site amplification. Uncertainties in observations can be used to generate a distribution of possible site amplifications. The biggest sources of uncertainty in site amplifications estimates are the uncertainties in (1) input ground motions, (2) shear-wave velocities (Vs), (3) dynamic soil properties, (4) soil response code used, and (5) dynamic pore pressure effects. A study of site amplification was conducted for the 1 km thick Mississippi embayment sediments beneath Memphis, Tennessee (see USGS OFR 04-1294 on the web). In this study, the first three sources of uncertainty resulted in a combined coefficient of variation of 10 to 60 percent. The choice of soil response computer program can lead to uncertainties in median estimates of +/- 50 percent. Dynamic pore pressure effects due to the passing of seismic waves in saturated soft sediments are normally not considered in site-amplification studies and can contribute further large uncertainties in site amplification estimates. The effects may range from dilatancy and high-frequency amplification (such as observed at some sites during the 1993 Kushiro-Oki, Japan and 2001 Nisqually, Washington earthquakes) or general soil failure and deamplification of ground motions (such as observed at Treasure Island during the 1989 Loma Prieta, California earthquake). Examples of two case studies using geotechnical data for downhole arrays in Kushiro, Japan and the Wildlife Refuge, California using one dynamic code, NOAH, will be presented as examples of modeling uncertainties associated with these effects. Additionally, an example of inversion for estimates of in-situ dilatancy-related geotechnical modeling parameters will be presented for the Kushiro, Japan site.

  5. The effects of metal ion PCR inhibitors on results obtained with the Quantifiler(®) Human DNA Quantification Kit. (United States)

    Combs, Laura Gaydosh; Warren, Joseph E; Huynh, Vivian; Castaneda, Joanna; Golden, Teresa D; Roby, Rhonda K


    Forensic DNA samples may include the presence of PCR inhibitors, even after extraction and purification. Studies have demonstrated that metal ions, co-purified at specific concentrations, inhibit DNA amplifications. Metal ions are endogenous to sample types, such as bone, and can be introduced from environmental sources. In order to examine the effect of metal ions as PCR inhibitors during quantitative real-time PCR, 2800 M DNA was treated with 0.0025-18.750 mM concentrations of aluminum, calcium, copper, iron, nickel, and lead. DNA samples, both untreated and metal-treated, were quantified using the Quantifiler(®) Human DNA Quantification Kit. Quantification cycle (Cq) values for the Quantifiler(®) Human DNA and internal PCR control (IPC) assays were measured and the estimated concentrations of human DNA were obtained. Comparisons were conducted between metal-treated and control DNA samples to determine the accuracy of the quantification estimates and to test the efficacy of the IPC inhibition detection. This kit is most resistant to the presence of calcium as compared to all metals tested; the maximum concentration tested does not affect the amplification of the IPC or quantification of the sample. This kit is most sensitive to the presence of aluminum; concentrations greater than 0.0750 mM negatively affected the quantification, although the IPC assay accurately assessed the presence of PCR inhibition. The Quantifiler(®) Human DNA Quantification Kit accurately quantifies human DNA in the presence of 0.5000 mM copper, iron, nickel, and lead; however, the IPC does not indicate the presence of PCR inhibition at this concentration of these metals. Unexpectedly, estimates of DNA quantity in samples treated with 18.750 mM copper yielded values in excess of the actual concentration of DNA in the samples; fluorescence spectroscopy experiments indicated this increase was not a direct interaction between the copper metal and 6-FAM dye used to label the probe that

  6. Technetium-99m ceftizoxime kit preparation

    Directory of Open Access Journals (Sweden)

    Simone Odília Fernandes Diniz


    Full Text Available The aim of this work was to prepare a kit of 99mTc-ceftizoxime (99mTc-CFT, with stability and biological activity preserved, able to identify a septic focus (E. coli in the experimental infection model in rats. The preparation of the CFT kit involved the use of lyophilized solutions containing the antibiotic ceftizoxime and the sodium dithionite reducing agent (6.0 mg/mL. After lyophilization, the kit was reconstituted with 1.0 mL of sodium 99mTc-pertechnetate solution (Na99mTcO4- with an activity of 370 MBq. The solution was boiled for 10 min and filtered through a cellulose ester filter. The labeling efficiency was on the order of 92%, remaining stable for six hours and the kit remained stable for two months. The biological activity of the 99mTc-CFT was evaluated by diffusion in agar impregnated with E.coli and S. aureus. Seven Wistar rats, weighing from 200 to 250 g, were used for the development of the septic focus. After 24 hours from the induction of the infectious site (E.coli, the animals were anesthetized and 0.1 mL of 99mTc-CFT (37 MBq was injected into the tail veins of the animals. The images were obtained with a gamma camera one, two and six hours after injection and the regions of interest (ROIs were calculated. The diameters of the inhibition halos for 99mTc-CFT were 27.16 ± 0.23 and 27.17 ± 0.20 for S.aureus and E.coli, respectively, while those for the unlabeled CFT were 30.4 ± 0.33 and 29.43 ± 0.26, respectively. The results for the biodistribution of 99mTc-CFT in infected animals furnished a ratio of 1.97 ± 0.31, 2.10 ± 0.42 and 2.01 ± 0.42 for cpm-target/cpm-no target for the one, two and six-hour periods, respectively. The images showed a clear uptake of labeled antibiotic (99mTc-CFT by the infectious site during the experiment. The results attest to the viability of producing a kit with 99m technetium-labeled ceftizoxime for the investigation of infectious processes.O objetivo deste trabalho foi preparar um kit de Tc

  7. Isothermal Amplification of Insect DNA (United States)

    The loop-mediated isothermal amplification of DNA (LAMP) method can amplify a target DNA sequence at a constant temperature in about one hour. LAMP has broad application in agriculture and medicine because of the need for rapid and inexpensive diagnoses. LAMP eliminates the need for temperature cycl...

  8. 14 CFR Appendix A to Part 121 - First Aid Kits and Emergency Medical Kits (United States)


    ... administer required drugs) 4 Needles (sizes necessary to administer required drugs) 6 50% Dextrose injection... dose ampule or equivalent 2 Nitroglycerin tablets 10 Basic instructions for use of the drugs in the kit... administer required medications) 4 Analgesic, non-narcotic, tablets, 325 mg 4 Antihistamine tablets, 25 mg...

  9. To Kit or Not to Kit? Evaluating and Implementing Science Materials and Resources (United States)

    Schiller, Ellen; Melin, Jacque; Bair, Mary


    With the release of the "Next Generation Science Standards," many schools are reexamining the science materials they are using. Textbook companies and kit developers are eager to meet the demand for "NGSS"-aligned teaching materials. Teacher may have been asked to serve on a science curriculum committee, or to evaluate current…

  10. 21 CFR 880.5960 - Lice removal kit. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lice removal kit. 880.5960 Section 880.5960 Food... § 880.5960 Lice removal kit. (a) Identification. The lice removal kit is a comb or comb-like device intended to remove and/or kill lice and nits from head and body hair. It may or may not be battery...

  11. Data Center Energy Efficiency Measurement Assessment Kit Guide and Specification

    Energy Technology Data Exchange (ETDEWEB)



    A portable and temporary wireless mesh assessment kit can be used to speed up and reduce the costs of a data center energy use assessment and overcome the issues with respect to shutdowns. The assessment kit is comprised of temperature, relative humidity, and pressure sensors. Also included are power meters that can be installed on computer room air conditioners (CRACs) without intrusive interruption of data center operations. The assessment kit produces data required for a detailed energy assessment of the data center.

  12. Forensic interlaboratory evaluation of the ForFLUID kit for vaginal fluids identification. (United States)

    Giampaoli, Saverio; Alessandrini, Federica; Berti, Andrea; Ripani, Luigi; Choi, Ajin; Crab, Roselien; De Vittori, Elisabetta; Egyed, Balazs; Haas, Cordula; Lee, Hwan Young; Korabecná, Marie; Noel, Fabrice; Podini, Daniele; Tagliabracci, Adriano; Valentini, Alessio; Romano Spica, Vincenzo


    Identification of vaginal fluids is an important step in the process of sexual assaults confirmation. Advances in both microbiology and molecular biology defined technical approaches allowing the discrimination of body fluids. These protocols are based on the identification of specific bacterial communities by microfloraDNA (mfDNA) amplification. A multiplex real time-PCR assay (ForFLUID kit) has been developed for identifying biological fluids and for discrimination among vaginal, oral and fecal samples. In order to test its efficacy and reliability of the assay in the identification of vaginal fluids, an interlaboratory evaluation has been performed on homogeneous vaginal swabs. All the involved laboratories were able to correctly recognize all the vaginal swabs, and no false positives were identified when the assay was applied on non-vaginal samples. The assay represents an useful molecular tool that can be easily adopted by forensic geneticists involved in vaginal fluid identification.

  13. Autometallography: tissue metals demonstrated by a silver enhancement kit

    DEFF Research Database (Denmark)

    Danscher, G; Nørgaard, J O; Baatrup, E


    In biological tissue, minute accumulations of gold, silver, mercury and zinc can be visualized by a technique whereby metallic silver is precipitated on tiny accumulations of the two noble metals, or on selenites or sulphides of all four metals. In the present study a silver enhancement kit...... silver enhancement kit (IntenSE, Janssen Pharmaceutica). It was found that the kit performs adequately to the silver lactate gum arabic developer and to the photographic emulsion technique. The kit can be used as a silver enhancement medium for the demonstration of zinc by the Neo-Timm and selenium...

  14. StochKit-FF: Efficient Systems Biology on Multicore Architectures

    CERN Document Server

    Aldinucci, Marco; Liò, Pietro; Sorathiya, Anil; Torquati, Massimo


    The stochastic modelling of biological systems is an informative, and in some cases, very adequate technique, which may however result in being more expensive than other modelling approaches, such as differential equations. We present StochKit-FF, a parallel version of StochKit, a reference toolkit for stochastic simulations. StochKit-FF is based on the FastFlow programming toolkit for multicores and exploits the novel concept of selective memory. We experiment StochKit-FF on a model of HIV infection dynamics, with the aim of extracting information from efficiently run experiments, here in terms of average and variance and, on a longer term, of more structured data.

  15. Desensitized Optimal Filtering and Sensor Fusion Tool Kit Project (United States)

    National Aeronautics and Space Administration — Research on desensitized optimal filtering techniques and a navigation and sensor fusion tool kit using advanced filtering techniques is proposed. Research focuses...

  16. The Virtual Physiological Human ToolKit. (United States)

    Cooper, Jonathan; Cervenansky, Frederic; De Fabritiis, Gianni; Fenner, John; Friboulet, Denis; Giorgino, Toni; Manos, Steven; Martelli, Yves; Villà-Freixa, Jordi; Zasada, Stefan; Lloyd, Sharon; McCormack, Keith; Coveney, Peter V


    The Virtual Physiological Human (VPH) is a major European e-Science initiative intended to support the development of patient-specific computer models and their application in personalized and predictive healthcare. The VPH Network of Excellence (VPH-NoE) project is tasked with facilitating interaction between the various VPH projects and addressing issues of common concern. A key deliverable is the 'VPH ToolKit'--a collection of tools, methodologies and services to support and enable VPH research, integrating and extending existing work across Europe towards greater interoperability and sustainability. Owing to the diverse nature of the field, a single monolithic 'toolkit' is incapable of addressing the needs of the VPH. Rather, the VPH ToolKit should be considered more as a 'toolbox' of relevant technologies, interacting around a common set of standards. The latter apply to the information used by tools, including any data and the VPH models themselves, and also to the naming and categorizing of entities and concepts involved. Furthermore, the technologies and methodologies available need to be widely disseminated, and relevant tools and services easily found by researchers. The VPH-NoE has thus created an online resource for the VPH community to meet this need. It consists of a database of tools, methods and services for VPH research, with a Web front-end. This has facilities for searching the database, for adding or updating entries, and for providing user feedback on entries. Anyone is welcome to contribute.

  17. Preparation of soluble and insoluble polymer supported IBX reagents. (United States)

    Reed, Neal N; Delgado, Mercedes; Hereford, Kristina; Clapham, Bruce; Janda, Kim D


    A series of soluble and insoluble polymer supported versions of the versatile oxidizing reagent IBX has been prepared. Each of the reagents were evaluated for their efficiency in the conversion of benzyl alcohol to benzaldehyde. Results from this study were that the soluble, non-crosslinked polystyrene supported IBX reagent gave the best rate of conversion to benzaldehyde, while the macroporous polymer supported IBX resin provided a superior rate of conversion to benzaldehyde when compared with a gel type resin. The macroporous IBX reagent was also shown to convert a series of alcohols to the corresponding aldehydes and ketones.

  18. Coat-nitrocarburizing using triazine polymer reagent (United States)

    Wen, Li.; Shi, J.; Smith, R. W.


    A chemico-thermal treatment process, coat-nitrocarburizing, has been developed for use on iron and steel. The process consists of treating the workpiece with a coat that forms on the surface from the gaseous products of sublimation and decomposition of a triazine polymer reagent in a closed volume. The process can be used over a wide range of temperatures, either below the eutectoid transformation temperature in the Fe-N-C system for low-temperature nitrocarburizing, or above this temperature for hightemperature nitrocarburizing in different applications. The process is very simple, easily controlled, and is economic. In addition, it is a nonpolluting process, unlike conventional chemico-thermal treatment processes that discharge harmful gases into the atmosphere.

  19. Supramolecular Tectonics for Enzyme-like Reagents

    Institute of Scientific and Technical Information of China (English)

    MAO; LuYuan


    The enzyme-likes and bioactive species were closely related with the life phenomena and served as the reagent of bioassy1,2. In present works, the flow cytometry (FCM) and rapid-scanning stopped-flow (RSSF) spectroscopy combine with the stopped-flow difference UV/Vis spectra, FT-IR and other methods of assay, being used to study the biomimetic reaction and enzyme mimic. Based on catalytic kinetics of enzyme reaction3,4, the reaction mechanisms of the enzyme-likes had been studied and some new methods of kinetic determination were proposed. The study and methods not only provided the basic theoretical models for the life science, but also widened the application fields of biomimetic and analytical chemistry. The main contents of our works and the supramolecular models can be described as follows:  ……

  20. The effects of different maceration techniques on nuclear DNA amplification using human bone. (United States)

    Lee, Esther J; Luedtke, Jennifer G; Allison, Jamie L; Arber, Carolyn E; Merriwether, D Andrew; Steadman, Dawnie Wolfe


    Forensic anthropologists routinely macerate human bone for the purposes of identity and trauma analysis, but the heat and chemical treatments used can destroy genetic evidence. As a follow-up to a previous study on nuclear DNA recovery that used pig ribs, this study utilizes human skeletal remains treated with various bone maceration techniques for nuclear DNA amplification using the standard Combined DNA Index System (CODIS) markers. DNA was extracted from 18 samples of human lower leg bones subjected to nine chemical and heat maceration techniques. Genotyping was carried out using the AmpFlSTR COfiler and AmpFlSTR Profiler Plus ID kits. Results showed that heat treatments via microwave or Biz/Na(2)CO(3) in sub-boiling water efficiently macerate bone and produce amplifiable nuclear DNA for genetic analysis. Long-term use of chemicals such as hydrogen peroxide is discouraged as it results in poor bone quality and has deleterious effects on DNA amplification.

  1. Spheromak Impedance and Current Amplification

    Energy Technology Data Exchange (ETDEWEB)

    Fowler, T K; Hua, D D; Stallard, B W


    It is shown that high current amplification can be achieved only by injecting helicity on the timescale for reconnection, {tau}{sub REC}, which determines the effective impedance of the spheromak. An approximate equation for current amplification is: dI{sub TOR}{sup 2}/dt {approx} I{sup 2}/{tau}{sub REC} - I{sub TOR}{sup 2}/{tau}{sub closed} where I is the gun current, I{sub TOR} is the spheromak toroidal current and {tau}{sub CLOSED} is the ohmic decay time of the spheromak. Achieving high current amplification, I{sub TOR} >> I, requires {tau}{sub REC} <<{tau}{sub CLOSED}. For resistive reconnection, this requires reconnection in a cold zone feeding helicity into a hot zone. Here we propose an impedance model based on these ideas in a form that can be implemented in the Corsica-based helicity transport code. The most important feature of the model is the possibility that {tau}{sub REC} actually increases as the spheromak temperature increases, perhaps accounting for the ''voltage sag'' observed in some experiments, and a tendency toward a constant ratio of field to current, B {proportional_to} I, or I{sub TOR} {approx} I. Program implications are discussed.

  2. Dr. E. Kits van Waveren (1906—1995)

    NARCIS (Netherlands)

    Bas, C.


    With the death on 3 September 1995 of Dr. E. Kits van Waveren, the Dutch mycologists lost one of their most prominent, internationally known amateurs, a specialist on the taxonomy of the genus Psathyrella and an ardent collector. Dr. Kits van Waveren, honorary staff member of the Rijksherbarium at L

  3. School-to-Work Jump-Start Equity Kit. (United States)

    Education Development Center, Inc., Newton, MA. Women's Educational Equity Act Dissemination Center.

    This starter kit is a resource for state and local school-to-work (STW) directors, educators, parents, students, business and community, and economic development organizations serving all students through STW. The kit begins with four articles: "STW and Gender Equity: Opportunity for or Barrier to Economic Parity?" (Katherine Hanson, Joyce…

  4. Energy Education Incentives: Evaluating the Impact of Consumer Energy Kits (United States)

    Kirby, Sarah D.; Guin, Autumn; Langham, Laura


    Measuring the energy and environmental impact of residential energy education efforts is difficult. The E-Conservation residential energy management program uses consumer energy kits to document the impact of energy-efficient improvements. The consumer energy kit provides an incentive for individuals attending energy education workshop, helps…

  5. Somatic mutations of KIT in familial testicular germ cell tumours

    NARCIS (Netherlands)

    Rapley, EA; Hockley, S; Warren, W; Johnson, L; Huddart, R; Crockford, G; Forman, D; Leahy, MG; Oliver, DT; Tucker, K; Friedlander, M; Phillips, KA; Hogg, D; Jewett, MAS; Lohynska, R; Daugaard, G; Richard, S; Heidenreich, A; Geczi, L; Bodrogi, [No Value; Olah, E; Ormiston, WJ; Daly, PA; Looijenga, LHJ; Guilford, P; Aass, N; Fossa, SD; Heimdal, K; Tjulandin, SA; Liubchenko, L; Stoll, H; Weber, W; Einhorn, L; Weber, BL; McMaster, M; Greene, MH; Bishop, DT; Easton, D; Stratton, M


    Somatic mutations of the KIT gene have been reported in mast cell diseases and gastrointestinal stromal tumours. Recently, they have also been found in mediastinal and testicular germ cell tumours (TGCTs), particularly in cases with bilateral disease. We screened the KIT coding sequence ( except exo

  6. ATP生物发光测定试剂研究进展%Reserach progress on ATP bioluminescence reagent

    Institute of Scientific and Technical Information of China (English)

    吴慧清; 李程思; 吴清平; 张菊梅


    Firefly luciferase is the key component of ATP bioluminescence reagent, gained from firefly lantern throuh extraction and purification or preparation through genetic engineering, the performance of ATP bioluminescence reagent was decided by the vitality and the purity of firefly luciferase.Up to now, many present advanced technology were applied on preparation the reagent such as genetic engineering, ATP amplification device, stabilization technology of luciferase protein and luminescence, and so on.Now research focus on improving detection sensitivity and luminescence performance of the ATP bioluminescence reagents, further raising the adaptability of ATP bioluminescence reagents.%萤火虫荧光素酶是ATP生物发光试剂的关键组成部分,可通过萤火虫尾提取纯化或基因工程技术制备,酶的活力和纯度决定了ATP生物发光试剂的性能.迄今许多先进技术在ATP生物发光试剂的制备中均有应用,包括酶基因工程改造技术、ATP循环的酶法放大技术、荧光素酶蛋白的活力及发光稳定技术,特异的细胞ATP提取技术等.ATP生物发光试剂的研究焦点主要集中在提高发光试剂的检测灵敏度和性能、增加产品的适应性等方面.

  7. Disruption of c-Kit Signaling in Kit(W-sh/W-sh) Growing Mice Increases Bone Turnover. (United States)

    Lotinun, Sutada; Krishnamra, Nateetip


    c-Kit tyrosine kinase receptor has been identified as a regulator of bone homeostasis. The c-Kit loss-of-function mutations in WBB6F1/J-Kit(W/W-v) mice result in low bone mass. However, these mice are sterile and it is unclear whether the observed skeletal phenotype is secondary to a sex hormone deficiency. In contrast, C57BL/6J-Kit(W-sh)/(W-sh) (W(sh)/W(sh)) mice, which carry an inversion mutation affecting the transcriptional regulatory elements of the c-Kit gene, are fertile. Here, we showed that W(sh)/W(sh) mice exhibited osteopenia with elevated bone resorption and bone formation at 6- and 9-week-old. The c-Kit W(sh) mutation increased osteoclast differentiation, the number of committed osteoprogenitors, alkaline phosphatase activity and mineralization. c-Kit was expressed in both osteoclasts and osteoblasts, and c-Kit expression was decreased in W(sh)/W(sh)osteoclasts, but not osteoblasts, suggesting an indirect effect of c-Kit on bone formation. Furthermore, the osteoclast-derived coupling factor Wnt10b mRNA was increased in W(sh)/W(sh) osteoclasts. Conditioned medium from W(sh)/W(sh) osteoclasts had elevated Wnt10b protein levels and induced increased alkaline phosphatase activity and mineralization in osteoblast cultures. Antagonizing Wnt10b signaling with DKK1 or Wnt10b antibody inhibited these effects. Our data suggest that c-Kit negatively regulates bone turnover, and disrupted c-Kit signaling couples increased bone resorption with bone formation through osteoclast-derived Wnt 10 b.

  8. Marshall Space Flight Center Telescience Resource Kit (United States)

    Wade, Gina


    Telescience Resource Kit (TReK) is a suite of software applications that can be used to monitor and control assets in space or on the ground. The Telescience Resource Kit was originally developed for the International Space Station program. Since then it has been used to support a variety of NASA programs and projects including the WB-57 Ascent Vehicle Experiment (WAVE) project, the Fast Affordable Science and Technology Satellite (FASTSAT) project, and the Constellation Program. The Payloads Operations Center (POC), also known as the Payload Operations Integration Center (POIC), provides the capability for payload users to operate their payloads at their home sites. In this environment, TReK provides local ground support system services and an interface to utilize remote services provided by the POC. TReK provides ground system services for local and remote payload user sites including International Partner sites, Telescience Support Centers, and U.S. Investigator sites in over 40 locations worldwide. General Capabilities: Support for various data interfaces such as User Datagram Protocol, Transmission Control Protocol, and Serial interfaces. Data Services - retrieve, process, record, playback, forward, and display data (ground based data or telemetry data). Command - create, modify, send, and track commands. Command Management - Configure one TReK system to serve as a command server/filter for other TReK systems. Database - databases are used to store telemetry and command definition information. Application Programming Interface (API) - ANSI C interface compatible with commercial products such as Visual C++, Visual Basic, LabVIEW, Borland C++, etc. The TReK API provides a bridge for users to develop software to access and extend TReK services. Environments - development, test, simulations, training, and flight. Includes standalone training simulators.

  9. Comparison of different protocols for DNA preparation and PCR amplification of mitochondrial genes of tardigrades

    Directory of Open Access Journals (Sweden)

    Ralph O. SCHILL


    Full Text Available Phylogenetic relationships and molecular taxonomy within the Tardigrada have been given a lot of attention in recent years. Here I present the first comparison of different protocols for DNA preparation by investigating six commercial available DNA extraction kits and the CTAB method. Successful extraction of DNA from tardigrades depends strongly on the life-stage (embryo, adult, and on the condition of the specimens, respectively on the preservation (anhydrobiotic, ethanol. Although the extraction kits showed differences in the amount of extracted DNA, in all cases fresh tissue of live animals or embryos resulted in the best quality and quantity of DNA. A lesser amount of DNA was extractable from anhydrobiotic animals and embryos and the results of specimens fixed in ethanol were unsatisfactory. All used commercially available DNA extraction kits and PCR cocktails have been focused on vertebrate tissues, blood, cultured cells, bacteria and yeast. However, I used successfully the kits according to the manufacturer’s instruction without changes in the protocols for DNA extraction of tardigrades. Commercial kits provide a simple and convenient way to isolate pure genomic DNA of high-quality from tardigrades. Furthermore I tested eight different Taq polymerase enzymes for PCR amplification of mitochondrial genes of tardigrades. Each of the enzymes resulted in a PCR product, and even if the amount of the PCR products was quite different, it was possible to use it successful for direct sequencing. Summarizing, the successful PCR of the target DNA depends on the purity and quality of the DNA template and for this the species preservation is more critical than the extraction method or the PCR cocktail which can be optimized.

  10. CCK-8试剂在血液毒理学检测中的应用%Application of CCK-8 reagent in detection of hematotoxicology

    Institute of Scientific and Technical Information of China (English)

    凌晓璇; 刘林华; 梁海荣; 程小广; 唐焕文


    Objective:To optimize the applicable condition for CCK -8 cell viability kit in hematotoxicology research. Methods: The CCK - 8 reagent kit and two optimized methods were used respectively to observe the influence of hydroquinone with different concentrations on proliferation of TK6 cells, then the results were compared with that by cell counting method to analyze the influencing factors. Results: Detection results based on the protocol demonstrated that HQ influenced the CCK - 8 reagent function, and led to discrepancy compared to the cell counting method, while results from the two optimized methods showed similarity with the cell counting method. Conclusion: HQ can influence the results with CCK - 8 reagent, so a detection method of cell proliferation was optimized in hematotoxicology for this kit.%目的:优化CCK-8试剂盒在血液毒理学检测中的应用条件.方法:以CCK-8试剂盒标准检测法和两种优化的方法分别检测不同浓度的氢醌(hydroquinone,HQ)对TK6细胞增殖速度的影响,分别与计数法比较,分析影响实验检测的原因.结果:试剂盒标准方法中,HQ对CCK-8试剂的检测结果有影响,结果出现偏差,优化后的两种方法与计数法检测结果较为相似.结论:HQ对CCK-8试剂检测结果有影响,优化出一种适用于HQ血液毒理学检测的细胞增殖方法.

  11. Portable kit for identification and detection of drugs in human urine using surface-enhanced Raman spectroscopy. (United States)

    Han, Zhenzhen; Liu, Honglin; Meng, Juan; Yang, Liangbao; Liu, Jing; Liu, Jinhuai


    A portable kit was demonstrated for rapid and reliable surface-enhanced Raman scattering (SERS) detection of drugs in human urine. This kit contains two sealed reagent tubes, a packet of standardized SERS substrates, and a mini Raman device. A 3 min pretreatment for separating amphetamines from human urine was developed with an extraction rate of >80% examined by ultraperformance liquid chromatography (UPLC). Simultaneously, highly reproducible two-dimensional (2D) gold nanorod (GNR) arrays were assembled by the use of methoxymercaptopoly(ethylene glycol) (mPEG-SH) capping. Thirty batches of GNR arrays produced the 1001 cm(-1) intensity of methamphetamine (MA) molecules with a relative standard deviation (RSD) of 7.9%, and a 21 × 21 μm(2) area mapping on a 2D GNR array produced a statistical RSD of amphetamines in human urine was at least 0.1 ppm. Moreover, the portable kit was successfully used for detecting MA, 3,4-methylenedioxymethamphetamine (MDMA), and methcathinone (MC) in 30 volunteers' urine samples with various clinical natures, and the dual-analyte detection of MA and MDMA implied a good capability of multiplex analysis. UPLC examination and the SERS recovery test clearly indicated that our pretreatment procedure was sufficient to lower the high background signals caused by complex components in urine and demonstrated the practicability and the resistance to false positives, which is a vital problem for law enforcement applications. The excellent performance of our portable kit promises a great prospective toward a rapid, reliable, and on-spot analyzer, especially for public safety and healthcare.

  12. Education kits for fiber optics, optoelectronics, and optical communications (United States)

    Hájek, Martin; Švrček, Miroslav


    Our company MIKROKOM, s.r.o. is engaged for many years in development of education equipment and kits for fiber optics, optoelectronics and optical communications. We would like to inform competitors of conference about results of this long-time development. Requirements on education kits and equipment in a modern and dynamic area as is optical communications and fiber optics are quite difficult. The education kits should to clearly introduce students to given issue - the most important physical principles and technical approaches, but it should to introduce also to new and modern technologies, which are quickly changing and developing. On the other hand should be these tools and kits reasonable for the schools. In our paper we would like to describe possible ways of development of this education kits and equipment and present our results of long-time work, which covers very wide range. On the one hand we developed equipment and kits for clear demonstration of physical effects using plastic optical fibers POF, next we prepare kits with a glass fibers, which are the most used fibers in practice and after as much as the kits, which covers broad range of passive and active elements of the optical networks and systems and which makes possible to create complex optical transmission connection. This kind of systems with using corresponding tools and equipment introduce the students to properties, manipulation, measurement and usage of optical fibers, traces and many active and passive components. Furthermore, with using different sorts of optical sources, photodetectors, fiber optics couplers etc., students can get acquainted with all optoelectronics transmission system, which uses different sorts of signals. Special part will be devoted also to effort mentioned before - to implement modern technologies such as e.g. Wavelength Division Multiplex (WDM) into the education kits. Our presentation will inform auditors about development of mentioned education kits and

  13. Oncogenic c-kit transcript is a target for binase. (United States)

    Mitkevich, Vladimir A; Petrushanko, Irina Y; Kretova, Olga V; Zelenikhin, Pavel V; Prassolov, Vladimir S; Tchurikov, Nickolai A; Ilinskaya, Olga N; Makarov, Alexander A


    Mutational activation of c-Kit receptor tyrosine kinase is common in acute myelogenous leukemia (AML). One such activating point mutation is the N822K replacement in the c-Kit protein. Here we investigate the selective cytotoxic effect of binase--RNase from Bacillus intermedius--on FDC-P1-N822K cells. These cells were derived from myeloid progenitor FDC-P1 cells, in which ectopic expression of N822K c-kit gene induces interleukin-3 independent growth. In order to determine whether the sensitivity of these cells to binase is caused by the expression of c-kit oncogene, the cytotoxicity of the RNase was studied in the presence of selective inhibitor of mutated c-Kit imatinib (Gleevec). Inhibition of mutated c-Kit protein leads to the loss of cell sensitivity to the apoptotic effect of binase, while the latter still decreases the amount of cellular RNA. Using green fluorescent protein as an expression marker for the c-Kit oncoprotein, we demonstrate that the elimination of c-Kit is the key factor in selective cytotoxicity of binase. Quantitative RT-PCR with RNA samples isolated from the binase-treated FDC-P1-N822K cells shows that binase treatment results in 41% reduction in the amount of с-kit mRNA. This indicates that the transcript of the activated mutant c-kit is the target for toxic action of binase. Thus, the combination of inhibition of oncogenic protein with the destruction of its mRNA is a promising approach to eliminating malignant cells.

  14. Synthesis and characterization of zwitterionic carbon dioxide fixing reagents

    DEFF Research Database (Denmark)

    Mikkelsen, Mette; Jørgensen, Mikkel; Krebs, Frederik C


    The synthesis of three amine-based carbon dioxide fixing reagents is presented. The reagents were designed so that they would be able to bind CO2 reversibly through the formation of the well known carbamates that was stabilized through forming a zwitterion. CO2 fixing experiments were performed...

  15. 21 CFR 866.3520 - Rubeola (measles) virus serological reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rubeola (measles) virus serological reagents. 866... Rubeola (measles) virus serological reagents. (a) Identification. Rubeola (measles) virus serological... to rubeola virus in serum. The identification aids in the diagnosis of measles and...

  16. 40 CFR 792.83 - Reagents and solutions. (United States)


    ... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Reagents and solutions. 792.83 Section... solutions. All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer... solutions shall not be used....

  17. 21 CFR 58.83 - Reagents and solutions. (United States)


    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Reagents and solutions. 58.83 Section 58.83 Food... solutions. All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer... solutions shall not be used....

  18. Investigation of Chemiluminescence with Electrogenerated Reagents and Its Analytical Application

    Institute of Scientific and Technical Information of China (English)

    章竹君; 李保新; 郑行望


    In this paper, studies on chemiluminescence (CL) systems with electrogenerated reagents, including BrO-, ClO-, Br2, [Cu-(HIO6)2]5-, H2O2, Mn3+, Co3+ and Ag2+, are described.The analytical applications of the CL system with electrogenerated reagents are reviewed.

  19. Use of Competition Kinetics with Fast Reactions of Grignard Reagents

    DEFF Research Database (Denmark)

    Holm, Torkil


    found for the four substrates do not differ significantly and it seems possible that there is a ceiling over the rate of reaction of this reagent, for example caused by diffusion control. This may explain that competition kinetics using allylmagnesium bromide have failed to show kinetic isotope effects......Competition kinetics are useful for estimation of the reactivities of Grignard reagents if the reaction rates do not differ widely and if exact rates are not needed. If the rate of mixing is slower than the rate of reaction the ratios between the rates of fast and slow reagents are found to be too...... small.This is concluded from experiments in which results obtained by competition kinetics are compared with results obtained directly by flow stream procedures. A clearer picture of the reactivity ratios is obtained when the highly reactive reagent is highly diluted with its competitor. A fast reagent...

