Sample records for blot hybridization spoligotyping


    Institute of Scientific and Technical Information of China (English)

    刘跃华; 王家璧; 司静懿


    This study differentiated pseudocondyloma of vulva from condyloma acunainata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases o{ pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the sttldy. The genital lesions were examined clinically and were biopsled. Each biopsy v-as subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82.6%) out of 23 cases of condyloma acuminata and 2(25%) out of 8 cases pseudocondyloma of vulvae(P<0. 05). PCR detected HPV DNA in 51(92.7%) our of 55 cases of eondyloma acuminata, compared with none in 23 cases of pseudocondylorna(P<0. 001). HPV DNA was present in the majority of condyloma acuminata specimens, HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful techntque for HPV DNA detection.

  2. The use of microbead-based spoligotyping for Mycobacterium tuberculosis complex to evaluate the quality of the conventional method: Providing guidelines for Quality Assurance when working on membranes

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    Garzelli Carlo


    Full Text Available Abstract Background The classical spoligotyping technique, relying on membrane reverse line-blot hybridization of the spacers of the Mycobacterium tuberculosis CRISPR locus, is used world-wide (598 references in Pubmed on April 8th, 2011. However, until now no inter-laboratory quality control study had been undertaken to validate this technique. We analyzed the quality of membrane-based spoligotyping by comparing it to the recently introduced and highly robust microbead-based spoligotyping. Nine hundred and twenty-seven isolates were analyzed totaling 39,861 data points. Samples were received from 11 international laboratories with a worldwide distribution. Methods The high-throughput microbead-based Spoligotyping was performed on CTAB and thermolyzate DNA extracted from isolated Mycobacterium tuberculosis complex (MTC strains coming from the genotyping participating centers. Information regarding how the classical Spoligotyping method was performed by center was available. Genotype discriminatory analyses were carried out by comparing the spoligotypes obtained by both methods. The non parametric U-Mann Whitney homogeneity test and the Spearman rank correlation test were performed to validate the observed results. Results Seven out of the 11 laboratories (63 %, perfectly typed more than 90% of isolates, 3 scored between 80-90% and a single center was under 80% reaching 51% concordance only. However, this was mainly due to discordance in a single spacer, likely having a non-functional probe on the membrane used. The centers using thermolyzate DNA performed as well as centers using the more extended CTAB extraction procedure. Few centers shared the same problematic spacers and these problematic spacers were scattered over the whole CRISPR locus (Mostly spacers 15, 14, 18, 37, 39, 40. Conclusions We confirm that classical spoligotyping is a robust method with generally a high reliability in most centers. The applied DNA extraction procedure (CTAB

  3. Application of a Dot Blot Hybridization Platform to Assess Streptococcus uberis Population Structure in Dairy Herds (United States)

    Albuquerque, Pedro; Ribeiro, Niza; Almeida, Alexandre; Panschin, Irena; Porfirio, Afonso; Vales, Marta; Diniz, Francisca; Madeira, Helena; Tavares, Fernando


    Streptococcus uberis is considered one of the most important pathogens associated with bovine mastitis. While traditionally acknowledged as an environmental pathogen, S. uberis has been shown to adopt a contagious epidemiological pattern in several dairy herds. Since different control strategies are employed depending on the mode of transmission, in-depth studies of S. uberis populations are essential to determine the best practices to control this pathogen. In this work, we optimized and validated a dot blot platform, combined with automatic image analysis, to rapidly assess the population structure of infective S. uberis, and evaluated its efficiency when compared to multilocus sequence analysis (MLSA) genotyping. Two dairy herds with prevalent S. uberis infections were followed in a 6 month period, in order to collect and characterize isolates from cows with persistent infections. These herds, located in Portugal (Barcelos and Maia regions), had similar management practices, with the herd from Barcelos being smaller and having a better milking parlor management, since infected cow segregation was immediate. A total of 54 S. uberis isolates were obtained from 24 different cows from the two herds. To overcome operator-dependent analysis of the dot blots and increase the technique's consistency and reliability, the hybridization signals were converted into probability values, with average probabilities higher than 0.5 being considered positive results. These data allowed to confirm the isolates' identity as S. uberis using taxa-specific markers and to determine the presence of virulence- and antibiotic resistance-related genes. In addition, MLSA allowed to disclose the most prevalent S. uberis clonal lineages in both herds. Seven different clusters were identified, with Barcelos showing a high clonal diversity and Maia a dominant lineage infecting most cows, suggesting distinct epidemiological patterns, with S. uberis displaying an environmental or contagious

  4. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization

    Institute of Scientific and Technical Information of China (English)

    GUO Qian; WAN Kang Lin; YU Yan; ZHU Yan Ling; ZHAO Xiu Qin; LIU Zhi Guang; ZHANG Yuan Yuan; LI Gui Lian; WEI Jian Hao; WU Yi Mou


    Objective A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in‘hot mutation region’ of Mycobacterium tuberculosis (M. tuberculosis). Methods 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay. Results The sensitivity and specificity of the RDBH assay were 91.2%(165/181) and 98.3%(117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7%(293/300), 98.2%(164/167), and 97.0%(129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively. Conclusion Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.

  5. Dot-Blot Hybridization for Detection of Five Cucurbit Viruses by Digoxigenin-Labelled cDNA Probes

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    MENG Juan; GU Qin-sheng; LIN Shi-ming; PENG Bin; LIU Li-feng; TIAN Yan-ping; LI Li


    Dot-blot hybridization was applied in this paper to detect five viruses infecting cucurbitaceous crops,Zuccini yellow mosaic virus(ZYMV),Watermelon mosaic virus(WMV),Cucumber mosaic virus(CMV),Papaya ringspot virus watermelon strain(PRSV-W)and Squash mosaic virus(SqMV),as a good alternative assay in seed health test and epidemiological and transgenic research.Digoxigenin-labelled cDNA probes of the five viruses were synthesized by PCR with the specific primers and applied in dot-blot hybridization to detect five viruses in crude extraction of the infected leaves.And three SqMV probes of different lengths(0.55,1.6,and 2.7 kb,respectively)were designed to investigate the effect of hybridization.The results showed that the sensitivity for detecting the crude extraction of infected leaves by ZYMV,WMV,CMV,PRSV-W,and SqMV was down to 1:160,1:160,1:320,1:160,and 1:320,respectively.Three SqMV probes of different length showed no differences on the sensitivity and specificity.The digoxigenin-labelled probes prepared by PCR could be used for accurate and rapid identification of 5 viruses infecting cucurbitaceous crops with good stabilities,sensitivities,specificity,and reproducibilities.

  6. A strip array for spoligotyping of Mycobacterium tuberculosis complex isolates. (United States)

    Tu, Yiling; Zeng, Xiaohong; Li, Hui; Zheng, Rongrong; Xu, Ye; Li, Qingge


    A novel strip array was developed for a nine-spacer spoligotyping scheme of Mycobacterium tuberculosis complex (MTBC). The new method was evaluated using 211 MTBC isolates and the results were fully concordant with the traditional spoligotyping approach. The strip array proved to be rapid and convenient for spoligotyping of MTBC.

  7. Northern blotting analysis

    DEFF Research Database (Denmark)

    Josefsen, Knud; Nielsen, Henrik


    is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing...

  8. Comparison of Southern blot analysis with isotopic and nonisotopic in situ hybridization for the detection of human papillomavirus sequences in invasive carcinoma of the uterine cervix. (United States)

    D'Amato, L; Pilotti, S; Rotola, A; Di Luca, D; Cassai, E; Rilke, F


    To compare the efficiency of hybridization methods for the detection of HPV genome, 22 cases of invasive squamous cell carcinoma of the uterine cervix were analyzed by Southern blot analysis and in situ hybridization carried out with 35S- and biotin-labeled probes. These cases contained from less than one to as many as 50 copies per cell of HPV 16 and 18 types. To increase the sensitivity of biotinylated probes, a silver enhancement procedure of the peroxidase reaction product was applied. Results showed that in situ hybridization performed with isotopic probes is as sensitive as Southern blot analysis and is more sensitive than that performed with biotin-labeled probe. However, the application of the silver enhancement procedure increases the percentage of HPV-positive cases from 27 to 50%.

  9. Mycobacterium bovis in Swine: Spoligotyping of Isolates from Argentina

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    Soledad Barandiaran


    Full Text Available A total of 143 Mycobacterium bovis isolates of pigs, from the most productive swine area in Argentina, were typed by spoligotyping. Twenty-two different spoligotypes were identified, and 133 (93% isolates were grouped into 12 clusters. One of them, designed SB0140, was the most frequent because it held 83 (58% isolates. This spoligotype also grouped 362 (43% out of 841 isolates from previously typed cattle and, thus, constitutes the most frequent in our country. In addition, 135 (94% isolates revealed spoligotypes identical to those of cattle, showing an epidemiological link. On the other hand, there were seven novel spoligotypes, six of which were also unique since they had only one isolate each. This study aimed to identify the spoligotypes of M. bovis isolated from pigs to contribute to a better understanding of the distribution of bovine tuberculosis in the main productive area of Argentina.

  10. A comparison of DNA extraction protocols from blood spotted on FTA cards for the detection of tick-borne pathogens by Reverse Line Blot hybridization. (United States)

    Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo


    An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic(®) cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex(®) resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent.

  11. The application of a photon-counting camera in very sensitive, bioluminescence-enhanced detection systems for protein blotting. Ultrasensitive detection systems for protein blotting and DNA hybridization, II. (United States)

    Hauber, R; Miska, W; Schleinkofer, L; Geiger, R


    A relatively simple, very sensitive bioluminescence-enhanced detection system for protein blots was described recently. This method utilizes antibodies conjugated with alkaline phosphatase. Alkaline phosphatase releases D-luciferin (Photinus pyralis) from D-luciferin-O-phosphate. Liberated D-luciferin reacts with luciferase, ATP and oxygen with light emission. The light produced is measured with a very sensitive photon counting camera (Argus 100), permitting visualization and localization of the alkaline phosphatase-conjugated antibodies on nitrocellulose sheets. Under non-optimized conditions the limit of detection is at present 5 to 500 fg of protein (rabbit immunoglobulin G), corresponding to 30 to 3 amol. The method is therefore 10(5) times more sensitive than other used at present.

  12. Detection of mutation in embB gene of Mycobacterium tuberculosis from clinical isolates of tuberculous patients in China by means of reverse-dot blot hybridization

    Institute of Scientific and Technical Information of China (English)



    The relationship between embB mutation of Mycobacterium tuberculosis and ethambutol (EMB) resistance of the clinical isolates of tuberculous patients in China was investigated by reversedot blot hybridization (RDBH) in addition to evaluating the clinical value with application of PCR-RD-BH technique to detect EMB resistance. In the present study, the genotypes of the 258 bp fragments of embB genes from 196 clinical isolates of M. tuberculosis were analysed with RDBH and DNA sequencing. It was demonstrated that 60 out of 91 phenotypically EMB-resistant isolates (65.9%) showed 5 types of missense mutations at codon 306 of embB gene, resulting in the replacement of the Met residue of the wild type strain with Val, Ile or Leu residues. In these mutations, the GTP mutation (38/91,41.8% ) and the ATA mutation ( 16/91, 17.6% ) were the most encountered genotypes. The embB mutation at codon 306 could also be found in 69 isolates of phenotypically EMB-sensitive but resistant to other anti-tuberculous drugs, but no such gene mutation could be found in 36 strains of drug-sensitive isolates. Meanwhile, the concordance with the results of DNA sequencing for one wide-type probe and 5 probes for specific mutations was 100%. It was concluded that the EMB-resistance occurring in most M. tuberculosis is due to appearance of embB mutation at codon 306, and the PCR-RDBH assay was proved to be a rapid, simple and reliable method for the detection of gene mutations, which might be a good alternative for the drug-resistance screening.

  13. Development of a Rapid Reverse Blot Hybridization Assay for Detection of Clinically Relevant Antibiotic Resistance Genes in Blood Cultures Testing Positive for Gram-Negative Bacteria (United States)

    Wang, Hye-young; Yoo, Gilsung; Kim, Juwon; Uh, Young; Song, Wonkeun; Kim, Jong Bae; Lee, Hyeyoung


    Rapid and accurate identification of the causative pathogens of bloodstream infections is crucial for the prompt initiation of appropriate antimicrobial therapy to decrease the related morbidity and mortality rates. The aim of this study was to evaluate the performance of a newly developed PCR-reverse blot hybridization assay (REBA) for the rapid detection of Gram-negative bacteria (GNB) and their extended-spectrum β-lactamase (ESBL), AmpC β-lactamase, and carbapenemase resistance genes directly from the blood culture bottles. The REBA-EAC (ESBL, AmpC β-lactamase, carbapenemase) assay was performed on 327 isolates that were confirmed to have an ESBL producer phenotype, 200 positive blood culture (PBCs) specimens, and 200 negative blood culture specimens. The concordance rate between the results of REBA-EAC assay and ESBL phenotypic test was 94.2%. The sensitivity, specificity, positive predictive value, and negative predictive value of the REBA-EAC assay for GNB identification in blood culture specimens were 100% (95% CI 0.938–1.000, P < 0.001), 100% (95% CI 0.986–1.000, P < 0.001), 100% (95% CI 0.938–1.000, P < 0.001), and 100% (95% CI 0.986–1.000, P < 0.001), respectively. All 17 EAC-producing GNB isolates from the 73 PBCs were detected by the REBA-EAC assay. The REBA-EAC assay allowed easy differentiation between EAC and non-EAC genes in all isolates. Moreover, the REBA-EAC assay was a rapid and reliable method for identifying GNB and their β-lactamase resistance genes in PBCs. Thus, this assay may provide essential information for accelerating therapeutic decisions to achieve earlier appropriate antibiotic treatment during the acute phase of bloodstream infection. PMID:28232823

  14. Development of a Rapid Reverse Blot Hybridization Assay for Detection of Clinically Relevant Antibiotic Resistance Genes in Blood Cultures Testing Positive for Gram-Negative Bacteria. (United States)

    Wang, Hye-Young; Yoo, Gilsung; Kim, Juwon; Uh, Young; Song, Wonkeun; Kim, Jong Bae; Lee, Hyeyoung


    Rapid and accurate identification of the causative pathogens of bloodstream infections is crucial for the prompt initiation of appropriate antimicrobial therapy to decrease the related morbidity and mortality rates. The aim of this study was to evaluate the performance of a newly developed PCR-reverse blot hybridization assay (REBA) for the rapid detection of Gram-negative bacteria (GNB) and their extended-spectrum β-lactamase (ESBL), AmpC β-lactamase, and carbapenemase resistance genes directly from the blood culture bottles. The REBA-EAC (ESBL, AmpC β-lactamase, carbapenemase) assay was performed on 327 isolates that were confirmed to have an ESBL producer phenotype, 200 positive blood culture (PBCs) specimens, and 200 negative blood culture specimens. The concordance rate between the results of REBA-EAC assay and ESBL phenotypic test was 94.2%. The sensitivity, specificity, positive predictive value, and negative predictive value of the REBA-EAC assay for GNB identification in blood culture specimens were 100% (95% CI 0.938-1.000, P < 0.001), 100% (95% CI 0.986-1.000, P < 0.001), 100% (95% CI 0.938-1.000, P < 0.001), and 100% (95% CI 0.986-1.000, P < 0.001), respectively. All 17 EAC-producing GNB isolates from the 73 PBCs were detected by the REBA-EAC assay. The REBA-EAC assay allowed easy differentiation between EAC and non-EAC genes in all isolates. Moreover, the REBA-EAC assay was a rapid and reliable method for identifying GNB and their β-lactamase resistance genes in PBCs. Thus, this assay may provide essential information for accelerating therapeutic decisions to achieve earlier appropriate antibiotic treatment during the acute phase of bloodstream infection.

  15. Simultaneous detection of Anaplasma and Ehrlichia species in ruminants and detection of Ehrlichia ruminantium in Amblyomma variegatum ticks by reverse line blot hybridization. (United States)

    Bekker, Cornelis P J; de Vos, Sander; Taoufik, Amar; Sparagano, Olivier A E; Jongejan, Frans


    The detection of Anaplasma and Ehrlichia species is usually based on species-specific PCR assays, since no assay is yet available which can detect and identify these species simultaneously. To this end, we developed a reverse line blot (RLB) assay for simultaneous detection and identification of Anaplasma and Ehrlichia species in domestic ruminants and ticks. In a PCR the hypervariable V1 region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for members of the genera Anaplasma and Ehrlichia [Int. J. Syst. Evol. Microbiol. 51 (2001) 2145]. Amplified PCR products from blood of domestic ruminants or Amblyomma variegatum tick samples were hybridized onto a membrane to which eight species-specific oligonucleotide probes and one Ehrlichia and Anaplasma catch-all oligonucleotide probe were covalently linked. No DNA was amplified from uninfected blood, nor from other hemoparasites such as Theileria annulata, or Babesia bigemina. The species-specific probes did not cross-react with DNA amplified from other species. E. ruminantium, A. ovis and another Ehrlichia were identified by RLB in blood samples collected from small ruminants in Mozambique. Finally, A. variegatum ticks were tested after feeding on E. ruminantium infected sheep. E. ruminantium could be detected in adult ticks even if feeding of nymphs was carried out 3.5 years post-infection. In conclusion, the developed species-specific oligonucleotide probes used in an RLB assay can simultaneously detect and identify several Ehrlichia and Anaplasma species. However, as no quantitative data for the detection limit are available yet, only positive results are interpretable at this stage.