  20. Spectrophotometric determination of tannins by phosphotungstic-phosphomolybdic reagent

    Energy Technology Data Exchange (ETDEWEB)

    Reicher, F.; Sierakowski, M.R.; Correa, J.B.C. (Parana Univ., Curitiba (Brazil). Dept. de Bioquimica)


    There are several colorimetric techniques to determine tannins in plant extracts. One frequently used is the Folin method (phosphotungstic acid reagent) that procedures a blue color with phenolic compounds. However, this coloured complex is unstable. With the Folin-Ciocalteau reagent, used in protein determination (Lowry et al. J.B.C. 193: 265, 1951) good results were obtained, even in the absence of cooper solution. Using phosphotungstic-phosphomolybdic reagent (Folin-Denis), it was obtained maximum color with 1,0 ml of the reagent in 20 minutes, after the additon of 10 ml 20% sodium carbonate solution. Tannins samples containing 10 to 200 were analysed. Absorbances are determined at 720 or 600 nm. Tannins of commercial preparations from Acacia negra were analysed by the phosphotungstic-phosphomolybdic reagent before (A) and after (B) treatment with chromate hyde powder. By this procedure hydrolysible tannins were determined (A-B).

  1. Target-Induced and Equipment-Free DNA Amplification with a Simple Paper Device. (United States)

    Liu, Meng; Hui, Christy Y; Zhang, Qiang; Gu, Jimmy; Kannan, Balamurali; Jahanshahi-Anbuhi, Sana; Filipe, Carlos D M; Brennan, John D; Li, Yingfu


    We report on a paper device capable of carrying out target-induced rolling circle amplification (RCA) to produce massive DNA amplicons that can be easily visualized. Interestingly, we observed that RCA was more proficient on paper than in solution, which we attribute to a significantly higher localized concentration of immobilized DNA. Furthermore, we have successfully engineered a fully functional paper device for sensitive DNA or microRNA detection via printing of all RCA-enabling molecules within a polymeric sugar film formed from pullulan, which was integrated with the paper device. This encapsulation not only stabilizes the entrapped reagents at room temperature but also enables colorimetric bioassays with minimal steps.

  2. Enhancing PCR Amplification of DNA from Recalcitrant Plant Specimens Using a Trehalose-Based Additive

    Directory of Open Access Journals (Sweden)

    Tharangamala Samarakoon


    Full Text Available Premise of the study: PCR amplification of DNA extracted from plants is sometimes difficult due to the presence of inhibitory compounds. An effective method to overcome the inhibitory effect of compounds that contaminate DNA from difficult plant specimens is needed. Methods and Results: The effectiveness of a PCR additive reagent containing trehalose, bovine serum albumin (BSA, and polysorbate-20 (Tween-20 (TBT-PAR was tested. PCR of DNA extracted from fresh, silica-dried, and herbarium leaf material of species of Achariaceae, Asteraceae, Lacistemataceae, and Samydaceae that failed using standard techniques were successful with the addition of TBT-PAR. Conclusions: The addition of TBT-PAR during routine PCR is an effective method to improve amplification of DNA extracted from herbarium specimens or plants that are known to contain PCR inhibitors.

  3. 3种国产化试剂盒与IdentifilerTM试剂盒检验结果比较%A comparative study on the typing difference between the IdentifilerTM kit and three domestic genotyping kits

    Institute of Scientific and Technical Information of China (English)

    吴微微; 郝宏蕾; 林锦锋; 应晓兰; 苏艳佳; 金敏贵


    目的 比较3种常用国产化PCR扩增试剂盒与IdentifilerTM试剂盒的检验结果.方法 对455份中国汉族无关个体血样,分别用IdentifilerTM、DNA TyperTM15、GoldenEye 16BT、AGCU 17+1 4种试剂盒进行检测,对其中共有的11个基因座位分型结果进行比较,并对有差异的样本进行测序分析.结果 3种国产化试剂盒在455份样本中,发现4例样本的分型结果有差异,差异率为0.88%.其中,DNA TyperTM15试剂盒CSF1PO基因座发现1例等位基因丢失,AGCU 17+1试剂盒D18S51基因座发现1例等位基因扩增不平衡,测序结果表明,2例均系点突变导致;GoldenEye16BT试剂盒D21S11基因座发现2例假3峰型,其中1例测序未发现碱基突变,分析3峰原因可能是stutter滑脱峰所致.结论 与IdentifilerTM试剂盒比较,3种国产化试剂盒均有分型差异现象,成因不同,在实践中应给予注意.%Objective To study the difference of DNA testing results between the IdentifilerTM kit and three domestic genotyping kits. Methods 455 unrelated individuals of Han population in China were typed using the ldentifilerTM, DNA TyperTM 15, GoldenEye 16BT and AGCU17 + 1 kits respectively. The genotypes were compared at 11 mutual loci among these four kits, and those samples with inconsistent genotypes were further analyzed by sequencing. Results Total 4 samples were observed to have inconsistent genotypes in all 455 individuals with three domestic genotyping kits, and the frequency was 0.88%. For these four samples, one allele dropout, one amplification imbalance, and two false tri-peak were observed at CSF1PO locus with the TyperTMl5 kit, D18S51 locus with the AGCU17 + 1 kit and D21S 11 locus with the 16BT kit correspondingly. The results of sequence analysis showed that all of these differences were caused by point mutation except a false tri-peak sample was caused by replication slippage. Conclusion Compared with the IdentifilerTM kit, occasional genotyping differences could

  4. Standardization of reagents and methods used in cytological and histological practice with emphasis on dyes, stains and chromogenic reagents

    DEFF Research Database (Denmark)

    Lyon, H O; De Leenheer, A P; Horobin, R W;


    The need for the standardization of reagents and methods used in the histology laboratory is demonstrated. After definitions of dyes, stains, and chromogenic reagents, existing standards and standards organizations are discussed. This is followed by practical instructions on how to standardize dy...

  5. Immobilized Bioluminescent Reagents in Flow Injection Analysis. (United States)

    Nabi, Abdul

    Available from UMI in association with The British Library. Bioluminescent reactions exhibits two important characteristics from an analytical viewpoint; they are selective and highly sensitive. Furthermore, bioluminescent emissions are easily measured with a simple flow-through detector based on a photomultiplier tube and the rapid and reproducible mixing of sample and expensive reagent is best achieved by a flow injection manifold. The two most important bioluminescent systems are the enzyme (luciferase)/substrate (luciferin) combinations extracted from fireflies (Photinus pyralis) and marine bacteria (Virio harveyi) which requires ATP and NAD(P)H respectively as cofactors. Reactions that generate or consume these cofactors can also be coupled to the bioluminescent reaction to provide assays for a wide range of clinically important species. A flow injection manifold for the study of bioluminescent reactions is described, as are procedures for the extraction, purification and immobilization of firefly and bacterial luciferase and oxidoreductase. Results are presented for the determination of ATP using firefly system and the determination of other enzymes and substrates participating in ATP-converting reactions e.g. creatine kinase, ATP-sulphurylase, pyruvate kinase, creatine phosphate, pyrophosphate and phophoenolypyruvate. Similarly results are presented for the determination of NAD(P)H, FMN, FMNH_2 and several dehydrogenases which produce NAD(P)H and their substrates, e.g. alcohol, L-lactate, L-malate, L-glutamate, Glucose-6-phosphate and primary bile acid.

  6. Detection of hazelnuts and almonds using commercial ELISA test kits. (United States)

    Garber, Eric A E; Perry, Jesse


    Three commercial sandwich enzyme-linked immunosorbent assay (ELISA) test kits for the detection of hazelnuts and almonds were evaluated. Limits of detection and dynamic ranges were determined for hazelnuts and almonds spiked into cooked oatmeal, dipping chocolate, and muffins (baked). The limit of detection values varied from 1 to 38 μg/g, depending on the food matrix and ELISA test kit. Percent recoveries based on the standards supplied with the test kits varied from 10% to 170%. It is impossible to ascertain whether the percent recoveries reflect the performance of the ELISAs or differences between the protein content of the nuts used to spike the samples and the test kit standards. Unfortunately, reference materials do not exist that can be used to compare the results from different test kits and standardize the test kit standards. Also, insufficient knowledge regarding the epitope specificity of the antibodies used in the ELISAs further hinders interpretation of the results generated by the different test kits.

  7. Label-free and highly sensitive electrochemical detection of E. coli based on rolling circle amplifications coupled peroxidase-mimicking DNAzyme amplification. (United States)

    Guo, Yuna; Wang, Yu; Liu, Su; Yu, Jinghua; Wang, Hongzhi; Wang, Yalin; Huang, Jiadong


    In this work, a simple, label-free, low cost electrochemical biosensor for highly sensitive and selective detection of Escherichia coli has been developed on the basis of rolling circle amplification (RCA) coupled peroxidase-mimicking DNAzyme amplification. A aptamer-primer probe (APP) containing anti-E. coli aptamer and a primer sequence complementary to a circular probe, which includes two G-quadruplex units, is used for recognizing target and triggering RCA-based polymerase elongation. Due to RCA coupled DNAzyme amplification strategy, the presence of target E. coli leads to the formation of numerous G-quadruplex oligomers on electrode, which folds into G-quadruplex/hemin complexs with the help of K(+) and hemin, thus generating extremely strong catalytic activity toward H2O2 and giving a remarkably strong electrochemical response. As far as we know, this work is the first time that RCA coupled peroxidase-mimicking DNAzyme amplification technique have been integrated into electrochemical assay for detecting pathogenic bacteria. Under optimal conditions, the proposed biosensor exhibits ultrahigh sensitivity toward E. coli with detection limits of 8cfumL(-1) and a detection range of 5 orders of magnitude. Besides, our biosensor also shows high selectivity toward target E. coli and has the advantages in its rapidness, low cost, simplified operations without the need of electrochemical labeling steps and additional labile reagents. Hence, the RCA coupled peroxidase-mimicking DNAzyme amplification-based electrochemical method might create a useful and practical platform for detecting E. coli and related food safety analysis and clinical diagnosis.

  8. Dynamics and Control of DNA Sequence Amplification

    CERN Document Server

    Marimuthu, Karthikeyan


    DNA amplification is the process of replication of a specified DNA sequence \\emph{in vitro} through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction (PCR) as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal tempe...

  9. Optimization of laser capture microdissection and RNA amplification for gene expression profiling of prostate cancer

    Directory of Open Access Journals (Sweden)

    Vasmatzis George


    Full Text Available Abstract Background To discover prostate cancer biomarkers, we profiled gene expression in benign and malignant cells laser capture microdissected (LCM from prostate tissues and metastatic prostatic adenocarcinomas. Here we present methods developed, optimized, and validated to obtain high quality gene expression data. Results RNase inhibitor was included in solutions used to stain frozen tissue sections for LCM, which improved RNA quality significantly. Quantitative PCR assays, requiring minimal amounts of LCM RNA, were developed to determine RNA quality and concentration. SuperScript II™ reverse transcriptase was replaced with SuperScript III™, and SpeedVac concentration was eliminated to optimize linear amplification. The GeneChip® IVT labeling kit was used rather than the Enzo BioArray™ HighYield™ RNA transcript labeling kit since side-by-side comparisons indicated high-end signal saturation with the latter. We obtained 72 μg of labeled complementary RNA on average after linear amplification of about 2 ng of total RNA. Conclusion Unsupervised clustering placed 5/5 normal and 2/2 benign prostatic hyperplasia cases in one group, 5/7 Gleason pattern 3 cases in another group, and the remaining 2/7 pattern 3 cases in a third group with 8/8 Gleason pattern 5 cases and 3/3 metastatic prostatic adenocarcinomas. Differential expression of alpha-methylacyl coenzyme A racemase (AMACR and hepsin was confirmed using quantitative PCR.

  10. Development of a Backpack Survival Kit for Ejection Seats. (United States)



  11. Dumpster Optics: teaching and learning optics without a kit (United States)

    Donnelly, Judy; Magnani, Nancy; Robinson, Kathleen


    The Next Generation Science Standards (NGSS) and renewed emphasis on STEM education in the U.S. have resulted in the development of many educational kits for teaching science in general and optics in particular. Many teachers do not have funding to purchase kits and practical experience has shown that even costly kits can have poorly written and misleading instructions and may include experiments that would not work in a classroom. Dumpster Optics lessons are designed to use inexpensive, commonly found materials. All lessons have been field-tested with students. We will describe the development of the lessons, provide examples of field testing experiences and outline possible future activities.

  12. Clinical usefulness of trypsin radioimmunoassay kit

    Energy Technology Data Exchange (ETDEWEB)

    Kanamori, Isao; Nakano, Satoshi; Hurukawa, Masakazu; Okumura, Yasuki; Higuchi, Chizuko; Yanase, Mikiko; Onogi, Michiteru


    From the clinical use of RIA-gnost trypsin kit, the following results were obtained. 1. Standard curve showed a steep and good curve was shown. 2. Incubation: The condition for the first incubation was set at the room temperature for 10-24 hours and that for the second incubation at the room temperature for 3-5 hours. With these settings, satisfactory results were obtained. 3. Reproducibility and recovery: The C.V. of the reproducibility and the recovery were considered superior, and the values were below 10 % and +-3 %, respectively. 4. Correlation between trypsin and serum elestase-1: An excellent positive correlation (coefficient of correlation r = 0.889) was shown. 5. Serum trypsin concentration of normal and pancreatic diseases: The normal range was from 100 to 500 ng/ml. Acute pancreatitis rose obviously. Diabetes mellitus and chronic pancreatitis was below 500 ng/ml and the pancreatic cancer showed a tendency to scatter in the range of 50-1 250 ng/ml. The above results indicated that serum trypsin can be easily measured with high precision by using this method. Thus the method is considered useful for the diagnosis of pancreatic diseases.

  13. Math Description Engine Software Development Kit (United States)

    Shelton, Robert O.; Smith, Stephanie L.; Dexter, Dan E.; Hodgson, Terry R.


    The Math Description Engine Software Development Kit (MDE SDK) can be used by software developers to make computer-rendered graphs more accessible to blind and visually-impaired users. The MDE SDK generates alternative graph descriptions in two forms: textual descriptions and non-verbal sound renderings, or sonification. It also enables display of an animated trace of a graph sonification on a visual graph component, with color and line-thickness options for users having low vision or color-related impairments. A set of accessible graphical user interface widgets is provided for operation by end users and for control of accessible graph displays. Version 1.0 of the MDE SDK generates text descriptions for 2D graphs commonly seen in math and science curriculum (and practice). The mathematically rich text descriptions can also serve as a virtual math and science assistant for blind and sighted users, making graphs more accessible for everyone. The MDE SDK has a simple application programming interface (API) that makes it easy for programmers and Web-site developers to make graphs accessible with just a few lines of code. The source code is written in Java for cross-platform compatibility and to take advantage of Java s built-in support for building accessible software application interfaces. Compiled-library and NASA Open Source versions are available with API documentation and Programmer s Guide at http:/ / prim e.jsc.n asa. gov.

  14. Dynamics and control of DNA sequence amplification

    Energy Technology Data Exchange (ETDEWEB)

    Marimuthu, Karthikeyan [Department of Chemical Engineering and Center for Advanced Process Decision-Making, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213 (United States); Chakrabarti, Raj, E-mail:, E-mail: [Department of Chemical Engineering and Center for Advanced Process Decision-Making, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213 (United States); Division of Fundamental Research, PMC Advanced Technology, Mount Laurel, New Jersey 08054 (United States)


    DNA amplification is the process of replication of a specified DNA sequence in vitro through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal temperature profile. Strategies for the optimal synthesis of the DNA amplification control trajectory are proposed. Analogous methods can be used to formulate control problems for more advanced amplification objectives corresponding to the design of new types of DNA amplification reactions.

  15. Regulatory role of kit ligand-c-kit interaction and oocyte factors in steroidogenesis by rat granulosa cells. (United States)

    Miyoshi, Tomoko; Otsuka, Fumio; Nakamura, Eri; Inagaki, Kenichi; Ogura-Ochi, Kanako; Tsukamoto, Naoko; Takeda, Masaya; Makino, Hirofumi


    Although kit ligand (KL)-c-kit interaction is known to be critical for oogenesis and folliculogenesis, its role in ovarian steroidogenesis has yet to be elucidated. We studied the impact of KL-c-kit interaction in regulation of steroidogenesis using rat oocyte/granulosa cell co-culture. In the presence of oocytes, soluble KL suppressed FSH-induced estradiol production and aromatase mRNA expression without affecting FSH-induced progesterone production. The KL effect on steroidogenesis was interrupted by an anti-c-kit neutralizing antibody, suggesting that KL-c-kit interaction is involved in suppression of estrogen by granulosa cells through oocyte c-kit action. The cAMP-PKA pathway activity was not directly involved in the estrogen regulation by KL-c-kit action. It was of note that KL treatment increased the expression levels of oocyte-derived FGF-8, GDF-9 and BMP-6, while it reduced the expression levels of oocyte-derived BMP-15 in the oocyte-granulosa cell co-culture. Given the findings that FGF-8, but not GDF-9, BMP-6 or -15, suppressed FSH-induced estrogen production by granulosa cells, oocyte-derived FGF-8 is linked to suppression of FSH-induced estrogen production through the KL-c-kit interaction. Furthermore, the suppression of FSH-induced estrogen production by KL in the co-culture was reversed by a FGF receptor kinase inhibitor and the effect of the inhibitor was enhanced in combination with extracellular-domain protein of BMPRII, which interferes with BMP-15 and GDF-9 activities. Thus, the actions of endogenous oocyte factors including FGF-8 and BMP-15/GDF-9 were involved in the KL activity that inhibited FSH-induced estradiol production. Collectively, the results indicate that KL-c-kit interaction plays a role in estrogenic regulation through oocyte-granulosa cell communication.


    Directory of Open Access Journals (Sweden)



    Full Text Available Used the polymeric reagents in oil wastewater treatment is an effective method of eliminate hydrocarbons. The present study aims to finding reagents that lead to lowering of extractible (EXT, suspended solids (SS and chemical oxygen demand (COD of industrial wastewater from washing cars in loading ramps petroleum products. For this purpose five reagents were tested, namely: polyamines, cationic polyacrylamides, polydiallydimethyl ammonium chloride (PolyDADMAC, melamine formaldehyde polymer resin and polydicyandiamide polymer resin. Obtaining removal degrees over 80 % justifies using this method in the industrial practice.

  17. Transition metal complexes with Girard reagents and their hydrazones

    Directory of Open Access Journals (Sweden)

    Vojinović-Ješić Ljiljana S.


    Full Text Available This is the first review dealing with the coordination chemistry of metal complexes with Girard's reagents and their hydrazones. The short introduction points out to chemical properties and significance of these organic compounds. The next section briefly describes synthetic methods for preparing complexes with Girard's reagents, as well as modes of coordination of these ligands. The last two extensive sections review the preparation, stereochemistry and structural characteristics of metal complexes with Girard's hydrazones, including some newer non-hydrazonic derivatives of Girard's reagents, also.[Acknowledgments. Projekat Ministarstva nauke Republike Srbije, br. 172014

  18. The effect of nest box temperature on kit growth rate and survival in the American mink (Neovison vison)

    DEFF Research Database (Denmark)

    Schou, Toke Munk; Malmkvist, Jens


    dying and kits growth. Instead parameters concerning litter composition did have significant effects on kit growth and survival. Litters with high number of Totborn and kit AliveD1 affected kit growth and kit viability negatively (increased number of live born kits dying), which indicates that factors...

  19. Comparison of amino acid derivatization reagents for LC-ESI-MS analysis. Introducing a novel phosphazene-based derivatization reagent. (United States)

    Rebane, Riin; Oldekop, Maarja-Liisa; Herodes, Koit


    Amino acid analysis with high performance liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS) is an emerging method. For more sensitive analysis, derivatization is used and next to commercially available derivatization reagents such as dansyl chloride (DNS), 9-fluorenylmethyl chloroformate (FMOC-Cl) and diethyl ethoxymethylenemalonate (DEEMM), new derivatization reagents are designed specially for LC-ESI-MS, like p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS) which provides very low limits of detection. In this work, a novel phosphazene based derivatization reagent (FOSF) that provides comparable limits of quantitation (LoQ) to TAHS is introduced. Moreover, a thorough comparison between FOSF, TAHS, DNS, FMOC-Cl and DEEMM is carried out for 7 different amino acids - Arg, Asp, Gly, β-Ala, Pro, Trp and Phe. This is a first time that thorough comparison is carried out on the same instrument for amino acid derivatization reagents. Results on the same instrument for five amino acid derivatization reagents show that novel reagents are sensitive with LoQ values around 80 fmol but have disadvantages such as problematic chromatographic separation. Next to novel reagents, DEEMM offers very good LoQ-s (average of 150 fmol) and wide dynamic linear range.

  20. Applications of homemade kit in mutation detection of genes

    Institute of Scientific and Technical Information of China (English)

    ZHAO Chunxia; XU Guowang; SHI Xianzhe; MA Jianmei; ZHANG Yan; L(U) Shen; YANG Qing


    Several methods of mutation detection, such as single-strand conformation polymorphism (SSCP), tandem SSCP/heteroduplex analysis and SNaPshot analysis were developed using homemade kit on ABI 310 genetic analyzer, and were successfully applied to mutation detection of 31 colorectal tumor samples. The sieving capability of homemade kit and commercial kit were compared, results demonstrate that homemade kit has higher resolution and shorter analysis time. In clinical tumor samples, 26% K-ras (exon 1) and 24% p53 (exons 7-8) were found to have mutations, and all mutations were single point variations. A majority of mutations occurred in one gene, only 1 tumor contained alterations in the two genes, which indicates that development of colorectal cancer lies on alternate pathways, and may correlate with different gene mutations.

  1. 21 CFR 872.3600 - Partially fabricated denture kit. (United States)


    ... (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3600 Partially fabricated denture kit. (a... mold, by partially polymerizing the resin denture base materials while the materials are in...

  2. Desensitized Optimal Filtering and Sensor Fusion Tool Kit Project (United States)

    National Aeronautics and Space Administration — It is proposed to develop desensitized optimal filtering techniques and to implement these algorithms in a navigation and sensor fusion tool kit. These proposed...

  3. 21 CFR 866.3370 - Mycobacterium tuberculosis immunofluorescent reagents. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycobacterium tuberculosis immunofluorescent... § 866.3370 Mycobacterium tuberculosis immunofluorescent reagents. (a) Identification. Mycobacterium... used to identify Mycobacterium tuberculosis directly from clinical specimens. The identification...

  4. Effect of polyamine reagents on exchange capacity in ion exchangers (United States)

    Petrova, T. I.; Dyachenko, F. V.; Bogatyreva, Yu. V.; Borodastov, A. K.; Ershova, I. S.


    Effect of compounds involved in complex reagents is described using Helamin 906H reagent as an example. The working exchange capacity of KU-2-8chs cation exchanger in hydrogen form and Amberlite IRA 900Cl anion exchanger in OH form remained almost unchanged when they were used repeatedly to purify water that contained Helamin 906H reagent; in addition, this capacity was the same upon filtration of water that did not contain this reagent. Leakage of total organic carbon was observed earlier than that of calcium ions upon filtration of the solution through the cation exchanger layer. The test results obtained in industrial conditions indicated that using H-OH filters to purify turbine condensate enables the decrease of the concentration of organic and other impurities therein.

  5. Characterizing fluidic seals for on-board reagent delivery (United States)

    Inamdar, Tejas; Anthony, Brian W.


    The reagent delivery mechanism in a point-of-care, HIV diagnostic, microfluidic device is studied. Reagents held in an aluminum blister pack are released on the opening of a fluidic seal. The fluidic seals, controlling the flow of reagents, are characterized to reduce anomalies in the desired flow pattern. The findings of this research can be divided into three categories - 1) bonding phenomenon 2) influence of seal pattern on flow and rupture mechanics and 3) process parameters which minimize flow anomalies. Four seal patterns - line hemisphere, line flat, chevron hemisphere and chevron flat were created and tested for reagent delivery using a flow sensor and a force gauge. Experiments suggest that one of the patterns - line-flat - inducted the fewest flow anomalies. A parameter scoping exercise of the seal manufacturing process parameters (temperature, time, pressure) was performed for the line flat seal. Temperature, time, pressure / gap and distance settings which minimize flow anomalies were found.

  6. Diagnosis of Spontaneous Bacterial Peritonitis in Children by Reagent Strips

    Directory of Open Access Journals (Sweden)

    Fatemeh Farahmand


    Full Text Available This study was aimed to evaluate the efficacy of dipstick tests (leukocyte esterase and nitrite in diagnosis of spontaneous bacterial peritonitis (SBP in cirrhotic patients. Forty six children with ascites hospitalized between 2009 and 2010 in Children Medical Center were enrolled in this study. Reagent strip assays for leukocyte esterase and nitrite were performed on ascetic fluid and the results were compared to manual cell counting and ascitic fluid culture. SBP was defined as having a polymorphonuclear ascites count of ≥ 250/mm3. Twenty children were female and twenty six were male with mean age of 3±3.9 years. The sensitivity specificity, positive and negative predictive values of the leukocyte esterase reagent strips were all 100%. The sensitivity, specificity, positive and negative predictive value of the nitrite reagent strip test were 100%, 97%, 90% and 100% respectively. Leukocyte esterase reagent strips may provide a rapid, bedside diagnostic test for the diagnosis of SBP.

  7. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification

    Directory of Open Access Journals (Sweden)

    Michael G. Mauk


    Full Text Available Microfluidic components and systems for rapid (<60 min, low-cost, convenient, field-deployable sequence-specific nucleic acid-based amplification tests (NAATs are described. A microfluidic point-of-care (POC diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of “lab on a chip” NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (polymerase-chain reaction-based tests in clinical laboratories can be realized with a disposable, palm-sized, plastic microfluidic chip in which: (1 nucleic acids (NAs are extracted from relatively large (~mL volume sample lysates using an embedded porous silica glass fiber or cellulose binding phase (“membrane” to capture sample NAs in a flow-through, filtration mode; (2 NAs captured on the membrane are isothermally (~65 °C amplified; (3 amplicon production is monitored by real-time fluorescence detection, such as with a smartphone CCD camera serving as a low-cost detector; and (4 paraffin-encapsulated, lyophilized reagents for temperature-activated release are pre-stored in the chip. Limits of Detection (LOD better than 103 virons/sample can be achieved. A modified chip with conduits hosting a diffusion-mode amplification process provides a simple visual indicator to readily quantify sample NA template. In addition, a companion microfluidic device for extracting plasma from whole blood without a centrifuge, generating cell-free plasma for chip-based molecular diagnostics, is described. Extensions to a myriad of related applications including, for example, food testing, cancer screening, and insect genotyping are briefly surveyed.

  8. Late development of homoeothermy in mink (Mustela vison) kits - a strategy for maximum survival rate

    DEFF Research Database (Denmark)

    Tauson, A-H; Chwalibog, André; Tygesen, M P


    of heat production (HE) by means of indirect calorimetry lasting 3 h were performed on neonatal kits and kits from 1 to 54 days of age. Both single kits and groups of 4-5 huddling kits were kept at 15 degrees C (L) or 30 degrees C (H) [from 35 days onwards at 25 degrees C (H)]. Animals were weighed before...

  9. Gel-forming reagents and uses thereof for preparing microarrays (United States)

    Golova, Julia; Chernov, Boris; Perov, Alexander


    New gel-forming reagents including monomers and cross-linkers, which can be applied to gel-drop microarray manufacturing by using co-polymerization approaches are disclosed. Compositions for the preparation of co-polymerization mixtures with new gel-forming monomers and cross-linker reagents are described herein. New co-polymerization compositions and cross-linkers with variable length linker groups between unsaturated C.dbd.C bonds that participate in the formation of gel networks are disclosed.

  10. Multiscale image contrast amplification (MUSICA) (United States)

    Vuylsteke, Pieter; Schoeters, Emile P.


    This article presents a novel approach to the problem of detail contrast enhancement, based on multiresolution representation of the original image. The image is decomposed into a weighted sum of smooth, localized, 2D basis functions at multiple scales. Each transform coefficient represents the amount of local detail at some specific scale and at a specific position in the image. Detail contrast is enhanced by non-linear amplification of the transform coefficients. An inverse transform is then applied to the modified coefficients. This yields a uniformly contrast- enhanced image without artefacts. The MUSICA-algorithm is being applied routinely to computed radiography images of chest, skull, spine, shoulder, pelvis, extremities, and abdomen examinations, with excellent acceptance. It is useful for a wide range of applications in the medical, graphical, and industrial area.

  11. Evaluation of novel derivatisation reagents for the analysis of oxysterols

    Energy Technology Data Exchange (ETDEWEB)

    Crick, Peter J., E-mail: [Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom); Aponte, Jennifer; Bentley, T. William [Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom); Matthews, Ian [College of Engineering, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom); Wang, Yuqin [Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom); Griffiths, William J., E-mail: [Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom)


    Graphical abstract: - Highlights: • New derivatisation reagents for LC–MS analysis of oxysterols. • New reagents based on Girard P give high ion-currents and informative LC–MS{sup n} spectra. • Permanent charge is vital for efficient MS{sup n} fragmentation. • New reagents offer greater scope for incorporation of isotope labels. - Abstract: Oxysterols are oxidised forms of cholesterol that are intermediates in the synthesis of bile acids and steroid hormones. They are also ligands to nuclear and G protein-coupled receptors. Analysis of oxysterols in biological systems is challenging due to their low abundance coupled with their lack of a strong chromophore and poor ionisation characteristics in mass spectrometry (MS). We have previously used enzyme-assisted derivatisation for sterol analysis (EADSA) to identify and quantitate oxysterols in biological samples. This technique relies on tagging sterols with the Girard P reagent to introduce a charged quaternary ammonium group. Here, we have compared several modified Girard-like reagents and show that the permanent charge is vital for efficient MS{sup n} fragmentation. However, we find that the reagent can be extended to include sites for potential stable isotope labels without a loss of performance.

  12. Development of a field kit for use by non-scientists for chemical tracking using 5-(4-nitrophenyl)-2,4-pentadien-1-al. (United States)

    Suzuki, Shinichi


    5-(4-Nitrophenyl)-2,4-pentadien-1-al (NPPD) can be used for chemical tracking in crime scene investigations. A color test kit for NPPD was developed for use by non-scientists, such as police officers, in the field. However, this kit had problems, including contact with concentrated HCl, and instability of the reagent (naphthoresorcinol methanol solution) used in the first step of color development. To overcome these problems, in the present study, a field kit was developed with the concentrated HCl sealed in a vial so it did not contact the operator. A glass tube with two compartments was used to separate the naphthoresorcinol and methanol before use. When the color test was conducted, a cotton swab was inserted into the tube. Before insertion, the cotton was used to collect a sample from a suspect that had been in contact with a surface sprayed with a 1% NPPD methanol solution. Insertion of the cotton swab broke the thin glass that separated the methanol and naphthoresorcinol, and any NPPD on the swab reacted with the naphthoresorcinol methanol solution. The cotton swab was then pushed further to break the glass separating the concentrated HCl. A red color then developed if NPPD was present on the cotton swab. For testing the kit, NPPD was sprayed in an area where a crime was expected to occur. This kit will be useful for detecting a contact with or near a crime scene, because samples do not require analysis in a forensic science laboratory. Instead, the results can be confirmed at the scene of crime.

  13. Amplification of cellular oncogenes in solid tumors

    Directory of Open Access Journals (Sweden)

    Ozkan Bagci


    Full Text Available The term gene amplification refers to an increase in copy number of a gene. Upregulation of gene expression through amplification is a general mechanism to increase gene dosage. Oncogene amplifications have been shown in solid human cancers and they are often associated with progression of cancer. Defining oncogene amplification is useful since it is used as a prognostic marker in clinical oncology nowadays, especially v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2 (HER2 targeted agents are used in breast cancer patients with high level of HER2 overexpression as a therapeutic approach. However, patients without HER2 overexpression do not appear to benefit from these agents. We concluded that determination of oncogene amplification in solid tumors is an important factor in treatment of human cancers with many unknowns. We have referred to PubMed and some databases to prepare this article.

  14. SAS molecular tests Escherichia coli O157 detection kit. Performance tested method 031203. (United States)

    Bapanpally, Chandra; Montier, Laura; Khan, Shah; Kasra, Akif; Brunelle, Sharon L


    The SAS Molecular tests Escherichia coli O157 Detection method, a loop-mediated isothermal amplification method, performed as well as or better than the U.S. Department of Agriculture, Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, bagged mixed lettuce, and fresh spinach. Ground beef (30% fat, 25 g test portion) was validated for 7-8 h enrichment, leafy greens were validated in a 6-7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16-20 h enrichment. The method performance for meat and leafy green matrixes was also shown to be acceptable under conditions of co-enrichment with Salmonella. Thus, after a short co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. The SAS Molecular tests Salmonella Detection Kit was validated using the same test portions as for the SAS Molecular tests E. coli O157 Detection Kit and those results are presented in a separate report. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 50 E. coli 0157 strains, including H7 and non-motile strains, and 30 non-E. coli O157 strains examined. Finally, the method was shown to be robust when variations to DNA extract hold time and DNA volume were varied. The method comparison and robustness data suggest a full 7 h enrichment time should be used for 25 g ground beef test portions.

  15. Pulse Compression And Raman Amplification In Optical Fibres (United States)

    Byron, Kevin C.


    Experimental and theoretical investigations on Raman amplification in fibres have been carried out and simultaneous amplification and pulse compression observed. With a fibre design optimised for amplification high gain may be obtained at practical pump power levels.

  16. Comparison of commercial DNA extraction kits and quantitative PCR systems for better sensitivity in detecting the causative agent of paratuberculosis in dairy cow fecal samples. (United States)

    Fock-Chow-Tho, D; Topp, E; Ibeagha-Awemu, E A; Bissonnette, N


    Mycobacterium avium ssp. paratuberculosis (MAP) causes ruminant paratuberculosis (Johne's disease) worldwide. Oral-fecal contamination is the most important mode of transmission of paratuberculosis, so eradicating MAP-shedding animals could prevent disease propagation. Fecal culture, a well-known method for MAP diagnosis, requires costly specialized media and a long incubation time that sometimes ends in disappointing bacterial contamination. To facilitate the efforts of control programs, we evaluated the performance of direct fecal quantitative PCR (qPCR) assays for their sensitivity and robustness for MAP detection. Commercial kits use different strategies for extracting DNA, combined with qPCR systems, to detect the presence of MAP in fecal samples. In this study, we compared the sensitivity of 3 commercially available DNA extraction kits (A, B, and C) combined with 2 qPCR systems (T and V) for the detection of MAP in infectious cows. A total of 49 dairy cows from 5 herds were sampled twice a year for 3 yr and diagnosed using fecal culture and ELISA. Eight replicates of their fecal samples from the first sampling were tested using each DNA extraction method and qPCR detection system. Although all 3 of the commercial DNA extraction kits have been previously described as very efficient for the diagnosis of paratuberculosis, kit B provided the highest sensitivity. Indeed, 89% of the cows declared positive for paratuberculosis by both fecal culture and ELISA were identified with kit B, whereas only 23 and 43% of the cows were identified with kits A and C, respectively. Interestingly, kit B was able to detect some low-MAP shedders. The qPCR detection system also played a critical role: system T yielded qPCR with the highest sensitivity. The results of this study suggest that DNA extraction kit B combined with detection system T provides the best amplification of MAP DNA from fecal samples with the highest sensitivity and specificity. Although 1 DNA extraction and q

  17. Validación y resultados preliminares del kit de AmpFlSTR® MinifilerTM en el Laboratorio de Genética Forense de la DIJIN, Policía Nacional de Colombia Validation and preliminary results of kit AmpFlSTR® MinifilerTM in the Laboratory of Forensic Genetics of DIJIN, National Police Colombia

    Directory of Open Access Journals (Sweden)

    M.L. Acevedo


    Full Text Available El laboratorio de la Dirección de Investigación Criminal e Interpol (DIJIN de la Policía Nacional implementó la utilización del kit AmpF l STR® MinifilerTM PCR Amplification de AppliedBiosystems con el fin de ayudar en la resolución de casos donde se sospechara una gran fragmentación del ADN. Metodología: Se evaluaron las alturas de los picos de los controles utilizando dos termocicladores diferentes, dos analizadores genéticos ABI Prism310® y empleando dos volúmenes finales de reacción de PCR. Se evaluó la sensibilidad utilizando muestras de concentraciones entre 0,1 ng/µL hasta 0,006 ng/µL y los datos fueron analizados con el software Genemapper ID v3.2. Se valoró la precisión y la reproducibilidad comparando 10 muestras donde se incluía sangre a diferentes concentraciones, saliva, prendas, evidencias traza, pelo, hueso y diente. Una vez realizada la validación se hicieron pruebas con muestras de 5 casos que habían presentado concentraciones bajas de ADN y amplificaciones parciales utilizando el kit AmpF l STR® Identifiler®. Resultados y discusión: Se pudo verificar que la concentración ideal para el uso del kit AmpF l STR® MinifilerTM está en un rango entre 0,05 y 0,075 ng/µl de ADN. Se encontró que es posible obtener resultados en muestras con concentraciones de 0,02 ng/µl; cuando la concentración es inferior, los resultados no son reproducibles. En 4 muestras previamente genotipificadas con el kit AmpF l STR® Identifiler®, se amplió el número de marcadores obtenidos en más del 48% con la utilización del kit AmpF l STR® MinifilerTM y se pudo llegar a la probabilidad exigida por la ley colombiana.The laboratory of the Bureau of Criminal Investigation and Interpol (DIJIN of the National Police implemented the use of the kit AMPF l STR®MinifilerTM PCR Amplification of AppliedBiosystems to assist in the resolution of cases where suspected large DNA fragmentation. Methods: We evaluated the peak heights of

  18. Dedifferentiation in Gastrointestinal Stromal Tumor to an Anaplastic KIT Negative Phenotype – a Diagnostic Pitfall. Morphologic and Molecular Characterization of 8 Cases Occurring either de-novo or after Imatinib Therapy (United States)

    Antonescu, Cristina R; Romeo, Salvatore; Zhang, Lei; Nafa, Khedoudja; Hornick, Jason L.; Nielsen, G. Petur; Mino-Kenudson, Mari; Huang, Hsuan-Ying; Mosquera, Juan-Miguel; Dei Tos, Paolo A; Fletcher, Christopher D.M.


    Most GISTs can be recognized by their monotonous cytologic features and overexpression of KIT oncoprotein. Altered morphology and loss of CD117 reactivity has been described previously after chronic imatinib treatment; however, this phenomenon has not been reported in imatinib-naïve tumors. Eight patients with abrupt transition from a classic CD117-positive spindle cell GIST to an anaplastic CD117-negative tumor were investigated for underlying molecular mechanisms of tumor progression. Pathologic and molecular analysis was performed on each of the two components. Genomic DNA PCR for KIT, PDGFRA, BRAF and KRAS hot spot-mutations and FISH for detecting KIT gene copy number alterations were performed. TP53 mutational analysis was performed in 5 cases. There were 7 males and 1 female, with an age range of 23–65 years. Five of the primary tumors were located in the stomach, while one case each originated in small bowel, colon and rectum. In 3 patients, the dedifferentiated component occurred in the setting of imatinib-resistance, while the remaining 5 occurred de novo. The dedifferentiated component had an anaplastic appearance, including one angiosarcomatous phenotype, with high mitotic activity and necrosis, and showed complete loss of CD117 (8/8) and CD34 (5/8) expression, and de novo expression of either cytokeratin (4/8) or desmin (1/8). There was no difference in the KIT genotype between the two components. However, two imatinib-resistant tumors showed co-existence of KIT exon 11 and exon 13 mutations. FISH showed loss of one KIT gene in 3 cases and low level amplification of KIT in 2 other cases in the CD117-negative component, compared to the CD117-positive area. TP53 mutation was identified in 1/5 cases tested, being present in both components. In summary, dedifferentiation in GIST may occur either de novo or after chronic imatinib exposure and can represent a diagnostic pitfall. This phenomenon is not related to additional KIT mutations, but might be

  19. A flavor kit for BSM models

    Energy Technology Data Exchange (ETDEWEB)

    Porod, Werner [Universitaet Wuerzburg, Institut fuer Theoretische Physik und Astronomie, Wuerzburg (Germany); Staub, Florian [BCTP und Physikalisches Institut, Universitaet Bonn, Bonn (Germany); Vicente, Avelino [Universite de Liege, IFPA, Liege (Belgium)


    We present a new kit for the study of flavor observables in models beyond the standard model. The setup is based on the public codes SARAH and SPheno and allows for an easy implementation of new observables. The Wilson coefficients of the corresponding operators in the effective lagrangian are computed by SPheno modules written by SARAH. New operators can also be added by the user in a modular way. For this purpose a handy Mathematica package called thePreSARAHhas been developed. This uses FeynArts and FormCalc to derive generic form factors at tree- and 1-loop levels and to generate the necessary input files for SARAH. This framework has been used to implement BR(l{sub α} → l{sub β}γ), BR(l{sub α} → 3l{sub β}), CR(μ - e, A), BR(τ → P l), BR(h → l{sub α}l{sub β}), BR(Z → l{sub α}l{sub β}), BR(B{sup 0}{sub s,d} → l anti l), BR(anti B → X{sub s}γ), BR(anti B → X{sub s}l anti l), BR(anti B → X{sub d,s}νanti ν), BR(K{sup +} → π{sup +}νanti ν), BR(K{sub L} → π{sup 0}νanti ν), ΔM{sub B{sub s,B{sub d}}}, ΔM{sub K}, ε{sub K}, BR(B → Kμanti μ), BR(B → lν), BR(D{sub s} → lν) and BR(K → lν) in SARAH. Predictions for these observables can now be obtained in a wide range of SUSY and non-SUSY models. Finally, the user can use the same approach to easily compute additional observables. (orig.)

  20. Telescience Resource Kit (TReK) (United States)

    Lippincott, Jeff


    Telescience Resource Kit (TReK) is one of the Huntsville Operations Support Center (HOSC) remote operations solutions. It can be used to monitor and control International Space Station (ISS) payloads from anywhere in the world. It is comprised of a suite of software applications and libraries that provide generic data system capabilities and access to HOSC services. The TReK Software has been operational since 2000. A new cross-platform version of TReK is under development. The new software is being released in phases during the 2014-2016 timeframe. The TReK Release 3.x series of software is the original TReK software that has been operational since 2000. This software runs on Windows. It contains capabilities to support traditional telemetry and commanding using CCSDS (Consultative Committee for Space Data Systems) packets. The TReK Release 4.x series of software is the new cross platform software. It runs on Windows and Linux. The new TReK software will support communication using standard IP protocols and traditional telemetry and commanding. All the software listed above is compatible and can be installed and run together on Windows. The new TReK software contains a suite of software that can be used by payload developers on the ground and onboard (TReK Toolkit). TReK Toolkit is a suite of lightweight libraries and utility applications for use onboard and on the ground. TReK Desktop is the full suite of TReK software -most useful on the ground. When TReK Desktop is released, the TReK installation program will provide the option to choose just the TReK Toolkit portion of the software or the full TReK Desktop suite. The ISS program is providing the TReK Toolkit software as a generic flight software capability offered as a standard service to payloads. TReK Software Verification was conducted during the April/May 2015 timeframe. Payload teams using the TReK software onboard can reference the TReK software verification. TReK will be demonstrated on-orbit running on

  1. The Role of c-KIT in Tumorigenesis: Evaluation in Canine Cutaneous Mast Cell Tumors

    Directory of Open Access Journals (Sweden)

    Joshua D. Webster


    Full Text Available The c-KIT proto-oncogene has been implicated in the pathogenesis of several neoplastic diseases, including gastrointestinal stromal tumors and mastocytosis in humans, and mast cell tumors (MCTs in canines. Cutaneous MCTs are common neoplasms in dogs and have a variable biologic behavior. The goal of this study was to define the prognostic significance of c-KIT mutations identified in canine MCTs and the associations between c-KIT mutations, KIT localization, and KIT expression levels. Microdissection and polymerase chain reaction were performed on 60 MCTs to identify c-KIT mutations. Anti-KIT antibodies were used for immunohistochemical evaluation of KIT localization. Forty-two MCTs were included in a tissue microarray, and KIT expression was quantified using immunofluorescence. Canine MCTs with c-KIT mutations were significantly associated with an increased incidence of recurrent disease and death. c-KIT mutations were also significantly associated with aberrant protein localization; however, the level of KIT expression did not correlate with either c-KIT mutations or changes in protein localization. Considering the high prevalence of canine MCTs and the central role of c-KIT in the tumorigenesis of certain tumors, canine MCTs are an excellent model for characterizing the role of c-KIT in neoplastic diseases and is a potential target for novel therapeutic agents in clinical trials.

  2. Creatine kinase B subunit as measured with a radioimmunoassay kit in detection of acute myocardial infarction. (United States)

    Witherspoon, L R; Shuler, S E; Genre, C F; Gilbert, S S; Moore, R J; Meihaus, V; Hurry, E K


    Results with a commercial radioimmunoassay (RIA) reagent kit for quantification of the creatine kinase B subunit (CK-B) (Nuclear-Medical Laboratories, Irving, TX 75061) were compared with results obtained by electrophoresis for patients consecutively admitted to our coronary care unit for suspected acute myocardial infarction. Analytical sensitivity, precision, and specificity of the RIA were satisfactory. Its clinical efficacy was assessed in 97 patients suspected of having had an acute myocardial infarction. Of 30 patients who had had an acute myocardial infarction, increased CK-B was detected by RIA in 30 and by electrophoresis in 27. The temporal relationship between CK-B by RIA and CK-MB by electrophoresis was similar. Of 66 admissions where infarction was not established, CK-B was negligibly increased in samples from four patients by RIA, and from one by electrophoresis. Although not abnormally increased (greater than 5 U/L), CK-MB was detected by electrophoresis in samples from another five of these 66 patients. We conclude that estimation of CK-B by this RIA is an excellent alternative to estimation of CK-MB by electrophoresis in patients suspected of having had an acute myocardial infarction.

  3. [Efficient extraction of transmembrane proteins using ProteoExtract Transmembrane Protein Extraction Kit]. (United States)

    Błachnio, Karina


    Detergents commonly used for solubilization of membrane proteins may be ionic or non-ionic. Exposing membrane proteins to detergents, however, can adversely affect their native structure, which can be a major hindrance for functional studies. This is especially true for proteins with multiple transmembrane domains. The ProteoExtract Transmembrane Protein Extraction Kit (TM-PEK), offered by Merck, provides a detergent-free novel reagents to enable the mild and efficient extraction of proteins containing seven transmembrane domains, such as GPCRs (G-Protein Coupled Receptors) e.g.: Frizzled-4 and CELSR-3, from mammalian cells. The fraction enriched in transmembrane proteins using TM-PEK is directly compatible with enzyme assays, non-denaturing gel electrophoresis, 1- and 2-D SDS-PAGE, MS analysis, Western blotting, immunoprecipitation and ELISA. Unlike many alternatives, TM-PEK extraction procedure does not require sonication, extended rigorous vortexing, ultracentrifugation, or incubation of samples at elevated temperatures--thus minimizing the risk of post-extraction degradation or modifications.

  4. LocoKit - A Construction Kit for Exploration of Morphology of Legged Robots

    DEFF Research Database (Denmark)

    Larsen, Jørgen Christian; Støy, Kasper


    Producing steady stable and energy efficient locomotion in legged robots with the ability to walk in unknown terrain is a big challenge in robotics. In addressing this challenge, it is often desirable to experiment with different morphologies and see how they influence on the way the robot walks....... This is however not always easy, since robots are often built as a fixed system with a limited possibility of changing the morphology without redesign a significant part of the robot. This work is focusing on the creation of a robotic construction kit specifically aimed at easing the process of constructing...... legged robots. This is accomplished by giving the creator the possibility to easily do morphological changes to the robot even after it have been build, to see how it effects the robot’s ability to walk in unknown terrain....

  5. LocoKit - A Construction Kit for Exploration of Morphology of Legged Robots

    DEFF Research Database (Denmark)

    Larsen, Jørgen Christian; Støy, Kasper


    Producing steady stable and energy efficient locomotion in legged robots with the ability to walk in unknown terrain is a big challenge in robotics. In addressing this challenge, it is often desirable to experiment with different morphologies and see how they influence on the way the robot walks....... This is however not always easy, since robots are often built as a fixed system with a limited possibility of changing the morphology without redesign a significant part of the robot. This work is focusing on the creation of a robotic construction kit specifically aimed at easing the process of constructing...... legged robots. This is accomplished by giving the creator the possibility to easily do morphological changes to the robot even after it have been build, to see how it effects the robot?s ability to walk in unknown terrain....

  6. Strategies to prepare and use functionalized organometallic reagents. (United States)

    Klatt, Thomas; Markiewicz, John T; Sämann, Christoph; Knochel, Paul


    Polyfunctional zinc and magnesium organometallic reagents occupy a central position in organic synthesis. Most organic functional groups are tolerated by zinc organometallic reagents, and Csp(2)-centered magnesium organometallic reagents are compatible with important functional groups, such as the ester, aryl ketone, nitro, cyano, and amide functions. This excellent chemoselectivity gives zinc- and magnesium-organometallic reagents a central position in modern organic synthesis. Efficient and general preparations of these organometallic reagents, as well as their most practical and useful reactions, are presented in this Perspective. As starting materials, a broad range of organic halides (iodides, bromides, and also to some extent chlorides) can be used for the direct insertion of magnesium or zinc powder; the presence of LiCl very efficiently promotes such insertions. Alternatively, aromatic or heterocyclic bromides also undergo a smooth bromine-magnesium exchange when treated with i-PrMgCl·LiCl. Alternative precursors of zinc and magnesium reagents are polyfunctionalized aryl and heteroaryl molecules, which undergo directed metalations with sterically hindered TMP bases (TMP = 2,2,6,6-tetramethylpiperide) of magnesium and zinc. This powerful C-H functionalization method gives access to polyfunctional heterocyclic zinc and magnesium reagents, which undergo efficient reactions with numerous electrophiles. The compatibility of the strong TMP-bases with BF3·OEt2 (formation of frustrated Lewis pairs) dramatically increases the scope of these metalations, giving for example, a practical access to magnesiated pyridines and pyrazines, which can be used as convenient building blocks for the preparation of biologically active molecules.

  7. Linking Arctic amplification and local feedbacks (United States)

    Balcerak, Ernie


    Climate simulations show that as the Earth warms, the Arctic warms more than the average global warming. However, models differ on how much more the Arctic warms, and although scientists have proposed a variety of mechanisms to explain the Arctic warming amplification, there is no consensus on the main reasons for it. To shed light on this issue, Hwang et al. investigated the relationship between Arctic amplification and poleward energy transport and local Arctic feedbacks, such as changes in cloud cover or ice loss, across a group of models. The researchers noted that differences in atmospheric energy transport did not explain the ranges of polar amplification; rather, models with more amplification showed less energy transport into high latitudes. The authors found that decreasing energy transport is due to a coupled relationship between Arctic amplification and energy transport: Arctic amplification reduces the equator-to-pole temperature gradient, which strongly decreases energy transport. They suggest that this coupled relationship should be taken into account in studies of Arctic amplification. (Geophysical Research Letters, doi:10.1029/2011GL048546, 2011)

  8. Quantum Amplitude Amplification and Estimation

    CERN Document Server

    Brassard, G; Mosca, M; Tapp, A; Brassard, Gilles; Hoyer, Peter; Mosca, Michele; Tapp, Alain


    Consider a Boolean function $\\chi: X \\to \\{0,1\\}$ that partitions set $X$ between its good and bad elements, where $x$ is good if $\\chi(x)=1$ and bad otherwise. Consider also a quantum algorithm $\\mathcal A$ such that $A \\ket{0} = \\sum_{x\\in X} \\alpha_x \\ket{x}$ is a quantum superposition of the elements of $X$, and let $a$ denote the probability that a good element is produced if $A \\ket{0}$ is measured. If we repeat the process of running $A$, measuring the output, and using $\\chi$ to check the validity of the result, we shall expect to repeat $1/a$ times on the average before a solution is found. *Amplitude amplification* is a process that allows to find a good $x$ after an expected number of applications of $A$ and its inverse which is proportional to $1/\\sqrt{a}$, assuming algorithm $A$ makes no measurements. This is a generalization of Grover's searching algorithm in which $A$ was restricted to producing an equal superposition of all members of $X$ and we had a promise that a single $x$ existed such tha...

  9. Raman amplification in optical communication systems

    DEFF Research Database (Denmark)

    Kjær, Rasmus


    Fiber Raman amplifiers are investigated with the purpose of identifying new applications and limitations for their use in optical communication systems. Three main topics are investigated, namely: New applications of dispersion compensating Raman amplifiers, the use Raman amplification to increase...

  10. Can Anomalous Amplification be Attained Without Postselection?

    CERN Document Server

    Martínez-Rincón, Julián; Viza, Gerardo I; Howell, John C


    We present a parameter estimation technique based on performing joint measurements of a weak interaction away from the weak-value-amplification approximation. Two detectors are used to collect full statistics of the correlations between two weakly entangled degrees of freedom. Without the need of postselection, the protocol resembles the anomalous amplification of an imaginary-weak-value-like response. The amplification is induced in the difference signal of both detectors allowing robustness to different sources of technical noise, and offering in addition the advantages of balanced signals for precision metrology. All of the Fisher information about the parameter of interest is collected, and a phase controls the amplification response. We experimentally demonstrate the proposed technique by measuring polarization rotations in a linearly polarized laser pulse. The effective sensitivity and precision of a split detector is increased when compared to a conventional continuous-wave balanced detection technique...

  11. Mass and Volume Optimization of Space Flight Medical Kits (United States)

    Keenan, A. B.; Foy, Millennia Hope; Myers, Jerry


    Resource allocation is a critical aspect of space mission planning. All resources, including medical resources, are subject to a number of mission constraints such a maximum mass and volume. However, unlike many resources, there is often limited understanding in how to optimize medical resources for a mission. The Integrated Medical Model (IMM) is a probabilistic model that estimates medical event occurrences and mission outcomes for different mission profiles. IMM simulates outcomes and describes the impact of medical events in terms of lost crew time, medical resource usage, and the potential for medically required evacuation. Previously published work describes an approach that uses the IMM to generate optimized medical kits that maximize benefit to the crew subject to mass and volume constraints. We improve upon the results obtained previously and extend our approach to minimize mass and volume while meeting some benefit threshold. METHODS We frame the medical kit optimization problem as a modified knapsack problem and implement an algorithm utilizing dynamic programming. Using this algorithm, optimized medical kits were generated for 3 mission scenarios with the goal of minimizing the medical kit mass and volume for a specified likelihood of evacuation or Crew Health Index (CHI) threshold. The algorithm was expanded to generate medical kits that maximize likelihood of evacuation or CHI subject to mass and volume constraints. RESULTS AND CONCLUSIONS In maximizing benefit to crew health subject to certain constraints, our algorithm generates medical kits that more closely resemble the unlimited-resource scenario than previous approaches which leverage medical risk information generated by the IMM. Our work here demonstrates that this algorithm provides an efficient and effective means to objectively allocate medical resources for spaceflight missions and provides an effective means of addressing tradeoffs in medical resource allocations and crew mission success

  12. The Quality Lighting Teaching Kit: enlightening our future (United States)

    Walker, Constance E.; Pompea, Stephen M.


    The U.S. National Optical Astronomy Observatory's Education and Public Outreach group has produced a Quality Lighting Teaching (QLT) Kit, as an outcome of the International Year of Light 2015. The kits are designed around problem-based learning scenarios. The kit's six activities allow students to address real lighting problems that relate to wildlife, sky glow, aging eyes, energy consumption, safety, and light trespass. The activities are optimized for 11-14 year olds but can be expanded to younger and older. Most of the activities can be done within in a few minutes with the exception of the Energy Activity. The activities can be done during class or afterschool and as stations (that the students rotate through) or as stand-alones (one at a time). All aspects of the program are as ready-for-use. Everything you need for the six activities is included in the kit. Tutorial videos (on the program's webpage) have been created on how to do the activities. They can be found on the webpage, Fourteen Google+ Hangouts on Air have been offered, addressing questions on the activities and logistics. Assessments (in the form of pre- and post-surveys for the students and as post-surveys for the instructors) provide learning outcomes and improvements. Eighty-nine out of 100 kits have been distributed to SPIE, OSA, CIE, IDA and the IAU in 31 countries. The QLT Kit is a stepping-stone to bring awareness to the (younger) public on how quality lighting locally can redress issues like light pollution globally.

  13. Oncogenic signaling by Kit tyrosine kinase occurs selectively on the Golgi apparatus in gastrointestinal stromal tumors. (United States)

    Obata, Y; Horikawa, K; Takahashi, T; Akieda, Y; Tsujimoto, M; Fletcher, J A; Esumi, H; Nishida, T; Abe, R


    Gastrointestinal stromal tumors (GISTs) are caused by gain-of-function mutations in the Kit receptor tyrosine kinase. Most primary GIST patients respond to the Kit inhibitor imatinib, but this drug often becomes ineffective because of secondary mutations in the Kit kinase domain. The characteristic intracellular accumulation of imatinib-sensitive and -resistant Kit protein is well documented, but its relationship to oncogenic signaling remains unknown. Here, we show that in cancer tissue from primary GIST patients as well as in cell lines, mutant Kit accumulates on the Golgi apparatus, whereas normal Kit localizes to the plasma membrane (PM). In imatinib-resistant GIST with a secondary Kit mutation, Kit localizes predominantly on the Golgi apparatus. Both imatinib-sensitive and imatinib-resistant Kit (Kit(mut)) become fully auto-phosphorylated only on the Golgi and only if in a complex-glycosylated form. Kit(mut) accumulates on the Golgi during the early secretory pathway, but not after endocytosis. The aberrant kinase activity of Kit(mut) prevents its export from the Golgi to the PM. Furthermore, Kit(mut) on the Golgi signals and activates the phosphatidylinositol 3-kinase-Akt (PI3K-Akt) pathway, signal transducer and activator of transcription 5 (STAT5), and the Mek-Erk pathway. Blocking the biosynthetic transport of Kit(mut) to the Golgi from the endoplasmic reticulum inhibits oncogenic signaling. PM localization of Kit(mut) is not required for its signaling. Activation of Src-family tyrosine kinases on the Golgi is essential for oncogenic Kit signaling. These results suggest that the Golgi apparatus serves as a platform for oncogenic Kit signaling. Our study demonstrates that Kit(mut)'s pathogenicity is related to its mis-localization, and may offer a new strategy for treating imatinib-resistant GISTs.Oncogene advance online publication, 13 February 2017; doi:10.1038/onc.2016.519.

  14. Rh(II)-catalyzed Reactions of Diazoesters with Organozinc Reagents (United States)

    Panish, Robert; Selvaraj, Ramajeyam; Fox, Joseph M.


    Rh(II)-catalyzed reactions of diazoesters with organozinc reagents are described. Diorganozinc reagents participate in reactions with diazo compounds by two distinct, catalyst-dependent mechanisms. With bulky diisopropylethylacetate ligands, the reaction mechanism is proposed to involve initial formation of a Rh-carbene and subsequent carbozincation to give a zinc enolate. With Rh2(OAc)4, it is proposed that initial formation of an azine precedes 1,2-addition by an organozinc reagent. This straightforward route to the hydrazone products provides a useful method for preparing chiral quaternary α-aminoesters or pyrazoles via the Paul-Knorr condensation with 1,3-diketones. Crossover and deuterium labeling experiments provide evidence for the mechanisms proposed. PMID:26241081

  15. Preparation and Purification of Zinc Sulphinate Reagents for Organic Synthesis (United States)

    O’Hara, Fionn; Baxter, Ryan D.; O’Brien, Alexander G.; Collins, Michael R.; Dixon, Janice A.; Fujiwara, Yuta; Ishihara, Yoshihiro; Baran, Phil S.


    SUMMARY The present protocol details the synthesis of zinc bis(alkanesulphinate)s that can be used as general reagents for the formation of radical species. The zinc sulphinates described herein have been generated from the corresponding sulphonyl chlorides by treatment with zinc dust. The products may be used crude, or a simple purification procedure may be performed to minimize incorporation of water and zinc chloride. Elemental analysis has been conducted in order to confirm the purity of the zinc sulphinate reagents; reactions with caffeine have also been carried out to verify the reactivity of each batch that has been synthesized. Although the synthesis of the zinc sulphinate salts generally proceeds within 3 h, workup can take up to 24 h and purification can take up to 3 h. Following the steps in this protocol would enable the user to generate a small toolkit of zinc sulphinate reagents over the course of one week. PMID:23640168

  16. Elution of lead from vermiculite with environmentally benign reagents

    Institute of Scientific and Technical Information of China (English)


    The elution of lead from vermiculite was investigated by using a novel biodegradable chelating reagent, L-asparagic-N,N-diacetic acid (ASDA) and water soluble depolymerized pectic acid and comparing with a conventional chelating reagent, EDTA, as well as acetic acid. The influences of the reagent concentration, equilibrium pH and the suspension contact time on Pb extraction were examined. It is concluded that the acetic acid is not effective for Pb removal in any case due to its weak complexing ability with Pb. Although Pb is easier to be released by EDTA with stoichiometric amount, it is by no means the preferable alternative for the purpose because of its low biodegradability. On the other hand, ASDA and depolymerized pectic acid have the potential application because they are not only effective for Pb elution but also environmentally friendly.

  17. Rolling circle amplification of metazoan mitochondrialgenomes

    Energy Technology Data Exchange (ETDEWEB)

    Simison, W. Brian; Lindberg, D.R.; Boore, J.L.


    Here we report the successful use of rolling circle amplification (RCA) for the amplification of complete metazoan mt genomes to make a product that is amenable to high-throughput genome sequencing techniques. The benefits of RCA over PCR are many and with further development and refinement of RCA, the sequencing of organellar genomics will require far less time and effort than current long PCR approaches.

  18. Viability Reagent, PrestoBlue, in Comparison with Other Available Reagents, Utilized in Cytotoxicity and Antimicrobial Assays

    Directory of Open Access Journals (Sweden)

    Namrita Lall


    Full Text Available This study compared different commercially available viability reagents. The growth indicator reagents include p-iodonitrotetrazolium violet (INT, PrestoBlue, and Alamar Blue which were used for antimicrobial analysis against Streptococcus mutans, Prevotella intermedia, Propionibacterium acnes, and Mycobacterium tuberculosis. PrestoBlue and Alamar Blue are resazurin based reagents that resulted in a quick and easily distinguishable colour change that allowed for visual readings. INT and Sodium 3′-[1-(phenyl amino-carbonyl-3,4-tetrazolium]-bis-[4-methoxy-6-nitro] benzene sulfonic acid hydrate (XTT are tetrazolium based reagents which are converted to a formazan dye in the presence of metabolically active mitochondria enzyme. For cell viability analysis, reagents XTT and PrestoBlue were compared. PrestoBlue was able to clearly indicate the minimum inhibitory concentration (MIC of various positive drug controls on various microbial strains. PrestoBlue was also a good indicator of the 50% inhibitory concentration (IC50 of positive drug controls on various cell lines.

  19. Kit de pràctiques basat en microcontrolador PIC


    Verdaguer Estarlich, Meritxell


    Aquest treball consisteix en el disseny i prototipat d‟un kit de pràctiques com a suport a la docència, basat en l‟arquitectura PIC. El microcontrolador escollit per a la implementació és el PIC18F4550, el qual reuneix les característiques necessàries per a la necessitat requerida. El kit compta amb diversos dispositius, els de sortida com són díodes LED, visualitzador 7 segments, display LCD; i d‟altres d‟entrada: teclat hexadecimal, interruptors, optoacobladors i un convertidor analòg...

  20. GossipKit: A Framework of Gossip Protocol Family


    Lin, Shen; Taiani, Francois; Blair, Gordon S.; European Science Foundation


    A large number of gossip protocols have been developed in the last few years to address a wide range of functionalities. So far, however, very few software frameworks have been proposed to ease the development and deployment of these gossip protocols. To address this issue, this paper presents GossipKit, an event-driven framework that provides a generic and extensible architecture for the development of (re)configurable gossip-oriented middleware. GossipKit is based on a generic interaction m...

  1. The Basics of Line Balancing and JIT Kitting

    CERN Document Server

    Townsend, Beverly


    Accessible to the Lean novice and shop floor employee, The Basics of Line Balancing and JIT Kitting explores line balancing and the pre-assembly of components into a finished product in a just-in-time fashion (JIT Kitting). It explains how to use time studies, develop yamazumi charts, discover and eliminate waste, balance your line, and create new standard work content for the shop floor. The book facilitates a clear understanding of the seven deadly wastes (muda) as well as what you can do to eliminate them from your facility. Describing the purpose and use of standard work, it explains how t

  2. The effect of nest box temperature on kit growth rate and survival in the American mink (Neovison vison)

    DEFF Research Database (Denmark)

    Schou, Toke Munk; Malmkvist, Jens


    growth. Instead parameters concerning litter composition did have significant effects on kit growth and survival. Litters with high number of Totborn and kit AliveD1 affected kit growth and kit viability negatively (increased number of live born kits dying), which indicates that factors acting...

  3. KIT competence center for decommissioning. Innovation and promotion of trainees; Kompetenzzentrum Rueckbau am KIT. Nachwuchsfoerderung und Innovationen fuer den Rueckbau

    Energy Technology Data Exchange (ETDEWEB)

    Gentes, Sascha [Karlsruher Institut fuer Technologie (KIT), Karlsruhe (Germany). Inst. fuer Technologie und Management im Baubetrieb


    The safe decommissioning of nuclear installations is technically feasible, but is also still a challenge for science, technology and industry. The expertise and know how for decommissioning must be ensured because it will be needed for further decades. Already in 2008 the Karlsruhe Institute of Technology (KIT) had identified this challenge that later emerged through the closure of nuclear power plants in Germany. The KIT opened the professorship Technology and Management of the Decommissioning of Nuclear Installations. In 2014, this section was extended through the dismantling of conventional installations.