  16. Northern blotting analysis

    DEFF Research Database (Denmark)

    Josefsen, Knud; Nielsen, Henrik


    Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membran...

  17. Spoligotype patterns of Mycobacterium tuberculosis isolated from extra pulmonary tuberculosis patients in Puducherry, India

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    G Kandhakumari


    Full Text Available Purpose: Genotyping studies like spoligotyping are valuable tools in understanding the genetic diversity and epidemiology of Mycobacterium tuberculosis. Though there are reports of spoligotyping of M. tuberculosis isolates from pulmonary specimens from different parts of India, spoligotyping of extra pulmonary tuberculosis isolates are very few. Puducherry has not yet recorded spoligopatterns of M. tuberculosis from either pulmonary or extra pulmonary (EPTB specimens. The aim of this study is to analyze the spoligotype patterns of EPTB strains circulating in Puducherry and neighboring districts of Tamil Nadu. Materials and Methods: During June 2011 to December 2013, 570 EPTB specimens were processed by culturing on to Lowenstein Jensen (LJ medium and automated Mycobacterium Growth Indicator Tube system (MGIT960. Identification of M. tuberculosis was carried out as per standard procedures, and MPT 64 antigen positivity in a commercial immunochromatography kit. Spoligotyping was carried out at National Institute of Research in Tuberculosis (ICMR, Chennai. Results: M. tuberculosis was isolated from 67 single EPTB specimens (11.8% like pus/cold abscess (34, TB spine (10, pleural fluid (10, urine (5, tissue bit (2, lymph nodes (2, ascitic fluid (2, synovial fluid (1 and endometrial curetting (1. Among 67 isolates with 41 spoligopatterns, EAI lineage with 28 isolates (41.8% predominated followed by 18 orphans (26.9%, 10 Beijing (14.9% and 8 U (11.9%. BOVIS1_BCG (ST482, T1-T2 (ST78 and H3 (ST50 were represented by one strain each (1.5%. C onclusions: Spoligotyping plays a significant role in the epidemiology of tuberculosis. Three spoligotypes, T1-T2 (ST78, EAI6 (ST292 and U (ST1429 are reported for the first time in India.

  18. Simultaneous identification of Aspergillus and Mucorales species by reverse line blot (RLB) hybridization assay%应用反向线点杂交技术鉴定临床常见曲霉属和毛霉目真菌

    Institute of Scientific and Technical Information of China (English)

    赵作涛; 李丽丽; 王晓阳; 万喆; 陈伟; 李若瑜


    Objective To Rapidly differentiate Aspergillus and Mucorales species by reverse line blot hybridization (RLB). Methods Totally 98 isolates including five Aspergillus strains ( Aspergillus fumigatus, Aspergillus flavus,Aspergillus niger, Aspergillus terreus and Aspergillus nidulans ) and seven Mucorales species ( Mucor heimalis ,Mucor racemosus ,Mucor cercinelloidea,Rhizopus arrhizus,Rhizopus microsporus,Rhizomucor pusillus and Absidia corymbifera ) were obtained from Research Center for Medical Mycology and Mycoses, Peking University. ITS1 and ITS4 fungal universal primers were chosen for PCR amplification, and the amplified products were used for reverse line blot hybridization with 12 fungal specie-specific probes. RLB data were compared with those by traditional fungal morphology and ITS sequencing methods. Results RLB showed high sensitivity and specificity , with 100% correct identification percentage of all the isolates and no cross hybridization between the species-specific probes. Eight negative control stains ( Candida albicans ,Fusarium solani ,Scedosporium apiospermum ,Penicillium marneffei ,Exophiala verrucosa,Aspergillus clavatus,Aspergillus japonicus and Cunninghamella elegans ) also showed negative by RLB. The analytical sensitivity of RLB was 1. 8 x 10-3 ng/μL by 10-fold serial dilution of Aspergillus fumigatus genomic DNA. Conclusions The RLB assay provides a rapid and reliable option for laboratory diagnosis and identification of Aspergillus and Mucorales species.%目的 应用反向线点杂交技术(reverse line blot hybridization,RLB)快速鉴定临床常见的曲霉属和毛霉目真菌.方法 收集我院真菌和真菌病研究中心保存的5种曲霉菌(烟曲霉、黄曲霉、黑曲霉、土曲霉、构巢曲霉)和7种毛霉目真菌(冻土毛霉菌、总状毛霉菌、卷枝毛霉菌、少根根霉、小孢根霉、微小根毛霉、伞状犁头霉),共计98株菌株.利用真菌通用引物ITS1和ITS4对菌株进行PCR扩增,用12个

  19. Characterization of Mycobacterium tuberculosis complex DNAs from Egyptian mummies by spoligotyping. (United States)

    Zink, Albert R; Sola, Christophe; Reischl, Udo; Grabner, Waltraud; Rastogi, Nalin; Wolf, Hans; Nerlich, Andreas G


    Bone and soft tissue samples from 85 ancient Egyptian mummies were analyzed for the presence of ancient Mycobacterium tuberculosis complex DNA (aDNA) and further characterized by spoligotyping. The specimens were obtained from individuals from different tomb complexes in Thebes West, Upper Egypt, which were used for upper social class burials between the Middle Kingdom (since ca. 2050 BC) and the Late Period (until ca. 500 BC). A total of 25 samples provided a specific positive signal for the amplification of a 123-bp fragment of the repetitive element IS6110, indicating the presence of M. tuberculosis DNA. Further PCR-based tests for the identification of subspecies failed due to lack of specific amplification products in the historic tissue samples. Of these 25 positive specimens, 12 could be successfully characterized by spoligotyping. The spoligotyping signatures were compared to those in an international database. They all show either an M. tuberculosis or an M. africanum pattern, but none revealed an M. bovis-specific pattern. The results from a Middle Kingdom tomb (used exclusively between ca. 2050 and 1650 BC) suggest that these samples bear an M. africanum-type specific spoligotyping signature. The samples from later periods provided patterns typical for M. tuberculosis. This study clearly demonstrates that spoligotyping can be applied to historic tissue samples. In addition, our results do not support the theory that M. tuberculosis originated from the M. bovis type but, rather, suggest that human M. tuberculosis may have originated from a precursor complex probably related to M. africanum.

  20. Clustering of Mycobacterium tuberculosis Cases in Acapulco: Spoligotyping and Risk Factors

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    Elizabeth Nava-Aguilera


    Full Text Available Recurrence and reinfection of tuberculosis have quite different implications for prevention. We identified 267 spoligotypes of Mycobacterium tuberculosis from consecutive tuberculosis patients in Acapulco, Mexico, to assess the level of clustering and risk factors for clustered strains. Point cluster analysis examined spatial clustering. Risk analysis relied on the Mantel Haenszel procedure to examine bivariate associations, then to develop risk profiles of combinations of risk factors. Supplementary analysis of the spoligotyping data used SpolTools. Spoligotyping identified 85 types, 50 of them previously unreported. The five most common spoligotypes accounted for 55% of tuberculosis cases. One cluster of 70 patients (26% of the series produced a single spoligotype from the Manila Family (Clade EAI2. The high proportion (78% of patients infected with cluster strains is compatible with recent transmission of TB in Acapulco. Geomatic analysis showed no spatial clustering; clustering was associated with a risk profile of uneducated cases who lived in single-room dwellings. The Manila emerging strain accounted for one in every four cases, confirming that one strain can predominate in a hyperendemic area.

  1. Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

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    Patrícia Martins Parreiras


    Full Text Available We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16 or high (0.6, MIRU26 discriminatory index (h. Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86 and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

  2. Contribution of spoligotyping to the characterization of the population structure of Mycobacterium tuberculosis isolates in Portugal. (United States)

    David, Suzana; Ribeiro, Diana Raposo; Antunes, Abílio; Portugal, Clara; Sancho, Luísa; de Sousa, José Germano


    Tuberculosis is a major health problem in Portugal. To begin characterizing the population structure of Mycobacterium tuberculosis, spoligotyping was used for the systematic typing, through consecutive sampling, of patient isolates from the Amadora-Sintra area of Greater Lisbon. Distribution amongst major spoligotype families, including the Latin American Mediterranean (LAM), T, Haarlem and Beijing, was compared to that of the international spoligotype database SpolDB4 and to the European countries of traditional Portuguese immigration represented in SpolDB4. Spoligotypes from 665 isolates were analyzed and 97 shared international types (SITs) identified. In SpolDB4 Portugal is represented by part of the spoligotypes from this study explaining the reduced number of unidentified patterns. The importance of the LAM family, and especially of LAM1 and LAM9 sub-families that alone represented 38% of all the isolates in this study as compared to 8% relative to the European sub group, led us to believe that at least in this respect the population structure was closer to that of Africa and South America than to Europe. Spoligotypes characteristic of Portugal or Portuguese related settings were identified. These included SIT244 a T1 sub-family predominant in Portugal and Bangladesh, SIT64 a LAM 6 sub-family common to Portugal and Brazil, and SIT1106 a LAM 9 sub-family. These studies were the first in Portugal stressing the importance of monitoring the population structure of M. tuberculosis isolates, an important step towards gaining an understanding of tuberculosis and the dynamics of this disease.

  3. Comparison of multiplex real-time PCR and PCR-reverse blot hybridization assay for the direct and rapid detection of bacteria and antibiotic resistance determinants in positive culture bottles. (United States)

    Wang, Hye-Young; Kim, Seoyong; Kim, Jungho; Park, Soon Deok; Kim, Hyo Youl; Uh, Young; Lee, Hyeyoung


    The aim of this study was to evaluate the performance of a commercially available multiplex real-time PCR assay and a PCR-reverse blot hybridization assay (PCR-REBA) for the rapid detection of bacteria and identification of antibiotic resistance genes directly from blood culture bottles and to compare the results of these molecular assays with conventional culture methods. The molecular diagnostic methods were used to evaluate 593 blood culture bottles from patients with bloodstream infections. The detection positivity of multiplex real-time PCR assay for Gram-positive bacteria, Gram-negative bacteria and Candida spp. was equivalent to PCR-REBA as 99.6 %, 99.1 % and 100 %, respectively. Using conventional bacterial cultures as the gold standard, the sensitivity, specificity, positive predictive value and negative predictive value of these two molecular methods were 99.5 % [95 % confidence interval (CI), 0.980-1.000; PReal-methicillin-resistant Staphylococcusaureus multiplex real-time PCR assay targeting the mecA gene to detect methicillin resistance was lower than that of the PCR-REBA method, detecting an overall positivity of 98.4 % (n=182; 95 % CI, 0.964-1.000; P<0.009) and 99.5 % (n=184; 95 % CI, 0.985-1.000; P<0.0001), respectively. The entire two methods take about 3 h, while results from culture can take up to 48-72 h. Therefore, the use of these two molecular methods was rapid and reliable for the characterization of causative pathogens in bloodstream infections.

  4. A comparison of real-time PCR and reverse line blot hybridization in detecting feline haemoplasmas of domestic cats and an analysis of risk factors associated with haemoplasma infections

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    Georges Karla


    Full Text Available Abstract Background Three species of feline haemoplasma are recognised: Mycoplasma haemofelis (Mhf, ‘Candidatus Mycoplasma haemominutum’ (CMhm and ‘Candidatus Mycoplasma turicensis (CMt. This study compared a reverse line blot hybridization (RLB assay for simultaneous detection of Mhf, CMhm with three separate quantitative real-time polymerase chain reaction (qPCR assays used for diagnosis of Mhf, CMhm and CMt. The RLB and qPCR assays were applied to DNA extracted from blood samples collected from 154 cats from Trinidad and Tobago. Results CMhm and Mhf DNA were detected using both RLB and qPCR. CMt DNA was detected by qPCR only. Comparing RLB and qPCR for the detection of CMhm DNA, 40 (26.3% and 48 (31.6% cats, respectively, were positive. The difference was more marked for Mhf, with RLB detecting a total of only 11 (7.2% positive cats whereas qPCR detected 41 (27.0% positive cats. Using qPCR as a gold standard, haemoplasma infected cats were more likely to be retrovirus positive (OR = 5.68, P = 0.02 and older (median age 5.5 years, than non-infected cats. In addition, CMhm positive cats were more likely to be male (OR = 3.4, P = 0.04. Conclusions Overall the qPCR was more sensitive than RLB. In addition, age (median 5.5 years and retrovirus positivity were risk factors for infection with the feline haemoplasmas in this study population. Further studies on feline haemoplasma infections in cats are needed to determine the significance of detecting small amounts of haemoplasma DNA, feline retrovirus infection and other associated risk factors on the clinical manifestation of disease.

  5. Uso da técnica de Southern Blot/Hibridização associada à reação em cadeia da polimerase para aumentar a sensibilidade no diagnóstico das infecções por hemoplasmas em gatos domésticos: Use of Southern Blot/Hybridization technique associated to polymerase chain reaction to improve the sensitivity in the diagnosis of hemoplasma infections in domestic cats

    Directory of Open Access Journals (Sweden)

    Daniel B. Macieira


    Full Text Available O objetivo deste trabalho foi verificar se a técnica de Southern Blot/Hibridização (SB em associação à reação de polimerização em cadeia (PCR aumenta a sensibilidade na detecção de DNA de hemoplasmas em gatos domésticos (Felis catus. O sangue total foi coletado em tubos contendo o anticoagulante ácido etilenodiamino tetra-acético, o DNA extraído a partir de 149 animais e a PCR realizada com o uso de sequências iniciadoras espécie-específicas, para amplificar subunidade 16S do RNA ribossomal de Mycoplasma haemofelis e 'Candidatus M. haemominutum' dessas amostras. Para a hibridização, foram utilizadas sondas específicas quimicamente marcadas, e os resultados visualizados por meio da adição de substrato quimiluminescente seguida de autoradiografia. Dezoito (12,1% das 149 amostras testadas apresentaram resultado PCR-positivo para o DNA de hemoplasmas. A técnica de SB mostrou que 24/149 (16,1% amostras apresentaram resultado positivo para hemoplasmas, confirmando os 18 resultados PCR-positivos, além de revelar seis outros adicionais (p The aim of this study was to determine whether Southern Blot/Hybridization (SB associated to Polymerase Chain Reaction (PCR improves the sensitivity in the detection of hemoplasma DNA in domestic cats (Felis catus. Whole blood was collected in tubes containing the anticoagulant ethylenediamine tetra-acetic acid and DNA extracted from 149 animals. PCR was performed using species specific primers to amplify the 16S ribosomal RNA subunit of Mycoplasma haemofelis and 'Candidatus M. haemominutum' from these samples. Hybridization was performed using a 16S rDNA probes chemically labeled and the results were visualized using a chemiluminescent substrate addition followed by autoradiography. Eighteen (12.1% of the 149 tested samples had a positive PCR result for hemoplasma species DNA. SB/hybridization technique showed that 24/149 (16.1% samples were positive for hemoplasmas, confirming the 18 PCR

  6. 实时PCR和PCR-RDB法检测人乳头瘤病毒的比对研究%Comparison of real time PCR and PCR¯reverse dot blot hybridization for detection of Human papillomavirus

    Institute of Scientific and Technical Information of China (English)

    肖克林; 严泽浩; 罗茗月; 麦光兴; 陈熙; 熊礼宽


    目的:比较实时 PCR 法和 PCR-反向点杂交法(PCR-RDB)检测 HPV 的一致性。方法收集109份女性生殖道样本,利用实时 PCR 法和 PCR-RDB 法分别检测 HPV 感染和基因型分布情况,不一致样本采用 PCR-悬浮芯片杂交法复检。结果83.5%(91/109)的样本两种方法结果一致(kappa=0.671),18例不一致样本 PCR-悬浮芯片杂交法复检显示7例与实时 PCR相符,11例与 PCR-RDB 相符;高、低病毒载量组间 PCR-RDB 法的检测结果差异无统计学意义(χ2=1.476,P =0.224)。结论实时 PCR 和 PCR-RDB 两法用于 HPV 检测一致性一般;HPV 病毒载量在103~108范围内时 PCR-RDB 法的阳性率较稳定。%Objective To compare real time PCR with PCR-reverse dot blot hybridization (PCR-RDB)for detecting human pap-illomavirus (HPV)infection in women.Methods A total of 109 genital specimens from women were collected in the study.All specimens were tested HPV by using real time PCR and PCR-RDB,discrepant samples were tested again by PCR-xMAP.Results The concordant rate was 83.5%(91/109)between real time PCR and PCR-RDB (kappa=0.671),the other 18 discrepant samples were retested by PCR-xMAP,7 of those were identical with real time PCR and 11 with PCR-RDB.No differences of PCR-RDB pos-itive rates were found between the high and low viral load groups (χ2 =1.476,P =0.224).Conclusion It demonstrated moderate consistency between real time PCR and PCR-RDB.The HPV positive rates of PCR-RDB were stable when the viral loads were 103-108 .

  7. Automated design of genomic Southern blot probes

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    Komiyama Noboru H


    Full Text Available Abstract Background Sothern blotting is a DNA analysis technique that has found widespread application in molecular biology. It has been used for gene discovery and mapping and has diagnostic and forensic applications, including mutation detection in patient samples and DNA fingerprinting in criminal investigations. Southern blotting has been employed as the definitive method for detecting transgene integration, and successful homologous recombination in gene targeting experiments. The technique employs a labeled DNA probe to detect a specific DNA sequence in a complex DNA sample that has been separated by restriction-digest and gel electrophoresis. Critically for the technique to succeed the probe must be unique to the target locus so as not to cross-hybridize to other endogenous DNA within the sample. Investigators routinely employ a manual approach to probe design. A genome browser is used to extract DNA sequence from the locus of interest, which is searched against the target genome using a BLAST-like tool. Ideally a single perfect match is obtained to the target, with little cross-reactivity caused by homologous DNA sequence present in the genome and/or repetitive and low-complexity elements in the candidate probe. This is a labor intensive process often requiring several attempts to find a suitable probe for laboratory testing. Results We have written an informatic pipeline to automatically design genomic Sothern blot probes that specifically attempts to optimize the resultant probe, employing a brute-force strategy of generating many candidate probes of acceptable length in the user-specified design window, searching all against the target genome, then scoring and ranking the candidates by uniqueness and repetitive DNA element content. Using these in silico measures we can automatically design probes that we predict to perform as well, or better, than our previous manual designs, while considerably reducing design time. We went on to

  8. Molecular Typing of Mycobacterium tuberculosis Based on Variable Number of Tandem DNA Repeats Used Alone and in Association with Spoligotyping (United States)

    Filliol, Ingrid; Ferdinand, Severine; Negroni, Laetitia; Sola, Christophe; Rastogi, Nalin


    Fingerprinting based on variable numbers of tandem DNA repeats (VNTR), a recently described methodology, was evaluated for molecular typing of Mycobacterium tuberculosis in an insular setting. In this study, VNTR fingerprinting was used alone or as a second-line test in association with spoligotyping, double-repetitive-element PCR (DRE-PCR), and IS6110 restriction fragment length polymorphism (RFLP) analysis, and the discriminatory power for each method or the combination of methods was compared by calculating the Hunter-Gaston discriminative index (HGI). The results obtained showed that in 6 out of 12 (50%) cases, VNTR-defined clusters were further subdivided by spoligotyping, compared to 7 out of 18 (39%) cases where spoligotyping-defined clusters were further subdivided by VNTR. When used alone, VNTR was the least discriminatory method (HGI = 0.863). Although VNTR was significantly more discriminatory when used in association with spoligotyping (HGI = 0.982), the combination of spoligotyping and DRE-PCR (HGI = 0.992) was still the most efficient among rapid, PCR-based methodologies, giving results comparable to IS6110 RFLP analysis. Nonetheless, VNTR typing may provide additional phylogenetical information that may be helpful to trace the molecular evolution of tubercle bacilli. PMID:10878036

  9. Spoligotype profile of Mycobacterium tuberculosis complex strains from HIV-positive and -negative patients in Nigeria: a comparative analysis.

    NARCIS (Netherlands)

    Cadmus, S.; Hill, V.; Soolingen, D. van; Rastogi, N.


    We ran a comparative analysis of all patients for whom a positive culture of Mycobacterium tuberculosis complex was available between April 2004 and October 2005 and whose HIV serology results were known, with spoligotyping results (n = 163) split into 49 HIV-positive patients and 114 HIV-negative p

  10. Genotypic characterization by spoligotyping and VNTR typing of Mycobacterium bovis and Mycobacterium caprae isolates from cattle of Tunisia. (United States)

    Lamine-Khemiri, Hela; Martínez, Remigio; García-Jiménez, Waldo Luis; Benítez-Medina, Jose Manuel; Cortés, Maria; Hurtado, Inés; Abassi, Mohammed Salah; Khazri, Imed; Benzarti, Mohammed; Hermoso-de-Mendoza, Javier


    This work is an approach to the molecular epidemiology of Mycobacterium tuberculosis complex (MTBC) bovine infections in Tunisia. A total of 35 MTBC isolates from both lateral retropharyngeal lymph node samples of cattle slaughtered in different Tunisian regions were genotyped by spoligotyping and variable number tandem repeat typing (VNTR)-typing. Spoligotyping allowed to identify two profiles not previously registered, namely SB2024, a Mycobacterium caprae isolate from Nabeul Region (North East Tunisia), the first description of this species in the country, and SB2025 (Mycobacterium bovis) from Sfax Region (Southern Tunisia). A second M. caprae isolate with a spoligotyping profile previously described in Europe mainland, SB0418, was also isolated from a bovine of Sfax region. Both isolates suggest the possibility of a widespread distribution of this species in the country. The predominant spoligotype was SB0120, present in all Tunisian regions selected for the study but Nabeul. Molecular typing also allowed to describe a mixed infection caused by two different M. bovis isolates (SB0120 and SB0848) in the same animal. VNTR typing was highly discriminant by testing a panel of six loci. Loci QUB3232 and QUB11b were the most discriminant, whereas ETR-D and QUB11a had the lower diversity index. The value of allelic diversity can significantly vary among countries; thus, it is important to standardize a panel of loci for future inter-laboratory comparisons. Although VNTR typing proved to be useful for an efficient discrimination among MTBC isolates, especially in combination with spoligotyping, further studies are needed in order to assess the genetic diversity of the MTBC in Tunisia.

  11. Genotyping of Mycobacterium tuberculosis Isolates from Hormozgan Province of Iran Based on 15-Locus MIRU-VNTR and Spoligotyping (United States)

    Zamani, Samin; Haeili, Mehri


    Background. Considering that Hormozgan province in Iran (southern part of Iran on the Persian Gulf) is among the areas with high prevalence of MDR-MTB and attracts so many sailors and tourists, genetic diversity of MTB isolates circulating in this part of Iran was evaluated. Pattern of TB transmission was also examined. Methods and Material. A total of 38 isolates of MTB were cultured from TB patients from Hormozgan province of Iran and standard MIRU-VNTR typing and spoligotyping were applied to genotype these isolates. Drug susceptibility testing was performed using proportion method. Results. There were 28 VNTR profiles comprising 5 clusters and 23 unique isolates compared to 21 spoligotyping profiles, which contained 9 clusters and 12 unique isolates. Latin American-Mediterranean (n = 9, 23.6%) was found to be the most predominant lineage. MIRU-VNTR analysis, with an HGDI of 0.975, was more discriminating than spoligotyping, which had an HGDI of 0.955. The estimated proportion of TB cases due to recent transmission was 26.3% and 44.7% by MIRU-VNTR and spoligotyping, respectively. The rates of monodrug resistance and MDR were 15.8% and 7.9%, respectively. Two of 3 MDR strains were found to be related to MIRU-VNTR and belonged to the same spoligotyping cluster characterized with T1/SIT53 genotype. Conclusions. The high genetic diversity among MTB isolates suggests that transmission occurred from different sources to this area. Reactivation of a priori, latent MTB infection was found to contribute mainly to TB cases in this geographic region. PMID:27819023

  12. Usefulness of restriction fragment length polymorphism and spoligotyping for epidemiological studies of Mycobacterium bovis in Madagascar: description of new genotypes. (United States)

    Rasolofo Razanamparany, Voahangy; Quirin, René; Rapaoliarijaona, Andriampenomaka; Rakotoaritahina, Hugues; Vololonirina, Elie Jeanne; Rasolonavalona, Tiana; Ferdinand, Séverine; Sola, Christophe; Rastogi, Nalin; Ramarokoto, Herimanana; Chanteau, Suzanne


    Tuberculosis is highly prevalent in cattle in Madagascar. An epidemiological study based on genotyping of Mycobacterium bovis and its transmission to humans was carried out. The restriction fragment length polymorphism (IS6110 and DR markers) and spoligotyping were used to assess the genetic diversity of strains from different regions of Madagascar. One of these strains was isolated from goat, the other strains were isolated from zebu cattle. Nine IS6110 profiles, 20 DR profiles and 12 spoligotypes were obtained. About 90% of all isolates gave a single IS6110 band at about 1.8 kb. Most strains had the same spoligotype. M. bovis strains commonly lack spacers 39-43, and all Malagasy strains also lacked spacers 3-5, 8-10 and 16. This pattern has not been reported elsewhere. DR was the most discriminatory of the three markers. The patterns obtained with the three markers were combined to identify 34 different genotypes, one of which was found in 35% of the strains. No region-specific M. bovis genotype was identified, but the genotyping of 18 M. bovis strains isolated from patients showed that the human and bovine strains were identical, suggesting possible human contamination from zebu cattle.

  13. Spoligotyping of clinical Mycobacterium tuberculosis isolates from the state of Minas Gerais, Brazil

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    Silvana Spíndola de Miranda


    Full Text Available We performed spoligotyping on 114 strains of the Mycobacterium tuberculosis (Mtb complex that had been isolated from patients in Minas Gerais Health Units during 2004. A total of 82/114 (72% clinical isolates were clustered and 32/114 (28% were unique. Seven shared types containing nine strains were newly created. A total of nine patterns corresponded to unreported orphan strains, as evaluated against all of the strains recorded in the SITVIT2 proprietary database in the Institut Pasteur de la Guadeloupe. The major clades were composed of isolates that belong to the following genotypes: Latin-America and Mediterranean (63/114, 55.3% (the ill-defined T superfamily (12/114, 10.5%, Haarlem (8/114, 7%, X clade (6/114, 5.3%, S clade (3/114, 2.6% and the East-African Indian and Manu types, each with 1/114 (0.9% isolates. A considerable number of strains (n = 20, 17.5% showed patterns that did not fall within any of the previously described major clades. We conclude the bulk of tuberculosis (TB (92/114, 80.7% in our location is recent evolutionary strains that belong to the principal genetic groups 2/3. Further studies on epidemiology of TB are required to understand Mtb biodiversity and TB transmission in this region.

  14. The study of multiple RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses%多重RT-PCR结合反向杂交技术检测多种流感病毒方法的研究

    Institute of Scientific and Technical Information of China (English)

    杨亮; 钱永华; 曾毅; 张晓梅; 张晓光; 马晶; 王敏; 温乐英; 王大燕; 白天; 舒跃龙


    目的 建立一种基于多重RT-PCR结合反向斑点杂交技术的多种流感病毒核酸快速检测方法.方法 对5种亚型流感病毒的HA基因序列进行比对分析,选择合适的保守区域设计引物序列,并在扩增产物序列内部选择高变区设计探针序列.首先,使用生物素标记的引物扩增出同样带标记的PCR产物;然后,产物与膜上的探针杂交,通过生物素-亲和素反应,最后以斑点显色的方式来检测和鉴别不同亚型的流感病毒.实验中还通过摸索不同的引物浓度和探针浓度来优化反应条件.结果 成功建立了一种基于多重RT-PCR结合反向斑点杂交技术的检测方法,该方法能够成功检测和鉴别5种不同亚型的流感病毒,检测灵敏度比传统的RT-PCR方法高100~1000倍.结论 成功建立了一种可高通量快速检测多种流感病毒的多重RT-PCR结合反向斑点杂交技术的检测方法.%Objective To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses. Methods Obtain the HA nucleotide sequences of seasonal influenza H1 N1, seasonal influenza H3N2, influenza H1 N1 and human avian influenza H5 N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance. Results Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more

  15. Studying protein-protein interactions via blot overlay/far western blot. (United States)

    Hall, Randy A


    Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or simply visualized with a specific antibody. When the probe is visualized via antibody detection, this technique is often referred to as "Far Western blot." Many different kinds of protein-protein interactions can be studied via blot overlay, and the method is applicable to screens for unknown protein-protein interactions as well as to the detailed characterization of known interactions.

  16. Single cell-resolution western blotting. (United States)

    Kang, Chi-Chih; Yamauchi, Kevin A; Vlassakis, Julea; Sinkala, Elly; Duncombe, Todd A; Herr, Amy E


    This protocol describes how to perform western blotting on individual cells to measure cell-to-cell variation in protein expression levels and protein state. Like conventional western blotting, single-cell western blotting (scWB) is particularly useful for protein targets that lack selective antibodies (e.g., isoforms) and in cases in which background signal from intact cells is confounding. scWB is performed on a microdevice that comprises an array of microwells molded in a thin layer of a polyacrylamide gel (PAG). The gel layer functions as both a molecular sieving matrix during PAGE and a blotting scaffold during immunoprobing. scWB involves five main stages: (i) gravity settling of cells into microwells; (ii) chemical lysis of cells in each microwell; (iii) PAGE of each single-cell lysate; (iv) exposure of the gel to UV light to blot (immobilize) proteins to the gel matrix; and (v) in-gel immunoprobing of immobilized proteins. Multiplexing can be achieved by probing with antibody cocktails and using antibody stripping/reprobing techniques, enabling detection of 10+ proteins in each cell. We also describe microdevice fabrication for both uniform and pore-gradient microgels. To extend in-gel immunoprobing to gels of small pore size, we describe an optional gel de-cross-linking protocol for more effective introduction of antibodies into the gel layer. Once the microdevice has been fabricated, the assay can be completed in 4-6 h by microfluidic novices and it generates high-selectivity, multiplexed data from single cells. The technique is relevant when direct measurement of proteins in single cells is needed, with applications spanning the fundamental biosciences to applied biomedicine.

  17. TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics. (United States)

    Taki, Takao


    A simple method for transfer of lipids-including phospholipids, glycolipids, and neutral lipids-from a high performance thin layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, TLC-Blot (Far-Eastern Blot), and its biochemical applications are presented. This chapter presents the conventional procedures for separating lipid from tissue samples, cultured cells, and serum and the subsequent development of TLC. Individual lipids separated on an HPTLC plate can be transferred to the PVDF membrane quantitatively and also isolated from the lipid-blotted membrane by a one-step purification procedure. Immunodetection with monoclonal antibodies and treatment with lipid-metabolizing enzymes on the lipid-blotted membrane are possible. The method for identification of individual lipids transferred on the PVDF membrane using matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (TLC-Blot/MALDI-TOF MS) is shown as a functional lipidomics application.

  18. Mycobacterium tuberculosis spoligotypes and drug susceptibility pattern of isolates from tuberculosis patients in South-Western Uganda

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    Bwanga Freddie


    Full Text Available Abstract Background Determination of the prevalence and drug susceptibility of the M. tuberculosis strains is important in tuberculosis control. We determined the genetic diversity and susceptibility profiles of mycobacteria isolated from tuberculosis patients in Mbarara, South Western Uganda. Methods We enrolled, consecutively; all newly diagnosed and previously treated smear-positive TB patients aged ≥ 18 years. The isolates were characterized using regions of difference (RD analysis and spoligotyping. Drug resistance against rifampicin and isoniazid were tested using the Genotype® MDRTBplus assay and the indirect proportion method on Lowenstein-Jensen media. HIV-1 testing was performed using two rapid HIV tests. Results A total of 125 isolates from 167 TB suspects (60% males with a mean age 33.7 years and HIV prevalence of 67.9% (55/81 were analyzed. Majority (92.8% were new cases while only 7.2% were retreatment cases. All the 125 isolates were identified as M. tuberculosis strict sense with the majority (92.8% of the isolates being modern strains while seven (7.2% isolates were ancestral strains. Spoligotyping revealed 79 spoligotype patterns, with an overall diversity of 63.2%. Sixty two (49.6% of the isolates formed 16 clusters consisting of 2-15 isolates each. A majority (59.2% of the isolates belong to the Uganda genotype group of strains. The major shared spoligotypes in our sample were SIT 135 (T2-Uganda with 15 isolates and SIT 128 (T2 with 3 isolates. Sixty nine (87% of the 79 patterns had not yet been defined in the SpolDB4.0.database. Resistance mutations to either RIF or INH were detected in 6.4% of the isolates. Multidrug resistance, INH and RIF resistance was 1.6%, 3.2% and 4.8%, respectively. The rpoβ gene mutations seen in the sample were D516V, S531L, H526Y H526D and D516V, while one strain had a Δ1 mutation in the wild type probes. There were three strains with katG (codon 315 gene mutations only while one strain

  19. Miniaturized fluorescent RNA dot blot method for rapid quantitation of gene expression

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    Yadetie Fekadu


    Full Text Available Abstract Background RNA dot blot hybridization is a commonly used technique for gene expression assays. However, membrane based RNA dot/slot blot hybridization is time consuming, requires large amounts of RNA, and is less suited for parallel assays of more than one gene at a time. Here, we describe a glass-slide based miniaturized RNA dot blot (RNA array procedure for rapid and parallel gene expression analysis using fluorescently labeled probes. Results RNA arrays were prepared by simple manual spotting of RNA onto amino-silane coated microarray glass slides, and used for two-color fluorescent hybridization with specific probes labeled with Cy3 and 18S ribosomal RNA house-keeping gene probe labeled with Cy5 fluorescent dyes. After hybridization, arrays were scanned on a fluorescent microarray scanner and images analyzed using microarray image analysis software. We demonstrate that this method gives comparable results to Northern blot analysis, and enables high throughput quantification of transcripts from nanogram quantities of total RNA in hundreds of samples. Conclusion RNA array on glass slide and detection by fluorescently labeled probes can be used for rapid and parallel gene expression analysis. The method is particularly well suited for gene expression assays that involve quantitation of many transcripts in large numbers of samples.