  4. Heat induces gene amplification in cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Bin, E-mail: [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Mercy Cancer Center, Mercy Medical Center-North Iowa, Mason City, IA 50401 (United States); Ouyang, Ruoyun [Department of Respiratory Medicine, The Second Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410011 (China); Huang, Chenghui [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Department of Oncology, The Third Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410013 (China); Liu, Franklin [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Neill, Daniel [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Li, Chuanyuan [Dermatology, Duke University Medical Center, Durham, NC 27710 (United States); Dewhirst, Mark [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States)


    Highlights: Black-Right-Pointing-Pointer This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. Black-Right-Pointing-Pointer Hyperthermia induces DNA double strand breaks. Black-Right-Pointing-Pointer DNA double strand breaks are considered to be required for the initiation of gene amplification. Black-Right-Pointing-Pointer The underlying mechanism of heat-induced gene amplification is generation of DNA double strand breaks. -- Abstract: Background: Hyperthermia plays an important role in cancer therapy. However, as with radiation, it can cause DNA damage and therefore genetic instability. We studied whether hyperthermia can induce gene amplification in cancer cells and explored potential underlying molecular mechanisms. Materials and methods: (1) Hyperthermia: HCT116 colon cancer cells received water-submerged heating treatment at 42 or 44 Degree-Sign C for 30 min; (2) gene amplification assay using N-(phosphoacetyl)-L-aspartate (PALA) selection of cabamyl-P-synthetase, aspartate transcarbarmylase, dihydro-orotase (cad) gene amplified cells; (3) southern blotting for confirmation of increased cad gene copies in PALA-resistant cells; (4) {gamma}H2AX immunostaining to detect {gamma}H2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 Degree-Sign C for 30 min induces gene amplification. The frequency of cad gene amplification increased by 2.8 and 6.5 folds respectively; (2) heat exposure at both 42 and 44 Degree-Sign C for 30 min induces DNA double strand breaks in HCT116 cells as shown by {gamma}H2AX immunostaining. Conclusion: This study shows that heat exposure can induce gene amplification in cancer cells, likely through the generation of DNA double strand breaks, which are believed to be required for the initiation of gene amplification. This process may be promoted by heat when cellular proteins that are responsible for checkpoints, DNA replication, DNA repair and

  5. Forensic genetic value of a 27 Y-STR loci multiplex (Yfiler(®) Plus kit) in an Italian population sample. (United States)

    Rapone, Cesare; D'Atanasio, Eugenia; Agostino, Alessandro; Mariano, Martina; Papaluca, Maria Teresa; Cruciani, Fulvio; Berti, Andrea


    The analysis of Y chromosome short tandem repeat (Y-STR) haplotypes provides important information that can be used for investigative purposes and in population studies. The Yfiler(®) Plus PCR Amplification kit (Yfiler(®) Plus, Thermo Fisher Scientific, Waltham, MA, USA) allows the multiplex amplification of 27 Y-STRs, including 7 rapidly mutating markers (RM Y-STRs). In this study, 203 unrelated males from Italy, which were subdivided into 4 different geographical groups (North, Center, South and Sardinia) were analyzed. Several intra-population diversity indexes were computed and compared to those obtained using only loci either from the minimal haplotype or the 17-plex (Yfiler(®), Thermo Fisher Scientific, Waltham, MA, USA). In addition, inter-population diversity analysis (RST) among the four Italian samples was performed. The same analysis was also used to compare the Italian sub-sets to other European populations where the Yfiler(®) Plus haplotype frequency data were available. The Sardinians were significantly differentiated from the other three Italian groups, thus requiring a specific sub-national Y-STR haplotype database. The Yfiler(®) Plus kit showed a high power of discrimination which is useful for criminal investigations, principally due to the inclusion of RM Y-STRs.

  6. Evaluation of the kinase domain of c-KIT in canine cutaneous mast cell tumors

    Directory of Open Access Journals (Sweden)

    Kiupel Matti


    Full Text Available Abstract Background Mutations in the c-KIT proto-oncogene have been implicated in the progression of several neoplastic diseases, including gastrointestinal stromal tumors and mastocytosis in humans, and cutaneous mast cell tumors (MCTs in canines. Mutations in human mastocytosis patients primarily occur in c-KIT exon 17, which encodes a portion of its kinase domain. In contrast, deletions and internal tandem duplication (ITD mutations are found in the juxtamembrane domain of c-KIT in approximately 15% of canine MCTs. In addition, ITD c-KIT mutations are significantly associated with aberrant KIT protein localization in canine MCTs. However, some canine MCTs have aberrant KIT localization but lack ITD c-KIT mutations, suggesting that other mutations or other factors may be responsible for aberrant KIT localization in these tumors. Methods In order to characterize the prevalence of mutations in the phospho-transferase portion of c-KIT's kinase domain in canine MCTs exons 16–20 of 33 canine MCTs from 33 dogs were amplified and sequenced. Additionally, in order to determine if mutations in c-KIT exon 17 are responsible for aberrant KIT localization in MCTs that lack juxtamembrane domain c-KIT mutations, c-KIT exon 17 was amplified and sequenced from 18 canine MCTs that showed an aberrant KIT localization pattern but did not have ITD c-KIT mutations. Results No mutations or polymorphisms were identified in exons 16–20 of any of the MCTs examined. Conclusion In conclusion, mutations in the phospho-transferase portion of c-KIT's kinase domain do not play an important role in the progression of canine cutaneous MCTs, or in the aberrant localization of KIT in canine MCTs.

  7. Src-like-adaptor protein (SLAP) differentially regulates normal and oncogenic c-Kit signaling. (United States)

    Kazi, Julhash U; Agarwal, Shruti; Sun, Jianmin; Bracco, Enrico; Rönnstrand, Lars


    The Src-like-adaptor protein (SLAP) is an adaptor protein sharing considerable structural homology with Src. SLAP is expressed in a variety of cells and regulates receptor tyrosine kinase signaling by direct association. In this report, we show that SLAP associates with both wild-type and oncogenic c-Kit (c-Kit-D816V). The association involves the SLAP SH2 domain and receptor phosphotyrosine residues different from those mediating Src interaction. Association of SLAP triggers c-Kit ubiquitylation which, in turn, is followed by receptor degradation. Although SLAP depletion potentiates c-Kit downstream signaling by stabilizing the receptor, it remains non-functional in c-Kit-D816V signaling. Ligand-stimulated c-Kit or c-Kit-D816V did not alter membrane localization of SLAP. Interestingly oncogenic c-Kit-D816V, but not wild-type c-Kit, phosphorylates SLAP on residues Y120, Y258 and Y273. Physical interaction between c-Kit-D816V and SLAP is mandatory for the phosphorylation to take place. Although tyrosine-phosphorylated SLAP does not affect c-Kit-D816V signaling, mutation of these tyrosine sites to phenylalanine can restore SLAP activity. Taken together the data demonstrate that SLAP negatively regulates wild-type c-Kit signaling, but not its oncogenic counterpart, indicating a possible mechanism by which the oncogenic c-Kit bypasses the normal cellular negative feedback control.

  8. Genoviz Software Development Kit: Java tool kit for building genomics visualization applications

    Directory of Open Access Journals (Sweden)

    Chervitz Stephen A


    Full Text Available Abstract Background Visualization software can expose previously undiscovered patterns in genomic data and advance biological science. Results The Genoviz Software Development Kit (SDK is an open source, Java-based framework designed for rapid assembly of visualization software applications for genomics. The Genoviz SDK framework provides a mechanism for incorporating adaptive, dynamic zooming into applications, a desirable feature of genome viewers. Visualization capabilities of the Genoviz SDK include automated layout of features along genetic or genomic axes; support for user interactions with graphical elements (Glyphs in a map; a variety of Glyph sub-classes that promote experimentation with new ways of representing data in graphical formats; and support for adaptive, semantic zooming, whereby objects change their appearance depending on zoom level and zooming rate adapts to the current scale. Freely available demonstration and production quality applications, including the Integrated Genome Browser, illustrate Genoviz SDK capabilities. Conclusion Separation between graphics components and genomic data models makes it easy for developers to add visualization capability to pre-existing applications or build new applications using third-party data models. Source code, documentation, sample applications, and tutorials are available at

  9. Evaluation of the Illumina(®) Beta Version ForenSeq™ DNA Signature Prep Kit for use in genetic profiling. (United States)

    Churchill, Jennifer D; Schmedes, Sarah E; King, Jonathan L; Budowle, Bruce


    While capillary electrophoresis-based technologies have been the mainstay for human identity typing applications, there are limitations with this methodology's resolution, scalability, and throughput. Massively parallel sequencing (MPS) offers the capability to multiplex multiple types of forensically-relevant markers and multiple samples together in one run all at an overall lower cost per nucleotide than traditional capillary electrophoresis-based methods; thus, addressing some of these limitations. MPS also is poised to expand forensic typing capabilities by providing new strategies for mixture deconvolution with the identification of intra-STR allele sequence variants and the potential to generate new types of investigative leads with an increase in the overall number and types of genetic markers being analyzed. The beta version of the Illumina ForenSeq DNA Signature Prep Kit is a MPS library preparation method with a streamlined workflow that allows for targeted amplification and sequencing of 63 STRs and 95 identity SNPs, with the option to include an additional 56 ancestry SNPs and 22 phenotypic SNPs depending on the primer mix chosen for amplification, on the MiSeq desktop sequencer (Illumina). This study was divided into a series of experiments that evaluated reliability, sensitivity of detection, mixture analysis, concordance, and the ability to analyze challenged samples. Genotype accuracy, depth of coverage, and allele balance were used as informative metrics for the quality of the data produced. The ForenSeq DNA Signature Prep Kit produced reliable, reproducible results and obtained full profiles with DNA input amounts of 1ng. Data were found to be concordant with current capillary electrophoresis methods, and mixtures at a 1:19 ratio were resolved accurately. Data from the challenged samples showed concordant results with current DNA typing methods with markers in common and minimal allele drop out from the large number of markers typed on these

  10. A Simple DNA Preparation Method for PCR Amplifications in Marker-Assisted Selection of Wheat

    Institute of Scientific and Technical Information of China (English)

    WANG Shu; R E Knox; R M DePauw; J M Clarke; WANG Bo-lun


    An important, but often limiting step in marker-assisted breeding is the efficient isolation of plant DNA for polymerase chain reaction (PCR) amplification. A simple method using an alkali treatment to extract wheat DNA for marker-assisted selection (MAS) in wheat breeding programs was compared to a commercial kit and cetyltrimethylammonium bromide (CTAB) extraction. DNA concentration from the alkali extraction was higher than the other two methods but purity was lower than CTAB extraction. The alkali extraction method was used on breeding lines to determine its usefulness. The alkali-extracted DNA samples were suitable for several PCR-based procedures, including random amplified polymorphic DNA (RAPD), microsatellite (simple sequence repeat, i.e., SSR) and sequence characterized amplified region (SCAR)analyses.

  11. Rapid and economic DNA extraction from a single salmon egg for real-time PCR amplification. (United States)

    Yang, Jing-Iong; Huang, Hsiao-Yun; Chou, Yii-Cheng; Chen, Chien-Cheng; Lee, Guo-Chi; Chang, Hsueh-Wei


    Salmon eggs are common in Japanese sushi and other seafood products; however, certain fish eggs are used as counterfeit salmon eggs which are found in foods and processed products. This study develops a simple, rapid, and cost-effective method for DNA extraction, filtration (FT) and dilution (DL) protocols from a single salmon egg with good DNA quality for real-time PCR amplification. The DNA amount, DNA quality, and real-time PCR performance for different dilutions and different lengths of PCR amplicons were evaluated and compared with the common Qiagen tissue kit (QTK) and Chelex-100-based (CX) protocols. The extracted DNA from a single salmon egg using the FT or DL protocol can be applied in phylogenic research, food authentication and post-marketing monitoring of genetically modified (GM) food products.

  12. Documentation for MeshKit - Reactor Geometry (&mesh) Generator

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Rajeev [Argonne National Lab. (ANL), Argonne, IL (United States); Mahadevan, Vijay [Argonne National Lab. (ANL), Argonne, IL (United States)


    This report gives documentation for using MeshKit’s Reactor Geometry (and mesh) Generator (RGG) GUI and also briefly documents other algorithms and tools available in MeshKit. RGG is a program designed to aid in modeling and meshing of complex/large hexagonal and rectilinear reactor cores. RGG uses Argonne’s SIGMA interfaces, Qt and VTK to produce an intuitive user interface. By integrating a 3D view of the reactor with the meshing tools and combining them into one user interface, RGG streamlines the task of preparing a simulation mesh and enables real-time feedback that reduces accidental scripting mistakes that could waste hours of meshing. RGG interfaces with MeshKit tools to consolidate the meshing process, meaning that going from model to mesh is as easy as a button click. This report is designed to explain RGG v 2.0 interface and provide users with the knowledge and skills to pilot RGG successfully. Brief documentation of MeshKit source code, tools and other algorithms available are also presented for developers to extend and add new algorithms to MeshKit. RGG tools work in serial and parallel and have been used to model complex reactor core models consisting of conical pins, load pads, several thousands of axially varying material properties of instrumentation pins and other interstices meshes.

  13. World War II Spy Kit: "The Great Nazi Intelligence Coup." (United States)

    Haight, David

    This instructional packet is designed to introduce students to primary source material by having them participate in an historical "what might have been." Students engage in critical thinking and document analysis, and through the process learn about Operation OVERLORD and World War II in general. This spy kit centers on Operation…

  14. Influence of nest box environment on kit survival

    DEFF Research Database (Denmark)

    Lund, V.H.; Malmkvist, Jens


    . In conclusion, access to different nesting materials reduced the risk of dying in farm mink in relation with factors as e.g. nest quality and maternal behaviour. A decrease in kit mortality from 20% to 5% would give the Danish mink production a profound number of Danish kroner extra per year....

  15. [Kit of Materials for Needs Assessment and Evaluation. (United States)

    California State Dept. of Education, Sacramento. Bureau of Intergroup Relations.

    The items included in this kit represent a variety of needs assessment instruments and evaluation designs and methods offered by school districts in their plans for implementation of Article 3.3, Education Code Sections 13344-13344.4, school staff preparation in the history, culture, and current problems of racial and ethnic minorities. Some are…

  16. Language Coordinators Resource Kit. Section Two: Teacher Supervision. (United States)

    Peace Corps, Washington, DC. Information Collection and Exchange Div.

    The guide is one section of a resource kit designed to assist Peace Corps language instruction coordinators in countries around the world in understanding the principles underlying second language learning and teaching and in organizing instructional programs. This section focuses on selection and supervision of language teachers. An introductory…

  17. National High Blood Pressure 12-Month Kit. May 1988. (United States)

    National Heart and Lung Inst. (DHHS/NIH), Bethesda, MD. National High Blood Pressure Education Program.

    Part I of this kit provides information for program planners and health professionals on ways to overcome barriers to health care among the medically underserved, promote high blood pressure control through the media and other community channels, and improve adherence to treatment among hypertensive patients. It lists additional resources for…

  18. Programming with the KIBO Robotics Kit in Preschool Classrooms (United States)

    Elkin, Mollie; Sullivan, Amanda; Bers, Marina Umaschi


    KIBO is a developmentally appropriate robotics kit for young children that is programmed using interlocking wooden blocks; no screens or keyboards are required. This study describes a pilot KIBO robotics curriculum at an urban public preschool in Rhode Island and presents data collected on children's knowledge of foundational programming concepts…

  19. Improvement in carbofuran degradation by different Fenton's reagent dosing processes. (United States)

    Ma, Ying-Shih


    Attempts were made in this study to examine the efficiency of Fenton's reagent with different dosing processes and H(2)O(2) and Fe(2+) concentrations for the treatment of carbofuran wastewater. Carbofuran degradation, total organic carbon (TOC) removal and H(2)O(2) consumption were determined during the experiments. Increases in H(2)O(2) and Fe(2+) concentrations led to an increase in the degradation of carbofuran. Almost 100% of carbofuran could be degraded at pH 3, 120 mg L(-1) H(2)O(2), 24 mg L(-1) Fe(2+) and 30 minutes reaction time; removals of TOC were among 48.8%-53.3% under different dosing processes. A continuous dosing process was beneficial to improve the removal of TOC by Fenton's reagent. Rate constants of carbofuran degradation could be calculated by the first-order kinetics; increase in the Fenton's reagent generally increased the rate constants. Gas chromatography-mass spectrometry analysis found five degradation products by hydroxyl radicals attack. Thus, this study might offer an effective dosing way for carbofuran wastewater treatment by Fenton's reagent.

  20. Protein-Protein Interaction Reagents | Office of Cancer Genomics (United States)

    The CTD2 Center at Emory University has a library of genes used to study protein-protein interactions in mammalian cells. These genes are cloned in different mammalian expression vectors. A list of available cancer-associated genes can be accessed below. Emory_CTD^2_PPI_Reagents.xlsx Contact: Haian Fu

  1. Selective Flotation of Calcite from Fluorite: A Novel Reagent Schedule

    Directory of Open Access Journals (Sweden)

    Zhiyong Gao


    Full Text Available Fluorite is an important strategic mineral. In general, fluorite ores will contain a certain amount of calcite gangue mineral. Thus, they need to be separated from each other. For an economic separation, a reverse flotation process is used to float calcite gangue from fluorite. However, little information on the separation is available. In this study, a novel reagent schedule using citric acid (CA as the depressant, sodium fluoride (NaF as the regulator and sulfoleic acid (SOA as the collector, was developed to separate calcite from fluorite. The results demonstrated a high selectivity for the flotation of calcite from fluorite using this new reagent schedule. The best selective separation for a single mineral and mixed binary minerals was obtained when 200 mg/L of NaF, 50 mg/L of CA, and 6 mg/L of SOA were used at pH 9. In addition, a batch flotation experiment was carried out using a run-of-mine feed material. Selective separation was achieved with 85.18% calcite removal while only 11.2% of fluorite was lost. An attempt was made to understand the effect of the new reagent schedule on the flotation of calcite. The results from both microflotation and bench scale flotation demonstrated a great potential for industrial application using this novel reagent schedule to upgrade fluorite ore.

  2. Improved amine spray reagent for the detection of sugars

    NARCIS (Netherlands)

    Niemann, G.J.


    In the course of our investigations on naturally occurring flavonoid glycosides, the sugars obtained after acid hydrolysis were mainly analysed by gas-liquid chromatography and/or paper chromatography, using p-anisidine phosphate as the spray reagent. Often only very small amounts of the isolated co

  3. Toward a dry reagent immunoassay of progesterone in bovine milk

    NARCIS (Netherlands)

    Posthuma-Trumpie, Geertruida Afina


    This thesis is aimed at the development of a dry reagent immunoassay of progesterone in cow's milk. Progesterone is a steroid hormone and regulates ovulation in female mammals. The concentration of progesterone in blood and in milk is in accordance with the reproductive cycle of the individual femal

  4. One New Method of Nucleic Acid Amplification-Loop-mediated Isothermal Amplification of DNA

    Institute of Scientific and Technical Information of China (English)

    Xue-en FANG; Jian LI; Qin CHEN


    Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method, which amplifies DNA with high specificity, sensitivity, rapidity and efficiency under isothermal conditions using a set of four specially designed primers and a Bst DNA polymerase with strand displacement activity. The basic principle, characteristics, development of LAMP and its applications are summarized in this article.

  5. Pharmacological inhibitors of c-KIT block mutant c-KIT mediated migration of melanocytes and melanoma cells in vitro and in vivo (United States)

    Posch, Christian; Moslehi, Homayoun; Sanlorenzo, Martina; Green, Gary; Vujic, Igor; Panzer-Grümayer, Renate; Rappersberger, Klemens; Ortiz-Urda, Susana


    Mutations in the receptor tyrosine kinase c-KIT (KIT) are frequent oncogenic alterations in melanoma and are predominantly detected in tumors of acral, mucosal, and chronically sun-damaged skin. Research indicates that melanocytes with aberrant KIT signaling can be found in the distant periphery of the primary tumor; However, it is hitherto unknown whether KIT might confer a migratory advantage, thereby enabling genetically abnormal cells to populate a distal area. In this study, we investigated the role of mutant KIT in melanocyte- and melanoma cell migration using KIT mutant lines as well as genetically manipulated murine and primary human melanocytes. Our results revealed that melanocytes, stably transduced with mutant KIT closed a gap inflicted on cell monolayers faster than wild-type controls. Similarly, KIT mutant human melanoma lines were able to populate a larger area in a 3D in vitro skin model compared to KIT wild type and BRAF mutant lines. Genomic profiling revealed that genes associated with increased cell-dispersal of KIT mutant variants were linked to a statistically significant up-regulation of 60 migratory genes (z-score 1.334; p=0.0001). In addition, in vivo experiments harnessing a mouse xenograft model of early melanoma development demonstrated rapid lateral migration of KIT mutant cells compared to respective controls. The specific kinase inhibitors imatinib and nilotinib, could abrogate this migratory advantage in vitro and in vivo. Our work suggests that KIT inhibition might help to target migratory active, KIT mutant melanoma cells, thus representing a potential strategy to reduce spread and local recurrence. PMID:27322141

  6. The Use of Kits in the Analysis of Tissue Lipids Requires Validation. (United States)

    Rider, T; LeBoeuf, R C; Tso, Patrick; Jandacek, R J


    The ready availability and ease of use of kits for the measurement of serum lipids has greatly facilitated these measurements. In many cases it would be convenient to use these kits in the determination of lipid concentrations in tissues. The successful application of serum kits in tissue analysis requires that two important issues be considered. First, the solvent system for the extraction of the lipids and the solvent used for analysis by the kit must be compatible with the reactions in the kit. Second, the concentration range in the analyzed solution must be within the range for which the kit is used. We report here that lipids in liver and adipose tissues may be significantly underestimated by the use of some kits. We recommend that the use of kits for tissue analysis of lipids be validated for the specific analysis.

  7. Stem cell factor and c-Kit in human primordial germ cells and fetal ovaries

    DEFF Research Database (Denmark)

    Høyer, Poul Erik; Byskov, Anne Grete; Møllgård, Kjeld


    Prenatal ovary (human), Primordial germ cells, Folliculogenesis, c-Kit, Stem cell factor, immunohistochemistry......Prenatal ovary (human), Primordial germ cells, Folliculogenesis, c-Kit, Stem cell factor, immunohistochemistry...

  8. C-kit gene mutation in human gastrointestinal stromal tumors

    Institute of Scientific and Technical Information of China (English)

    Ying-Yong Hou; Ai-Hua Zheng; Tai-Ming Zhang; Wen-Zhong Hou; Jian Wang; Xiang Du; Xiong-Zeng Zhu; Yun-Shan Tan; Meng-Hong Sun; Yong-Kun Wei; Jian-Fang Xu; Shao-Hua Lu; Su-Jie A-Ke-Su; Yan-Nan Zhou; Feng Gao


    AIM: To investigate the significance of c-kit gene mutation in gastrointestinal stromal tumors (GIST).METHODS: Fifty two cases of GIST and 28 cases of other tumors were examined. DNA samples were extracted from paraffin sections and fresh blocks. Exons 11, 9 and 13 of the c-kit gene were amplified by PCR and sequenced.RESULTS: Mutations of exon 11 were found in 14 of 25 malignant GISTs (56%), mutations of exon 11 of the c-kit gene were revealed in 2 of 19 borderline GISTs (10.5%),and no mutation was found in benign tumors. The mutation rate showed significant difference (X2=14.39, P<0.01)between malignant and benign GISTs. Most of mutations consisted of the in-frame deletion or replication from 3 to 48 bp in heterozygous and homozygous fashions, None of the mutations disrupted the downstream reading frame of the gene. Point mutations and frame deletions were most frequently observed at codons 550-560, but duplications were most concentrated at codons 570-585. No mutations of exons 9 and 13 were revealed in GISTs, Neither c-kit gene expression nor gene mutations were found in 3 leiomyomas, 8 leiomyosarcomas, 2 schwannomas, 2malignant peripheral nerve sheath tumors, 2 intraabdominal fibromatoses, 2 malignant fibrous histiocytomas and 9 adenocarcinomas.CONCLUSION: C-kit gene mutations occur preferentially in malignant GISTs and might be a clinically useful adjunct marker in the evaluation of GISTs and can help to differentiate GISTs from other mesenchymal tumors of gastrointestinal tract, such as smooth muscle tumors,schwannomas, etc.

  9. Detection and identification of opportunistic Exophiala species using the rolling circle amplification of ribosomal internal transcribed spacers. (United States)

    Najafzadeh, M J; Dolatabadi, S; Saradeghi Keisari, M; Naseri, A; Feng, P; de Hoog, G S


    Deep infections by melanized fungi deserve special attention because of a potentially fatal, cerebral or disseminated course of disease in otherwise healthy patients. Timely diagnostics are a major problem with these infections. Rolling circle amplification (RCA) is a sensitive, specific and reproducible isothermal DNA amplification technique for rapid molecular identification of microorganisms. RCA-based diagnostics are characterized by good reproducibility, with few amplification errors compared to PCR. The method is applied here to species of Exophiala known to cause systemic infections in humans. The ITS rDNA region of five Exophiala species (E. dermatitidis, E. oligosperma, E. spinifera, E. xenobiotica, and E. jeanselmei) was sequenced and aligned in view of designing specific padlock probes to be used for the detection of single nucleotide polymorphisms (SNPs) of the Exophiala species concerned. The assay proved to successfully amplify DNA of the target fungi at the level of species; while no cross-reactivity was observed. Amplification products were visualized on 1% agarose gels to verify the specificity of probe-template binding. Amounts of reagents were minimized to avoid the generation of false positive results. The sensitivity of RCA may help to improve early diagnostics of these difficult to diagnose infections.

  10. The Comparison of different Procedures for DNA extraction from paraffin-embedded Tissues: A commercial kit and a traditional method based on heating

    Directory of Open Access Journals (Sweden)

    Davoodi, H


    Full Text Available Background and objectives: Paraffin-embedded tissues and clinicalsamples are a valuable resource for molecular genetic studies, but theextraction of high-quality genomic DNA from this tissues is still aproblematic issue. In the Present study, the efficiency of two DNAextraction protocols, a commercial kit and a traditional method based onheating and K Proteinase was compared.Material and Methods: Fifty paraffin-embedded blocks of colon cancertissues (more than 5 years old were used to compare two methods ofDNA extraction. DNA was extracted by traditional method using heat andcommercial DNA extraction (Qiagen kit method. Then the purity andconcentration of extracted DNA were measured by Spectrophotometer.Two sequences of TLR4 “The most important receptors in innateimmunity” were amplified by polymerase chain reaction. SH-1 ‘188bp’and SH-2 ‘124bp’ were amplified and then the products were separated ona 2% agarose gel.Results: The results show that the yield of DNA by traditional method(297 mg/ml is significantly (p<0.01 higher than Commercial kit(176mg/ ml. But traditional method has the lower OD ratio (1.2Compared to Commercial method. The Amplification of the TLR4 genesequences is more successful by the traditional method (p<0.01compared with commercial method. The length of the sequence affectson the results of PCR in that short sequence is amplified more successfulcompared to the long sequence.Conclusion: The traditional method is more successful in PCRamplification and also simple and cheap. Therefore, we recommendusing this method for DNA extraction taken from the paraffin-embeddedblocks with more than 5 years old and selecting shorter sequence forbetter amplification in PCR.Key words: DNA Extraction, paraffin embedded tissue, PCR

  11. Extending Mechanical Construction Kits to Incorporate Passive and Compliant Elements for Educational Robotics

    DEFF Research Database (Denmark)

    Assaf, Dorit; Larsen, Jørgen Christian; Reichardt, Markus


    artificial intelligence and biomechanics to students with different backgrounds. The robots we use both for research and education are usually built incorporating compliant materials as well as passive dynamics. These kind of properties are often not available in classical robot kits or mechanical...... and mechanical construction kits we describe interface solutions between several kits. Further we show some solutions to incorporate compliant materials and passive dynamics to traditional mechanical construction kits by using cheap and widely available materials....

  12. Loop-mediated isothermal amplification targeting 18S ribosomal DNA for rapid detection of Acanthamoeba. (United States)

    Yang, Hye-Won; Lee, Yu-Ran; Inoue, Noboru; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Kim, Hong-Kyun; Lee, Junhun; Goo, Youn-Kyoung; Kong, Hyun-Hee; Chung, Dong-Il; Hong, Yeonchul


    Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

  13. Take-Home Art Appreciation Kits for Kindergartners and Their Families. (United States)

    Mulcahey, Christine


    Describes collaborative project between an art specialist and kindergarten teacher to develop take-home art appreciation kits for kindergartners. Discusses benefits of the collaboration for teachers, children, and families. Focuses on aesthetic inquiry approach to the art kits to develop inquiry and critical thinking skills. Describes the kits'…

  14. 33 CFR 149.323 - What are the requirements for first aid kits? (United States)


    ... first aid kits? 149.323 Section 149.323 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Lifesaving Equipment Manned Deepwater Port Requirements § 149.323 What are the requirements for first aid kits? (a) Each manned deepwater port must have an industrial first aid kit, approved by an...

  15. 21 CFR 866.2480 - Quality control kit for culture media. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Quality control kit for culture media. 866.2480... control kit for culture media. (a) Identification. A quality control kit for culture media is a device...-dried, viable microorganism, intended for medical purposes to determine if a given culture medium...

  16. Pharmacological targeting of the KIT growth factor receptor: a therapeutic consideration for mast cell disorders

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Akin, C; Gilfillan, A M


    KIT is a member of the tyrosine kinase family of growth factor receptors which is expressed on a variety of haematopoietic cells including mast cells. Stem cell factor (SCF)-dependent activation of KIT is critical for mast cell homeostasis and function. However, when KIT is inappropriately activa...

  17. Gene Duplication of the zebrafish kit ligand and partitioning of melanocyte development functions to kit ligand a.

    Directory of Open Access Journals (Sweden)

    Keith A Hultman


    Full Text Available The retention of particular genes after the whole genome duplication in zebrafish has given insights into how genes may evolve through partitioning of ancestral functions. We examine the partitioning of expression patterns and functions of two zebrafish kit ligands, kit ligand a (kitla and kit ligand b (kitlb, and discuss their possible coevolution with the duplicated zebrafish kit receptors (kita and kitb. In situ hybridizations show that kitla mRNA is expressed in the trunk adjacent to the notochord in the middle of each somite during stages of melanocyte migration and later expressed in the skin, when the receptor is required for melanocyte survival. kitla is also expressed in other regions complementary to kita receptor expression, including the pineal gland, tail bud, and ear. In contrast, kitlb mRNA is expressed in brain ventricles, ear, and cardinal vein plexus, in regions generally not complementary to either zebrafish kit receptor ortholog. However, like kitla, kitlb is expressed in the skin during stages consistent with melanocyte survival. Thus, it appears that kita and kitla have maintained congruent expression patterns, while kitb and kitlb have evolved divergent expression patterns. We demonstrate the interaction of kita and kitla by morpholino knockdown analysis. kitla morphants, but not kitlb morphants, phenocopy the null allele of kita, with defects for both melanocyte migration and survival. Furthermore, kitla morpholino, but not kitlb morpholino, interacts genetically with a sensitized allele of kita, confirming that kitla is the functional ligand to kita. Last, we examine kitla overexpression in embryos, which results in hyperpigmentation caused by an increase in the number and size of melanocytes. This hyperpigmentation is dependent on kita function. We conclude that following genome duplication, kita and kitla have maintained their receptor-ligand relationship, coevolved complementary expression patterns, and that

  18. Time varying arctic climate change amplification

    Energy Technology Data Exchange (ETDEWEB)

    Chylek, Petr [Los Alamos National Laboratory; Dubey, Manvendra K [Los Alamos National Laboratory; Lesins, Glen [DALLHOUSIE U; Wang, Muyin [NOAA/JISAO


    During the past 130 years the global mean surface air temperature has risen by about 0.75 K. Due to feedbacks -- including the snow/ice albedo feedback -- the warming in the Arctic is expected to proceed at a faster rate than the global average. Climate model simulations suggest that this Arctic amplification produces warming that is two to three times larger than the global mean. Understanding the Arctic amplification is essential for projections of future Arctic climate including sea ice extent and melting of the Greenland ice sheet. We use the temperature records from the Arctic stations to show that (a) the Arctic amplification is larger at latitudes above 700 N compared to those within 64-70oN belt, and that, surprisingly; (b) the ratio of the Arctic to global rate of temperature change is not constant but varies on the decadal timescale. This time dependence will affect future projections of climate changes in the Arctic.

  19. Amplification, Redundancy, and Quantum Chernoff Information (United States)

    Zwolak, Michael; Riedel, C. Jess; Zurek, Wojciech H.


    Amplification was regarded, since the early days of quantum theory, as a mysterious ingredient that endows quantum microstates with macroscopic consequences, key to the "collapse of the wave packet," and a way to avoid embarrassing problems exemplified by Schrödinger's cat. Such a bridge between the quantum microworld and the classical world of our experience was postulated ad hoc in the Copenhagen interpretation. Quantum Darwinism views amplification as replication, in many copies, of the information about quantum states. We show that such amplification is a natural consequence of a broad class of models of decoherence, including the photon environment we use to obtain most of our information. This leads to objective reality via the presence of robust and widely accessible records of selected quantum states. The resulting redundancy (the number of copies deposited in the environment) follows from the quantum Chernoff information that quantifies the information transmitted by a typical elementary subsystem of the environment.

  20. On Arbitrary Phases in Quantum Amplitude Amplification

    CERN Document Server

    Hoyer, P


    We consider the use of arbitrary phases in quantum amplitude amplification which is a generalization of quantum searching. We prove that the phase condition in amplitude amplification is given by $\\tan(\\phi/2)=\\tan(\\phi/2)(1-2a)$, where $\\phi$ and $\\phi$ are the phases used and where $a$ is the success probability of the given algorithm. Thus the choice of phases depends nontrivially and nonlinearly on the success probability. Utilizing this condition, we give methods for constructing quantum algorithms that succeed with certainty and for implementing arbitrary rotations. We also conclude that phase errors of order up to $\\frac{1}{\\sqrt{a}}$ can be tolerated in amplitude amplification.

  1. Continuous phase amplification with a Sagnac interferometer

    CERN Document Server

    Starling, David J; Williams, Nathan S; Jordan, Andrew N; Howell, John C


    We describe a weak value inspired phase amplification technique in a Sagnac interferometer. We monitor the relative phase between two paths of a slightly misaligned interferometer by measuring the average position of a split-Gaussian mode in the dark port. Although we monitor only the dark port, we show that the signal varies linearly with phase and that we can obtain similar sensitivity to balanced homodyne detection. We derive the source of the amplification both with classical wave optics and as an inverse weak value.

  2. Effective Privacy Amplification for Secure Classical Communications

    CERN Document Server

    Horvath, Tamas; Scheuer, Jacob


    We study the effectiveness of privacy amplification for classical key-distribution schemes. We find that, unlike quantum key distribution schemes, the high fidelity of the raw key in classical systems allow the users to always sift a secure shorter key, given that they have an upper bound of eavesdropper probability to correctly guess the exchanged key-bits. We establish the number of privacy amplification iterations needed to achieve information leak of 10^-8 in several classical systems and highlight the inherent tradeoff between the number of iterations and the security of the raw key.