  20. Molecular characterization of Mycobacterium tuberculosis isolates from Tehran, Iran by restriction fragment length polymorphism analysis and spoligotyping. (United States)

    Feyisa, Seifu Gizaw; Haeili, Mehri; Zahednamazi, Fatemeh; Mosavari, Nader; Taheri, Mohammad Mohammad; Hamzehloo, Gholamreza; Zamani, Samin; Feizabadi, Mohammad Mehdi


    INTRODUCTION Characterization of Mycobacterium tuberculosis (MTB) isolates by DNA fingerprinting has contributed to tuberculosis (TB) control. The aim of this study was to determine the genetic diversity of MTB isolates from Tehran province in Iran. METHODS MTB isolates from 60 Iranian and 10 Afghan TB patients were fingerprinted by standard IS6110-restriction fragment length polymorphism (RFLP) analysis and spoligotyping. RESULTS The copy number of IS6110 ranged from 10-24 per isolate. The isolates were classified into 22 clusters showing ≥ 80% similarity by RFLP analysis. Fourteen multidrug-resistant (MDR) isolates were grouped into 4 IS6110-RFLP clusters, with 10 isolates [71% (95% CI: 45-89%)] in 1 cluster, suggesting a possible epidemiological linkage. Eighteen Iranian isolates showed ≥ 80% similarity with Afghan isolates. There were no strains with identical fingerprints. Spoligotyping of 70 isolates produced 23 distinct patterns. Sixty (85.7%) isolates were grouped into 13 clusters, while the remaining 10 isolates (14.2%) were not clustered. Ural (formerly Haarlem4) (n = 22, 31.4%) was the most common family followed by Central Asian strain (CAS) (n = 18, 25.7%) and T (n = 9, 12.8%) families. Only 1strain was characterized as having the Beijing genotype. Among 60 Iranian and 10 Afghan MTB isolates, 25% (95% CI: 16-37) and 70% (95% CI: 39-89) were categorized as Ural lineage, respectively. CONCLUSIONS A higher prevalence of Ural family MTB isolates among Afghan patients than among Iranian patients suggests the possible transmission of this lineage following the immigration of Afghans to Iran.

  1. Use of a Combined Duplex PCR/Dot Blot Assay for more sensitive genetic characterization

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    Erin Curry


    Full Text Available A reliable and sensitive method of genetic analysis is necessary to detect multiple specific nucleic acid sequences from samples containing limited template. The most widely utilized method of specific gene detection, polymerase chain reaction (PCR, imparts inconsistent results when assessing samples with restricted template, especially in a multiplex reaction when copies of target genes are unequal. This study aimed to compare two methods of PCR product analysis, fluorescent detection following agarose gel electrophoresis or dot blot hybridization with chemiluminescent evaluation, in the detection of a single copy gene (SRY and a multicopy gene (β-actin. Bovine embryo sex determination was employed to exploit the limited DNA template available and the target genes of unequal copies. Primers were used either independently or together in a duplex reaction with purified bovine genomic DNA or DNA isolated from embryos. When used independently, SRY and β-actin products were detected on a gel at the equivalent of 4-cell or 1-cell of DNA, respectively; however, the duplex reaction produced visible SRY bands at the 256 cell DNA equivalent and β-actin products at the 64 cell DNA equivalent. Upon blotting and hybridization of the duplex PCR reaction, product was visible at the 1–4 cell DNA equivalent. Duplex PCR was also conducted on 186 bovine embryos and product was subjected to gel electrophoresis or dot-blot hybridization in duplicate. Using PCR alone, sex determination was not possible for 22.6% of the samples. Using PCR combined with dot blot hybridization, 100.0% of the samples exhibited either both the male specific and β-actin products or the β-actin signal alone, indicating that the reaction worked in all samples. This study demonstrated that PCR amplification followed by dot blot hybridization provided more conclusive results in the evaluation of samples with low DNA concentrations and target genes of unequal copies.

  2. PCR-反向点杂交基因分型与实时荧光定量PCR检测人乳头瘤病毒的研究%Use of a PCR-based reverse blot hybridization assay for subtyping and real-time quantitative PCR to detect human papilloma virus

    Institute of Scientific and Technical Information of China (English)

    向华国; 曾锦婷; 何婉意; 黎国


    Objective To evaluate the significance of a PCR-based reverse blot hybridization (PCR-RDB) assay and realtime quantitative PCR for detecting human papilloma virus in female outpatients. Methods A total of 121 female outpatients were checked for 23 HFV DNA types by PCR-RDB and 13 high-risk HPV genotypes by real-time quantitative PCR. Results According to PCR-RDB, 28.10% of the women(34/121) tested positive while 16. 53%(20/121) tested positive according to real-time quantitative PCR. HPV was detected more often with PCR-RDB than with real-time quantitative PCR (P<0.05). The concordance rate for the two techniques was 93. 39%(113/121). Conclusion PCR-RDB can be used to screen for HPV infection while real-time quantitative PCR facilitates evaluation of the effectiveness of treatment and the prognosis for cervical carcinoma. Combining the two should increase the specificity and sensitivity of HPV detection.%目的 评价PCR-反向点杂交基因分型与实时荧光定量PCR在检测人乳头瘤病毒(HPV)的意义.方法 同时采用PCR-反向点杂交基因分型和实时荧光定量PCR对121例女性官颈脱离细胞标本进行HPV检测.其中PCR-反向点杂交基因分型能检测23种HPV亚型,实时荧光定量PCR定量检测常见的13种高危HPV亚型.结果 PCR-反向点杂交基因分型检测HPV的阳性率为28.10%(34/121),实时荧光定量PCR检测HPV的阳性率为16.53%(20/121),差异有统计学意义(P<0.05);二者检测的符合率为93.39%(113/121).结论 PCR-反向杂交基因分型适用于HPV感染的筛查,而实时荧光定量PCR适用于HPV感染相关疾病的疗效与预后的判断.PCR-反向杂交基因分型与实时荧光定量PCR联合检测可提高HPV检测的特异性和敏感度,对于生殖道HPV感染以及子宫颈癌的早期发现、预防和治疗具有重要意义.

  3. Molecular Typing of Mycobacterium Tuberculosis Complex by 24-Locus Based MIRU-VNTR Typing in Conjunction with Spoligotyping to Assess Genetic Diversity of Strains Circulating in Morocco.

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    Nada Bouklata

    Full Text Available Standard 24-locus Mycobacterial Interspersed Repetitive Unit Variable Number Tandem Repeat (MIRU-VNTR typing allows to get an improved resolution power for tracing TB transmission and predicting different strain (sub lineages in a community.During 2010-2012, a total of 168 Mycobacterium tuberculosis Complex (MTBC isolates were collected by cluster sampling from 10 different Moroccan cities, and centralized by the National Reference Laboratory of Tuberculosis over the study period. All isolates were genotyped using spoligotyping, and a subset of 75 was genotyped using 24-locus based MIRU-VNTR typing, followed by first line drug susceptibility testing. Corresponding strain lineages were predicted using MIRU-VNTRplus database.Spoligotyping resulted in 137 isolates in 18 clusters (2-50 isolates per cluster: clustering rate of 81.54% corresponding to a SIT number in the SITVIT database, while 31(18.45% patterns were unique of which 10 were labelled as "unknown" according to the same database. The most prevalent spoligotype family was LAM; (n = 81 or 48.24% of isolates, dominated by SIT42, n = 49, followed by Haarlem (23.80%, T superfamily (15.47%, >Beijing (2.97%, > U clade (2.38% and S clade (1.19%. Subsequent 24-Locus MIRU-VNTR typing identified 64 unique types and 11 isolates in 5 clusters (2 to 3isolates per cluster, substantially reducing clusters defined by spoligotyping only. The single cluster of three isolates corresponded to two previously treated MDR-TB cases and one new MDR-TB case known to be contact a same index case and belonging to a same family, albeit residing in 3 different administrative regions. MIRU-VNTR loci 4052, 802, 2996, 2163b, 3690, 1955, 424, 2531, 2401 and 960 were highly discriminative in our setting (HGDI >0.6.24-locus MIRU-VNTR typing can substantially improve the resolution of large clusters initially defined by spoligotyping alone and predominating in Morocco, and could therefore be used to better study tuberculosis

  4. Selected Reaction Monitoring (SRM) frente a Western Blot


    Heredia Ponce, Zahira Maria; Urbano Gámez, Jose Alberto


    We propose the SRM technology as a complementary method to the Western Blot for the detection and quantification of proteins in a sample. The technique Western Blot has its own limitations: i) only a protein-of-choice is detected, ignoring any non-relevant proteins, ii) the sensitivity of the technique depends on the specificity of the antibody and iii) Western Blot is expensive and time-consuming. The advantages of SRM with respect Western Blot are remarkable: i) you can detect up to h...


    Hubbard, Katherine; Hegarty, Peter


    Despite the easily recognizable nature of the Rorschach ink blot test very little is known about the history of the test in Britain. We attend to the oft-ignored history of the Rorschach test in Britain and compare it to its history in the US. Prior to the Second World War, Rorschach testing in Britain had attracted advocates and critiques. Afterward, the British Rorschach Forum, a network with a high proportion of women, developed around the Tavistock Institute in London and The Rorschach Newsletter. In 1968, the International Rorschach Congress was held in London but soon after the group became less exclusive, and fell into decline. A comparative account of the Rorschach in Britain demonstrates how different national institutions invested in the 'projective hypothesis' according to the influence of psychoanalysis, the adoption of a nationalized health system, and the social positioning of 'others' throughout the twentieth century. In comparing and contrasting the history of the Rorschach in Britain and the US, we decentralize and particularize the history of North American Psychology.

  6. Fast and Simple micro-RNA Northern Blots

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    Nham Tran


    Full Text Available RNA northern blots provide robust measurements of gene expression. The simple northern blot technique described in this report has been optimised to provide rapid, reproducible detection and analysis of mature and precursor forms of microRNAs. This protocol economises on the use of commercially available components and secondly reduces the hybridisation step to 2 hours.

  7. The Design of a Quantitative Western Blot Experiment

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    Sean C. Taylor


    Full Text Available Western blotting is a technique that has been in practice for more than three decades that began as a means of detecting a protein target in a complex sample. Although there have been significant advances in both the imaging and reagent technologies to improve sensitivity, dynamic range of detection, and the applicability of multiplexed target detection, the basic technique has remained essentially unchanged. In the past, western blotting was used simply to detect a specific target protein in a complex mixture, but now journal editors and reviewers are requesting the quantitative interpretation of western blot data in terms of fold changes in protein expression between samples. The calculations are based on the differential densitometry of the associated chemiluminescent and/or fluorescent signals from the blots and this now requires a fundamental shift in the experimental methodology, acquisition, and interpretation of the data. We have recently published an updated approach to produce quantitative densitometric data from western blots (Taylor et al., 2013 and here we summarize the complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting.

  8. Multidrug-Resistant Mycobacterium tuberculosis of the Latin American Mediterranean Lineage, Wrongly Identified as Mycobacterium pinnipedii (Spoligotype International Type 863 [SIT863]), Causing Active Tuberculosis in South Brazil

    KAUST Repository

    Dalla Costa, Elis R.


    We recently detected the spoligotype patterns of strains of Mycobacterium pinnipedii, a species of the Mycobacterium tuberculosis complex, in sputum samples from nine cases with pulmonary tuberculosis residing in Porto Alegre, South Brazil. Because this species is rarely encountered in humans, we further characterized these nine isolates by additional genotyping techniques, including 24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) typing, verification of the loci TbD1, RD9, pks15/1, RDRio, and fbpC, the insertion of IS6110 at a site specific to the M. tuberculosis Latin American Mediterranean (LAM) lineage, and whole-genome sequencing. The combined analysis of these markers revealed that the isolates are in fact M. tuberculosis and more specifically belong to the LAM genotype. Most of these isolates (n = 8) were shown to be multidrug resistant (MDR), which prompted us to perform partial sequencing of the rpoA, rpoB, rpoC, katG, and inhA genes. Seven isolates (77.8%) carried the S315T mutation in katG, and one of these (11%) also presented the C(−17)T single-nucleotide polymorphism (SNP) in inhA. Interestingly, six of the MDR isolates also presented an undescribed insertion of 12 nucleotides (CCA GAA CAA CCC) in codon 516 of rpoB. No putative compensatory mutation was found in either rpoA or rpoC. This is the first report of an M. tuberculosis LAM family strain with a convergent M. pinnipedii spoligotype. These spoligotypes are observed in genotype databases at a modest frequency, highlighting that care must be taken when identifying isolates in the M. tuberculosis complex on the basis of single genetic markers.

  9. Different domains of Bacillus thuringiensis delta-endotoxins can bind to insect midgut membrane proteins on ligand blots

    NARCIS (Netherlands)

    Maagd, de R.A.; Klei, van der H.; Bakker, P.L.; Stiekema, W.J.; Bosch, D.


    We investigated the role of the constituent domains of the CryIA(b) and CryIA(c) δ-endotoxins in binding to midgut epithelial cell membrane proteins of Spodoptera exigua and Manduca sexta on ligand blots. A collection of wild- type and CryIC-CryIA hybrid toxins was used for this purpose. As demonstr

  10. Mixed lubrication after rewetting of blotted pleural mesothelium. (United States)

    Bodega, Francesca; Sironi, Chiara; Porta, Cristina; Pecchiari, Matteo; Zocchi, Luciano; Agostoni, Emilio


    Coefficient of kinetic friction (μ) of pleural mesothelium blotted with filter paper, and rewetted with Ringer solution markedly increases; this increase is removed if a sufficient amount of sialomucin or hyaluronan is added to Ringer (Bodega et al., 2012. Respiratory Physiology and Neurobiology 180, 34-39). In this research we found that μ of pleural mesothelium blotted, rewetted, and sliding at physiological velocities and loads, decreased with increase of velocity, mainly at low velocities. Despite this decrease, μ at highest velocity was still double that before blotting. With small concentration of sialomucin or hyaluronan μ was markedly smaller at each velocity, decreased less with increase of velocity, and at highest velocity approached preblotting value. These findings indicate a regime of mixed lubrication in post-blotting Ringer, at variance with boundary lubrication occurring before blotting or postblotting with sufficient macromolecule addition. Greater roughness of mesothelial surface, caused by blotting, likely induces zones of elastohydrodynamic lubrication, which increase with velocity, while contact area decreases.

  11. The line blot assay: problems with titrating first and second antibodies for Western blot and immunohistochemistry assays? (United States)

    Rojas-Espinosa, O; Silva-Miranda, M; Wek-Rodriguez, K; Arce-Paredes, P


    We describe a technique designed to assess the optimal dilution of primary and secondary antibodies, to be used in Western blot, dot blot, the multi-antigen print immunoassay (MAPIA) and immunohistochemistry assays. The method that we call "line blot" is not an alternative but a practical, complementary tool for the above techniques that assures definitive results are obtained from single assays, so there is no need to repeat the assay. As with most immunoenzymatic assays, the line blot assay is very sensitive, allowing the detection of absolute amounts of antigen as low as 2.5 ng in the 0.5 cm-long segment line (see Results), depending on the strength of the secondary, enzyme-labelled antibody.

  12. Routine Western blot to check autophagic flux : Cautions and recommendations

    NARCIS (Netherlands)

    Gomez-Sanchez, Ruben; Pizarro-Estrella, Elisa; Yakhine-Diop, Sokhna M. S.; Rodriguez-Arribas, Mario; Bravo-San Pedro, Jose M.; Fuentes, Jose M.; Gonzalez-Polo, Rosa A.


    At present, the analysis of autophagic flux by Western blotting (WB), which measures two of the most important markers of autophagy, i.e., microtubule-associated protein 1 light chain 3 (LC3) and p62, is widely accepted in the scientific community. In this study, we addressed the possible disadvanta

  13. Finer characterization of Mycobacterium tuberculosis using spoligotyping and 15-loci MIRU-VNTRs reveals phylogeographical specificities of isolates circulating in Guyana and Suriname. (United States)

    Streit, Elisabeth; Baboolal, Shirematee; Akpaka, Patrick E; Millet, Julie; Rastogi, Nalin


    In this study we used spoligotyping and 15-loci MIRU-VNTRs for a finer characterization of Mycobacterium tuberculosis strains isolated from patients residing in Guyana (n=74) and Suriname (n=80). The mean age of the patients was 38.5years (36.5 and 40.2years for Guyana vs. Suriname), with a male-to-female sex-ratio of 2.25 for Guyana vs. 4.27 in Suriname. Spoligotyping and 15-loci MIRU-VNTRs led to a total of 41 and 65 different patterns respectively, with an overall clustering rate of 83.8% vs. 68.8%. Combined spoligotyping and VNTR analysis led to the detection of 18 clusters of 2-41 isolates, with an overall clustering of 67.5% and a recent "n-1" transmission rate of 55.8%. Importantly, Guyana was characterized by a significantly higher percentage of clustered isolates than Suriname (79.7% vs. 56.3%; p=0.0019). Three big spoligo/MIRU (SIT/MIT) clusters containing >10 isolates each were shared between the 2 countries, and concerned: T1 sublineage cluster 53/861 (n=41, 37 in Guyana vs. 4 in Suriname); EAI6-BGD1 sublineage cluster 1340/860 (n=16, 3 in Guyana vs. 13 in Suriname); and T1 sublineage cluster 131/146 (n=11, 6 in Guyana vs. 5 in Suriname); as well as 2 smaller clusters of 2 and 3 isolates respectively. However, the relative phylogeographical specificities of strains in circulation as well as a lack of drug-resistance observed among strains from Suriname suggested that trans-border transmission of drug-resistant isolates occurred less frequently than thought. Tracing and interrupting transmission channels of a specific clone (SIT53/15-MIT861) should become a priority in Guyana, not only because it is by far most abundant but also because it accounts for almost half of the drug resistant isolates (n=8/17, 47.1%) in our study, and clustered 5/12 (41.7%) MDR isolates.

  14. Use of cluster-graphs from spoligotyping data to study genotype similarities and a comparison of three indices to quantify recent tuberculosis transmission among culture positive cases in French Guiana during a eight year period

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    Brudey Karine


    Full Text Available Abstract Background French Guiana has the highest tuberculosis (TB burden among all French departments, with a strong increase in the TB incidence over the last few years. It is now uncertain how best to explain this incidence. The objective of this study was to compare three different methods evaluating the extent of recent TB transmission in French Guiana. Methods We conducted a population-based molecular epidemiology study of tuberculosis in French Guiana based on culture-positive TB strains (1996 to 2003, n = 344 to define molecular relatedness between isolates, i.e. potential transmission events. Phylogenetic relationships were inferred by comparing two methods: a "cluster-graph" method based on spoligotyping results, and a minimum spanning tree method based on both spoligotyping and variable number of tandem DNA repeats (VNTR. Furthermore, three indices attempting to reflect the extent of recent TB transmission (RTIn, RTIn-1 and TMI were compared. Results Molecular analyses showed a total amount of 120 different spoligotyping patterns and 273 clinical isolates (79.4% that were grouped in 49 clusters. The comparison of spoligotypes from French Guiana with an international spoligotype database (SpolDB4 showed that the majority of isolates belonged to major clades of M. tuberculosis (Haarlem, 22.6%; Latin American-Mediterranean, 23.3%; and T, 32.6%. Indices designed to quantify transmission of tuberculosis gave the following values: RTIn = 0.794, RTIn-1 = 0.651, and TMI = 0.146. Conclusion Our data showed a high number of Mycobacterium tuberculosis clusters, suggesting a high level of recent TB transmission, nonetheless an estimation of transmission rate taking into account cluster size and mutation rate of genetic markers showed a low ongoing transmission rate (14.6%. Our results indicate an endemic mode of TB transmission in French Guiana, with both resurgence of old spatially restricted genotypes, and a significant importation of new TB

  15. Fluorescent detection of Southern blots and PCR-based genetic typing tests

    Energy Technology Data Exchange (ETDEWEB)

    Mansfield, E.S.; Worley, J.M. [Molecular Dynamics, Inc., Sunnyvale, CA (United States); Zimmerman, P.A. [Laboratory of Parasitic Diseases, Bethesda, MD (United States)] [and others


    The Southern blot is used to study gene organization, to identify disease-causing genomic rearrangements, or for typing RFLP markers in forensic, paternity, or prenatal diagnostic testing. Fluorescence offers a much greater dynamic range and a more linear response than film used in radioactive or chemiluminescent detection of RFLPs. We therefore investigated using the Fluorimager{trademark} 575 (Molecular Dynamics, Inc.) for analyzing Southern blots. Using a single-locus probe to D2S44 (YNH24) (Promega Corp.), we detect as little as 100 ng (0.05 attomole) genomic DNA. The alkaline phosphatase-labeled probe is detected using AttoPhos (JBL Scientific), and the developed membrane is scanned with the Fluorimager. Biotinylated hybridization probes can also be developed using a streptavidin-alkaline phosphatase conjugate and AttoPhos. The instrument scan parameters can be adjusted to prevent overexposure and accompanying loss of resolution in images of blots, gels, or 96-well microplates. We have used these other sample formats in PCR-based genetic typing assays. We use FluorKit DQS (Molecular Dynamics) to accurately quantify PCR template DNA (1-500 ng) in 96-well microplates scanned using the same instrument. Mutation detection assays run include heteroduplex gels (5% polyacrylamide, 2.7 M urea), short tandem repeat (STR) markers, amplified fragment length polymorphisms (AmpFLP), competitive priming PCR, and allele-specific oligotyping. These assays are run using either 1- or 2-color labeling. We detect unlabeled PCR products, such as the AmpFLP marker D1S80 (Perkin-Elmer) by post-staining gels for 10 minutes with SYBR Green 1 (Molecular Probes) and scanning the wet gel. The Fluorimager scans a 20 x 25 cm sample within three minutes, allowing rapid optimization of fluorescent protocols and high sample throughput.

  16. Determination of Diagnostic Antigens in Cattle Amphistomiasis Using Western Blotting

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    A Halajian


    Full Text Available "nBackground: Mixed infection with amphistomes seems common in native cattle of Iran. The aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis."nMethods: Specific antigens of Cotylophoron cotylophorum, Gastrothylax crumenifer and Paramphisto­mum cervi (mixed infection, the most common species, were collected from cattle was deter­mined. Adult trematodes were collected from the rumen of naturally infected cattle at meat inspec­tion. After their homogenization and centrifugation, somatic antigens were prepared and ana­lyzed by SDS-PAGE. Specific antigens were determinated by western blot with homologous and heterolo­gous sera. SDS-PAGE of whole worms extract was performed at different concentrations and subse­quent gels staining. Immunoblotting analysis using sera from cattle naturally infected with am­phistomes, Dicrocoelium dendriticum, Fasciola spp. and hydatid cyst was performed."nResults: Electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 µgr/ml and 8 µgr/ml with molecular weights ranging from 25-120 and 25-150 kDa, respectively. The best result was taken at 8 mg/ml concentration. Although western blot of these proteins demon­strate 5 major antigenic polypeptides ranging from 50 to 100 kDa which were recognized by serum of cat­tle naturally infected with mixed amphistomes.

  17. Simultaneous preparation of RNA and nuclei for Northern blot and flow cytometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Tesfaigzi, J.; Jaramillo, R. [Inhalation Toxicology Research Inst., Albuquerque, NM (United States)


    Several methods have been developed to quantify RNA synthesis during the progression of the cell cycle. The rate of RNA synthesis can be detected during different stages of the cell cycle by staining cells with agents that intercalate with nucleic acids. For example, following staining of mammalian cells with acridin orange, the green and red fluorescence that correlates with DNA and RNA content, respectively, can be analyzed by flow cytometry. Increase in RNA content during the progression of cells through the cell cycle can be measured after staining with acridin orange. RNA synthesis resulting from the stimulation of quiescent cells with various growth factors has also been demonstrated by labeling cells with bromo-uridine and using the anti-bromo-deoxyuridine antibody. These methods allow measurement of the overall RNA content in cells; however, they do not allow the measurement of the levels of specific mRNAs throughout the cell cycle. Current methods to quantify specific mRNAs generally require the preparation of a large number of cells (5--10 {times} 10{sup 6} cells) to carry out flow cytometric analyses and to isolate RNA for Northern blot analysis or solution hybridization. In this report, the authors describe a method of simultaneously preparing RNA and nuclei for Northern blot and flow cytometric analyses, respectively. The minimum number of nuclei required to obtain flow cytometric data and the effect of conserving nuclei in methanol for several days are also presented.

  18. Antibody performance in western blot applications is context-dependent. (United States)

    Algenäs, Cajsa; Agaton, Charlotta; Fagerberg, Linn; Asplund, Anna; Björling, Lisa; Björling, Erik; Kampf, Caroline; Lundberg, Emma; Nilsson, Peter; Persson, Anja; Wester, Kenneth; Pontén, Fredrik; Wernérus, Henrik; Uhlén, Mathias; Ottosson Takanen, Jenny; Hober, Sophia


    An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13 000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data.

  19. Cy5 total protein normalization in Western blot analysis. (United States)

    Hagner-McWhirter, Åsa; Laurin, Ylva; Larsson, Anita; Bjerneld, Erik J; Rönn, Ola


    Western blotting is a widely used method for analyzing specific target proteins in complex protein samples. Housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. We present a new, more reliable method for normalization using Cy5-prelabeled total protein as a loading control. We used a prelabeling protocol based on Cy5 N-hydroxysuccinimide ester labeling that produces a linear signal response. We obtained a low coefficient of variation (CV) of 7% between the ratio of extracellular signal-regulated kinase (ERK1/2) target to Cy5 total protein control signals over the whole loading range from 2.5 to 20.0μg of Chinese hamster ovary cell lysate protein. Corresponding experiments using actin or tubulin as controls for normalization resulted in CVs of 13 and 18%, respectively. Glyceraldehyde-3-phosphate dehydrogenase did not produce a proportional signal and was not suitable for normalization in these cells. A comparison of ERK1/2 signals from labeled and unlabeled samples showed that Cy5 prelabeling did not affect antibody binding. By using total protein normalization we analyzed PP2A and Smad2/3 levels with high confidence.

  20. Characterization of Nora Virus Structural Proteins via Western Blot Analysis

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    Brad L. Ericson


    Full Text Available Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs. The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles and ~1.33 g/mL (complete virions. Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses.

  1. The Recent-Transmission of Mycobacterium tuberculosis Strains among Iranian and Afghan Relapse Cases: a DNA-fingerprinting using RFLP and spoligotyping

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    Zarifi Abolhasan


    Full Text Available Abstract Background Relapse of tuberculosis (TB may develop as the result of reactivation of the endogenous primary infection, or as a result of a exogenous reinfection. This survey evaluated the rate of reactivation versus recent transmission among Iranian and Afghan relapse cases. Methods The sputum specimens were digested, examined microscopically for acid-fast bacilli, and inoculated into Löwenstein-Jensen slants by standard procedures. Thereafter, the susceptibility and identification tests were performed on culture positive specimens. Subsequently, the strains that were identified as Mycobacterium tuberculosis (258 isolates were subjected to IS6110 restriction fragment length polymorphism (RFLP and spoligotyping. Additional patient's information was collected for further epidemiological analysis. Patients whose isolates had identical genotyping patterns were considered a cluster with recent transmission episode. Results Out of 258 available isolates, 72(28% had multi-drug resistant (MDR-TB in ratio and 42 (16.2% had other resistant. Notably, 38 of MDR-TB cases (52% were isolated from Afghan patients. By IS6110-RFLP typing method, 65 patients (25% were clustered in 29 clusters. In cluster cases, the intra-community transmissions between Iranian and Afghan patients were 41%. All MDR-TB patients in clusters had either Haarlem I or Beijing characteristic. The risk factors like sex, family history, close contact, living condition, PPD test result and site of TB infection were not associated with clustering. Although, the MDR-TB strains were more frequent in non-cluster cases (31% than cluster one(18% (P M. tuberculosis strains isolated from non-cluster cases were belong to EAI3 (51; 30% and CASI(32;18.6% superfamilies. Conclusion During the studied period, reactivation of a previous infection remain the more probable cause of recurrence. Although, the evidence of intra- community transmission between Iranian and Afghan TB cases, highlighted

  2. Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

    Institute of Scientific and Technical Information of China (English)

    Jian-Ming Xing; Su Zhang; Ying Du; Dan Bi; Li-Hui Yao


    AIM:To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples.METHODS:The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7,Clostridium botulinum, Bacillus cereus,Clostridium perfringens, Vibrio parahaemolyticus,Shigella spp., Yersinia enterocolitica, Vibrio cholerae,Listeria monocytogenes and Staphylococcus aureus.The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay.RESULTS:The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%.CONCLUSION:The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.


    Institute of Scientific and Technical Information of China (English)

    LIU Lun-xu; ZHOU Qing-hua; SHI Ying-kang; QIN Yang; SUN Zhi-lin; SUN Ze-fang


    Objective: To investigate the role of nm23 gene expression in human lung cancer. Methods: Forty human lung cancer tissues and 19 non-cancer pulmonary tissues were studied for their nm23-H1 and nm23-H2 mRNA expression with non-radioactive Northern blot hybridization. The correlation of nm23 mRNA expression with clinical features of lung cancer was analyzed. Results: The mRNA expression of nm23-H2 gene in poorly differentiated squamous cell carcinoma was significantly decreased compared to that in moderate-high differentiated squamous cell carcinoma. The mRNA expression of nm23-H1 and nm23-H2 gene in small cell lung cancer was significantly decreased compared to that in squamous cell carcinoma. No significant difference in nm23 mRNA expression was observed between lung cancer with and without lymph node metastasis, nor was there significant difference between tumor stage. Conclusion: The mRNA expression of nm23 gene is correlated with the degree of differentiation of lung cancer, but there is no evidence of metastasis suppression effect by nm23 gene.

  4. RT-PCR and Northern blot analysis in search for a putative Paramecium beta-adrenergic receptor. (United States)

    Płatek, A; Wiejak, J; Wyroba, E


    RT-PCR and Northern blot analysis were performed in order to search for a putative beta-adrenergic receptor (beta-AR) in Paramecium using several beta2-adrenergic-specific molecular probes. Under strictly defined RT-PCR conditions DNA species of expected molecular size about 360 bp were generated with the primers corresponding to the universal mammalian beta2-AR sequence tagged sites (located within the 4th and the 6th transmembrane regions of the receptor). This RT-PCR product hybridized in Southern blot analysis with the oligonucleotide probe designed to the highly conservative beta2-AR region involved in G-proteins interaction and located within the amplified region. Northern hybridization was performed on Paramecium total RNA and mRNA with human beta2-AR cDNA and two oligonucleotide probes: the first included Phe 290 involved in agonist binding (Strader et al., 1995) and the second was the backward RT-PCR primer. All these probes revealed the presence of about 2 kb mRNA which is consistent with the size of beta2-AR transcripts found in higher eukaryotes.

  5. RNase protection assays and RNA gel blots: a direct comparison of sensitivity. (United States)

    Higgs, D C; Colbert, J T


    RNase protection assays are commonly thought to be a more sensitive means of detecting and quantitating specific mRNAs than are RNA gel blots (Northern blots). We have directly compared the sensitivity of these two approaches by assaying for known amounts of in vitro synthesized beta-glucuronidase mRNA. With the probes and protocols employed here, the ability to detect a specific mRNA was similar whether RNase protection or RNA gel blot analyses were performed.

  6. Evaluation of an indigenous western blot kit for human immunodeficiency virus

    Directory of Open Access Journals (Sweden)

    Lakshmi V


    Full Text Available PURPOSE: The Western Blot test is considered a gold standard test for the confirmation of an ELISA and/or rapid assay screened reactive sample in the diagnosis of HIV infection, especially in the low risk population. In this study, an indigenously developed HIV W. Blot kit (J.Mitra & Co., New Delhi, India was compared for its performance characteristics with a widely used Western Blot kit, HIV Blot 2.2 (Genelabs, Singapore. Antigens of both HIV-1 and the indicator antigen gp36 of HIV-2 are included in the strips. METHODS: A panel of 150 clinical serum samples was used in the evaluation. All the sera were tested simultaneously by both the kits. RESULTS: The HIV W. Blot kit had high performance characteristics (100% sensitivity and 100% specificity, like the HIV Blot 2.2. The test procedure was easy to perform. There was clear delineation of the bands. CONCLUSIONS: The interpretation of the results on the HIV W. Blot was less prone to subjective errors. The test gave positive bands at even very high serum dilutions in the test kit. This fact indicates that HIV W. Blot probably has a potential application in early phases of infection, when the antibody concentrations are still very low.