  3. Parametric Amplification For Detecting Weak Optical Signals (United States)

    Hemmati, Hamid; Chen, Chien; Chakravarthi, Prakash


    Optical-communication receivers of proposed type implement high-sensitivity scheme of optical parametric amplification followed by direct detection for reception of extremely weak signals. Incorporates both optical parametric amplification and direct detection into optimized design enhancing effective signal-to-noise ratios during reception in photon-starved (photon-counting) regime. Eliminates need for complexity of heterodyne detection scheme and partly overcomes limitations imposed on older direct-detection schemes by noise generated in receivers and by limits on quantum efficiencies of photodetectors.

  4. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification. (United States)

    Mauk, Michael G; Liu, Changchun; Song, Jinzhao; Bau, Haim H


    Microfluidic components and systems for rapid (microfluidic point-of-care (POC) diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of "lab on a chip" NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (polymerase-chain reaction)-based tests in clinical laboratories can be realized with a disposable, palm-sized, plastic microfluidic chip in which: (1) nucleic acids (NAs) are extracted from relatively large (~mL) volume sample lysates using an embedded porous silica glass fiber or cellulose binding phase ("membrane") to capture sample NAs in a flow-through, filtration mode; (2) NAs captured on the membrane are isothermally (~65 °C) amplified; (3) amplicon production is monitored by real-time fluorescence detection, such as with a smartphone CCD camera serving as a low-cost detector; and (4) paraffin-encapsulated, lyophilized reagents for temperature-activated release are pre-stored in the chip. Limits of Detection (LOD) better than 10³ virons/sample can be achieved. A modified chip with conduits hosting a diffusion-mode amplification process provides a simple visual indicator to readily quantify sample NA template. In addition, a companion microfluidic device for extracting plasma from whole blood without a centrifuge, generating cell-free plasma for chip-based molecular diagnostics, is described. Extensions to a myriad of related applications including, for example, food testing, cancer screening, and insect genotyping are briefly surveyed.

  5. A community standard format for the representation of protein affinity reagents

    DEFF Research Database (Denmark)

    Gloriam, David Erik Immanuel; Orchard, Sandra; Bertinetti, Daniela


    Protein affinity reagents (PARs), most commonly antibodies, are essential reagents for protein characterization in basic research, biotechnology, and diagnostics as well as the fastest growing class of therapeutics. Large numbers of PARs are available commercially; however, their quality is often...

  6. Controlling the Orientation and Alignment of Reagent Molecules by a Polarized Laser

    Institute of Scientific and Technical Information of China (English)

    丛书林; 韩克利; 楼南泉


    The expressions used for controlling the alignment and orientation of reagent molecules are derived. The problem to the control of the orientation and alignment of reagent molecules by the polarization direction and propagation direction of laser is discussed.

  7. Molecular characterization of metastatic exon 11 mutant gastrointestinal stromal tumors (GIST) beyond KIT/PDGFRα genotype evaluated by next generation sequencing (NGS). (United States)

    Saponara, Maristella; Urbini, Milena; Astolfi, Annalisa; Indio, Valentina; Ercolani, Giorgio; Del Gaudio, Massimo; Santini, Donatella; Pirini, Maria Giulia; Fiorentino, Michelangelo; Nannini, Margherita; Lolli, Cristian; Mandrioli, Anna; Gatto, Lidia; Brandi, Giovanni; Biasco, Guido; Pinna, Antonio Daniele; Pantaleo, Maria Abbondanza


    About 85% of GISTs are associated with KIT and PDGFRα gene mutations, which predict response to tyrosine kinase inhibitors. Although the outcomes in patients affected by GIST have dramatically improved, tumor progression control still remains a challenge. The aim of this study is the genomic characterization of individual metastatic KIT-exon 11-mutant GIST to identify additional aberrations and simultaneous molecular events representing potential therapeutic targets.Seven patients with metastatic GIST were studied with whole transcriptome sequencing and copy number analysis. Somatic single nucleotide variations were called; however, no shared mutated genes were detected except KIT. Almost all patients showed loss of genomic regions containing tumor suppressor genes, sometimes coupled with single nucleotide mutation of the other allele. Additionally, six fusion transcripts were found and three patients showed amplifications involving known oncogenes.Evaluating the concordance between CN status and mRNA expression levels, we detected overexpression of CCND2 and EGFR and silencing of CDKN2A, CDKN2C, SMARCB1, PTEN and DMD. Altered expression of these genes could be responsible for aberrant activation of signaling pathways that support tumor growth. In this work, we assessed the effect of Hedgehog pathway inhibition in GIST882 cells, which causes decrement of cell viability associated with reduction of KIT expression.Additional genomic alterations not previously reported in GIST were found even if not shared by all samples. This contributes to a more detailed molecular understanding of this disease, useful for identification of new targets and novel therapeutics and representing a possible point of departure for a truly individualized clinical approach.

  8. A dual amplification fluorescent strategy for sensitive detection of DNA methyltransferase activity based on strand displacement amplification and DNAzyme amplification. (United States)

    Cui, Wanling; Wang, Lei; Jiang, Wei


    DNA methyltransferase (MTase) plays a critical role in many biological processes and has been regarded as a predictive cancer biomarker and a therapeutic target in cancer treatment. Sensitive detection of DNA MTase activity is essential for early cancer diagnosis and therapeutics. Here, we developed a dual amplification fluorescent strategy for sensitive detection of DNA MTase activity based on strand displacement amplification (SDA) and DNAzyme amplification. A trifunctional double-stranded DNA (dsDNA) probe was designed including a methylation site for DNA MTase recognition, a complementary sequence of 8-17 DNAzyme for synthesizing DNAzyme, and a nicking site for nicking enzyme cleavage. Firstly, the trifunctional dsDNA probe was methylated by DNA MTase to form the methylated dsDNA. Subsequently, HpaII restriction endonuclease specifically cleaved the residue of unmethylated dsDNA. Next, under the action of polymerase and nicking enzyme, the methylared dsDNA initiated SDA, releasing numbers of 8-17 DNAzymes. Finally, the released 8-17 DNAzymes triggered DNAzyme amplification reaction to induce a significant fluorescence enhancement. This strategy could detect DNA MTase activity as low as 0.0082U/mL. Additionally, the strategy was successfully applied for evaluating the inhibitions of DNA MTase using two anticancer drugs, 5-azacytidine and 5-aza-2'-deoxycytidine. The results indicate the proposed strategy has a potential application in early cancer diagnosis and therapeutics.

  9. Polyclonal antibody against an insect excitatory toxin BmKIT from Buthus Martensii karsch and detection of BmKIT expressed in transgenic cotton

    Institute of Scientific and Technical Information of China (English)

    Hao Chanjuan; Xu Chenggang; Zhang Zhiyun; Liang Aihua


    An insect excitatory toxin gene from Buthus martensii Karsch (BmKIT) was cloned into the expression vector, pET-28a. BmKIT was expressed as inclusion bodies in Escherichia coli BL21 (DE3) host cells. The authenticity of in vitro expressed protein was confirmed by Western blot. The inclusion body protein band in SDS-PAGE was excised and the protein, BmKIT, was extracted. Polyclonal antibodies to the purified protein were raised in rabbits. The antibody reacted specifically with the expressed BmKIT and was used to quantify its presence in transgenic cotton.

  10. Inhibition of KIT RNAi mediated with adenovirus in gastrointestinal stromal tumor xenograft

    Institute of Scientific and Technical Information of China (English)


    AIM: To investigate a therapeutic method for gastrointestinal stromal tumor (GIST) based on KIT RNA interference (RNAi) with AdMax adenovirus. METHODS: KIT short hairpin RNA (shRNA), whose lateral sides were decorated with restriction endonuclease sequences, was designed. T 4 DNA ligase catalyzed the joint of the KIT shRNA and the green fluorescent protein-containing PDC316-EGFP-U6 to form PDC316EGFP-U6-KIT. Homologous recombination of AdEGFPU6-KIT was performed with the AdMax system. Heterotopically transp...

  11. Discovery of amido-benzisoxazoles as potent c-Kit inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Kunz, Roxanne K.; Rumfelt, Shannon; Chen, Ning; Zhang, Dawei; Tasker, Andrew S.; Bürli, Roland; Hungate, Randall; Yu, Violeta; Nguyen, Yen; Whittington, Douglas A.; Meagher, Kristin L.; Plant, Matthew; Tudor, Yanyan; Schrag, Michael; Xu, Yang; Ng, Gordon Y.; Hu, Essa (Amgen)


    Deregulation of the receptor tyrosine kinase c-Kit is associated with an increasing number of human diseases, including certain cancers and mast cell diseases. Interference of c-Kit signaling with multi-kinase inhibitors has been shown clinically to successfully treat gastrointestinal stromal tumors and mastocytosis. Targeted therapy of c-Kit activity may provide therapeutic advantages against off-target effects for non-oncology applications. A new structural class of c-Kit inhibitors is described, including in vitro c-Kit potency, kinase selectivity, and the observed binding mode.

  12. Catalytic enantioselective addition of Grignard reagents to aromatic silyl ketimines (United States)

    Rong, Jiawei; Collados, Juan F.; Ortiz, Pablo; Jumde, Ravindra P.; Otten, Edwin; Harutyunyan, Syuzanna R.


    α-Chiral amines are of significant importance in medicinal chemistry, asymmetric synthesis and material science, but methods for their efficient synthesis are scarce. In particular, the synthesis of α-chiral amines with the challenging tetrasubstituted carbon stereocentre is a long-standing problem and catalytic asymmetric additions of organometallic reagents to ketimines that would give direct access to these molecules are underdeveloped. Here we report a highly enantioselective catalytic synthesis of N-sulfonyl protected α-chiral silyl amines via the addition of inexpensive, easy to handle and readily available Grignard reagents to silyl ketimines. The key to this success was our ability to suppress any unselective background addition reactions and side reduction pathway, through the identification of an inexpensive, chiral Cu-complex as the catalytically active structure.

  13. Anionic reagents with silicon-containing double bonds. (United States)

    Scheschkewitz, David


    E=Si transfer: Anionic compounds capable of transferring a silicon-containing double bond are reviewed (see figure), particularly reagents with Si=Si moieties (Tip=2,4,6-iPr(3)C(6)H(2), M=Li, Na, K) and their applications towards main-group and transition-metal electrophiles, as well as their reactivity towards organic compounds. A few recently reported derivatives with Si=C (Ad=1-adamantyl) and Si=P moieties are included for completeness.Anionic compounds capable of transferring a silicon double bond are summarized following an introduction to the differences between alkenes and their heavier homologues. The main focus is on reagents with Si=Si moieties and their applications towards main-group and transition-metal electrophiles, as well as their reactivity towards organic compounds, but a few recently reported derivatives with Si=C and Si=P bonds are also included.

  14. Recent advances in trifluoromethylation of organic compounds using Umemoto's reagents. (United States)

    Zhang, Cai


    The incorporation of fluorine-containing moieties into organic compounds is of great importance in pharmaceutical, agricultural, and materials science. Within these organofluorides, the trifluoromethyl group is one of the most important motifs. In recent years, the trifluoromethyl group has attracted more and more attention, and many trifluoromethylated compounds have been found to possess special activities. However, until now, only a few methods have been developed to achieve this efficiently using Umemoto's reagents. This review highlights recent developments in the direct introduction of a trifluoromethyl group into organic compounds with Umemoto's reagents. Seven approaches to the trifluoromethylation of organic compounds are summarized: (i) trifluoromethylation of arenes, (ii) trifluoromethylation of alkenes, (iii) trifluoromethylation of terminal alkynes, (iv) deoxygenative trifluoromethylation of benzylic xanthates, (v) trifluoromethylation of ketoesters, (vi) trifluoromethylation of aryl boronic acids and aromatic amines (synthesis of ArCF3) and (vii) trifluoromethylation of biphenyl isocyanide derivatives.

  15. Treatment of Wastewater Containing RDX by Fenton's Reagent

    Institute of Scientific and Technical Information of China (English)

    XU Hang; ZHANG Dong-xiang; XU Wen-guo


    Fenton's reagent was employed to treat the wastewater containing RDX. The effects of FeSO4 concentration, H2O2 concentration, pH value, reaction time, temperature and initial COD of wastewater on residual COD of wastewater were investigated. The results show that the optimum FeSO4 concentration and pH are 700mg/L and 2.5, respectively, and the residual COD of wastewater decreases with the rise in H2O2 concentration, but increases with the rise in temperature. After Fenton's reagent treatment, the initial COD of less than 874 mg/L wastewater can meet effluent standard. Under conditions of 100 mg/L H2O2, 437 mg/L initial COD and 15 ℃ temperature, the lowest residual COD is obtained at 83.80 mg/L in 5 min.

  16. Shankar: reproductive health teaching aids kit for adolescent boys. (United States)


    Currently being developed by the Thoughtshop Foundation in collaboration with the Child In Need Institute in West Bengal, this kit is intended to help peer educators in generating awareness about reproductive health issues among rural adolescent boys and men through discussions and other activities. The central theme is men's responsibility for their sexual behavior and the reproductive and sexual health of their partners. In five modules, the kit tells the story of Shankar, a boy 13 years old. Each module consists of a flip chart and accompanying visual aids/activities. The modules are as follows: 1) Puberty, self-esteem, responsibility; 2) Knowledge of changes which girls experience during puberty; 3) Childbirth; 4) Contraception--why/how; 5) Safer sex, hygiene, STD/HIV/AIDS.

  17. Assessment of RFID Read Accuracy for ISS Water Kit (United States)

    Chu, Andrew


    The Space Life Sciences Directorate/Medical Informatics and Health Care Systems Branch (SD4) is assessing the benefits Radio Frequency Identification (RFID) technology for tracking items flown onboard the International Space Station (ISS). As an initial study, the Avionic Systems Division Electromagnetic Systems Branch (EV4) is collaborating with SD4 to affix RFID tags to a water kit supplied by SD4 and studying the read success rate of the tagged items. The tagged water kit inside a Cargo Transfer Bag (CTB) was inventoried using three different RFID technologies, including the Johnson Space Center Building 14 Wireless Habitat Test Bed RFID portal, an RFID hand-held reader being targeted for use on board the ISS, and an RFID enclosure designed and prototyped by EV4.

  18. 3D vision upgrade kit for TALON robot (United States)

    Edmondson, Richard; Vaden, Justin; Hyatt, Brian; Morris, James; Pezzaniti, J. Larry; Chenault, David B.; Tchon, Joe; Barnidge, Tracy; Kaufman, Seth; Pettijohn, Brad


    In this paper, we report on the development of a 3D vision field upgrade kit for TALON robot consisting of a replacement flat panel stereoscopic display, and multiple stereo camera systems. An assessment of the system's use for robotic driving, manipulation, and surveillance operations was conducted. The 3D vision system was integrated onto a TALON IV Robot and Operator Control Unit (OCU) such that stock components could be electrically disconnected and removed, and upgrade components coupled directly to the mounting and electrical connections. A replacement display, replacement mast camera with zoom, auto-focus, and variable convergence, and a replacement gripper camera with fixed focus and zoom comprise the upgrade kit. The stereo mast camera allows for improved driving and situational awareness as well as scene survey. The stereo gripper camera allows for improved manipulation in typical TALON missions.

  19. 碱性磷酸酶试剂开瓶的稳定性研究%Study on stability of alkaline phosphatase reagent after the packaging was opened

    Institute of Scientific and Technical Information of China (English)

    段爱军; 段爱华


    Objective:To study the stability of alkaline phosphatase(ALP) after the packaging was opened and to provide the data for purchasing right amount reageng. Methods:The method of synchronous measurement was used and five kits(S0, S1, S2, S3, S4) were chosen. ALP routine test was performed using the kit S4 at eight a.m. in first day. The remaining reagent was covered tightly and preserved at 2-8 ℃. Then S3, S2, S1 and S0 kits were opened at the same time in second, third, fourth and fifth day using the same method, respectively. Five of the same parameters for ALP detection channel (ALP0, ALP1, ALP2, ALP3, ALP4, corresponding to S0, S1, S2, S3, S4) were also set in the instrument according to the reagent instruction. The detection channels were calibrated using the calibration materials(BioSino Bio-technology and Science Inc) in Selectra-E fully automatic biochemical analyzer. Results: Compared with the results obtained by S0, the results from S3 and S4 showed statistically significance(P0.05). Conclusion: The ALP reagent is stable in 48 hours after the packaging was opened and the laboratory should choose suitable packing size of reagent kit according to the dosage to use.%目的:研究碱性磷酸酶(alkaline phosphatase, ALP)试剂开瓶后的稳定性,为实验室合理购买试剂包规格提供依据。方法:采用同步测量设计方案,首先选择5个试剂盒,分别记为S0、S1、S2、S3、S4,第1天上午8点启用试剂S4进行ALP常规检测,未用完试剂盖紧盖,置2~8℃保存备用;第2天、第3天、第4天上午相同时间、相同方式启用S3、S2、S1;第5天启用试剂S0;在仪器上按试剂说明书设置5个相同参数的ALP检测通道,记为ALP0、ALP1、ALP2、ALP3、ALP4,对应试剂为S0、S1、S2、S3、S4;在荷兰威图Selectra-E全自动生化仪上用校准品(中生北控生物科技有限公司)校准各个通道;测量样品并进行结果比较分析。结果:试剂 S3、S4与试剂 S0

  20. Development of Ammonia Gas Sensor Using Optimized Organometallic Reagent


    Aubrecht, J.; Kalvoda, L.


    Reliable, continuous, and spatially distributed monitoring of dangerous or irritating chemical substances belongs to standard functions of contemporary industrial and public security systems. Fiber-optic-based detection provides feasible platform to fulfill such aims. This paper deals with characterization of ammonia sensing elements based on multimode polysiloxane-clad silica-core optical fibers sensitized with 5-(4′-dioctylamino phenylimino) quinoline-8-1 cobalt bromide complex reagent immo...

  1. Functionalization of heterocyclic compounds using polyfunctional magnesium and zinc reagents

    Directory of Open Access Journals (Sweden)

    Paul Knochel


    Full Text Available In this review we summarize the most important procedures for the preparation of functionalized organzinc and organomagnesium reagents. In addition, new methods for the preparation of polyfunctional aryl- and heteroaryl zinc- and magnesium compounds, as well as new Pd-catalyzed cross-coupling reactions, are reported herein. Experimental details are given for the most important reactions in the Supporting Information of this article.

  2. Supramolecular Architecture of Enzyme-like Reagents ( I )

    Institute of Scientific and Technical Information of China (English)

    MAO; LuYuan


    Metalloporphyrin has been widely used in the enzymatic method of analysis as a mimesis of peroxidase [1]. It is interesting work to utilize functional polymer to not only serve as a carrier of biomimetic catalyst[2] , but also as a biochemical reagent or mimic the spacial structure of enzyme. In this paper, we studied properties of recently synthesized β-CDEP resin, a cyclodextrin polymer, for the inclusion and adsorption of indissoluble porphyrin.……

  3. New reagents for detecting free radicals and oxidative stress. (United States)

    Barzegar Amiri Olia, Mina; Schiesser, Carl H; Taylor, Michelle K


    Free radicals and oxidative stress play important roles in the deterioration of materials, and free radicals are important intermediates in many biological processes. The ability to detect these reactive species is a key step on the road to their understanding and ultimate control. This short review highlights recent progress in the development of reagents for the detection of free radicals and reactive oxygen species with broad application to materials science as well as biology.

  4. Intramolecular cyclization of steroidal semicarbazones to pyrazoles using Vilsmeier reagent

    Institute of Scientific and Technical Information of China (English)

    Mahboob Alam; M.Mushfiq


    The preparation of hitherto unknown steroidal heterocycles containing pyrazole fused to 6,7-position of the steroidal nucleus is described.These heterocycles were prepared by the action of Vilsmeier reagent with steroidal semicarbazones in DMF.The slructure of the compounds has been established on the basis of their elemental analysis and spectral data.A general mechanistic scheme for these reactions is also suggested based on current and previous results.

  5. Reaction rates between water and some modified rapidly-reacting Karl Fischer reagents. (United States)

    Cedergren, A


    Rate constants were determined for the reaction between water and various modified Karl Fischer reagents containing formamide, dimethylformamide or N-methylformamide. It was shown that the reaction rate can be increased by a factor of 100 by using a reagent containing 40% v/v formamide in pyridine compared to that obtained by use of the conventional methanolic reagent.

  6. 21 CFR 866.3332 - Reagents for detection of specific novel influenza A viruses. (United States)


    ... A viruses. 866.3332 Section 866.3332 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF... Reagents § 866.3332 Reagents for detection of specific novel influenza A viruses. (a) Identification. Reagents for detection of specific novel influenza A viruses are devices that are intended for use in...

  7. Improving Situation Awareness with the Android Team Awareness Kit (ATAK) (United States)


    video. This paper describes the Android Team Awareness Kit (ATAK), an advanced, distributed tool for commercial- o -the-shelf (COTS) mobile devices...interrelated technologies : increasingly powerful mobile computing devices and increasingly available and rich georeferenced data. ATAK is designed to...50 different apps in the Trans Apps marketplace. Trans Apps uses a modified version of the Android operating system with enhanced security features

  8. Wong Leung Kit-Wah and Her Exhibition Tour

    Institute of Scientific and Technical Information of China (English)


    BEAUTIFUL ladies painted by beautiful woman Annie Wong Leung Kit-Wah, a renowned painter from Hong Kong recently came to Beijing. Upon the first anniversary of Hong Kong’s return to its motherland, Leung held an exhibition of her work at the China Art Gallery. Each piece conveyed her ability to use the unique techniques of Chinese painting to manifest the beauty of women, and her own admiration for women’s qualities: grace and poise, modesty and

  9. Software Development Kit for Internet Payment Gateway Service


    Larjomaa, Jonni


    In this thesis a Software Development Kit (SDK) was developed which simplifies the integration work of an Internet payment gateway service called the Payment Highway to a PHP based electronic commerce solution. The Payment Highway is an Internet payment gateway solution developed by a software company called Solinor Oy. The payment gateway acts as middleware for authorizations, credit and debit transactions securely storing and handling the consumers’ credit card information. PHP is a...

  10. Usefulness of ED036 kit for measuring serum PIVKA-II levels in small hepatocellular carcinoma. (United States)

    Kuromatsu, R; Tanaka, M; Shimauchi, Y; Shimada, M; Tanikawa, K; Watanabe, K; Yokoo, T


    As a tumor marker for hepatocellular carcinoma (HCC), serum protein induced by vitamin K absence or antagonist-II (PIVKA-II) has high specificity, yet its sensitivity is relatively low, marking it less suitable to serve as an adjunct in the diagnosis of small HCC. Recently, the ED036 kit (Eisai, Tokyo, Japan), whose detection limit is approximately ten times superior to that of a conventional kit (Eitest MONOP II, Eisai) has been developed. In this study, serum PIVKA-II levels in serum samples from 83 patients with benign chronic liver diseases (CLD) and 129 patients with HCC were measured with those two kits. With the ED036 kit, the cut-off value was set at 40 mAU/ml. For PIVKA-II measured with the ED036 kit, sensitivity was 45.0%, specificity 92.8%, and accuracy 63.7%, when we discriminated patients with HCC from those with CLD without HCC. While maintaining a high specificity, of 92.8%, the ED036 kit showed a significantly higher sensitivity than the conventional kit (45.0% versus 27.9%; P PIVKA-II with the ED036 kit was significantly greater than the rate with the conventional kit (21.4% versus 9.5%; P < 0.005). Thus, the ED036 kit was thought to be more useful than the conventional kit as a tumor marker for small HCC.

  11. Changes in c-Kit expression levels during the course of radiation therapy for nasopharyngeal carcinoma (United States)

    Jiang, Feng; Hu, Wei; Zhang, Bicheng; Xu, Jing; Shui, Yongjie; Zhou, Xiaofeng; Ren, Xiaoqiu; Chen, Xiaozhong; Shen, Li; Wei, Qichun


    In the era of intensity-modulated radiotherapy, distant metastasis is currently the main cause of treatment failure for nasopharyngeal carcinoma (NPC). Additional therapeutic strategies are required to control the metastasis and improve survival. One strategy is targeted therapy, for example against c-Kit. In the current study, the frequency of c-Kit expression was determined immunohistochemically in 106 NPC patients. c-Kit expression changes during the course of radiation therapy were detected in 41 cases via weekly biopsy. Twelve cases (11.3%) had c-Kit expression scores of 3+ and 16 (15.1%) had scores of 2+. Thus, c-Kit overexpression (2+ or 3+) was observed in 28 (26.4%) patients. There were 35 (33.0%) and 43 (40.6%) patients with c-Kit expression scores of 1+ and 0, respectively. Furthermore, a trend of decreased c-Kit expression was observed after commencing radiotherapy according to the 41 NPC patients who were biopsied weekly. Therefore, c-Kit overexpression was identified to be common in NPC, and evaluating c-Kit as a therapeutic target for metastatic NPC via c-Kit overexpression subsequent to first line treatment may be of interest. To the best of our knowledge, the present study is the first to demonstrate a trend of decreased c-Kit expression during the course of radiotherapy. PMID:27699010

  12. Identification of mimotopes of Mycobacterium leprae as potential diagnostic reagents

    Directory of Open Access Journals (Sweden)

    Alban Silvana M


    Full Text Available Abstract Background An early diagnostic test for detecting infection in leprosy is fundamental for reducing patients’ sequelae. The currently used lepromin is not adequate for disease diagnosis and, so far, no antigen to be used in intradermoreaction has proved to be sensitive and specific for that purpose. Aiming at identifying new reagents to be used in skin tests, candidate antigens were investigated. Methods Random peptide phage display libraries were screened by using antibodies from leprosy patients in order to identify peptides as diagnostic reagents. Results Seven different phage clones were identified using purified antibodies pooled from sera of leprosy patients. When the clones were tested with serum samples by ELISA, three of them, 5A, 6A and 1B, allowed detecting a larger number of leprosy patients when compared to controls. The corresponding peptides expressed by selected phage clones were chemically synthesized. A pilot study was undertaken to assess the use of peptides in skin tests. The intradermal challenge with peptides in animals previously sensitized with Mycobacterium leprae induced a delayed-type hypersensitivity with peptide 5A (2/5 and peptide 1B (1/5. In positive controls, there was a 3/5 reactivity for lepromin and a 4/5 reactivity of the sensitized animals with soluble extract of M. leprae. Conclusions The preliminary data suggest that may be possible to develop reagents with diagnostic potential based on peptide mimotopes selected by phage display using polyclonal human antibodies.

  13. Fabrication of CANFLEX bundle kit for irradiation test in NRU

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Moon Sung; Kwon, Hyuk Il; Ji, Chul Goo; Chang, Ho Il; Sim, Ki Seob; Suk, Ho Chun


    CANFLEX bundle kit was prepared at KAERI for the fabrication of complete bundle at AECL. Completed bundle will be used for irradiation test in NRU. Provisions in the `Quality Assurance Manual for HWR Fuel Projects,` `Manufacturing Plan` and `Quality Verification, Inspection and Test Plan` were implemented as appropriately for the preparation of CANFLEX kit. A set of CANFLEX kit consist of 43 fuel sheath of two different sizes with spacers, bearing pads and buttons attached, 2 pieces of end plates and 86 pieces of end caps with two different sizes. All the documents utilized as references for the fabrication such as drawings, specifications, operating instructions, QC instructions and supplier`s certificates are specified in this report. Especially, suppliers` certificates and inspection reports for the purchased material as well as KAERI`s inspection report are integrated as attachments to this report. Attached to this report are supplier`s certificates and KAERI inspection reports for the procured materials and KAERI QC inspection reports for tubes, pads, spacers, buttons, end caps, end plates and fuel sheath. (author). 37 refs.

  14. Intelligence amplification framework for enhancing scheduling processes

    NARCIS (Netherlands)

    Dobrkovic, Andrej; Liu, Luyao; Iacob, Maria-Eugenia; Hillegersberg, van Jos


    The scheduling process in a typical business environment consists of predominantly repetitive tasks that have to be completed in limited time and often containing some form of uncertainty. The intelligence amplification is a symbiotic relationship between a human and an intelligent agent. This partn

  15. Social amplification of risk: a conceptual framework

    Energy Technology Data Exchange (ETDEWEB)

    Kasperson, R.E.; Renn, O.; Slovic, P.; Brown, H.S.; Emel, J.; Goble, R.; Kasperson, J.X.; Ratick, S.


    One of the most perplexing problems in risk analysis is why some relatively minor risks or risk events, as assessed by technical experts, often elicit strong public concerns and result in substantial impacts upon society and economy. This article sets forth a conceptual framework that seeks to link systematically the technical assessment of risk with psychological, sociological, and cultural perspectives of risk perception and risk-related behavior. The main thesis is that hazards interact with psychological, social, institutional, and cultural processes in ways that may amplify or attenuate public responses to the risk or risk event. A structural description of the social amplification of risk is now possible. Amplification occurs at two stages: in the transfer of information about the risk, and in the response mechanisms of society. Signals about risk are processed by individual and social amplification stations, including the scientist who communicates the risk assessment, the news media, cultural groups, interpersonal networks, and others. Key steps of amplifications can be identified at each stage. The amplified risk leads to behavioral responses, which, in turn, result in secondary impacts. Models are presented that portray the elements and linkages in the proposed conceptual framework.

  16. Desert Amplification in a Warming Climate (United States)

    Zhou, Liming


    Here I analyze the observed and projected surface temperature anomalies over land between 50°S-50°N for the period 1950–2099 by large-scale ecoregion and find strongest warming consistently and persistently seen over driest ecoregions such as the Sahara desert and the Arabian Peninsula during various 30-year periods, pointing to desert amplification in a warming climate. This amplification enhances linearly with the global mean greenhouse gases(GHGs) radiative forcing and is attributable primarily to a stronger GHGs-enhanced downward longwave radiation forcing reaching the surface over drier ecoregions as a consequence of a warmer and thus moister atmosphere in response to increasing GHGs. These results indicate that desert amplification may represent a fundamental pattern of global warming associated with water vapor feedbacks over land in low- and mid- latitudes where surface warming rates depend inversely on ecosystem dryness. It is likely that desert amplification might involve two types of water vapor feedbacks that maximize respectively in the tropical upper troposphere and near the surface over deserts, with both being very dry and thus extremely sensitive to changes of water vapor.

  17. Fluorogenic Detection of Duck Tembusu Virus( DTMUV ) by Loop-mediated Isothermal Amplification(LAMP)

    Institute of Scientific and Technical Information of China (English)

    Zhang; Lin; Wang; Bin; Zhang; Wei; Zhang; Xiumei


    This study was to develop an efficient and simple method for the detection of duck Tembusu virus( DTMUV) by loop-mediated isothermal amplification( LAMP). Six pairs of LAMP primers were designed according to the conserved region of the DTMUV E gene sequence in Gen Bank,which were then used for the optimization of various reaction components and reaction system of specific LAMP for DTMUV. Further the fluorescent reagent SYBR Green I and a certain proportion of calcium and manganese ion were used to determin the color development of products for visible analysis instead of agarose gel electrophoresis. The results showed that the sensitivity SYBR Green I as the fluorescent reagent was 10 copies viruses per μL,which is 100 times higher than normal PCR method,while the detection limit of combined use of calcium and manganese ion was 1 000 copies viruses per μL. Although the sensitivity of mixture of calcium and manganese ion is lower than SYBR Green I,it can avoid the aerosol contamination. The fluorogenic analysis-based LAMP system established in our study has a high sensitivity and avoid the cross contamination,which is of huge potential in research institutions,grass-roots laboratories and field testing and can provide effective means to completely curb the occurrence and spreading of DTMUV.

  18. Electrical detection of dsDNA and polymerase chain reaction amplification. (United States)

    Salm, Eric; Liu, Yi-Shao; Marchwiany, Daniel; Morisette, Dallas; He, Yiping; Razouk, Laila; Bhunia, Arun K; Bashir, Rashid


    Food-borne pathogens and food safety-related outbreaks have come to the forefront over recent years. Estimates on the annual cost of sicknesses, hospitalizations, and deaths run into the billions of dollars. There is a large body of research on detection of food-borne pathogens; however, the widely accepted current systems are limited by costly reagents, lengthy time to completion, and expensive equipment. Our aim is to develop a label-free method for determining a change in DNA concentration after a PCR assay. We first used impedance spectroscopy to characterize the change in concentration of purified DNA in deionized water within a microfluidic biochip. To adequately measure the change in DNA concentration in PCR solution, it was necessary to go through a purification and precipitation step to minimize the effects of primers, PCR reagents, and excess salts. It was then shown that the purification and precipitation of the fully amplified PCR reaction showed results similar to the control tests performed with DNA in deionized water. We believe that this work has brought label free electrical biosensors for PCR amplification one step closer to reality.

  19. A multicenter international evaluation of single-tube amplification protocols for sequencing-based typing of HLA-DRB1 and HLA-DRB3,4,5. (United States)

    Sayer, D C; Whidborne, R; De Santis, D; Rozemuller, E H; Christiansen, F T; Tilanus, M G


    We have described previously a novel single-tube polymerase chain reaction (PCR) amplification (STAmp) protocol for the efficient sequencing-based typing (SBT) of human leukocyte antigen (HLA)-DRB1. The PCR amplification mix includes primers to each of seven allele group-sequence motifs. We have applied this principle to the simultaneous SBT of HLA-DRB3, -DRB4, and -DRB5 using locus specific primers. We report here a multicenter international evaluation of the STAmp protocols performed as a component of the 13th International Histocompatibility Workshop. Identical amplification primer mixes, sequencing primers, and DNA were sent to participating laboratories. The primer mixes contained the amplification primers and the PCR buffer. Each laboratory was requested to amplify the DNA with the primer mixes and perform SBT on the resulting PCR protocols, using their own protocols, and return the typing results for analysis. The reported results indicated that the expected sequence could be obtained with a variety of PCR amplification and sequencing platforms and protocols. There were difficulties but these seemed unrelated to STAmp reagents and suggest that optimal SBT results can be obtained if bi-directional sequencing is performed and software is used for sequence verification and editing. This indicates that SBT by STAmp can be applied in many laboratories for high-throughput HLA-DRB1 and HLA-DRB3,4,5 SBT.