  7. Immunochemische detectiemethoden na western blotting van cytochroom P-450 iso-enzymen

    NARCIS (Netherlands)

    Laan CA; Jansen EHJM


    In this report a number of staining techniques on Western blots have been compared with respect to sensitivity, background staining, practical applicability and cost aspects. After electrophoresis of a rat microsomal liver sample followed by blotting, an incubation was performed of a primary antibo

  8. Zinc Blotting Assay for Detection of Zinc-Binding Prolamin in Barley (Hordeum vulgare) Grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Langkilde, Ane; Vincze, Éva


    In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol-soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. 65Zn overlay and blotting techniques have been widely used for detecting zinc......-binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (65ZnCl2) blotting technique was optimized to detect zinc-binding prolamins, followed by development of an easy-to-follow nonradioactive colorimetric...... zinc blotting method with a zinc-sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS-PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc-binding specificity of certain proteins was detected either by autoradiography or color...

  9. GTP-blot analysis of small GTP-binding proteins. The C-terminus is involved in renaturation of blotted proteins. (United States)

    Klinz, F J


    Recombinant c-Ha-ras, ralA and rap2, but not rap1A or rap1B proteins retained their ability to bind [alpha-32P]GTP after SDS/PAGE and transfer to nitrocellulose. Recombinant c-Has-ras missing the C-terminal 23 amino acid residues failed to bind [alpha-32P]GTP after the blot, and the ability of recombinant ralA missing the C-terminal 28 amino acid residues to bind [alpha-32P]GTP was decreased many-fold. The presence of nonionic detergents of the polyoxyethylene type such as Tween 20, Triton X-100, Nonidet P40 or Lubrol PX in the incubation buffer was necessary to induce renaturation of blotted recombinant c-Ha-ras protein, whereas other types of detergents were ineffective. We propose that detergents of the polyoxyethylene type induce the refolding of some types of blotted small GTP-binding proteins and that the C-terminus is involved in the refolding process. Membranes from NIH3T3 fibroblasts overexpressing c-Ha-ras protein showed much weaker binding of [alpha-32P]GTP as expected from the level of ras immunoreactivity. Treatment of fibroblasts with lovastatin, an inhibitor of hydroxymethylglutaryl-coenzyme A reductase, caused the accumulation of the unfarnesylated form of c-Ha-ras in the cytosol. Examination of [alpha-32P]GTP-binding and immunoreactivity for cytosolic and membrane-bound c-Ha-ras revealed that binding of [alpha-32P]GTP to unprocessed c-Ha-ras was increased about threefold compared to the same amount of processed c-Ha-ras. Our results demonstrate that detection and quantification of small GTP-binding proteins in eukaryotic cells by GTP-blot analysis is hampered by the fact that these proteins differ strongly in their ability to renature after blotting to nitrocellulose.

  10. First insight into genetic diversity of the Mycobacterium tuberculosis complex in Albania obtained by multilocus variable-number tandem-repeat analysis and spoligotyping reveals the presence of beijing multidrug-resistant isolates. (United States)

    Tafaj, Silva; Zhang, Jian; Hauck, Yolande; Pourcel, Christine; Hafizi, Hasan; Zoraqi, Grigor; Sola, Christophe


    We characterized a set of 100 Mycobacterium tuberculosis complex clinical isolates from tuberculosis (TB) patients in Albania, typing them with a 24-locus variable-number tandem-repeat-spoligotyping scheme. Depending on the cluster definition, 43 to 49 patients were distributed into 15 to 16 clusters which were likely to be epidemiologically linked, indicative of a recent transmission rate of 28 to 34%. This result suggests that TB is under control in Albania. However, two multidrug-resistant (MDR) Beijing genotypes harboring the same S531A mutation on the rpoB gene were also found, suggesting a potential recent transmission of MDR TB. Three brand new genotypes, Albania-1 to Albania-3, are also described.

  11. Accuracy of Reverse Dot-Blot PCR in Detection of Different β-Globin Gene Mutations. (United States)

    El-Fadaly, N; Abd-Elhameed, A; Abd-Elbar, E; El-Shanshory, M


    Prevention programs for β-thalassemia based on molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation screening methods. The aim of this study was to compare between direct DNA sequencing, and reverse dot-blot PCR in detection of different β-globin gene mutations in Egyptian children with β-thalassemia. Forty children with β-thalassemia were subjected to mutation analysis, performed by both direct DNA sequencing and β-globin Strip Assay MED™ (based on reverse dot-blot PCR). The most frequent mutant alleles detected by reverse dot-blot PCR were; IVSI-110 G>A (31.25 %), IVS I-6 T > C (21.25 %), and IVS I-1 G>A (20 %). Relatively less frequent mutant alleles detected by reverse dot-blot PCR were "IVSII-1 G>A (5 %), IVSII-745 C>G (5 %), IVSII-848 C>A (2.5 %), IVSI-5 G>C (2.5 %), -87 C>G(2.5 %), and cd39 C>T (2.5 %)", While the genotypes of three patients (6 alleles 7.5 %) were not detected by reverse dot-blot PCR. Mutant alleles detected by direct DNA sequencing were the same as reverse dot-blot PCR method except it revealed the genotypes of 3 undetected patients (one patient was homozygous IVSI-110 G>A, and two patients were homozygous IVS I-1 G>A. Sensitivity of the reverse dot-blot PCR was 92.5 % when compared to direct DNA sequencing for detecting β-thalassemia mutations. Our results therefore suggest that, direct DNA sequencing may be preferred over reverse dot-blot PCR in critical diagnostic situations like genetic counseling for prenatal diagnosis.


    Directory of Open Access Journals (Sweden)

    V G Barskova


    Full Text Available There are currently no accepted criteria for positive Western blots in Russian patients with Lyme borreliosis. The purpose of the current study was to develop criteria for a positive IgG westem-blot to aid particularly in the diagnosis of patients with joint manifestation of the disorder. Patients: 97 with Lyme disease, 145 - control subjects. IgG antibody responses were determined to 3 species ofB.burgdorferi sensu lato by Western blotting, using blots prepared by manufacturer. The best discriminatory ability of test criteria was chained by requiring any 3 of 11 IgG bands, a definition that could be used with B. burgdorferi sensu stricto, B.garinii and B.afzelii strains. With these 3 antigen preparation, positive IgG blots were found in 0 to 18% of patients with localized erythema migrans of < 4 weeks duration, 23 to 39% of those with disseminated infection < 20 weeks duration, and in 39 to 46% of those with late arthritis/arthralgia of >6 months duration the specificity was 93 to 99%. Thus, IgG Western blotting may bring greater specificity to serologic testing in Lyme borreliosis, but the sensitivity is limited.

  13. Imprinting mutations in Angelman syndrome detected by Southern blotting using a probe containing exon {alpha} of SNRPN

    Energy Technology Data Exchange (ETDEWEB)

    Beuten, J.; Sutcliffe, J.S.; Nakao, M. [Baylor College of Medicine, Houston, TX (United States)] [and others


    Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are associated with paternal and maternal deficiencies respectively, of gene expression within human chromosome 15q11-q13, and are caused by deletion, uniparental disomy (UPD), or other mutations. The SNRPN gene maps in this region, is paternally expressed, and is a candidate gene for PWS. Southern blotting using methylation-sensitive enzymes and a genomic DNA probe from the CpG island containing exon {alpha} of the SNRPN gene reveals methylation specific for the maternal allele. In cases of the usual deletions or UPD, the probe detects absence of an unmethylated allele in PWS and absence of a methylated allele in AS. We have analyzed 21 nondeletion/nonUPD AS patients with this probe and found evidence for an imprinting mutation (absence of a methylated allele) in 3 patients. Southern blotting with methylation-sensitive enzymes using the exon {alpha} probe, like use of the PW71 probe, should detect abnormalities in all known PWS cases and in 3 of the 4 forms of AS: deletion, UPD and imprinting mutations. This analysis provides a valuable diagnostic approach for PWS and AS. In efforts to localize the imprinting mutations in AS, one patient was found with failure to inherit a dinucleotide repeat polymorphism near probe 189-1 (D15S13). Analysis of this locus in AS families and CEPH families demonstrates a polymorphism that impairs amplification and a different polymorphism involving absence of hybridization to the 189-1 probe. The functional significance, if any, of deletion of the 189-1 region is unclear.


    Institute of Scientific and Technical Information of China (English)

    Bao-qing Sun; Adrian Wu; Albert Chan; Stanley Chik; Dorothy Wong; Nan-shan Zhong


    Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P< 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P< 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.

  15. Total protein or high-abundance protein: Which offers the best loading control for Western blotting? (United States)

    Thacker, Jonathan S; Yeung, Derrick H; Staines, W Richard; Mielke, John G


    Western blotting routinely involves a control for variability in the amount of protein across immunoblot lanes. Normalizing a target signal to one found for an abundantly expressed protein is widely regarded as a reliable loading control; however, this approach is being increasingly questioned. As a result, we compared blotting for two high-abundance proteins (actin and glyceraldehyde 3-phosphate dehydrogenase [GAPDH]) and two total protein membrane staining methods (Ponceau and Coomassie Brilliant Blue) to determine the best control for loading variability. We found that Ponceau staining optimally balanced accuracy and precision, and we suggest that this approach be considered as an alternative to normalizing with a high-abundance protein.

  16. Indeterminate HIV-1 Western Blots: Etiology, Natural History, and Psychological Reactions (United States)


    systemic lupus erythematosus, rheumatoid arthritis, juvenile onset diabetes mellitus , Sjogren’s disease, thyroiditis, and Crohn’s disease 4 TABLE 8 45...Cat wound ply 0-no; 1-scratched only; 2-bitten only; 3-both BE.2Bb Owned other pets 0-no;l- dog ; 2-rodent;3-other A/< . 2 # BE.3 SMOKING HISTORY 53 BE...blot. Sera from 26 cases with a pet cat were tested for reactivity on Western blots for feline leukemia virus (FeLV) and for feline immunodeficiency

  17. A Streamlined Western Blot Exercise: An Efficient and Greener Approach in the Laboratory Classroom (United States)

    Ness, Traci L.; Robinson, Rebekah L.; Mojadedi, Wais; Peavy, Lydia; Weiland, Mitch H.


    SDS-PAGE and western blotting are two commonly taught protein detection techniques in biochemistry and molecular biology laboratory classrooms. A pitfall associated with incorporating these techniques into the laboratory is the significant wait times that do not allow students to obtain timely results. The waiting associated with SDS-PAGE comes…

  18. A Study of Rubisco through Western Blotting and Tissue Printing Techniques (United States)

    Ma, Zhong; Cooper, Cynthia; Kim, Hyun-Joo; Janick-Buckner, Diane


    We describe a laboratory exercise developed for a cell biology course for second-year undergraduate biology majors. It was designed to introduce undergraduates to the basic molecular biology techniques of Western blotting and immunodetection coupled with the technique of tissue printing in detecting the presence, relative abundance, and…

  19. Ferulic acid enhances IgE binding to peanut allergens in western blots. (United States)

    Phenolic compounds at high concentrations are known to form insoluble complexes with proteins. We hypothesized that this complex formation could interfere with Western blot and ELISA assays for peanut allergens. To verify this, three simple phenolic compounds (ferulic, caffeic, and chlorogenic acids...

  20. PET-blot analysis contributes to BSE strain recognition in C57Bl/6 mice. (United States)

    Lezmi, Stéphane; Bencsik, Anna; Baron, Thierry


    Identification of the strain of agent responsible for bovine spongiform encephalopathy (BSE) can be made histologically through the analysis of both distribution and intensity of brain vacuolar lesions after BSE transmission to mouse. Another useful way to distinguish the BSE agent from other prion strains is the study of the distribution of the abnormal prion protein (PrP(res)). For that purpose, paraffin-embedded tissue blot (PET-blot) method was applied on brains from C57Bl/6 mice infected with cattle BSE, experimental sheep BSE, or feline spongiform encephalopathy (FSE) from a cheetah. PrP(res) distribution was comparable, whichever of the three BSE agent sources was considered and was distinct from the PrP(res) distribution in C57Bl/6 mice inoculated with a French scrapie isolate or with a mouse-adapted scrapie strain (C506M3). These data confirm a common origin of infectious agent responsible for the British and French cattle BSE. They also indicate that PET-blot method appears as a precise complementary tool in prion strain studies because it offers easy and quick assessment of the PrP(res) mapping. Advantages and limits of the PET-blot method are discussed and compared with other established and validated methods of strain typing.

  1. Combined use of Western blot/ELISA to improve the serological diagnosis of human tuberculosis

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    S. T. Beck


    Full Text Available Two recombinant antigens and a crude bacterial antigen of a wild M. tuberculosis strain were used to detect specific IgG antibodies in sera from 52 patients with pulmonary tuberculosis, confirmed by an acid-fast smear and serum culture of these patients and that of 25 contacts. The patients were not infected with HIV. We evaluated the sensitivity and specificity of ELISA, based on the recombinant TbF6® and TbF6/DPEP antigen and a search for reactivity patterns in the Western blot technique, using whole mycobacterium antigen. Serum samples from 22 healthy individuals and from 30 patients with lung diseases other than tuberculosis were used as controls. The best ELISA results were obtained with the TbF6/DPEP antigen combination, which gave 85% sensitivity and 91% specificity. ELISA sensitivity improved from 85% to 92% when the Western blot results were used. Western blot specificity was 100% when antibody reactivity with different antigenic bands was analyzed and associated. The association of TbF6/DPEP antigens used in ELISA with specific patterns of reactivity determined by Western blot can help make an identification when classic methods for the diagnosis of pulmonary tuberculosis are not sufficient.

  2. Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting (United States)

    Omotola, Oluwabukola B.; Heda, Rajiv P.; Avery, Jamie


    SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin’s transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting. PMID:27582639

  3. Western blotting as a method for studying cell-biomaterial interactions : The role of protein collection

    NARCIS (Netherlands)

    van Kooten, TG; Klein, CL; Kirkpatrick, CJ


    Research of cell-biomaterial interactions is building on knowledge and methods available in cell and molecular biology. Western blotting is one of the options to characterize protein expression in cell populations. Method transfer to biomaterial model systems is not trivial because of the structure

  4. Biofilm detection by wound blotting can predict slough development in pressure ulcers: A prospective observational study. (United States)

    Nakagami, Gojiro; Schultz, Gregory; Gibson, Daniel J; Phillips, Priscilla; Kitamura, Aya; Minematsu, Takeo; Miyagaki, Tomomitsu; Hayashi, Akitatsu; Sasaki, Sanae; Sugama, Junko; Sanada, Hiromi


    Bacteria have been found to form multicellular aggregates which have collectively been termed "biofilms." It is hypothesized that biofilm formation is a means to protect bacterial cells including protection form the immune response of humans. This protective mechanism is believed to explain persistent chronic wound infections. At times, the biofilms are abundant enough to see, and remove by simple wiping. However, recent evidence has shown that the removal of these visible portions are not sufficient, and that biofilms can continue to form even with daily wiping. In this work, we tested an approach to detect the biofilms which are present after clinically wiping or sharp wound debridement. Our method is based on a variation of impression cytology in which a nitrocellulose membrane was used to collect surface biofilm components, which were then differentially stained. In this prospective study, members of an interdisciplinary pressure ulcer team at a university hospital tested our method's ability to predict the generation of wound slough in the week that followed each blotting. A total of 70 blots collected from 23 pressure ulcers produced 27 wounds negative for staining and 43 positive. In the negative blots 55.6% were found to have decreased wound slough, while 81.4% with positive staining had either increase or unchanged wound slough generation. These results lead to an odds ratio of positive blotting cases of 9.37 (95% confidence intervals: 2.47-35.5, p = 0.001) for slough formation; suggesting that the changes in wound slough formation can be predicted clinically using a non-invasive wound blotting method.

  5. Design, preparation and use of ligated phosphoproteins: a novel approach to study protein phosphatases by dot blot array, ELISA and Western blot assays. (United States)

    Sun, Luo; Ghosh, Inca; Barshevsky, Tanya; Kochinyan, Samvel; Xu, Ming-Qun


    The study of substrate specificity of protein phosphatases (PPs) is very challenging since it is difficult to prepare a suitable phosphorylated substrate. Phosphoproteins, phosphorylated by a protein kinase, or chemically synthesized phosphopeptides are commonly used substrates for PPs. Both types of these substrates have their advantages and limitations. Phosphoproteins mimic more closely the physiologically relevant PP substrates, but their preparation is technically demanding. Synthetic phosphopeptides present advantages over proteins because they can be easily produced in large quantity and their amino acid sequence can be designed to contain potential determinants of substrate specificity. However, short peptides are less optimal compared to in vivo PP substrates and often display poor and variable binding to different matrices, resulting in low sensitivity in analysis of PP activity on solid support. In this work we utilize the intein-mediated protein ligation (IPL) technique to generate substrates for PPs, combining the advantages of proteins and synthetic peptides in one molecule. The ligation of a synthetic phosphopeptide to an intein-generated carrier protein (CP) with a one-to-one stoichiometry results in the formation of a ligated phosphoprotein (LPP). Three widely used assays, dot blot array, Western blot and ELISA were employed to study the PP activity on LPP substrates. Dephosphorylation was measured by detection of the remaining phosphorylation, or lack of it, with a phospho-specific antibody. The data show the advantage of LPPs over free peptides in assays on solid supports. LPPs exhibited enhanced binding to the matrices used in the study, which significantly improved sensitivity and consistency of the assays. In addition, saturation of the signal was circumvented by serial dilution of the assay samples. This report describes detailed experimental procedures for preparation of LPP substrates and their use in PP assays based on immobilization on


    Directory of Open Access Journals (Sweden)

    Gusti Ngurah Permana


    Full Text Available Pemahaman tentang penentuan jenis kelamin dalam populasi induk merupakan hal yang sangat penting bagi keberhasilan program pembenihan. Pengukuran reaksi antibodi dan aktivitas hormon testosterone, serta estradiol adalah metode dengan potensi yang secara akurat dapat menentukan jenis kelamin ikan tanpa mematikan ikan. Tujuan penelitian ini adalah untuk mengetahui akurasi metode dot blot dan ELISA dengan 11-ketotestorsterone (11-KT yang tersedia secara komersial EIA-kit untuk membedakan jenis kelamin ikan tuna sirip kuning. Hasil analisis menunjukkan bahwa metode dot blot menghasilkan ekspresi vitelogenin tampak jelas pada individu betina dan efek plasma terlihat transparan, jika dibandingkan dengan individu jantan. Interpretasi dari metode ini memerlukan pengalaman dan keahlian dalam akurasi pembacaan hasil. Aktivitas hormon 11-KT dengan sampel klip sirip dan plasma memberikan hasil yang baik dengan aktivitas hormon terlihat jelas.

  7. Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite


    Son, Eui-Sun; Kim, Tong Soo; Nam, Ho-Woo


    Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce ...

  8. Positive IgG Western Blot for Borrelia burgdorferi in Colombia

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    Palacios Ricardo


    Full Text Available In order to evaluate the presence of specific IgG antibodies to Borrelia burgdorferi in patients with clinical manifestations associated with Lyme borreliosis in Cali, Colombia, 20 serum samples from patients with dermatologic signs, one cerebrospinal fluid (CSF sample from a patient with chronic neurologic and arthritic manifestations, and twelve serum samples from individuals without clinical signs associated with Lyme borreliosis were analyzed by IgG Western blot. The results were interpreted following the recommendations of the Centers for Diseases Control and Prevention (CDC for IgG Western blots. Four samples fulfilled the CDC criteria: two serum specimens from patients with morphea (localized scleroderma, the CSF from the patient with neurologic and arthritic manifestations, and one of the controls. Interpretation of positive serology for Lyme disease in non-endemic countries must be cautious. However these results suggest that the putative "Lyme-like" disease may correlate with positivity on Western blots, thus raising the possibility that a spirochete genospecies distinct from B. burgdorferi sensu stricto, or a Borrelia species other than B. burgdorferi sensu lato is the causative agent. Future work will focus on a survey of the local tick and rodent population for evidence of spirochete species that could be incriminated as the etiologic agent.

  9. Banding pattern indicative of echinococcosis in a commercial cysticercosis western blot

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    Tappe D


    Full Text Available Abstract Objective A commercial cysticercosis Western blot was evaluated for serological cross-reactivity of sera from patients with alveolar (AE and cystic echinococcosis (CE. Methods A total of 161 sera were examined, including 31 sera from AE-patients, 11 sera from CE-patients, 9 sera from patients with other parasitic diseases and 109 sera from patients with unrelated medical conditions. All AE-and CE-sera were also examined by the echinococcosis Western blot. Results More sera from patients with AE than with CE showed cross-reactivity in the form of ladder-like patterns ("Mikado aspect" and untypical bands at 6-8 kDa (71% and 77.4% versus 27.3% and 45.5%, respectively. In contrast, triplets of bands in the area above 50 kDa and between 24 and 39-42 kDa were more frequent in CE than in AE sera. The fuzzy band at 50-55 kDa typical for cysticercosis was absent in all AE and CE sera. Conclusions Atypical banding patterns in the cysticercosis Western blot should raise the suspicion of a metacestode infection different from Taenia solium, i.e. Echinococcus multilocularis or E. granulosus, especially when the Mikado aspect and an altered 6-8 kDa band is visible in the absence of a fuzzy 50-55 kDa band.

  10. Standardization of Licorice and TCM Formulations Using Eastern Blot Fingerprinting Analysis

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    Yukihiro Shoyama


    Full Text Available To prepare the antiglycyrrhizin (GC monoclonal antibody (MAb, GC was treated with NaIO4 resulting in aldehyde which can be combined with carrier protein. An antigen conjugate was performed by a matrix-assisted laser desorption/ionization TOF mass spectrometry to determine the hapten numbers in the conjugate. Anti-GC MAb was prepared from a hybridoma which was fixed from the spleen cells producing anti-GC MAb and the myeloma cells after immunization. The TCM and licorice extract were developed by TLC and blotted to a polyvinylidene difluoride (PVDF membrane. The membrane was treated by NaIO4 and protein, enzyme labeled secondary MAb, and finally substrate was added. Clear spot appeared on PVDF membrane identifying GC against a background containing large amount of impurities. In eastern blotting, the GC molecule was divided into two functions. The aglycone part is recognized as an epitope and the sugar moiety can be combined to membrane. The specific reactivity of sugar moiety in the GC molecule against anti-GC MAb might be modified by the NaIO4 treatment on the membrane because glycyrrhetic acid 3-O-glucuronide can be stained although the cross-reactivity is only 4.3%. Eastern blotting for GC can not only apply for the standardization of licorice and TCM, but also it can open for the other bioactive products.

  11. Use of a Western blot technique for the serodiagnosis of glanders

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    de Souza Marcilia MA


    Full Text Available Abstract Background The in vivo diagnosis of glanders relies on the highly sensitive complement fixation test (CFT. Frequently observed false positive results are troublesome for veterinary authorities and cause financial losses to animal owners. Consequently, there is an urgent need to develop a test with high specificity. Hence, a Western blot assay making use of a partly purified lipopolysaccaride (LPS containing antigen of three Burkholderia mallei strains was developed. The test was validated investigating a comprehensive set of positive and negative sera obtained from horses and mules from endemic and non endemic areas. Results The developed Western blot assay showed a markedly higher diagnostic specificity when compared to the prescribed CFT and therefore can be used as a confirmatory test. However, the CFT remains the test of choice for routine testing of glanders due to its high sensitivity, its feasibility using standard laboratory equipment and its worldwide distribution in diagnostic laboratories. Conclusions The CFT should be amended by the newly validated Western blot to increase the positive likelihood ratio of glanders serodiagnosis in non endemic areas or areas with low glanders prevalence. Its use for international trade of horses and mules should be implemented by the OIE.

  12. Comparison of Linear Array and Line Blot Assay for Detection of Human Papillomavirus and Diagnosis of Cervical Precancer and Cancer in the Atypical Squamous Cell of Undetermined Significance and Low-Grade Squamous Intraepithelial Lesion Triage Study▿


    Castle, Philip E.; Gravitt, Patti E.; Solomon, Diane; Wheeler, Cosette M.; Schiffman, Mark


    We evaluated Linear Array (LA), a newly commercialized PGMY09/11 L1 consensus primer PCR test that detects 37 human papillomavirus (HPV) genotypes by reverse line blot hybridization, for the detection of individual HPV genotypes and carcinogenic HPV and its clinical performance for detecting 2-year cumulative cervical precancer and cancer using archived specimens from the Atypical Squamous Cell of Undetermined Significance (ASCUS) and Low-Grade Squamous Intraepithelial Lesion Triage Study. LA...

  13. Hybrid Baryons

    CERN Document Server

    Page, P R


    We review the status of hybrid baryons. The only known way to study hybrids rigorously is via excited adiabatic potentials. Hybrids can be modelled by both the bag and flux-tube models. The low-lying hybrid baryon is N 1/2^+ with a mass of 1.5-1.8 GeV. Hybrid baryons can be produced in the glue-rich processes of diffractive gamma N and pi N production, Psi decays and p pbar annihilation.

  14. Correlations of mutations in katG, oxyR-ahpC and inhA genes and in vitro susceptibility in Mycobacterium tuberculosis clinical strains segregated by spoligotype families from tuberculosis prevalent countries in South America

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    Suffys Philip N


    Full Text Available Abstract Background Mutations associated with resistance to rifampin or streptomycin have been reported for W/Beijing and Latin American Mediterranean (LAM strain families of Mycobacterium tuberculosis. A few studies with limited sample sizes have separately evaluated mutations in katG, ahpC and inhA genes that are associated with isoniazid (INH resistance. Increasing prevalence of INH resistance, especially in high tuberculosis (TB prevalent countries is worsening the burden of TB control programs, since similar transmission rates are noted for INH susceptible and resistant M. tuberculosis strains. Results We, therefore, conducted a comprehensive evaluation of INH resistant M. tuberculosis strains (n = 224 from three South American countries with high burden of drug resistant TB to characterize mutations in katG, ahpC and inhA gene loci and correlate with minimal inhibitory concentrations (MIC levels and spoligotype strain family. Mutations in katG were observed in 181 (80.8% of the isolates of which 178 (98.3% was contributed by the katG S315T mutation. Additional mutations seen included oxyR-ahpC; inhA regulatory region and inhA structural gene. The S315T katG mutation was significantly more likely to be associated with MIC for INH ≥2 μg/mL. The S315T katG mutation was also more frequent in Haarlem family strains than LAM (n = 81 and T strain families. Conclusion Our data suggests that genetic screening for the S315T katG mutation may provide rapid information for anti-TB regimen selection, epidemiological monitoring of INH resistance and, possibly, to track transmission of INH resistant strains.

  15. Proteínas inmunodominantes de Brucella Melitensis evaluadas por Western Blot

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    Elizabeth Anaya


    Full Text Available Se separaron extractos de proteínas totales de Brucella melitensis en gel 15% SDS-PAGE. Su seroreactividad fue analizada por Western Blot con resultados satisfactorios. Para éste propósito sueros controles negativos (n=03, sueros de pacientes con brucelosis (n=34, cólera (n=12, tifoidea (n=02 y tuberculosis (n=02 fueron usados. Esta prueba inmunodiagnóstica detectó bandas seroreactivas altamente específicas (100% correspondientes a 8,14,18, un complejo de 25-48 y 58kDa. La sensibilidad del test fue del 90% usando los sueros antes mencionados.

  16. A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot

    DEFF Research Database (Denmark)

    Albers, Eliene; Sbroggiò, Mauro; Martin Gonzalez, Javier


    The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific...... that match exons flanking the targeting construct. The detection of the expected modification in this PCR product confirms the integration at the correct genomic location and shows that the mutant mRNA is expressed. This is a simple and sensitive strategy to screen locus-specific recombination of targeting...

  17. A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot. (United States)

    Albers, Eliene; Sbroggiò, Mauro; Martin-Gonzalez, Javier; Avram, Alexandra; Munk, Stephanie; Lopez-Contreras, Andres J


    The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific genomic locus, but limits the detection of its correct genomic integration by standard PCR methods. Genomic Long Range PCR (LR-PCR), using primers adjacent to the homology arms, has been used as an alternative to radioactive-based Southern blot screenings. However, LR-PCRs are often difficult and render many false positive and false negative results. Here, we propose an alternative screening method based on the detection of a genetic modification at the mRNA level, which we successfully optimized in two mouse models. This screening method consists of a reverse-transcription PCR (RT-PCR) using primers that match exons flanking the targeting construct. The detection of the expected modification in this PCR product confirms the integration at the correct genomic location and shows that the mutant mRNA is expressed. This is a simple and sensitive strategy to screen locus-specific recombination of targeting constructs which can also be useful to screen KO and KI mutant mice or cell lines including those generated by CRISPR/Cas9.

  18. A sensitive non-radioactive northern blot method to detect small RNAs. (United States)

    Kim, Sang Woo; Li, Zhihua; Moore, Patrick S; Monaghan, A Paula; Chang, Yuan; Nichols, Mark; John, Bino


    The continuing discoveries of potentially active small RNAs at an unprecedented rate using high-throughput sequencing have raised the need for methods that can reliably detect and quantitate the expression levels of small RNAs. Currently, northern blot is the most widely used method for validating small RNAs that are identified by methods such as high-throughput sequencing. We describe a new northern blot-based protocol (LED) for small RNA (approximately 15-40 bases) detection using digoxigenin (DIG)-labeled oligonucleotide probes containing locked nucleic acids (LNA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide for cross-linking the RNA to the membrane. LED generates clearly visible signals for RNA amounts as low as 0.05 fmol. This method requires as little as a few seconds of membrane exposure to outperform the signal intensity using overnight exposure of isotope-based methods, corresponding to approximately 1000-fold improvement in exposure-time. In contrast to commonly used radioisotope-based methods, which require freshly prepared and hazardous probes, LED probes can be stored for at least 6 months, facilitate faster and more cost-effective experiments, and are more environmentally friendly. A detailed protocol of LED is provided in the Supplementary Data.

  19. Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

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    N.S. Sato


    Full Text Available Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples, from healthy blood donors (50 samples, individuals with sexually transmitted disease other than syphilis (3 samples, and from individuals with other spirochetal diseases such as Lyme disease (20 samples and leptospirosis (3 samples. The assay had sensitivity of 95.1% (95% CI, 86.1 to 98.7% and a specificity of 94.7% (95% CI, 87.0 to 98.7%; a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

  20. Gel electrophoresis of polyphenol oxidase with instant identification by in situ blotting. (United States)

    Cheng, Tsai-Mu; Huang, Pei-Chen; Pan, Ju-Pin; Lin, Kuan-Yu; Mao, Simon J T


    Polyphenol oxidase (PPO) or tyrosinase is an important and ubiquitous enzyme responsible for browning in plants and melanization in animals. The molecular size of the plant PPO is varied among the species and its activity can be enhanced by a variety of anionic detergents. In the present study, we developed a simple method for the first-step identification of PPO in fruit and vegetable extracts. First, 3mm chromatographic paper was immersed in 0.5% (w/v) catechol solution as an immobilized PPO substrate. After running the extract with 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), one side of the glass plate was removed. The plate was immediately laid on top of the dried catechol-paper. A dark-brown band corresponding to PPO was visualized within 1 min and was further confirmed by a conventional Western blot using an antibody prepared against mushroom PPO. It also reveals that some vegetation (such as tomato, radish, and oriental melon) with low or no detectable activity in a conventional enzyme assay actually possessed marked levels of PPO activity when assessed by PAGE-blot. We propose that an inhibitor is associated with PPO in some plants; the inhibitor, however, is dissociated during the electrophoresis. Therefore, in addition to identify the molecular form of PPO, the present technique may explore the existence of PPO inhibitor(s) in plants. The detail of the method with respect to its relevance for searching a natural PPO inhibitor is described and discussed.

  1. Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein (United States)

    Yang, Zhaoshou; Lee, Jihoo; Ahn, Hye-Jin; Chong, Chom-Kyu; Dias, Ronaldo F.; Nam, Ho-Woo


    Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection. PMID:27180586

  2. Analysis of sperm membrane antigens relevant to antisperm antibody using Western blot

    Institute of Scientific and Technical Information of China (English)



    Objective:To identify the sperm membrane antigens associated with antisperm antibody.Methods:The antisperm antibody in serum was tested by ELISA.Antisperm antibody positive sera from 18 infertile men and 15 infertile women were used.The molecular weight(MW) of sperm membrane antigens associated with the antisperm antibody was analyzed with antisperm antibody positive serum using Western blot.Results:Eight kinds of MW of sperm membrane antigens were identified.The ratio of identification on the 78 KD(60.7%),60KD(71.4%),51KD(14.9%) and 23KD(14.29%)sperm antigen was higher than other.Conclusion:sperm membrane antigens with MW of 78KD,60KD,51KD and 23KD were associated with antisperm antibody and immunological infertility.

  3. Electrospun nitrocellulose and nylon: Design and fabrication of novel high performance platforms for protein blotting applications

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    Bowlin Gary L


    Full Text Available Abstract Background Electrospinning is a non-mechanical processing strategy that can be used to process a variety of native and synthetic polymers into highly porous materials composed of nano-scale to micron-scale diameter fibers. By nature, electrospun materials exhibit an extensive surface area and highly interconnected pore spaces. In this study we adopted a biological engineering approach to ask how the specific unique advantages of the electrospinning process might be exploited to produce a new class of research/diagnostic tools. Methods The electrospinning properties of nitrocellulose, charged nylon and blends of these materials are characterized. Results Nitrocellulose electrospun from a starting concentration of Conclusion The flexibility afforded by electrospinning process makes it possible to tailor blotting membranes to specific applications. Electrospinning has a variety of potential applications in the clinical diagnostic field of use.

  4. Rapid detection and differentiation of important Campylobacter spp. in poultry samples by dot blot and PCR. (United States)

    Fontanot, Marco; Iacumin, Lucilla; Cecchini, Francesca; Comi, Giuseppe; Manzano, Marisa


    The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively.