  20. A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method). (United States)

    Visvanathan, Rizliya; Jayathilake, Chathuni; Liyanage, Ruvini


    For the first time, a reliable, simple, rapid and high-throughput analytical method for the detection and quantification of α-amylase inhibitory activity using the glucose assay kit was developed. The new method facilitates rapid screening of a large number of samples, reduces labor, time and reagents and is also suitable for kinetic studies. This method is based on the reaction of maltose with glucose oxidase (GOD) and the development of a red quinone. The test is done in microtitre plates with a total volume of 260μL and an assay time of 40min including the pre-incubation steps. The new method is tested for linearity, sensitivity, precision, reproducibility and applicability. The new method is also compared with the most commonly used 3,5-dinitrosalicylic acid (DNSA) method for determining α-amylase activity.

  1. A new evolutionary theory deduced mathematically from entropy amplification

    Institute of Scientific and Technical Information of China (English)


    A new evolutionary theory which is able to unite the present evolutionary debates is deduced mathematically from the principle of entropy amplification.It suggests that the extensive evolution is driven by the amplification of entropy,or microscopic diversity,and the biological evolution is driven by the amplification of biodiversity.Forming high hierarchies is the most important way for the amplification and brings out spontaneously three kinds of selection.This theory has some positive cultural meanings.

  2. 罗氏尿微量清蛋白检测试剂盒性能评价%Performance evaluation on Roche urine trace albumin detection reagent kit

    Institute of Scientific and Technical Information of China (English)

    黄勤烽; 黄少铃; 王美珠; 陈敏


    Objective To evaluate the detection performance of the cobas8000 c702 fully automatic biochemical analyzer for de‐tecting the second generation Roche urine trace albumin (ALBU2) .Methods (1) The precise evaluation :with the allowable error stipulated by CLIA 88 as the basis ,the requirements were the repeat precision <1/4TEa ,and intermediate precision <1/3TEa;(2) the linear range and the evaluation of the reportable range :the EP6‐A scheme was adopted ,and extend to calculate the average re‐covery rate of dilution ,the clinical reportable range was evaluated by the average dilution recovery of 90% -110% ;(3) the carry o‐ver pollution assessment :the carry over pollution of serum albumin on urine trace albumin detection was evaluated by the judgment standard of carry over pollution rate of 0 .5% ;(4)the methodological comparative analysis :with SIEMENS BN Ⅱas the reference system ,the Roche Cobas 8000 C702 and the BN2 results were performed the correlation contrastive analysis .Results The repeat precision :low concentration CV=1 .98% .high concentration CV=1 .64% ;intermediate precision :low concentration CV=4 .35% , high concentration CV=1 .20% ;the linear range verification :the measurement range 5 .6-413 .55 mg/L ;clinical reportable range :in the maximum diluted multiples of 30 times ,the clinical reportable range was 5 .6-12 406 .5 mg/L ;the carry over pollution rate :serum albumin (42 .6 g/L) on urine trace albumin(6 .9 mg/L) ,the carry over pollution rate was 0 .28% ;the indoor comparison :in the concentration within 200 mg/L ,the regression line was Y=0 .896 X+5 .049 ,the correlation coefficient r2 =0 .994 4 ,the system shift was passed at the medical decision level .When the specimen concentration within 201-413 .55 mg/L ,the regression line was Y=0 .848X-10 .44 ,the correlation coefficient r2 =0 .917 ,the system shift was not passed at the medical decision level .Conclusion The detection of the Roche ALBU2 in the Cobas 8000 C702 platform can meet the clinical needs ,the comparison among different instruments has difference in different concentration ranges ,therefore the independent reference ranges should be established ac‐cording to the each instrument system .%目的:评价罗氏第2代尿微量清蛋白(ALBU2)在Cobas 8000 C702全自动生化分析仪上的检测性能。方法(1)精密度评价:以CLIA′88规定的允许误差TEa为依据,要求重复精密度<1/4TEa ,中间精密度<1/3TEa;(2)线性范围及可报告范围评价:采用EP6‐A方案,并延伸计算平均稀释回收率,以平均稀释回收率在90%~110%,评价临床可报告范围;(3)携带污染评估:评估血清清蛋白对尿量微量清蛋白检测的携带污染,以携带污染率≤0.5%标准判定;(4)方法比对分析:以Siemens BNⅡ为参考系统,将罗氏Cobas 8000 C702与BN2结果进行相关性比对分析。结果重复精密度:低浓度 CV为1.98%,高浓度 CV为1.64%;中间精密度:低浓度 CV为4.35%,高浓度 CV为1.20%。线性范围验证:测量范围为5.6~413.55 mg/L ;临床可报告范围:最大稀释倍数为30倍,临床可报告范围为5.6~12406.5mg/L。携带污染率:血清清蛋白(42.6g/L)对尿液微量清蛋白(6.9 mg/L )的携带污染为0.28%。室内比对:标本浓度在200 mg/L时,回归直线为Y =0.896 X+5.049,r2=0.9944,医学决定水平处系统偏移通过;当标本浓度在201~413.55 m g/L时,回归直线为Y=0.848 X -10.44,r2=0.917,医学决定水平处系统偏移未通过。结论罗氏ALBU2在Cobas 8000 C702平台上检测能满足临床的需求,不同仪器间的比对,在不同浓度范围内有差异,应根据各自仪器的系统建立独立的参考范围。

  3. Quantitation of viral load using real-time amplification techniques

    NARCIS (Netherlands)

    Niesters, H G


    Real-time PCR amplification techniques are currently used to determine the viral load in clinical samples for an increasing number of targets. Real-time PCR reduces the time necessary to generate results after amplification. In-house developed PCR and nucleic acid sequence-based amplification (NASBA

  4. Isothermal DNA amplification in bioanalysis: strategies and applications. (United States)

    Kim, Joonyul; Easley, Christopher J


    Isothermal DNA amplification is an alternative to PCR-based amplification for point-of-care diagnosis. Since the early 1990s, the approach has been refined into a simple, rapid and cost-effective tool by means of several distinct strategies. Input signals have been diversified from DNA to RNA, protein or small organic molecules by translating these signals into input DNA before amplification, thus allowing assays on various classes of biomolecules. In situ detection of single biomolecules has been achieved using an isothermal method, leveraging localized signal amplification in an intact specimen. A few pioneering studies to develop a homogenous isothermal protein assay have successfully translated structure-switching of a probe upon target binding into input DNA for isothermal amplification. In addition to the detection of specific targets, isothermal methods have made whole-genome amplification of single cells possible owing to the unbiased, linear nature of the amplification process as well as the large size of amplified products given by ϕ29 DNA polymerase. These applications have been devised with the four isothermal amplification strategies covered in this review: strand-displacement amplification, rolling circle amplification, helicase-dependent amplification and recombinase polymerase amplification.

  5. Bioanalytical applications of isothermal nucleic acid amplification techniques. (United States)

    Deng, Huimin; Gao, Zhiqiang


    The most popular in vitro nucleic acid amplification techniques like polymerase chain reaction (PCR) including real-time PCR are costly and require thermocycling, rendering them unsuitable for uses at point-of-care. Highly efficient in vitro nucleic acid amplification techniques using simple, portable and low-cost instruments are crucial in disease diagnosis, mutation detection and biodefense. Toward this goal, isothermal amplification techniques that represent a group of attractive in vitro nucleic acid amplification techniques for bioanalysis have been developed. Unlike PCR where polymerases are easily deactivated by thermally labile constituents in a sample, some of the isothermal nucleic acid amplification techniques, such as helicase-dependent amplification and nucleic acid sequence-based amplification, enable the detection of bioanalytes with much simplified protocols and with minimal sample preparations since the entire amplification processes are performed isothermally. This review focuses on the isothermal nucleic acid amplification techniques and their applications in bioanalytical chemistry. Starting off from their amplification mechanisms and significant properties, the adoption of isothermal amplification techniques in bioanalytical chemistry and their future perspectives are discussed. Representative examples illustrating the performance and advantages of each isothermal amplification technique are discussed along with some discussion on the advantages and disadvantages of each technique.

  6. Plasmonic Terahertz Amplification in Graphene-Based Asymmetric Hyperbolic Metamaterial

    Directory of Open Access Journals (Sweden)

    Igor Nefedov


    Full Text Available We propose and theoretically explore terahertz amplification, based on stimulated generation of plasmons in graphene asymmetric hyperbolic metamaterials (AHMM, strongly coupled to terahertz radiation. In contrast to the terahertz amplification in resonant nanocavities, AHMM provides a wide-band THz amplification without any reflection in optically thin graphene multilayers.

  7. Plasmonic Terahertz Amplification in Graphene-Based Asymmetric Hyperbolic Metamaterial


    Igor Nefedov; Leonid Melnikov


    We propose and theoretically explore terahertz amplification, based on stimulated generation of plasmons in graphene asymmetric hyperbolic metamaterials (AHMM), strongly coupled to terahertz radiation. In contrast to the terahertz amplification in resonant nanocavities, AHMM provides a wide-band THz amplification without any reflection in optically thin graphene multilayers.

  8. Stability study for magnetic reagent assaying Hb and HbA1c (United States)

    Hsieh, Wen-Pin; Chieh, J. J.; Yang, C. C.; Yang, S. Y.; Chen, Po-Yu; Huang, Yu-Hao; Hong, Y. W.; Horng, H. E.


    Reagents for magnetically labeled immunoassay on human Hb and human HbA1c have been synthesized. The reagents consist of Fe3O4 magnetic particles biofunctionalized with antibodies against Hb and HbA1c. It has been demonstrated that the reagents can be applied to quantitatively detect Hb and HbA1c by using immunomagnetic reduction assay. In addition to characterizing the assay properties, such as the standard curve and the low-detection limit, the stability of reagents is investigated. To do this, the temporal dependence of particle sizes and the bio-activity of reagents are monitored. The results show that the reagents are highly stable when stored at 2-8 °C. This means that the reagents synthesized in this work are promising for practical applications.

  9. RNA amplification for successful gene profiling analysis

    Directory of Open Access Journals (Sweden)

    Wang Ena


    Full Text Available Abstract The study of clinical samples is often limited by the amount of material available to study. While proteins cannot be multiplied in their natural form, DNA and RNA can be amplified from small specimens and used for high-throughput analyses. Therefore, genetic studies offer the best opportunity to screen for novel insights of human pathology when little material is available. Precise estimates of DNA copy numbers in a given specimen are necessary. However, most studies investigate static variables such as the genetic background of patients or mutations within pathological specimens without a need to assess proportionality of expression among different genes throughout the genome. Comparative genomic hybridization of DNA samples represents a crude exception to this rule since genomic amplification or deletion is compared among different specimens directly. For gene expression analysis, however, it is critical to accurately estimate the proportional expression of distinct RNA transcripts since such proportions directly govern cell function by modulating protein expression. Furthermore, comparative estimates of relative RNA expression at different time points portray the response of cells to environmental stimuli, indirectly informing about broader biological events affecting a particular tissue in physiological or pathological conditions. This cognitive reaction of cells is similar to the detection of electroencephalographic patterns which inform about the status of the brain in response to external stimuli. As our need to understand human pathophysiology at the global level increases, the development and refinement of technologies for high fidelity messenger RNA amplification have become the focus of increasing interest during the past decade. The need to increase the abundance of RNA has been met not only for gene specific amplification, but, most importantly for global transcriptome wide, unbiased amplification. Now gene

  10. Identification of a novel subgroup of melanomas with KIT/cyclin-dependent kinase-4 overexpression. (United States)

    Smalley, Keiran S M; Contractor, Rooha; Nguyen, Thiennga K; Xiao, Min; Edwards, Robin; Muthusamy, Viswanathan; King, Alastair J; Flaherty, Keith T; Bosenberg, Marcus; Herlyn, Meenhard; Nathanson, Katherine L


    Although many melanomas harbor either activating mutations in BRAF or NRAS, there remains a substantial, yet little known, group of tumors without either mutation. Here, we used a genomic strategy to define a novel group of melanoma cell lines with co-overexpression of cyclin-dependent kinase 4 (CDK4) and KIT. Although this subgroup lacked any known KIT mutations, they had high phospho-KIT receptor expression, indicating receptor activity. Quantitative PCR confirmed the existence of a similar KIT/CDK4 subgroup in human melanoma samples. Pharmacologic studies showed the KIT/CDK4-overexpressing subgroup to be resistant to BRAF inhibitors but sensitive to imatinib in both in vitro and in vivo melanoma models. Mechanistically, imatinib treatment led to increased apoptosis and G(1) phase cell cycle arrest associated with the inhibition of phospho-ERK and increased expression of p27(KIP). Other melanoma cell lines, which retained some KIT expression but lacked phospho-KIT, were not sensitive to imatinib, suggesting that KIT expression alone is not predictive of response. We suggest that co-overexpression of KIT/CDK4 is a potential mechanism of oncogenic transformation in some BRAF/NRAS wild-type melanomas. This group of melanomas may be a subpopulation for which imatinib or other KIT inhibitors may constitute optimal therapy.

  11. Increased DNA amplification success of non-invasive genetic samples by successful removal of inhibitors from faecal samples collected in the field

    DEFF Research Database (Denmark)

    Hebert, Louise; Darden, Safi K.; Pedersen, Bo Vest;


    The use of non-invasive genetic sampling (NGS) is becoming increasingly important in the study of wild animal populations. Obtaining DNA from faecal samples is of particular interest because faeces can be collected without deploying sample capture devices. However, PCR amplification of DNA...... extracted from faeces is problematic because of high concentrations of inhibitors. Here we present a method for increasing the successful application of donor DNA extracted from faecal samples through inhibitor reduction. After standard extraction with a DNA stool kit we used a ‘Concentrated Chelex...... Treatment’ (CCT) that increased the amplification success from 31.7 to 61.4% of loci. Our results suggest that darker supernatant and samples with more precipitate contain more inhibitors than lighter samples and samples with little or no precipitate. We expect the use of this technique to have wide...

  12. The development of new diagnostic test-systems and improvement of the existed radio immunochemical kits in the industrial production conditions

    CERN Document Server

    Abdukayumov, A M


    completed the laboratory and clinical testing of the kits of reagents 'IRMA-M-HBsAg- sup 1 sup 2 sup 5 I', 'ELISA-HBsAg', 'Recombinant-anti-HCV-strip' that had shown the high specificity and reproduction of the analysis results. Immunoenzymetic kit 'ELISA-HBsAg' of the third generation for HBsAg detection in human blood serum with the sensitivity not less than 0.5 ng/ml was introduced into industrial production. It had been developed and introduced into industrial production the highly sensitive immunoenzymetic test-system 'Recombinant-anti-HCV-strip' of the third generation for antibodies to hepatitis C virus detection in human blood serum. The highly effective method had been developed for HBsAg preparing and purification that allowed to use as the raw material a plasma of donors with low content of the target product and based on the application of the stepped fractionation by Ammonium Sulphate and affinity chromatography. The use efficiency of the produced HBsAg preparations had been shown for laboratory ...

  13. Gravito-magnetic amplification in cosmology

    CERN Document Server

    Tsagas, Christos G


    Magnetic fields interact with gravitational waves in various ways. We consider the coupling between the Weyl and the Maxwell fields in cosmology and study the effects of the former on the latter. The approach is fully analytical and the results are gauge-invariant. We show that the nature and the outcome of the gravito-magnetic interaction depends on the electric properties of the cosmic medium. When the conductivity is high, gravitational waves reduce the standard (adiabatic) decay rate of the B-field, leading to its superadiabatic amplification. In poorly conductive environments, on the other hand, Weyl-curvature distortions can result into the resonant amplification of large-scale cosmological magnetic fields. Driven by the gravitational waves, these B-fields oscillate with an amplitude that is found to diverge when the wavelengths of the two sources coincide. We present technical and physical aspects of the gravito-magnetic interaction and discuss its potential implications.

  14. Parametric nanomechanical amplification at very high frequency. (United States)

    Karabalin, R B; Feng, X L; Roukes, M L


    Parametric resonance and amplification are important in both fundamental physics and technological applications. Here we report very high frequency (VHF) parametric resonators and mechanical-domain amplifiers based on nanoelectromechanical systems (NEMS). Compound mechanical nanostructures patterned by multilayer, top-down nanofabrication are read out by a novel scheme that parametrically modulates longitudinal stress in doubly clamped beam NEMS resonators. Parametric pumping and signal amplification are demonstrated for VHF resonators up to approximately 130 MHz and provide useful enhancement of both resonance signal amplitude and quality factor. We find that Joule heating and reduced thermal conductance in these nanostructures ultimately impose an upper limit to device performance. We develop a theoretical model to account for both the parametric response and nonequilibrium thermal transport in these composite nanostructures. The results closely conform to our experimental observations, elucidate the frequency and threshold-voltage scaling in parametric VHF NEMS resonators and sensors, and establish the ultimate sensitivity limits of this approach.

  15. Amplification of postwildfire peak flow by debris (United States)

    Kean, J. W.; McGuire, L. A.; Rengers, F. K.; Smith, J. B.; Staley, D. M.


    In burned steeplands, the peak depth and discharge of postwildfire runoff can substantially increase from the addition of debris. Yet methods to estimate the increase over water flow are lacking. We quantified the potential amplification of peak stage and discharge using video observations of postwildfire runoff, compiled data on postwildfire peak flow (Qp), and a physically based model. Comparison of flood and debris flow data with similar distributions in drainage area (A) and rainfall intensity (I) showed that the median runoff coefficient (C = Qp/AI) of debris flows is 50 times greater than that of floods. The striking increase in Qp can be explained using a fully predictive model that describes the additional flow resistance caused by the emergence of coarse-grained surge fronts. The model provides estimates of the amplification of peak depth, discharge, and shear stress needed for assessing postwildfire hazards and constraining models of bedrock incision.

  16. Silver nanoparticles incorporated onto ordered mesoporous silica from Tollen's reagent (United States)

    Zienkiewicz-Strzałka, M.; Pasieczna-Patkowska, S.; Kozak, M.; Pikus, S.


    Noble metal nanostructures supported on mesoporous silica are bridge between traditional silica adsorbents and modern catalysts. In this work the Ag/SBA-15 mesoporous materials were synthesized and characterized. Various forms of nanosilver supported on ordered mesoporous template have been successfully obtained via proposed procedures. In all synthesized materials, Tollen's reagent (diammine silver complex [Ag(NH3)2]+) was used as a silver source. Silver nanoparticles were prepared by reduction of ammoniacal silver complex by formaldehyde in the solution of stabilizer. After reduction, Ag nanoparticles could be deposited on SBA-15, or added during traditional synthesis of SBA-15 giving silver or silver chloride nanoparticles in the combination with porous silica. Silver nanostructures as nanoparticles or nanowires were also embedded onto the SBA-15 by incipient wetness impregnation of silver ions. Absorbed silver ions were next reduced under hydrogen at high temperature. There are many advantages of utilized ammoniacal silver complex as a silver source. Proposed method is capable to synthesis of various metal nanostructures with controlled composition and morphology. The silver ammonia complex is composed of two ions surrounding and protecting the central silver ion, so it is possible to obtain very small nanoparticles using simple approach without any functionalization of external and internal surface of SBA-15. This approach allows obtaining greatly small silver nanoparticles on SBA-15 (4 nm) or nanowires depending on the metal loading amount. Moreover, the colloidal silver solution prepared from Tollen's reagent, in the presence of triblock copolymer, remains stable for a long time. Reduction of Tollen's reagent to silver colloidal solution seems to be efficient, fast and interesting approach for the preparation of supported silver nanostructures Obtained samples were characterized by powder X-ray diffraction, small angle X-ray scattering (SAXS), UV

  17. Internal entanglement amplification by external interactions



    We propose a scheme to control the level of entanglement between two fixed spin-1/2 systems by interaction with a third particle. For specific designs, entanglement is shown to be "pumped" into the system from the surroundings even when the spin-spin interaction within the system is small or nonexistent. The effect of the external particle on the system is introduced by including a dynamic spinor in the Hamiltonian. Controlled amplification of the internal entanglement to its maximum value is...

  18. Usage of fly ash as a coal desulphurization reagent

    Energy Technology Data Exchange (ETDEWEB)

    Yaman, S.; Kuecuekbayrak, S. [Istanbul Technical Univ. (Turkey). Chemical and Metallurgical Engineering Faculty


    This paper covers the direct usage of fly ash to remove sulphur from coal. Experiments were carried out on a high sulphur Turkish lignite. 5 g of fly ash was extracted in 200 ml of water under pressure and the dilute solution containing water extractable parts of fly ash was used as desulphurization reagent. Oxygen pressure was created over desulphurization medium during the extraction period by which dissolved oxygen was concentrated in the solution. Effects of temperature, partial pressure of oxygen, and time were investigated in the ranges of 403--498 K, 0.0--1.5 MPa and 15--90 min, respectively.

  19. Nucleophilic Addition of Organozinc Reagents to 2-Sulfonyl Cyclic Ethers (United States)

    Kim, Hyoungsu; Kasper, Amanda C.; Moon, Eui Jung; Park, Yongho; Wooten, Ceshea M.; Dewhirst, Mark W.; Hong, Jiyong


    A convergent route to the synthesis of manassantins A and B, potent inhibitors of HIF-1, is described. Central to the synthesis is a stereoselective addition of an organozinc reagent to a 2-benzenesulfonyl cyclic ether to achieve the 2,3-cis-3,4-trans-4,5-cis-tetrahydrofuran of the natural products. Preliminary structure—activity relationships suggested that the (R)-configuration at C-7 and C-7″′ is not critical for HIF-1 inhibition. In addition, the hydroxyl group at C-7 and C-7″′ can be replaced with carbonyl group without loss of activity. PMID:19111058

  20. Dosing of Reagents and Solid Supports as Tablets

    Institute of Scientific and Technical Information of China (English)

    T. Ruhland; P. Holm; K Andersen


    @@ 1Introduction During the latest decade, the intensive investigation into the solid-phase synthesis of small organic molecules, as well as the use of polymer-supported reagents and catalysts for solution-phase organic synthesis has lead to paradigm shifts in many areas of chemistry. This has particularly been the case within the fields of biological and medicinal chemistry where the parallel synthesis of discrete molecules (in series or larger libraries), either by manual or automated methods, has been implemented as a key technology/methodology in the preparation of compounds for biological evaluation[1a-c].

  1. Current Status on Stress Diagnostic Kit and Detection Technology

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sang Hyun; Choi, Mi Hee; Ko, Kyong Cheol


    The accurate measurement of a stress level is one of the most important issues in a stress diagnosis and its measurement could be of great value in clinical medicine. Stress has a potent effect on the spirit and physical condition of an individual. There are various methods available for its measurement. Some of the commonly used techniques for the diagnosis of a stress level include analysis of the body fluids, questionnaire assessments, psychophysiological evaluations and by determining heart rate variability (HRV) of subjects. However, the existing diagnostic methods have several defects like, a low sensitivity, inaccuracy and long of operation time. In this report, we present a diagnostic technology to detect a stress level which is the origin of various diseases. This method can be of great help in providing an early diagnosis through a biosensor and might play a vital role in preventing diseases like hypochondria and hypertension. Majority of the human population is exposed to stress in one way or another and hence developing a convenient stress diagnosis kit will be of great use to all. This stress diagnostic kit and detection technology dose not involve simple a mechanical measurement or questionnaires, but is based on developing a detection kit with a high sensitivity, which will mean an easy use for common man. Individuals can undergo regular check ups and can personally diagnose their present situation of health by determining their stress levels, thus enabling them to diagnose the early onset of several stress disorders. This might help them take precautionary measures and thereby lead to a healthy life.

  2. Embryo sexing and sex chromosomal chimerism analysis by loop-mediated isothermal amplification in cattle and water buffaloes. (United States)

    Hirayama, Hiroki; Kageyama, Soichi; Moriyasu, Satoru; Sawai, Ken; Minamihashi, Akira


    In domestic animals of the family Bovidae, sex preselection of offspring has been demanded for convenience of milk/beef production and animal breeding. Development of the nonsurgical embryo transfer technique and sexing methods of preimplantation embryos made it possible. Sexing based on detection of Y chromosome-specific DNA sequences is considered the most reliable method to date. PCR enables amplification of a target sequence from a small number of blastomeres. However, it requires technical skill and is time consuming. Furthermore, PCR has the risk of false positives because of DNA contamination during handling of the PCR products in duplicate PCR procedures and/or electrophoresis. Therefore, for embryo sexing to become widely used in the cattle embryo transfer industry, a simple, rapid and precise sexing method needs to be developed. Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification method, and the reaction is carried out under isothermal conditions (range, 60 to 65 C) using DNA polymerase with strand displacement activity. When the target DNA is amplified by LAMP, a white precipitate derived from magnesium pyrophosphate (a by-product of the LAMP reaction) is observed. It is noteworthy that LAMP does not need special reagents or electrophoresis to detect the amplified DNA. This review describes the development and application of an embryo sexing method using LAMP in cattle and water buffaloes.

  3. A prototype of the direct agglutination test kit (DAT-Canis) for the serological diagnosis of canine visceral leishmaniasis. (United States)

    Oliveira, Edward; Saliba, Juliana Wilke; Oliveira, Diana; Dias, Edelberto Santos; Paz, Gustavo Fontes


    This report describes the stege I/II development of a new direct agglutination test (DAT) for the diagnosis of canine visceral leishmaniasis (CVL) using freeze-dried antigen produced Coomassie blue-stained Leishmania (Leishmania) infantum promastigotes. In stage I, 16 canine serum samples, collected from eight dogs carrying CVL and eight healthy dogs, were assessed with the DAT using 2-mercaptoethanol (2-ME), N-acetyl-cysteine (NAC), kaolin or NAC plus urea (NAC+U) to improve the assay conditions. Stage II assessed the diagnostic accuracy with 100 serum samples collected from dogs with symptomatic CVL and clinically healthy dogs, comparing the four different sample diluents. The CVL-DAT prototype kit showed equivalent performances when 2-ME, NAC or NAC+U were used: 97.1% sensitivity (CI: 83-99.8%), 97% specificity (CI: 88.5-99.5%) and a 97% diagnostic accuracy (CI: 90.8-99.2). With kaolin, a 94.1% sensitivity (CI: 79-99%), 97% specificity (CI: 88.5-99.5%) and 96% diagnostic accuracy were observed (CI: 89.5-98.7), with no statistically significant differences among the four reagents (p=1.0). The NAC plus urea in sample diluent decreased non-specific agglutination, promoted a better defined sharp-edged blue spot and was thus chosen as a component for the new DAT prototype to diagnose canine VL, designated DAT-Canis.

  4. Office 2010 eLearning Kit For Dummies

    CERN Document Server

    Wempen, Faithe


    Create and work with Microsoft Office 2010 with this learning package Microsoft Office 2010 is the most commonly used office productivity suite and if you're eager to get started using all it has to offer, this value-packed eLearning kit is essential to your learning process. This complete Microsoft Office 2010 course includes a full-color printed book and a Dummies interactive eLearning course on CD. You'll discover the basics of the Office interface, how to navigate it, and how to use the features common to all Office programs. Then you'll get detailed instruction in working with Word, Excel

  5. Windows 7 eLearning Kit For Dummies

    CERN Document Server

    Fulton, Jennifer


    Self-motivators will get moving with Windows 7 using this interactive eLearning course! Windows 7 is the number one operating system in the world and if you're eager to get started using all it has to offer, this value-packed eLearning kit is essential to your learning process. A complete Microsoft Windows 7 course, it includes a full-color printed book and a Dummies interactive eLearning course on CD. Each lesson opens with an introduction to the content and explains the importance and potential uses for every task described. Featuring both written and animated step-by-step how-tos, practice

  6. A Safety-Critical Java Technology Compatibility Kit

    DEFF Research Database (Denmark)

    Søndergaard, Hans; Korsholm, Stephan Erbs; Ravn, Anders P.


    In order to claim conformance with a given Java Specification Request (JSR), a Java implementation has to pass all tests in an associated Technology Compatibility Kit (TCK). This paper presents development of test cases and tools for the draft Safety-Critical Java (SCJ) specification. In previous...... work we have shown how the Java Modeling Language (JML) is applied to specify conformance constraints for SCJ, and how JML-related tools may assist in generating and executing tests. Here we extend this work with a layout for concrete test cases including checking of results in a simplified version...

  7. A safety-critical java technology compatibility kit

    DEFF Research Database (Denmark)

    Søndergaard, Hans; Korsholm, Stephan E.; Ravn, Anders Peter


    In order to claim conformance with a given Java Specification Request (JSR), a Java implementation has to pass all tests in an associated Technology Compatibility Kit (TCK). This paper presents development of test cases and tools for the draft Safety-Critical Java (SCJ) specification. In previous...... work we have shown how the Java Modeling Language (JML) is applied to specify conformance constraints for SCJ, and how JML-related tools may assist in generating and executing tests. Here we extend this work with a layout for concrete test cases including checking of results in a simplified version...

  8. Engineering Customized TALENs Using the Platinum Gate TALEN Kit. (United States)

    Sakuma, Tetsushi; Yamamoto, Takashi


    Among various strategies for constructing customized transcription activator-like effector nucleases (TALENs), the Golden Gate assembly is the most widely used and most characterized method. The principle of Golden Gate assembly involves cycling reactions of digestion and ligation of multiple plasmids in a single tube, resulting in PCR-, fragmentation-, and purification-free concatemerization of DNA-binding repeats. Here, we describe the protocols for Golden Gate assembly-based TALEN construction using the Platinum Gate TALEN Kit, which allows generation of highly active Platinum TALENs.

  9. Evaluation of Two Anti-Hepatitis E Virus IgM Kits

    Institute of Scientific and Technical Information of China (English)


    Objective To evaluate two commercial anti-hepatitis E virus (HEV) IgM kits used for differential diagnosis of acute enteric viral hepatitis. Methods The kit for IgM capture assay, was produced with a recombinant HEV structural protein protecting primates against experimental infection by different HEV genotypes, while the other kit for indirect ELISA was produced with recombinant structural proteins from different HEV genotypes. The serum specimens were taken from 241cases with a confirmed or presumptive diagnosis of hepatitis A and 74 cases with a confirmed or presumptive diagnosis of hepatitis E. Results The sensitivity and specificity of the IgM capture assay kit were 97% and 100%, respectively, and the corresponding values for the other kit were 70% and 78%, respectively. Conclusion The IgM capture assay kit has higher sensitivity and specificity in diagnosing acute enteric viral hepatitis E.

  10. [Sensitivity and specificity of the ELISA Kit for the detection of antidobies to Junin virus]. (United States)

    Pirozhkov, A P; Timofeev, M A; Borisevich, I V; Syromiatnikova, S I; Shatokhina, I V; Pantyukhov, V B; Kovalchuk, A V; Borisevich, S V


    The goal of this work was to describe methodological approaches to determination of sensitivity and specificity of the enzyme-linked immunosorbent assay kit (ELISA Kit) for detection of the specific anti-Junin virus (JV) antibody. Comparison of ELISA to plaque reduction neutralization test (PRNT) showed direct relationship between antibody titers in the samples of serum of immunized animals, determined by either PRNT or ELISA methods. The obtained results provided an opportunity to form the panels of positive and negative serum samples to determine the sensitivity and specificity of the ELISA Kit. Sensitivity of the ELISA Kit was at least 98% when studying the samples of serum of immunized guinea pigs and rabbits (determined as positive in PRNT). The sensitivity of the ELISA Kit was at least 68% when studying the samples determined by PNRT as uncertain positive. The specificity was 98%. The specificity of the ELISA Kit was 98%.

  11. Prolonged expression of the c-kit receptor in germ cells of intersex fetal testes. (United States)

    Rajpert-De Meyts, E; Jørgensen, N; Müller, J; Skakkebaek, N E


    Stem cell factor (SCF) and its receptor Kit encoded by the c-kit proto-oncogene are crucial for the development and migration of primordial germ cells in rodents. The expression of Kit has been examined immunohistochemically in gonads obtained from five specimens of fetal tissues with intersex conditions which included 45,X/46,XY mosaicism; androgen insensitivity syndrome; and 46,XY/iso(p)Y mosaicism. Individuals with such disorders of sexual differentiation and Y-chromosome material carry a very high risk of developing testicular neoplasms. Fetal testicular germ cells of the intersex subjects expressed Kit at a later developmental age than controls, in which no Kit protein was detectable beyond the 15th week of gestation. This finding may indicate a disturbance of the chronology of germ cell development, or it may suggest a change of the regulation of c-kit expression in subjects with disorders of gonadal development.

  12. Prolonged expression of the c-kit receptor in germ cells of intersex fetal testes

    DEFF Research Database (Denmark)

    Rajpert-De Meyts, Ewa; Jørgensen, N; Müller, Jørn


    Stem cell factor (SCF) and its receptor Kit encoded by the c-kit proto-oncogene are crucial for the development and migration of primordial germ cells in rodents. The expression of Kit has been examined immunohistochemically in gonads obtained from five specimens of fetal tissues with intersex...... conditions which included 45,X/46,XY mosaicism; androgen insensitivity syndrome; and 46,XY/iso(p)Y mosaicism. Individuals with such disorders of sexual differentiation and Y-chromosome material carry a very high risk of developing testicular neoplasms. Fetal testicular germ cells of the intersex subjects...... expressed Kit at a later developmental age than controls, in which no Kit protein was detectable beyond the 15th week of gestation. This finding may indicate a disturbance of the chronology of germ cell development, or it may suggest a change of the regulation of c-kit expression in subjects with disorders...

  13. Validity of HydraTrend reagent strips for the assessment of hydration status. (United States)

    Abbey, Bryce M; Heelan, Kate A; Brown, Gregory A; Bartee, Rodrick T


    Hydration is used by athletic governing organizations for weight class eligibility. The measurement of urine specific gravity (USG) as a measure of hydration by reagent strips is a controversial issue. The purpose of this study was to determine the validity of HydraTrend reagent strips that facilitate the correction of USG for alkaline urine samples against refractometry for the assessment of USG. Fifty-one participants (33 males, age = 22.3 ± 1.3 years; 18 females, age = 22.4 ± 1.2 years) provided 84 urine samples. The samples were tested for USG using refractometry and reagent strips and for pH using reagent strips and a digital pH meter. Strong correlation coefficients were found between refractometry and reagent strips for USG (rs(82) = 0.812, p refractometry with USG >1.020, pass reagent strips with USG ≤1.020) occurred 39% (33/84) of the time and false negative results for National Federation of State High School Association (NFHS) requirements (fail refractometry with USG >1.025, pass reagent strips with USG ≤1.025) occurred 14% (12/84) of the time. There were no false positives (pass refractometry and fail reagent strips) for NCAA or NFHS requirements. These data show that refractometry and reagent strips have strong positive correlations. However, the risk of a false negative result leading to incorrect certification of euhydration status outweighs the benefits of the HydraTrend reagent strips for the measurement of USG.

  14. Activated c-Kit receptor in the heart promotes cardiac repair and regeneration after injury (United States)

    Di Siena, S; Gimmelli, R; Nori, S L; Barbagallo, F; Campolo, F; Dolci, S; Rossi, P; Venneri, M A; Giannetta, E; Gianfrilli, D; Feigenbaum, L; Lenzi, A; Naro, F; Cianflone, E; Mancuso, T; Torella, D; Isidori, A M; Pellegrini, M


    The role of endogenous c-Kit receptor activation on cardiac cell homeostasis and repair remains largely unexplored. Transgenic mice carrying an activating point mutation (TgD814Y) in the kinase domain of the c-Kit gene were generated. c-KitTgD814Y receptor was expressed in the heart during embryonic development and postnatal life, in a similar timing and expression pattern to that of the endogenous gene, but not in the hematopoietic compartment allowing the study of a cardiac-specific phenotype. c-KitTgD814Y mutation produced a constitutive active c-Kit receptor in cardiac tissue and cells from transgenic mice as demonstrated by the increased phosphorylation of ERK1/2 and AKT, which are the main downstream molecular effectors of c-Kit receptor signaling. In adult transgenic hearts, cardiac morphology, size and total c-Kit+ cardiac cell number was not different compared with wt mice. However, when c-KitTgD814Y mice were subjected to transmural necrotic heart damage by cryoinjury (CI), all transgenic survived, compared with half of wt mice. In the sub-acute phase after CI, transgenic and wt mice showed similar heart damage. However, 9 days after CI, transgenic mice exhibited an increased number of c-Kit+CD31+ endothelial progenitor cells surrounding the necrotic area. At later follow-up, a consistent reduction of fibrotic area, increased capillary density and increased cardiomyocyte replenishment rate (as established by BrdU incorporation) were observed in transgenic compared with wt mice. Consistently, CD45−c-Kit+ cardiac stem cells isolated from transgenic c-KitTgD814Y mice showed an enhanced endothelial and cardiomyocyte differentiation potential compared with cells isolated from the wt. Constitutive activation of c-Kit receptor in mice is associated with an increased cardiac myogenic and vasculogenic reparative potential after injury, with a significant improvement of survival. PMID:27468693

  15. Total Synthesis of Natural Products Using Hypervalent Iodine Reagents (United States)

    Maertens, Gaetan; L'homme, Chloe; Canesi, Sylvain


    We present a review of natural product syntheses accomplished in our laboratory during the last five years. Each synthetic route features a phenol dearomatization promoted by an environmentally benign hypervalent iodine reagent. The dearomatizations demonstrate the “aromatic ring umpolung” concept, and involve stereoselective remodeling of the inert unsaturations of a phenol into a highly functionalized key intermediate that may contain a quaternary carbon center and a prochiral dienone system. Several new oxidative strategies were employed, including transpositions (1,3-alkyl shift and Prins-pinacol), a polycyclization, an ipso rearrangement, and direct nucleophilic additions at the phenol para position. Several alkaloids, heterocyclic compounds, and a polycyclic core have been achieved, including sceletenone (a serotonin reuptake inhibitor), acetylaspidoalbidine (an antitumor agent), fortucine (antiviral and antitumor), erysotramidine (curare-like effect), platensimycin (an antibiotic), and the main core of a kaurane diterpene (immunosuppressive agent and stimulator of apoptosis). These concise and in some cases enantioselective syntheses effectively demonstrate the importance of hypervalent iodine reagents in the total synthesis of bioactive natural products.

  16. Total Synthesis of Natural Products Using Hypervalent Iodine Reagents

    Directory of Open Access Journals (Sweden)

    Gaetan eMaertens


    Full Text Available We present a review of natural product syntheses accomplished in our laboratory during the last five years. Each synthetic route features a phenol dearomatization promoted by an environmentally benign hypervalent iodine reagent. The dearomatizations demonstrate the aromatic ring umpolung concept, and involve stereoselective remodeling of the inert unsaturations of a phenol into a highly functionalized key intermediate that may contain a quaternary carbon center and a prochiral dienone system. Several new oxidative strategies were employed, including transpositions (1,3-alkyl shift and Prins-pinacol, a polycyclization, an ipso rearrangement, and direct nucleophilic additions at the phenol para position. Several alkaloids, heterocyclic compounds, and a polycyclic core have been achieved, including sceletenone (a serotonin reuptake inhibitor, acetylaspidoalbidine (an antitumor agent, fortucine (antiviral and antitumor, erysotramidine (curare-like effect, platensimycin (an antibiotic, and the main core of a kaurane diterpene (immunosuppressive agent and stimulator of apoptosis. These concise and in some cases enantioselective syntheses effectively demonstrate the importance of hypervalent iodine reagents in the total synthesis of bioactive natural products.

  17. Fenton’s reagent application in the domestic sewers disinfection

    Directory of Open Access Journals (Sweden)

    Juacyara Cabonelli Campos


    Full Text Available This paper investigated the application of advanced oxidative processes – Fenton’s reagent - in wastewater disinfection. The treatments included the variation of the hydrogen peroxide and ferrous ions concentrations (Fe2+/H2O2 and pH values. The sewage samples were collected at Ilha do Governador Wastewater Treatment Plant (ETIG in Rio de Janeiro, Brazil, before the biological treatment with activated sludge. The average pH fluctuated from 6.5 to 7.2 and the most common value was 6.7. The reactions with the Fenton´s reagents, as well as the beginning of the analysis occurred within 24 hours after the sewage sample`s collection. The oxidative process, its behavior and the treatment effectiveness have been monitored by microorganism counting, COD, BOD, ammoniacal nitrogen and others. The results have shown a total elimination of the fecal coliforms in the wastewater samples when treated with H2O2 and Fe2+ in concentrations of 200 mg/L of 50 mg/L, respectively.

  18. Development of Ammonia Gas Sensor Using Optimized Organometallic Reagent

    Directory of Open Access Journals (Sweden)

    J. Aubrecht


    Full Text Available Reliable, continuous, and spatially distributed monitoring of dangerous or irritating chemical substances belongs to standard functions of contemporary industrial and public security systems. Fiber-optic-based detection provides feasible platform to fulfill such aims. This paper deals with characterization of ammonia sensing elements based on multimode polysiloxane-clad silica-core optical fibers sensitized with 5-(4′-dioctylamino phenylimino quinoline-8-1 cobalt bromide complex reagent immobilized into the cross-linked polymer matrix from a proper mixture of organic solvents and a radical scavenger contributing to the desired long-term stability of optical properties. The applied sensing mechanism combines optical detection principle with chemical reaction of the reagent and ammonia resulting in changes in the visible near-infrared optical absorption spectrum of the cladding layer, influencing via evanescent optical field interactions the spectral distribution of the guided light intensity. Reaction kinetics of short fiber sections exposed to ammonia/nitrogen mixture of various ammonia concentrations is tested and evaluated. The obtained sensitivity, limit of detection, and forward response time of the prepared sensors amount to 1.52⁎10-5 ppm−1, 31 ppm, and 25 s, respectively. The obtained results are promising for fabrication of distributed fiber-optic sensors applicable to detection and location of ammonia gas leaks in industrial as well as general public premises.

  19. Evaluation of an automated urine chemistry reagent-strip analyzer. (United States)

    Lott, J A; Johnson, W R; Luke, K E


    We evaluated the Miles Inc., Clinitek Atlas Automated Urine Chemistry Analyzer for 11 tests: bilirubin, color, glucose, ketones, leukocyte esterase, nitrite, occult blood, pH, protein, specific gravity, and urobilinogen. The instrument uses a roll of reagent strips affixed to a clear plastic support; urine specimens are automatically pipetted onto these strips. The instrument measures the pads' color using reflectance colorimetry. Specific gravity is measured using a fiberoptic refractive index method. Four hospitals participated in the evaluation, and tests were performed only on fresh urine samples. We found the instrument easy to use; it has walk-away capability with up to 40-specimen loading capacity plus spaces for STATs, calibrators and controls. We found good comparability with chemical tests and other nonreagent strip procedures, as well as good agreement with the Miles Inc. Clinitek 200+ urine chemistry analyzer and visual reading of the Miles Inc. Multistix Reagent Strips. The Clinitek Atlas is rugged and reliable, and is suitable for a high-volume urinalysis laboratory.

  20. Reaction rates between water and the Karl Fischer reagent. (United States)

    Cedergren, A


    Reaction rates between water and the Karl Fischer reagent have been determined by potentiometric measurement for various compositions of the Karl Fischer reagent. The study has been made with an iodine complex concentration of 0.3-1.2 mM and sulphur dioxide complex at 0.01-0.5M. The concentration of excess of pyridine had no measurable effect on the rate of the main reaction. The reaction was found to be first-order with respect to iodine complex, to sulphur dioxide complex, and to water. The rate constant was (1.2+/-0.2) x 10(3) 1(2). mole(-2). sec(-1). In an ordinary titration it is therefore essential to keep the sulphur dioxide concentration high for the reaction to go to completion within a reasonable time. The extent of side-reactions was found to be independent of the iodine concentration at low concentrations. The side-reactions increased somewhat with increasing sulphur dioxide pyridine concentrations and decreased to about 60% when the temperature was lowered from 24 degrees to 7 degrees.

  1. HPV16 detection by qPCR method in relation to quantity and quality of DNA extracted from archival formalin fixed and paraffin embedded head and neck cancer tissues by three commercially available kits. (United States)

    Biesaga, Beata; Janecka, Anna; Mucha-Małecka, Anna; Adamczyk, Agnieszka; Szostek, Sława; Słonina, Dorota; Halaszka, Krzysztof; Przewoźnik, Marcin


    The aim of the present study was to compare HPV16 detection by quantitative polymerase chain reaction (qPCR) in relation to the quantity and quality of DNA isolated from 21 formalin fixed and paraffin embedded (FFPE) head and neck cancer tissues by three commercially available kits: EX-WAX™ DNA Extraction Kit (M) (Merck Millipore, Darmstadt, Germany), QIAamp(®) DNA FFPE Tissue (Q) (Qiagen, Hilden, Germany) and ReliaPrep™ FFPE gDNA Miniprep System (P) (Promega, Madison, USA). Quantity of extracted DNA was assessed spectrophometrically and fluorometrically. Its quality was analyzed using A260/280 and A260/230 ratios and the β-actin fragment amplifiability in qPCR. HPV16 presence was detected by qPCR, using specific primers and TaqMan probe. HPV infection was found in 8 DNA samples extracted with M kit (38.1%) and in 7 (33.3%) isolated with Q and P kits. Three samples from M and Q kits were characterized by HPV16 positivity and lack of β-actin amplifiability. They had significantly lower A260/280 ratio (M: 1.6±0.0, p=0.044 and Q: 1.7±0.0, p=0.016) compared to samples with both fragments amplification (M: 1.7±0.0 and Q: 1.9±0.0). Therefore, for HPV detection by qPCR in FFPE tissues we recommend ReliaPrep™ FFPE gDNA Miniprep System.

  2. Diagnóstico das meningites através de fitas reagentes Diagnosis of meningitis with reagent strips

    Directory of Open Access Journals (Sweden)

    Roberta M.C. Romanelli


    Full Text Available OBJETIVO: determinar a utilidade de fitas reagentes para a avaliação liquórica de pleocitose, glicorraquia e proteinorraquia no diagnóstico precoce e rápido de meningites em crianças. MÉTODOS: Foram incluídas no estudo amostras de líquor provenientes de 164 crianças admitidas no ambulatório de doenças infecto-contagiosas do Centro Geral de Pediatria (CGP-FHEMIG, com suspeita clínica de meningite, no período diurno de Maio/97 à Maio/99. A faixa etária dos pacientes variou de um mês a 12 anos (mediana de 12 meses, sendo obtidos resultados da citobioquímica liquórica (celularidade, glicorraquia e proteinorraquia de 154 desses pacientes. Esses achados foram comparados com reações do líquor em fitas reagentes. RESULTADOS: Através da citobioquímica líquórica foram identificados 43 casos de provável meningite bacteriana, 19 provavelmente viróticas e 83 amostras sem alterações. Pelas fitas reagentes, detectaram-se 41 casos de provável meningite bacteriana, dois casos de infecção meníngea provavelmente virótica, e em 71 exames não se verificaram alterações. Comparando os resultados obtidos por meio das fitas reagentes com a citobioquímica convencional, observou-se sensibilidade, especificidade, valores preditivos positivo e negativo e acurácia (90,7; 98,1; 95,1; 96,4; 96,1%, respectivamente. Ademais, a análise estatística pelo teste de Mc Nemar não evidenciou discordância significativa no diagnóstico de meningite bacteriana obtido através de ambos os métodos (p=0,68 e, pela estatística Kappa, verificou-se elevado grau de concordância entre os testes (pOBJECTIVE: to determine the usefulness of reagent strips in the evaluation of pleocytosis, cerebrospinal fluid glucose and protein levels for early and rapid diagnosis of meningitis in children. METHODS: We included cerebrospinal fluid samples of 164 children admitted to the outpatient clinic of Communicable Diseases of the General Pediatric Center (Funda

  3. Analytical specificity and sensitivity of the novel dual-target GeneProof Neisseria gonorrhoeae PCR kit for detection of N. gonorrhoeae. (United States)

    Golparian, Daniel; Hellmark, Bengt; Unemo, Magnus


    Detection of Neisseria gonorrhoeae relies increasingly on nucleic acid amplification tests (NAATs). The specificity of many gonococcal NAATs has been suboptimal and supplementary testing remains recommended in Europe and several additional countries. The novel dual-target GeneProof Neisseria gonorrhoeae PCR kit, targeting porA pseudogene and 16S rRNA gene, showed a high specificity and sensitivity when isolates of non-gonococcal Neisseria and related species (n = 144), and gonococci (n = 104) were tested. However, rare gonococcal porA mutants were only detected in the 16S rRNA gene target and two non-gonococcal isolates showed a low-level cross-reactivity in the 16S rRNA gene target. The detection limit for both targets was 1.5 copies per reaction.

  4. Genetic data of 15 autosomal STRs (Identifiler kit) of three Mexican Mestizo population samples from the States of Jalisco (West), Puebla (Center), and Yucatan (Southeast). (United States)

    Rubi-Castellanos, R; Anaya-Palafox, M; Mena-Rojas, E; Bautista-España, D; Muñoz-Valle, J F; Rangel-Villalobos, H


    We report autosomal STR data (Identifiler PCR amplification kit) of a total sample of 884 unrelated Mestizos from three different regions of Mexico. The population sample included 309, 313 and 262 individuals from the states of Jalisco (West), Puebla (Center) and Yucatan (Southeast), respectively. Allele distribution and forensic statistical parameters are described. Genotype distribution by locus and two-loci combination was in agreement with Hardy-Weinberg expectations for all 15 STRs. Pairwise comparisons including Mexican populations reported in the literature demonstrated a significant differentiation, principally between North/West with regard to Center/Southeast Mexico. These results increase STR data from previously unreported regions of this country, and constitute a valuable guide in forensic casework for choosing an auxiliary STR database in states where it is not available.

  5. C-kit-targeted imaging of gastrointestinal stromal tumor using radiolabeled anti-c-kit monoclonal antibody in a mouse tumor model

    Energy Technology Data Exchange (ETDEWEB)

    Sogawa, Chizuru [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Tsuji, Atsushi B. [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan)], E-mail:; Sudo, Hitomi [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Department of Pathology and Oncology, Juntendo University School of Medicine, Tokyo 113-8421 (Japan); Sugyo, Aya [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Yoshida, Chisato [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Department of Molecular Imaging and Radiotherapy, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675 (Japan); Odaka, Kenichi [Molecular Probe Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan); Uehara, Tomoya; Arano, Yasushi [Department of Molecular Imaging and Radiotherapy, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675 (Japan); Koizumi, Mitsuru; Saga, Tsuneo [Diagnostic Imaging Group, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba 263-8555 (Japan)


    Introduction: Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor arising from the gastrointestinal tract and highly expresses mutated c-kit. We aimed to develop a specific and sensitive method for detecting GISTs using radiolabeled anti-c-kit monoclonal antibody. Methods: A mutated c-kit-expressing cell clone was established by transfecting an expressing vector of mutated c-kit gene into HEK293 human embryonic kidney cells. The tumors were developed by inoculating c-kit-expressing cells into nude mice. {sup 125}I- and {sup 111}In-labeled anti-c-kit antibodies (12A8 and 41A11) were evaluated in vitro by cell binding, competitive inhibition and cellular internalization assays, and in vivo by biodistribution and imaging studies in tumor-bearing mice. Results: Both {sup 125}I- and {sup 111}In-labeled antibodies showed specific binding with c-kit-expressing cells with high affinity (dissociation constants = 2.2-7.1x10{sup 9} M{sup -1}). Internalization assay showed that {sup 125}I-labeled antibodies were rapidly internalized and dehalogenated, with the release of {sup 125}I from the cells, resulting in reduction of cell-associated radioactivity with time. In contrast, {sup 111}In-labeled antibody was internalized but did not result in the reduced radioactivity associated with tumor cells. Reflecting this phenomenon, the in vivo tumor uptake of {sup 125}I-labeled antibody was low on Day 1, further decreasing with time, while tumor uptake of {sup 111}In-labeled antibody was high on Day 1, further increasing with time. The xenografted tumor was clearly visualized by scintigraphy after injection of {sup 111}In-labeled antibody. Conclusion: The anti-c-kit monoclonal antibody labeled with a metal radionuclide would be promising for c-kit-targeted imaging of GISTs.

  6. Kit signaling is required for development of coordinated motility patterns in zebrafish gastrointestinal tract. (United States)

    Rich, Adam; Gordon, Scott; Brown, Chris; Gibbons, Simon J; Schaefer, Katherine; Hennig, Grant; Farrugia, Gianrico


    Interstitial cells of Cajal (ICC) provide a pacemaker signal for coordinated motility patterns in the mammalian gastrointestinal (GI) tract. Kit signaling is required for development and maintenance of ICC, and these cells can be identified by Kit-like immunoreactivity. The zebrafish GI tract has two distinct ICC networks similar to mammals, suggesting a similar role in the generation of GI motility; however, a functional role for Kit-positive cells in zebrafish has not been determined. Analysis of GI motility in intact zebrafish larvae was performed during development and after disruption of Kit signaling. Development of coordinated motility patterns occurred after 5 days post-fertilization (dpf) and correlated with appearance of Kit-positive cells. Disruptions of Kit signaling using the Kit antagonist imatinib mesylate, and in Sparse, a null kita mutant, also disrupted development of coordinated motility patterns. These data suggest that Kit signaling is necessary for development of coordinated motility patterns and that Kit-positive cells in zebrafish are necessary for coordinated motility patterns.

  7. Temporal expression of c-kit in spermatogenesis of two grasshopper species

    Institute of Scientific and Technical Information of China (English)



    Two species of grasshoppers,Calliptamus abbreviatus (Ikonn.) and Shirakiacris shirakii (I. Bol.),were collected randomly in the Siping area of Jilin Province,China. By using immunohistochemical methods and statistical analysis,we observed and compared the temporal expression of c-kit protein in four representative stages of spermatogenesis of the two grasshoppers,namely: spermatogonia; primary spermatocyte; secondary spermatocyte;and mature sperm. Results showed that there was c-kit positive temporal expression at each stage of spermatogenesis,but there were different positive expression levels: (i) weak positive expression of c-kit protein appeared in spermatogonia and the positive granules were thinner; (ii) strong positive expression of c-kit protein existed in primary spermatocyte and positive granules became biggest among all developmental stages; (iii) c-kit positive expression stayed stronger in secondary spermatocyte while positive granules became thinner; (iv) there was a strong positive expression of c-kit and thinner positive granules in mature sperm,which distributed on head and tail; (v) the biggest c-kit positive granules had been found massing at the end of spermary; and (vi) significant differences of c-kit positive expression existed in spermatogenesis between two species of grasshoppers. The results indicated that c-kit protein may play a crucial role in spermatogenesis and even retain the physiological action of sperms and fertilization in grasshoppers.

  8. Behaviour of mink kits and dams (Mustela vison) in the lactation period

    DEFF Research Database (Denmark)

    Brink, Anne-Line; Jeppesen, Leif Lau


    This study describes the development of the behaviour of mink kits and dams from the fourth to the eighth respectively seventh week after delivery. The study is based on scan observations of 72 mink dams and their kits at a conventional Danish mink farm. The kits started eating when they were about...... budget in the seventh week and the stereotypy frequency of the dams increased to about 4% of the time budget. It is suggested that some dams are frustrated by the forced cohabitation with their nutritionally independent kits already in the seventh week and that this should be taken into account when...

  9. Performances of Four Helicobacter pylori Serological Detection Kits Using Stool Antigen Test as Gold Standard (United States)


    The aim was to determine the performances of four Helicobacter pylori serological detection kits in different target groups, using Amplified IDEIA™ Hp StAR™ as gold standard. Kits studied were Rapid Immunochromatoghraphic Hexagon, Helicoblot 2.1, an EIA IgG kit and EIA IgA kit. Methods: Stool and blood samples were collected from 162 apparently healthy participants (control) and 60 Type 2 diabetes mellitus (T2DM) patients. Results: The performances of the four serological detection kits were found to be affected by gender, age, health status and ethnicity of the participants. In the control group, the Helicoblot 2.1 kit had the best performance (AUC = 0.85; p<0.05, accuracy = 86.4%), followed by EIA IgG (AUC = 0.75; p<0.05, accuracy = 75.2%). The Rapid Hexagon and EIA IgA kits had relatively poor performances. In the T2DM subgroup, the kits H2.1 and EIA IgG had best performances, with accuracies of 96.5% and 93.1% respectively. The performance of EIA IgG improved with adjustment of its cut-off value. Conclusion: The performances of the detection kits were affected by various factors which should be taken into consideration. PMID:27736910

  10. [Detection of the Zaire Subtype of the Ebola Virus by Isothermal Multiple Self-matching Initiated Amplification]. (United States)

    Li, Xinna; Nie, Kai; Wang, Ji; Zhang, Dan; Guan, Li; Liu, Jun; Ke, Yuehua; Zhou, Hangyu; Ma, Xuejun


    Given the Ebola outbreak in West Africa and the risks of spread to other regions, a rapid, sensitive and simple method for the detection of the Ebola virus (EBOV) is of great significance for the prevention and control of Ebola. We developed a simple colorimetric isothermal multiple self-matching initiated amplification (IMSA) for rapid detection of the Zaire subtype of the Ebola virus (EBOV-Z). This method employed six primers that recognized seven sites of the EBOV-Z nucleoprotein gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 1 h. Amplification products were detected through visual inspection of color change by pre-addition of hydroxyl naphthol blue dye. Relative sensitivity was validated by detection of serial tenfold dilutions of virus-like particles containing the partial EBOV-Z nucleoprotein gene and mock clinical sample. Specificity of IMSA was validated by detection of the plasma of 30 healthy volunteers, the dengue virus, and Japanese encephalitis virus. IMSA had comparable sensitivity to Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and cross-reaction with human plasma or other viruses was not observed. Reverse transcription-isothermal multiple self-matching initiated amplification (RT-IMSA) was also evaluated and compared in parallel with the commercial RT-qPCR kit for detection of EBOV-suspected samples of human blood in Sierra Leone. Sensitivity and specificity of the RT-IMSA was 91.4% and 100%, respectively. These data suggest that RT-IMSA is a valuable tool for the detection of the EBOV with the distinct advantages of simplicity and low cost compared with RT-qPCR.

  11. Isoquinolinium bromochromate: An efficient and stable reagent for bromination of hydroxylated aromatic compounds and oxidation of alcohols

    Institute of Scientific and Technical Information of China (English)

    Sandeep V. Khansole; Shivaji B. Patwari; Archana Y. Vibhute; Yeshwant B. Vibhute


    The new chromium (VI) oxidizing reagent isoquinolinium bromochromate (IQBC) was prepared and characterized. The IQBC has been found to be stable and an efficient solid reagent which can be easily prepared in good yield. It act as an efficient brominating reagent for hydroxylated aromatic compounds as well as good oxidizing reagent for the conversion of alcohols to carbonyl compounds in good to excellent yield. The synthesized isoquinolinium bromochromate is more ideal reagent, with number of specification including: higher yield, mild conditions and easy preparation. The results obtained with isoquinolinium bromochromate are satisfactory and suggest that the reagent has few advantages over the existing chromium (VI) reagents.

  12. Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases. (United States)

    Sahoo, Pravas Ranjan; Sethy, Kamadev; Mohapatra, Swagat; Panda, Debasis


    India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

  13. Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases

    Directory of Open Access Journals (Sweden)

    Pravas Ranjan Sahoo


    Full Text Available India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

  14. Transcriptome analysis of differentiating spermatogonia stimulated with kit ligand. (United States)

    Rossi, Pellegrino; Lolicato, Francesca; Grimaldi, Paola; Dolci, Susanna; Di Sauro, Annarita; Filipponi, Doria; Geremia, Raffaele


    Kit ligand (KL) is a survival factor and a mitogenic stimulus for differentiating spermatogonia. However, it is not known whether KL also plays a role in the differentiative events that lead to meiotic entry of these cells. We performed a wide genome analysis of difference in gene expression induced by treatment with KL of spermatogonia from 7-day-old mice, using gene chips spanning the whole mouse genome. The analysis revealed that the pattern of RNA expression induced by KL is compatible with the qualitative changes of the cell cycle that occur during the subsequent cell divisions in type A and B spermatogonia, i.e. the progressive lengthening of the S phase and the shortening of the G2/M transition. Moreover, KL up-regulates in differentiating spermatogonia the expression of early meiotic genes (for instance: Lhx8, Nek1, Rnf141, Xrcc3, Tpo1, Tbca, Xrcc2, Mesp1, Phf7, Rtel1), whereas it down-regulates typical spermatogonial markers (for instance: Pole, Ptgs2, Zfpm2, Egr2, Egr3, Gsk3b, Hnrpa1, Fst, Ptch2). Since KL modifies the expression of several genes known to be up-regulated or down-regulated in spermatogonia during the transition from the mitotic to the meiotic cell cycle, these results are consistent with a role of the KL/kit interaction in the induction of their meiotic differentiation.

  15. 三种HBV荧光定量PCR检测试剂的比较及结果分析%Comparison and evaluation of 3 diagnostic kits for quantification (PCR-Fluorescence Probing) of hepatitis B virus DNA

    Institute of Scientific and Technical Information of China (English)

    李兵; 王敏; 徐六妹; 刘赛云; 韩红星; 单万水; 陈心春


    Objective The goal of this clinic study is to evaluate the application performance for 2 new HBV DNA Quantitative Fluorescence Diagnostic Kits, which are recently emerged in the market.Methods Serial diluted HBV serum samples and 1001 clinical serum samples with random virus load were tested quantitatively with the 3 diagnostic kits A, B and C. By studying their linear range, specificity,precision and sensitivity, the two new reagents (A and B ) were used to test these samples and also to compare them with the quantitative results from the boiling method kit (C) which is of better quality and reliability than similar diagnostic kits in current market. Furthermore, the immunoassy results of these samples were evalued and compared with their quantitative results. Results The quantitative results of 767 samples showed that their average values of the 3 kits have no significant difference. However, in the low viral load group, the results of kit A showed the best sensitivity(1.00E + 01 IU/ml)and had much better sensitivity than kit B (1.00E + 02 IU/ml), while kit C kit ( 5.00E + 02 IU/ml ) failed to test positive for most of the low concentration samples. Conclusion The nucleic acid extraction-free method ( kit B) showed much better accuracy and much larger linear range than the conventional method. In this method, the nucleic acid templates extracted by lysis buffer all went into the PCR reaction, resulting in high extraction efficiency and minimum nucleic acid loss. With a simple procedure, great accuracy and good sensitivity, this new test kit is suitable for routine clinical lab usage.%目的 评估3种HBV DNA荧光定量PCR检测试剂的临床应用性能.方法 通过平行检测1001例临床血清样本及梯度稀释的阳性样本,从定量线性范围、准确性、灵敏度、特异性等方面,比较分析A试剂(磁珠分离法)、B试剂(免核酸提取的"一步法")与目前国内临床广泛应用的优质试剂C的相关性和差异,并与免

  16. Design and Development of Low Cost, Simple, Rapid and Safe, Modified Field Kits for the Visual Detection and Determination of Arsenic in Drinking Water Samples

    Directory of Open Access Journals (Sweden)

    Y. Anjaneyulu


    Full Text Available Arsenic is naturally found in surface and ground waters and the inorganic forms of arsenic are the most toxic forms. The adverse health effects of arsenic may involve the respiratory, gastrointestinal, cardiovascular, nervous, and haematopoietic systems. Arsenic contamination in drinking water is a global problem widely seen in Bangladesh and West Bengal of the Indian sub continent. As there is a great demand for field test kits due to the anticipated reduction of the US EPA arsenic standard from 50ppb to 10ppb a field kit which offers rapid, simple and safe method for precise estimation of arsenic at 10ppb in drinking water samples is developed. Field methods, based on the mercuric-bromide-stain, consist of three different major parts, which are carried out stepwise. The first part of the procedure is to remove serious interference caused by hydrogen sulphide. In commercially available kits either the sulphide is oxidized to sulphate and the excess oxidizing reagent removed prior to the hydride generation step or, the hydrogen sulphide is filtered out by passing the gas stream through a filter impregnated with lead acetate during the hydride generation step. The present method employs cupric chloride in combination with ferric chloride or Fenton’s reagent for the removal of hydrogen sulphide, which is rapid, simple and more efficient. Other interferences at this step of the analyses are normally not expected for drinking water analysis. In the second step, the generation of the arsine gas involves the classical way of using zinc metal and hydrochloric acid, which produce the ‘nascent’ hydrogen, which is the actual reducing agent. Hydrochloric acid can be replaced by sulfamic acid, which is solid and avoids a major disadvantage of having to handle a corrosive liquid in the field. The arsine gas produces a yellowish spot on the reagent paper. Depending on the arsenic content, either, Yellow – H

  17. Development and characterization of antibody reagents for detecting nanoparticles (United States)

    Ravichandran, Supriya; Sullivan, Mark A.; Callahan, Linda M.; Bentley, Karen L.; Delouise, Lisa A.


    The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in solution. We present a proof-of-concept for the generation of NProbes and their use for detecting quantum dots and titanium dioxide NPs in vitro and in an ex vivo human skin model. Continued development and refinement of NProbes to detect NPs that vary in composition, shape, size, and surface coating will comprise a powerful tool kit that can be used to advance nanotechnology research particularly in the nanotoxicology and nanotherapeutics fields.The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in solution. We present a proof-of-concept for the generation of NProbes and their use for detecting quantum dots and titanium dioxide NPs in vitro and in an ex vivo human skin model. Continued development and refinement of NProbes to detect NPs that vary in composition, shape, size, and surface coating will comprise a powerful tool kit that can be used to advance nanotechnology research particularly in the nanotoxicology and nanotherapeutics fields. Electronic supplementary information (ESI) available: Figures and detailed methods of various techniques

  18. Competitive amplification of differentially melting amplicons (CADMA improves KRAS hotspot mutation testing in colorectal cancer

    Directory of Open Access Journals (Sweden)

    Kristensen Lasse


    Full Text Available Abstract Background Cancer is an extremely heterogeneous group of diseases traditionally categorized according to tissue of origin. However, even among patients with the same cancer subtype the cellular alterations at the molecular level are often very different. Several new therapies targeting specific molecular changes found in individual patients have initiated the era of personalized therapy and significantly improved patient care. In metastatic colorectal cancer (mCRC a selected group of patients with wild-type KRAS respond to antibodies against the epidermal growth factor receptor (EGFR. Testing for KRAS mutations is now required prior to anti-EGFR treatment, however, less sensitive methods based on conventional PCR regularly fail to detect KRAS mutations in clinical samples. Methods We have developed sensitive and specific assays for detection of the seven most common KRAS mutations based on a novel methodology named Competitive Amplification of Differentially Melting Amplicons (CADMA. The clinical applicability of these assays was assessed by analyzing 100 colorectal cancer samples, for which KRAS mutation status has been evaluated by the commercially available TheraScreen® KRAS mutation kit. Results The CADMA assays were sensitive to at least 0.5% mutant alleles in a wild-type background when using 50 nanograms of DNA in the reactions. Consensus between CADMA and the TheraScreen kit was observed in 96% of the colorectal cancer samples. In cases where disagreement was observed the CADMA result could be confirmed by a previously published assay based on TaqMan probes and by fast COLD-PCR followed by Sanger sequencing. Conclusions The high analytical sensitivity and specificity of CADMA may increase diagnostic sensitivity and specificity of KRAS mutation testing in mCRC patients.

  19. Investigation of reagent distributions on glass fiber membrane filters used in air sampling. (United States)

    Tucker, Samuel P


    This project has arisen from the need to produce GFFs (glass fiber filters) bearing a thin and evenly distributed coating of a selected reagent in the equatorial plane for breakthrough studies. However, it has been discovered that today's two general techniques for coating GFFs (total immersion and application of reagent solution to GFFs) have usually produced unevenly distributed coatings of reagent in the equatorial plane. In addition, quantities of reagent on GFFs from commercial sources may vary widely in the same lot of coated GFFs. Consequences are variability in capacity of coated filters at the point of breakthrough and, perhaps, wasted reagent. Although today's reagent-coated filters may be satisfactory for routine air sampling, such filters may be unacceptable for precise breakthrough studies. Research has been conducted successfully to produce nearly evenly distributed coatings of reagents in the equatorial plane of GFFs by application of reagent solutions to the centers of GFFs which are resting on crisscrossing, fine, stainless-steel wire. Distributions of coatings have been determined by punching out twenty-one 5-mm circles from each GFF and analyzing each circle by flow-injection with a UV detector. Lowest achievable relative standard deviations of measurement (RSDs) for reagents in 5-mm circles have been 5 to 7%. Reagents studied have included 1-(2-pyridyl)piperazine (1-2PP), 2,4-dinitrophenylhydrazine (DNPH), and 1-(9-anthracenylmethyl)piperazine (MAP). Factors affecting the distribution of such coatings include choice of reagent and choice of solvent for the reagent solution.

  20. Silicon Amine Reagents for the Photocatalytic Synthesis of Piperazines from Aldehydes and Ketones. (United States)

    Hsieh, Sheng-Ying; Bode, Jeffrey W


    Silicon amine protocol (SLAP) reagents for photocatalytic cross-coupling with aldehydes and ketones to form N-unprotected piperazines have been developed. This blue light promoted process tolerates a wide range of heteroaromatic, aromatic, and aliphatic aldehydes and structurally and stereochemically complex SLAP reagents. It provides a tin-free alternative to SnAP (tin amine protocol) reagents for the synthesis of substituted piperazines.

  1. UP-TORR: online tool for accurate and Up-to-Date annotation of RNAi Reagents. (United States)

    Hu, Yanhui; Roesel, Charles; Flockhart, Ian; Perkins, Lizabeth; Perrimon, Norbert; Mohr, Stephanie E


    RNA interference (RNAi) is a widely adopted tool for loss-of-function studies but RNAi results only have biological relevance if the reagents are appropriately mapped to genes. Several groups have designed and generated RNAi reagent libraries for studies in cells or in vivo for Drosophila and other species. At first glance, matching RNAi reagents to genes appears to be a simple problem, as each reagent is typically designed to target a single gene. In practice, however, the reagent-gene relationship is complex. Although the sequences of oligonucleotides used to generate most types of RNAi reagents are static, the reference genome and gene annotations are regularly updated. Thus, at the time a researcher chooses an RNAi reagent or analyzes RNAi data, the most current interpretation of the RNAi reagent-gene relationship, as well as related information regarding specificity (e.g., predicted off-target effects), can be different from the original interpretation. Here, we describe a set of strategies and an accompanying online tool, UP-TORR (for Updated Targets of RNAi Reagents;, useful for accurate and up-to-date annotation of cell-based and in vivo RNAi reagents. Importantly, UP-TORR automatically synchronizes with gene annotations daily, retrieving the most current information available, and for Drosophila, also synchronizes with the major reagent collections. Thus, UP-TORR allows users to choose the most appropriate RNAi reagents at the onset of a study, as well as to perform the most appropriate analyses of results of RNAi-based studies.

  2. Novel Synthesis of α-Diketones from Bisbenzimidazolium Salt and Grignard Reagents

    Institute of Scientific and Technical Information of China (English)

    SHI,Zhen(史真); LU,Lin-Gang(卢林刚); YANG,Bing-Qin(杨秉勤); GUO,Yuan(郭媛)


    The reaction of bisbenzimidazolium salt with Grignard reagents and a new synthetic method of α-diketons from bisbenzimidazolium salt and Grignard reagents are reported. The structures urres of all a-diketones are characterized by elemental analyses, infrared spectroscopy and 1H NMR spectroscopy. The infuences of the various Grignard reagents on the yield of α-diketones and the mechanism are discussed, and a new convenient synthetic method for α-diketones is provid-ed.

  3. Visible spectrophotometric estimation of aceclofenac and indapamide from tablets using folin-ciocalteu reagent


    Singhvi I; Goyal Anju


    Two simple, accurate, fast and economical methods have been developed for the quantitative estimation of aceclofenac and indapamide from their respective tablet formulation using Folin-Ciocalteu reagent. Aceclofenac forms a blue colored chromogen with the reagent, which shows absorbance maxima at 642.6 nm and linearity in the concentration range of 80-160 µg/ml of drug. Indapamide forms a green colored chromogen with the reagent, which shows absorbance maxima at 783.2 nm and linearity ...

  4. Hiding inside? Intracellular expression of non-glycosylated c-kit protein in cardiac progenitor cells. (United States)

    Shi, Huilin; Drummond, Christopher A; Fan, Xiaoming; Haller, Steven T; Liu, Jiang; Malhotra, Deepak; Tian, Jiang


    Cardiac progenitor cells including c-kit(+) cells and cardiosphere-derived cells (CDCs) play important roles in cardiac repair and regeneration. CDCs were reported to contain only small subpopulations of c-kit(+) cells and recent publications suggested that depletion of the c-kit(+) subpopulation of cells has no effect on regenerative properties of CDCs. However, our current study showed that the vast majority of CDCs from murine heart actually express c-kit, albeit, in an intracellular and non-glycosylated form. Immunostaining and flow cytometry showed that the fluorescent signal indicative of c-kit immunostaining significantly increased when cell membranes were permeabilized. Western blots further demonstrated that glycosylation of c-kit was increased during endothelial differentiation in a time dependent manner. Glycosylation inhibition by 1-deoxymannojirimycin hydrochloride (1-DMM) blocked c-kit glycosylation and reduced expression of endothelial cell markers such as Flk-1 and CD31 during differentiation. Pretreatment of these cells with a c-kit kinase inhibitor (imatinib mesylate) also attenuated Flk-1 and CD31 expression. These results suggest that c-kit glycosylation and its kinase activity are likely needed for these cells to differentiate into an endothelial lineage. In vivo, we found that intracellular c-kit expressing cells are located in the wall of cardiac blood vessels in mice subjected to myocardial infarction. In summary, our work demonstrated for the first time that c-kit is not only expressed in CDCs but may also directly participate in CDC differentiation into an endothelial lineage.

  5. Features of the reaction of heterocyclic analogs of chalcone with lanthanide shift reagents

    Energy Technology Data Exchange (ETDEWEB)

    Turov, A.V.; Khilya, V.P. [Taras Shevchenko Kiev Univ. (Russian Federation)


    The PMR spectra of heterocyclic analogs of 2-hydroxychalcone containing thiazole, benzofuran, triazole, imidazole, benzodioxane, or pyridine rings in the presence of lanthanide shift reagents are studied. It is found that the most effective reagent for modifying the spectra of these compounds is Yb(fod)3. The broadening of the spectra of 2-hydroxy chalcones in the presence of lanthanide shift reagents is explained by the dynamic effects of complex formation. An example is given of the determination of the conformation of molecules of 2-hydroxychalcone by the simultaneous use of lanthanide shift reagents and the homonuclear Overhauser effect. 9 refs., 1 fig., 1 tab.

  6. Amplification Without Inversion in Semiconductor Quantum Dot (United States)

    Hajibadali, A.; Abbasian, K.; Rostami, A.

    In this paper, we have realized amplification without inversion (AWI) in quantum dot (QD). A Y-type four-level system of InxGa1-xN quantum dot has been obtained and investigated for AWI. It has been shown that, with proper setting of control fields' amplitude, we can obtain reasonable gain. With proper setting of phase difference of control fields and probe field, we can obtain considerable gain in resonant wavelength. We have designed this system by solving the Schrödinger-Poisson equations for InxGa1-xN quantum dot in GaN substrate, self-consistently.

  7. Amplification Effects and Unconventional Monetary Policies

    Directory of Open Access Journals (Sweden)



    Full Text Available Global financial crises trigger off amplification effects, which allow relatively small shocks to propagate through the whole financial system. For this reason, the range of Central banks policies is now widening beyond conventional monetary policies and lending of last resort. The aim of this paper is to establish a rule for this practice. The model is based on the formalization of funding conditions in various types of markets. We conduct a comprehensive analysis of the “unconventional monetary policies”, and especially quantify government bonds purchases by the Central bank.

  8. Amplification and characterization of eukaryotic structural genes. (United States)

    Maniatis, T; Efstratiadis, A; Sim, G K; Kafatos, F


    An approach to the study of eukaryotic structural genes which are differentially expressed during development is described. This approach involves the isolation and amplification of mRNA sequences by in vitro conversion of mRNA to double-stranded cDNA followed by molecular cloning in bacterial plasmids. This procedure provides highly specific hybridization probes that can be used to identify genes and their contiguous DNA sequences in genomic DNA, and to detect specific RNA transcripts during development. The nature of the method allows the isolation of individual mRNA sequences from a complex population of molecules at different stages of development.

  9. Advances in isothermal amplification: novel strategies inspired by biological processes. (United States)

    Li, Jia; Macdonald, Joanne


    Nucleic acid amplification is an essential process in biological systems. The in vitro adoption of this process has resulted in powerful techniques that underpin modern molecular biology. The most common tool is polymerase chain reaction (PCR). However, the requirement for a thermal cycler has somewhat limited applications of this classic nucleic acid amplification technique. Isothermal amplification, on the other hand, obviates the use of a thermal cycler because reactions occur at a single temperature. Isothermal amplification methods are diverse, but all have been developed from an understanding of natural nucleic acid amplification processes. Here we review current isothermal amplification methods as classified by their enzymatic mechanisms. We compare their advantages, disadvantages, efficiencies, and applications. Finally, we mention some new developments associated with this technology, and consider future possibilities in molecular engineering and recombinant technologies that may develop from an appreciation of the molecular biology of natural systems.

  10. Prenyl sulfates as alkylating reagents for mercapto amino acids. (United States)

    Maltsev, Sergey; Sizova, Olga; Utkina, Natalia; Shibaev, Vladimir; Chojnacki, Tadeusz; Jankowski, Wieslaw; Swiezewska, Ewa


    A new methodology for prenylation of thiol compounds has been developed. The approach is based on the use of prenyl sulfates as new reagents for S-prenylation of benzenethiol and cysteamine in aqueous systems. The C(10)-prenols geraniol and nerol that differ in the configuration (E or Z, correspondingly) of the alpha-isoprene unit were efficiently O-sulfated in the presence of a pyridine-SO(3') complex. The obtained geranyl and neryl sulfates were tested as alkylating agents. These compounds were chosen to reveal the influence of the alpha-isoprene unit configuration on their alkylation (prenylation) ability. S-Geranyl cysteine was prepared to demonstrate the applicability of this method for prenylation of peptides containing mercapto amino acids.

  11. Nanomechanical identification of liquid reagents in a microfluidic channel

    DEFF Research Database (Denmark)

    Khan, Faheem; Kim, Seonghwan; Lee, Dongkyu;


    Integration of promising technologies that can enhance sensitivity, selectivity, and throughput into micro total analysis systems (μTAS) are important in making them useful in precise screening of reaction byproducts in analytical chemistry, cellular biology and pharmaceutical industries. But unf......Integration of promising technologies that can enhance sensitivity, selectivity, and throughput into micro total analysis systems (μTAS) are important in making them useful in precise screening of reaction byproducts in analytical chemistry, cellular biology and pharmaceutical industries...... mechanical bending of the cantilever under infrared (IR) radiation. This technique also allows simultaneous physical characterization of the liquid reagent using variations in resonance frequency. It is useful in lab-on-a-chip devices and has a myriad of applications in drug screening, bioreactor monitoring...

  12. A Novel Fluorescent Reagent for Analysis of Hydrogen Peroxide

    Institute of Scientific and Technical Information of China (English)

    董素英; 苏美红; 聂丽华; 马会民


    8-(4,6-Dichloro-1,3,5-trazinoxy)quinoline(DTQ) was evaluated as a new fluorescent reagent for determining hydrogen peroxide.It was found that the fluorescence intensity of DTQ in alkaline medium could be dramatically enhanced upon addition of H2O2.Based on this effect,a simple and selective method for the spectrofluorimetric determination of hydrogen peroxide was estabhlished.The relative standard deviation of the method was found to be 1.1?for 9 replicate determinations of a 4.6×10-6mol/L hydrogen peroxide solution.The linear range was 2.3×10-7-2.3×10-5mol/L with a detection limit of 2.2×10-8mol/L(S/N=3).The ,method was attempted to determine hydrogen peroxide in synthetic human serum samples with satisfactory results.

  13. Mechanism of seasonal Arctic sea ice evolution and Arctic amplification


    Kim, Kwang-Yul; Hamlington, Benjamin D.; Na, Hanna; Kim, Jinju


    Sea ice loss is proposed as a primary reason for the Arctic amplification, although the physical mechanism of the Arctic amplification and its connection with sea ice melting is still in debate. In the present study, monthly ERA-Interim reanalysis data are analyzed via cyclostationary empirical orthogonal function analysis to understand the seasonal mechanism of sea ice loss in the Arctic Ocean and the Arctic amplification. While sea ice loss is widespread over much of the p...

  14. DNA Extraction from Formalin-fixed and Paraffin-embedded Tissues by Triton X-100 for Effective Amplification of EGFR Gene by Polymerase Chain Reaction

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-feng; DU Zhen-wu; WU Mei; ZHANG Yu-cheng; JIANG Yang; ZHANG Gui-zhen


    For first-line non-small-cell lung cancer(NSCLC) therapy,detecting mutation status of the epidermal growth factor receptor(EGFR) gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor(TKI) therapy.Now,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded(FFPE) tissues.DNA extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction(PCR) are two key steps for detecting EGFR gene mutation.We showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR gene.Extracting DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100(pH=10.0) was performed by heating at 95 C for 30min.Meanwhile,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for comparison.DNA extracted products were used as template for amplifying the exons 18,.19,20 and 21 of EGFR by PCR for different amplified fragments.Results show that DNA fragment size extracted from FFPE tissues with 1%Triton X was about 250-500 base pairs(bp).However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands bp.The DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial kit.For about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial kit.However,for about 200 bp fragment.This simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.

  15. An evaluation of the performance of five extraction methods: Chelex® 100, QIAamp® DNA Blood Mini Kit, QIAamp® DNA Investigator Kit, QIAsymphony® DNA Investigator® Kit and DNA IQ™. (United States)

    Ip, Stephen C Y; Lin, Sze-Wah; Lai, Kam-Ming


    DNA left at a crime scene was often limited in amount and far from pristine. To maximize the chance of recovering as much information as possible from such compromised samples, an appropriate extraction method using the available technologies needs to be devised. In this study, we used human blood, buffy coat and a total of 76 simulated touch DNA samples to test the effectiveness of the following five common DNA extraction methods, namely, Chelex® 100, QIAamp® DNA Blood Mini Kit, QIAamp® DNA Investigator Kit, QIAsymphony® DNA Investigator® Kit and DNA IQ™ system, in the recovery of such DNA. We demonstrated that the QIAamp® and QIAsymphony® DNA Investigator® Kits, and the DNA IQ™ system, exhibited a better effectiveness in DNA recovery amongst these methods and yielded extracts with higher success rate in subsequent DNA profiling. These extracts also generated profiles with better intra-colour signal balance. The findings in this work allowed us to propose an extraction approach as follows: 1) casework samples shall be extracted with the QIAamp®/QIAsymphony® DNA Investigator® Kits or the DNA IQ™ system, viz., QIAsymphony® DNA Investigator® Kit and DNA IQ™, due to their higher throughput, are for the touched DNA evidence from the volume crime, while QIAamp® DNA Investigator Kit is preferable for challenging bloodstain samples; and 2) control samples, such as buccal swab, with known identity can be extracted with the Chelex, due to their cheaper cost per sample.

  16. Does the prior application of the field kit bullet hole testing kit 3 on a suspected bullet hole bias the analysis of atomic absorption spectrophotometry? (United States)

    Seltenhammer, Monika H; Fitzl, Christine; Wieser, Ingo; Binder, Reinhard; Paula, Pia; Risser, Daniele U


    Forensic ballistics is the study of bullet trajectory and consists of determining gunshot residue (GSR) to identify bullet holes. Among several highly sensitive methods, atomic absorption spectrophotometry (AAS) is employed to analyze GSR in the laboratory. However, it is sometimes necessary to identify bullet holes immediately at a crime scene. The purpose of this examination was to determine whether the use of the field test Bullet Hole Testing Kit 3 (BTK3) on a suspected bullet hole would influence the outcome of AAS-analysis: Three commonly encountered firearms (Glock17, Tokarev, and Colt) were fired at skin, wood, and cloth. AAS-analysis was performed with and without previous BTK3 application. The results clearly indicate that there is no significant interaction on the grounds of BTK3 use (BTK3 vs. no-BTK3 [kit_nokit] [Pb: p = 0.1309; Sb: p = 0.9111], material*kit_nokit [Pb: p = 0.5960; Sb: p = 0.9930], distance*kit_nokit [Pb: p = 0.4014; Sb: p = 0.9184], and firearm type*kit_nokit [Pb: p = 0.9662; Sb: p = 0.9885]); hence, applying this field kit does not falsify later AAS outcomes.

  17. Adaptive Strategy for Online Gait Learning Evaluated on the Polymorphic Robotic LocoKit

    DEFF Research Database (Denmark)

    Johan Christensen, David; Larsen, Jørgen Christian; Støy, Kasper


    This paper presents experiments with a morphologyindependent, life-long strategy for online learning of locomotion gaits, performed on a quadruped robot build using the LockKit robotic building kit. The learning strategy applies a stochastic optimization algorithm to optimize eight open parameter...

  18. Pongase en accion! Materiales para actividades despues de la escuela (Get into Action! Afterschool Action Kit). (United States)

    Afterschool Alliance, Washington, DC.

    Noting that after-school programs are a critical link to helping children become successful adults, this Spanish-language kit explains what after-school programs can and should do for young people and how to locate or start an after-school program. The kit provides a rationale for developing after-school programs, noting the number of children…

  19. Expression and mutation of c-kit gene in gastrointestinal stromal tumors

    Institute of Scientific and Technical Information of China (English)

    Fei Feng; Xiao-Hong Liu; Qiang Xie; Wei-Qiang Liu; Cheng-Guang Bai; Da-Lie Ma


    AIM: To investigate the expression and mutation of c-kit gene and its correlation with the clinical pathology and prognosis of gastrointestinal stromal tumors (GISTs).METHODS: A total of 94 cases of GISTs, 10 leiomyomas and 2 schwannomas were studied for the expression of KIT by immunohistochemistry. The c-kit gene mutations in exon 11 of these specimens were detected by PCR-SSCP technique.RESULTS: Of the 94 cases of GISTs, 91 (96.8 %) expressed the KIT protein. Leiomyomas and schwannomas were negative for KIT. The c-kit gene mutations of exon 11 were found in 38 out of the 94 cases of GISTs (40.4 %). The mutations involved point mutations (Va1560-Asp, Ile563-Met),del 557-559 and 579ins12. No mutations were detectable in benign GISTs, leiomyomas or schwannomas. The patients with mutation-positive GISTs showed more frequent recurrences, invasion and metastasis in adjacent tissues than those with mutation-negative ones.CONCLUSION: KIT is a useful marker for diagnosis of GISTs.Mutation of the c-kit gene may play a significant role in the pathogenesis of GISTs and may be associated with poor prognosis in patients with GISTs.

  20. Acute interstitial pneumonia in mink kits inoculated with defined isolates of Aleutian mink disease parvovirus

    DEFF Research Database (Denmark)

    Alexandersen, Søren; Larsen, S; Aasted, B;


    The present study addressed the causal role of Aleutian mink disease parvovirus (ADV) in acute interstitial pneumonia in mink kits. All the examined isolates of ADV caused interstitial pneumonia in newborn kits, although the severity of disease and the mortality varied. These findings indicate...

  1. Procedures for Separations within Batches of Values, 1. The Orderly Tool Kit and Some Heuristics (United States)


    separations within batches of values, I. The orderly tool kit and some heuristics by Thu Hoang* and John W. Tukey** *Universite Rene Descartes ...separations with batches of values, . The orderly tool kit and heuristics Thu Hoang* and John W. Tukey** *Universite Rene Descartes Laboratoire de

  2. Evaluation of a radioimmunoassay (/sup 125/I) kit for cannabinoid metabolites in urine and whole blood

    Energy Technology Data Exchange (ETDEWEB)

    Childs, P.S.; McCurdy, H.H.

    The Abuscreen kit (Roche Diagnostics) for the analysis of 9/-tetrahydrocannabinol-9-carboxylic acid and other cannabinoids in urine was evaluated in terms of its accuracy, reproducibility, and sensitivity. A procedure is also presented for the analysis of total cannabinoids in whole blood using the RIA kit.

  3. Introductory Guide: How To Use This Kit. Job Match: Together for Good Business. (United States)

    Rood, Lois

    Intended for use with business and industry personnel, the guide is part of a kit designed to demonstrate how individuals with disabilities and older persons can be assimilated successfully into the labor force. Individual sections of the guide provide information on: (1) purpose of the training kit; (2) people with special needs defined (labels…

  4. Healthy Schools Campaign Pesticide Action Kit = Campana de Escuelas Saludables Paquete de Accion Pesticida. (United States)

    Arguello, Martha; Campbell, Kelly; Kegley, Susan; Olle, Teri; Porter, Catherine; Undem, Melanie

    This English/Spanish informational kit contains resource materials that school administrators and parents can use to take full advantage of the Healthy Schools Act of 2000 and help them eliminate hazardous pesticide use around their schools. The kit looks at how to organize community interest in least-toxic Integrated Pest Management policy, and…

  5. The Concurrent Validity of a New Intelligence Test: The Kahn Intelligence Test (KIT) (United States)

    Fenlon, Whatley; L'Abate, Luciano


    The concurrent validity of the KIT was evaluated by correlating it with other tests of intellectual, perceptual, social, linguistic and psycholinguistic functioning, using a sample of 106 retardates. Positive correlations between the KIT and these other measures attest to its potential usefulness as a measure of intelligence. (Author)

  6. Tickle Your Appetite: WIC/Team Nutrition Educator's Kit for Children. (United States)

    Food and Consumer Service (USDA), Washington, DC.

    Intended for use by WIC nutrition educators and other professionals, by day care and Head Start providers, and for home use, this educator's kit contains activities and information to improve nutrition experiences for preschool-age children. In addition to the educator's guide, the kit includes a short videotape and audiotape with three segments…

  7. The Safe Space Kit: Guide to Being an Ally to LGBT Students (United States)

    Gay, Lesbian and Straight Education Network (GLSEN), 2009


    "The Safe Space Kit" is designed to help educators create a safe space for LGBT (lesbian, gay, bisexual, and transgender) students. One of the most effective ways for an educator to create a safe space is to be a supportive ally to LGBT students. The hard copy of "The Safe Space Kit" includes the "Guide to Being an Ally," ten "Safe Space" stickers…

  8. How To Use Qualitative Methods in Evaluation. CSE Program Evaluation Kit, Volume 4. Second Edition. (United States)

    Patton, Michael Quinn

    The "CSE Program Evaluation Kit" is a series of nine books intended to assist people conducting program evaluations. This volume, the fourth in the kit, explains the basic assumptions underlying qualitative procedures, suggests evaluation situations where qualitative designs are useful, and provides guidelines for designing qualitative…

  9. W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity

    DEFF Research Database (Denmark)

    Huizinga, J D; Thuneberg, L; Klüppel, M;


    that the interstitial cells of Cajal express the Kit receptor tyrosine kinase. Furthermore, mice with mutations in the dominant white spotting (W) locus, which have cellular defects in haematopoiesis, melanogenesis and gametogenesis as a result of mutations in the Kit gene, also lack the network of interstitial cells...

  10. Targeting kit activation: a potential therapeutic approach in the treatment of allergic inflammation

    DEFF Research Database (Denmark)

    Jensen, Bettina M; Metcalfe, Dean D; Gilfillan, Alasdair M


    is necessary for optimal IgE/antigen-induced mast cell degranulation and cytokine production. Several drug candidates targeting SCF and/or Kit have been studied for their anti-allergic properties. These include anti-SCF antibodies, antisense oligonucleotides, Kit inhibitors, and inhibitors of downstream...

  11. DNA ploidy and c-Kit mutation in gastrointestinal stromal tumors

    Institute of Scientific and Technical Information of China (English)

    Ju Han Lee; Xianglan Zhang; Woon Yong Jung; Yang Seok Chae; Jong-Jae Park; Insun Kim


    AIM: To investigate the prognostic significance of c-Kit gene mutation and DNA ploidy in gastointestinal stromal tumors (GISTs).METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohistochemistry, and the c-Kit gene mutations in exons 9, 11, 13, and 17 were detected by polymerase chain reaction-single strand confirmation polymarphism (PCR-SSCP) and denaturing high performance liquid chromatography (D-HPLC)techniques. DNA ploidy was determined by flow cytometry.RESULTS: Of the 55 cases of GISTs, 53 cases (96.4%)expressed c-Kit protein. The c-Kit gene mutations of exons 11 and 9 were found in 30 (54.5%) and 7 cases (12.7%),respectively. No mutations were found in exons 13 and 17.DNA aneuploidy was seen in 10 cases (18.2%). The c-Kit mutation positive GISTs were larger in size than the negative GISTs. The aneuploidy tumors were statistically associated with large size, high mitotic counts, high risk groups, high cellularity and severe nuclear atypia, and epithelioid type.There was a tendency that c-Kit mutations were more frequently found in aneuploidy GISTs.CONCLUSION: DNA aneuploidy and c-Kit mutations can be considered as prognostic factors in GISTs.

  12. Things Really Exist? A Survival-Kit for Prospective Phenomenologists

    Directory of Open Access Journals (Sweden)

    Dirk Koob


    Full Text Available This article provides something of a survival kit for prospective phenomenologists. Starting with the main ideas of the founder of phenomenology Edmund HUSSERL, this article begins by articulating phenomenological thinking in abstract painting, existentialism, theology, and psychotherapy. On this background I discuss the theoretical and methodological development of phenomenology in the social sciences. Linking phenomenology and ordinary language philosophy rounds out this hopefully "easy-to-understand" introduction. After having read the text students should be familiar with the philosophical roots, the major application contexts, and some central reservations against phenomenology. Above all, students should be able to (a apply the term "phenomenology" appropriately (at least as far as this is possible and (b meet the requirements of phenomenologically oriented research methods in a reflective and mature way. URN: urn:nbn:de:0114-fqs0802202

  13. Shuttle performance enhancements using an OMS payload bay kit (United States)

    Templin, Kevin C.; Mallini, Charles J.


    The study focuses on the use of an orbital maneuvering system (OMS) payload bay kit (PBK) designed to utilize OMS tanks identical to those currently employed in the Orbiter OMS pods. Emphasis is placed on payload deployment capability and payload servicing/reboost capability augmentation from the point of view of payload mass, maximum deployment altitudes, and initial retrieval and final deployment altitudes. The deployment, servicing, and reboost requirements of the Hubble Space Telescope and Advanced X-ray and Astrophysics Facility are analyzed in order to show the benefits an OMS PBK can provide for these missions. It is shown that OMS PBKs can provide the required capability enhancement necessary to support deployment, reboost, and servicing of payloads requiring altitudes greater than 325 nautical miles.

  14. Word 2010 eLearning Kit For Dummies

    CERN Document Server

    Lowe, Lois


    Use this step-by-step learning package to master Word 2010 Word 2010 is one of the core applications of Microsoft Office and if you're eager to get started using all it has to offer, this value-packed eLearning Kit is essential to your learning process. This complete Word 2010 course includes a full-color printed book and a Dummies interactive eLearning course on CD. You'll discover the basics of the Word interface, how to navigate it, how to get comfortable with the terminology, and how to use its many features. Detailed instructions walk you through real-world exercises and help to make lear

  15. Office 2013 eLearning kit for dummies

    CERN Document Server

    Wempen, Faithe


    Unlock your new Office with this one-of-a-kind learning package! Whether you're meeting Office 2013 for the first time or upgrading your knowledge from an earlier version, this value-packed eLearning kit makes it easy to learn 2013 at your own pace. This complete learning package includes a full-color printed book and a Dummies interactive eLearning course on CD. You'll learn the basics of the Office interface, how to navigate it, and how to use the features common to all Office programs. Then you'll get detailed instructions for working with Word, Excel, PowerPoint, and Outlook. Follow the

  16. HTML5 eLearning kit for dummies

    CERN Document Server

    Boumphrey, Frank


    Helping self-directed learners of all levels learn HTML5 If you want to develop and structure pages for the web, HTML5 is one of the tools you need. This invaluable eLearning kit steps you through learning HTML5, CSS3, and JavaScript. With this dynamic combination of a full-color printed book and a Dummies interactive eLearning course on CD, you'll find a wealth of information on HTML5. Featuring both written and animated step-by-step how-tos, practice labs, helpful videos, numerous examples, and a host of Dummies hints and tips, this package makes your learning process easier. Follow the mate

  17. The Pandora software development kit for pattern recognition

    Energy Technology Data Exchange (ETDEWEB)

    Marshall, J.S.; Thomson, M.A. [University of Cambridge, Cavendish Laboratory, Cambridge (United Kingdom)


    The development of automated solutions to pattern recognition problems is important in many areas of scientific research and human endeavour. This paper describes the implementation of the Pandora software development kit, which aids the process of designing, implementing and running pattern recognition algorithms. The Pandora Application Programming Interfaces ensure simple specification of the building-blocks defining a pattern recognition problem. The logic required to solve the problem is implemented in algorithms. The algorithms request operations to create or modify data structures and the operations are performed by the Pandora framework. This design promotes an approach using many decoupled algorithms, each addressing specific topologies. Details of algorithms addressing two pattern recognition problems in High Energy Physics are presented: reconstruction of events at a high-energy e{sup +}e{sup -} linear collider and reconstruction of cosmic ray or neutrino events in a liquid argon time projection chamber. (orig.)

  18. Magnetic Field Amplification in Young Galaxies

    CERN Document Server

    Schober, Jennifer; Klessen, Ralf S


    The Universe at present is highly magnetized, with fields of the order of a few 10^-5 G and coherence lengths larger than 10 kpc in typical galaxies like the Milky Way. We propose that the magnetic field was amplified to this values already during the formation and the early evolution of the galaxies. Turbulence in young galaxies is driven by accretion as well as by supernova (SN) explosions of the first generation of stars. The small-scale dynamo can convert the turbulent kinetic energy into magnetic energy and amplify very weak primordial magnetic seed fields on short timescales. The amplification takes place in two phases: in the kinematic phase the magnetic field grows exponentially, with the largest growth on the smallest non-resistive scale. In the following non-linear phase the magnetic energy is shifted towards larger scales until the dynamo saturates on the turbulent forcing scale. To describe the amplification of the magnetic field quantitatively we model the microphysics in the interstellar medium ...

  19. Experimental noiseless linear amplification using weak measurements (United States)

    Ho, Joseph; Boston, Allen; Palsson, Matthew; Pryde, Geoff


    The viability of quantum communication schemes rely on sending quantum states of light over long distances. However, transmission loss can degrade the signal strength, adding noise. Heralded noiseless amplification of a quantum signal can provide a solution by enabling longer direct transmission distances and by enabling entanglement distillation. The central idea of heralded noiseless amplification—a conditional modification of the probability distribution over photon number of an optical quantum state—is suggestive of a parallel with weak measurement: in a weak measurement, learning partial information about an observable leads to a conditional back-action of a commensurate size. Here we experimentally investigate the application of weak, or variable-strength, measurements to the task of heralded amplification, by using a quantum logic gate to weakly couple a small single-optical-mode quantum state (the signal) to an ancilla photon (the meter). The weak measurement is carried out by choosing the measurement basis of the meter photon and, by conditioning on the meter outcomes, the signal is amplified. We characterise the gain of the amplifier as a function of the measurement strength, and use interferometric methods to show that the operation preserves the coherence of the signal.

  20. Loop-mediated isothermal amplification for detection of nucleic acids. (United States)

    Tanner, Nathan A; Evans, Thomas C


    Sequence-specific isothermal nucleic acid amplification techniques are ideally suited for use in molecular diagnostic applications because they do not require thermal cycling equipment and the reactions are typically fast. One of the most widely cited isothermal techniques is termed loop-mediated isothermal amplification (LAMP). This protocol allows amplification times as fast as 5 to 10 min. Furthermore, various methodologies to detect amplification have been applied to LAMP to increase its utility for the point-of-care market. Basic LAMP protocols are provided herein for detection of specific DNA and RNA targets, along with a method to perform multiplex LAMP reactions, permitting even greater flexibility from this powerful technique.