  5. Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite. (United States)

    Son, E S; Kim, T S; Nam, H W


    Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

  6. Total protein analysis as a reliable loading control for quantitative fluorescent Western blotting.

    Directory of Open Access Journals (Sweden)

    Samantha L Eaton

    Full Text Available Western blotting has been a key technique for determining the relative expression of proteins within complex biological samples since the first publications in 1979. Recent developments in sensitive fluorescent labels, with truly quantifiable linear ranges and greater limits of detection, have allowed biologists to probe tissue specific pathways and processes with higher resolution than ever before. However, the application of quantitative Western blotting (QWB to a range of healthy tissues and those from degenerative models has highlighted a problem with significant consequences for quantitative protein analysis: how can researchers conduct comparative expression analyses when many of the commonly used reference proteins (e.g. loading controls are differentially expressed? Here we demonstrate that common controls, including actin and tubulin, are differentially expressed in tissues from a wide range of animal models of neurodegeneration. We highlight the prevalence of such alterations through examination of published "-omics" data, and demonstrate similar responses in sensitive QWB experiments. For example, QWB analysis of spinal cord from a murine model of Spinal Muscular Atrophy using an Odyssey scanner revealed that beta-actin expression was decreased by 19.3±2% compared to healthy littermate controls. Thus, normalising QWB data to β-actin in these circumstances could result in 'skewing' of all data by ∼20%. We further demonstrate that differential expression of commonly used loading controls was not restricted to the nervous system, but was also detectable across multiple tissues, including bone, fat and internal organs. Moreover, expression of these "control" proteins was not consistent between different portions of the same tissue, highlighting the importance of careful and consistent tissue sampling for QWB experiments. Finally, having illustrated the problem of selecting appropriate single protein loading controls, we demonstrate

  7. Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis,

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    Jaqueline Dario Capobiango

    Full Text Available Abstract Objective: To evaluate the Western blotting method for the detection of IgG anti-Toxoplasma gondii (T. gondii (IgG-WB in the serum of children with suspected congenital toxoplasmosis. Methods: We accompanied 47 mothers with acquired toxoplasmosis in pregnancy and their children, between June of 2011 and June of 2014. The IgG-WB was done in house and the test was considered positive if the child had antibodies that recognized at least one band on IgG blots different from the mother's or with greater intensity than the corresponding maternal band, during the first three months of life. Results: 15 children (15.1% met the criteria for congenital toxoplasmosis and 32 (32.3% had the diagnosis excluded. The symptoms were observed in 12 (80.0% children and the most frequent were cerebral calcification in 9 (60.0%, chorioretinitis in 8 (53.3%, and hydrocephalus in 4 (26.6%. IgM antibodies anti-T. gondii detected by chemiluminescence (CL were found in 6 (40.0% children and the polymerase chain reaction (PCR for detection of T. gondii DNA was positive in 5 of 7 performed (71.4%. The sensitivity of IgG-WB was of 60.0% [95% confidence interval (CI 32.3-83.7%] and specificity 43.7% (95% CI 26.7-62.3%. The sensitivity of IgG-WB increased to 76.0 and 89.1% when associated to the research of IgM anti-T. gondii or PCR, respectively. Conclusions: The IgG-WB showed greater sensitivity than the detection of IgM anti-T. gondii; therefore, it can be used for the diagnosis of congenital toxoplasmosis in association with other congenital infection markers.

  8. Screening for hereditary fructose intolerance mutations by reverse dot-blot. (United States)

    Lau, J; Tolan, D R


    An assay is described which is useful for genetic screening of the two most prevalent mutations that cause hereditary fructose intolerance (HFI). Both mutations lie within exon 5 of the aldolase B gene. Amplification of exon 5 from genomic DNA isolated from peripheral lymphocytes using biotinylated aldolase B-specific primers yields a biotin-tagged probe. This probe is hybridized to complementary poly(dT)-tailed allele specific oligonucleotides (ASOs) that are bound to a nylon membrane. The length of the ASOs, the amount bound to the membrane and the time of hybridization are optimized for discrimination of all four alleles under the same hybridization conditions. Detection of biotinylated amplified DNA is performed by creating an avidin-alkaline phosphatase complex and visualization by chemiluminescence. This assay can rapidly detect the two mutations, A149P and A174D, which cause >70% of HFI worldwide, and offers a rapid and sensitive assay that is much less invasive for the diagnosis of this often difficult to diagnose disorder.

  9. HIV‑2 antibody detection after indeterminate or negative HIV‑1 Western blot in Cuba, 2005-2008. (United States)

    Díaz, Dervel F; Ortiz, Eva; Martín, Dayamí; Nibot, Carmen; Rizo, Adis; Silva, Eladio


    INTRODUCTION Differentiating between HIV-1 and HIV-2 infection is the first step to understanding HIV transmission, epidemiology and pathogenesis in geographical areas where both viruses circulate. In Cuba, positive results in mixed HIV-1/2 screening assays are confirmed by HIV-1 Western blot. Indeterminate results constitute the main limitation of this test and HIV-2 infection is among their possible causes; hence the importance of second-stage screening and confirmatory tests for HIV-2 infection. OBJECTIVE Investigate the contribution of HIV-2 antibodies to negative or indeterminate HIV-1 Western blot results in serum samples from 2005 through 2008 in Cuba. METHODS HIV-2 reactivity was studied using the ELISA DAVIH-VIH-2 diagnostic kit (Cuba) in 1723 serum samples with negative or indeterminate results for HIV-1 Western blot from January 2005 through December 2008. Duplicate sera reactive by ELISA were confirmed by HIV-2 Western blot, results interpreted according to WHO criteria. The epidemiological interview established by Cuba's National Program for Prevention and Control Sexually-Transmitted Diseases and HIV/AIDS was applied to HIV-2 Western blot-positive patients. RESULTS Among all sera studied, HIV-2 ELISA identified 12 reactive serum samples (0.70%) and 1711 non-reactive (99.30%). Western blot analysis of the 12 ELISA-reactive samples confirmed two positive samples (16.67%), 4 negative (33.33%) and 6 indeterminate (50%). Positive samples reacted against the p16, p26, gp36, p53, p56, p68 and gp105 proteins. All 12 ELISA-reactive samples belonged to the HIV-1 Western blot indeterminate group. The two HIV-2-positive samples showed well defined reactivity to gp160, p53, p55 and p34 of HIV-1. HIV-1 seroconversion was observed in all 10 remaining samples during serological followup. CONCLUSIONS Two new HIV-2 seropositive cases were diagnosed using DAVIH-VIH-2 and HIV-2 Western blot in indeterminate HIV-1 Western blot samples. Results support the recommendation

  10. Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting. (United States)

    Sudhakar, N R; Samanta, S; Sahu, Shivani; Raina, O K; Gupta, S C; Goswami, T K; Lokesh, K M; Kumar, Ashok


    Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

  11. The mouse prostaglandin E receptor EP2 subtype: cloning, expression, and northern blot analysis. (United States)

    Katsuyama, M; Nishigaki, N; Sugimoto, Y; Morimoto, K; Negishi, M; Narumiya, S; Ichikawa, A


    A functional cDNA clone for the mouse prostaglandin (PG) E receptor EP2 subtype was isolated from a mouse cDNA library. The mouse EP2 receptor consists of 362 amino acid residues with seven putative transmembrane domains. [3H]PGE2 bound specifically to the membrane of Chinese hamster ovary cells stably expressing the cloned receptor. This binding was displaced by unlabeled prostanoids in the order of PGE2 = PGE1 > iloprost, a stable PGI2 agonist > PGF2 alpha > PGD2. Binding was also inhibited by butaprost (an EP2 agonist) and to a lesser extent by M&B 28767 (an EP3 agonist), but not by sulprostone (an EP1 and EP3 agonist) or SC-19220 (an EP1 antagonist). PGE2 and butaprost increased the cAMP level in the Chinese hamster ovary cells in a concentration-dependent manner. Northern blot analysis revealed that EP2 mRNA is expressed most abundantly in the uterus, followed by the spleen, lung, thymus, ileum, liver, and stomach.

  12. A simple dot-blot-Sirius red-based assay for collagen quantification. (United States)

    Rodríguez-Rodríguez, Pilar; Arribas, Silvia M; de Pablo, Angel Luis López; González, M Carmen; Abderrahim, Fatima; Condezo-Hoyos, Luis


    The assessment of collagen content in tissues is important in biomedical research, since this protein is altered in numerous diseases. Hydroxyproline and Sirius red based assays are the most common methods for collagen quantification. However, these procedures have some pitfalls, such as the requirement of oxygen-free medium or expensive equipment and large sample size or being unsuitable for hydrolyzed collagen, respectively. Our objective was to develop a specific, versatile, and user-friendly quantitative method applicable to small tissue samples and extracts obtained from elastin purification, therefore, suitable for simultaneous quantification of elastin. This method is based on the binding of Sirius red to collagen present in a sample immobilized on a PVDF membrane, as in the dot-blot technique, and quantified by a scanner and image analysis software. Sample loading, Sirius red concentration, temperature and incubation time, type of standard substance, albumin interference, and quantification time are optimized. The method enabled the quantification of (1) intact collagen in several rat tissue homogenates, including small resistance-sized arteries, (2) partially hydrolyzed collagen obtained from NaOH extracts, compatible with elastin purification, and (3) for the detection of differences in collagen content between hypertensive and normotensive rats. We conclude that the developed technique can be widely used since it is versatile (quantifies intact and hydrolyzed collagen), requires small sample volumes, is user-friendly (low-cost, easy to use, minimum toxic materials, and reduced time of test), and is specific (minimal interference with serum albumin).

  13. Serological diagnosis of North American Paragonimiasis by Western blot using Paragonimus kellicotti adult worm antigen. (United States)

    Fischer, Peter U; Curtis, Kurt C; Folk, Scott M; Wilkins, Patricia P; Marcos, Luis A; Weil, Gary J


    Abstract. We studied the value of an IgG Western blot (WB) with Paragonimus kellicotti (Pk) antigen for diagnosis of North American paragonimiasis. The test was evaluated with sera from patients with Pk and Paragonimus westermani infections, with control sera from patients with other helminth infections, and sera from healthy Americans. All 11 proven Pk infection sera and two samples from suspected cases that were negative by P. westermani WB at the Centers for Disease Control and Prevention (CDC) contained antibodies to antigens at 34 kDa and at 21/23 kDa. Seven of 7 P. westermani sera contained antibodies to the 34 kDa antigen, but only 2 recognized the 21/23 kDa doublet. No control samples were reactive with these antigens. Antibody reactivity declined after praziquantel treatment. Thus, the P. kellicotti WB appears to be superior to P. westermani WB for diagnosing Pk infections, and it may be useful for assessing responses to treatment.

  14. Binding of 16S rRNA to chloroplast 30S ribosomal proteins blotted on nitrocellulose. (United States)

    Rozier, C; Mache, R


    Protein-RNA associations were studied by a method using proteins blotted on a nitrocellulose sheet. This method was assayed with Escherichia Coli 30S ribosomal components. In stringent conditions (300 mM NaCl or 20 degrees C) only 9 E. coli ribosomal proteins strongly bound to the 16S rRNA: S4, S5, S7, S9, S12, S13, S14, S19, S20. 8 of these proteins have been previously found to bind independently to the 16S rRNA. The same method was applied to determine protein-RNA interactions in spinach chloroplast 30S ribosomal subunits. A set of only 7 proteins was bound to chloroplast rRNA in stringent conditions: chloroplast S6, S10, S11, S14, S15, S17 and S22. They also bound to E. coli 16S rRNA. This set includes 4 chloroplast-synthesized proteins: S6, S11, S15 and S22. The core particles obtained after treatment by LiCl of chloroplast 30S ribosomal subunit contained 3 proteins (S6, S10 and S14) which are included in the set of 7 binding proteins. This set of proteins probably play a part in the early steps of the assembly of the chloroplast 30S ribosomal subunit.

  15. Renaturation of blotted allergens increases the sensitivity of specific IgE detection. (United States)

    Muro, M D; Fernández, C; Moneo, I


    Several authors have demonstrated that renaturation is an essential step for the appropriate recognition of blotted proteins. The use of nonionic detergents has been described as a useful alternative to enhance the antigenicity in immunoblotting, although elution from proteins by detergents has been observed. To measure the influence of different factors on the sensitivity of specific IgE by immunoblotting, we used twenty human sera from atopic patients who were allergic or nonallergic to a common, reliable allergen (grass pollen mixture). The use of Nonidet-P40 was found to be a useful alternative for the renaturation of the allergens. No elution from the membrane was found when employing this detergent, even at high concentrations (3%), and its use gave better sensitivity than methanol. On the other hand, we detected that methanol possessed renaturing properties. A transfer method using diffusion instead of electric transfer gave the best results and two membranes could be obtained from each gel. Using this method, we found that after NP-40 incubation of the membrane, the use of bovine albumin could be omitted as blocking agent and that its use had even deleterious effects.

  16. Glyceraldehyde-3-phosphate dehydrogenase: a universal internal control for Western blots in prokaryotic and eukaryotic cells. (United States)

    Wu, Yonghong; Wu, Min; He, Guowei; Zhang, Xiao; Li, Weiguang; Gao, Yan; Li, Zhihui; Wang, Zhaoyan; Zhang, Chenggang


    In the current study, we examined the expression level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein in a number of organisms and the stability of GAPDH under various conditions. Our results revealed that GAPDH is present in multiple Escherichia coli strains, the yeast strain GS115, Caenorhabditis elegans, rat PC12 cells, and both mouse and rat brain. Furthermore, GAPDH was stably expressed under different concentrations of inducer and at different times of induction in E. coli (BL21) cells and yeast GS115 cells. Stable expression of GAPDH protein was also observed in C.elegans and PC12 cells that were treated with different concentrations of paraquat or sodium sulfite, respectively. In addition, we were able to detect and identify the endogenous gapA protein in E.coli via immunoprecipitation and MALDI-TOF-MS analysis. Endogenous gapA protein and exogenously expressed (subcloned) GAPDH proteins were detected in E. coli BL21 but not for gapC. With the exception of gapC in E. coli, the various isoforms of GAPDH possessed enzymatic activity. Finally, sequence analysis revealed that the GAPDH proteins were 76% identical, with the exception of E. coli gapC. Taken together, our results indicate that GAPDH could be universally used as an internal control for the Western blot analysis of prokaryotic and eukaryotic samples.

  17. Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis

    Directory of Open Access Journals (Sweden)

    William H Roldán


    Full Text Available To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis. Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%, whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.

  18. Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis. (United States)

    Roldán, William H; Espinoza, Yrma A


    To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG) test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis). Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%), whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.

  19. Identification of toxocara canis antigens by Western blot in experimentally infected rabbits

    Directory of Open Access Journals (Sweden)

    MORALES Olga Lucía


    Full Text Available Toxocariasis is a frequent helminthiasis that can cause visceral and ocular damage in humans specially in children. The identification of specific antigens of Toxocara canis is important in order to develop better diagnostic techniques. Ten rabbits were infected orally with a dose of 5000 Toxocara canis embryonated eggs. Rabbits were bled periodically and an ELISA assay was performed to determine levels of specific Toxocara IgG antibodies. ELISA detected antibodies at day 15 after infection. Western blot (WB assay was performed using excretory/secretory antigens (E/S of T. canis second stage larvae. Different antigen concentrations were evaluated: 150, 200, 250 and 300 µg/mL. The concentration of 250 µg/mL was retained for analysis. Rabbit sera were diluted 1:100. Secondary antibody was used at a dilution of 1:1000. Results of WB indicated that in the first month after infection specific antibodies against the 200 KDa, 116 KDa, 92 KDa and 35 KDa antigens were detected; antibodies against the 92 KDa, 80 KDa, 66 KDa, 45 KDa, 31 KDa and 28 KDa antigens appeared later. All positive sera in the ELISA test were also positive in WB. Two antigen bands, 92 KDa and 35 KDa, were identified since the beginning and throughout the course of infection. These antigens merit further evaluation as candidates for use in diagnosis.

  20. Molecular analysis of Mycobacterium tuberculosis from tuberculous meningitis patients in Thailand. (United States)

    Yorsangsukkamol, Jutaporn; Chaiprasert, Angkana; Prammananan, Therdsak; Palittapongarnpim, Prasit; Limsoontarakul, Sunsern; Prayoonwiwat, Naraporn


    The first retrospective molecular characterization of Mycobacterium tuberculosis from cerebrospinal fluid of 158 tuberculous meningitis (TBM) in Thailand, collected between 1995 and 2005, was performed by Southern-blot hybridization with an IS6110 probe and spoligotyping on 152 and 147 isolates, respectively. Antituberculous drug susceptibility testing was performed in 149 patients, 118 of whom (79.2%) contained pan-sensitive strains and eight (5.4%) harbored multidrug-resistant M. tuberculosis. IS6110 RFLP typing revealed 110 RFLP patterns with 57.9% of patients infected with the Beijing genotype. This percentage was significantly higher than that in a previous report from pulmonary tuberculosis patients. Fifteen of 18 TBM patients (83%) aged <15 years were infected with Beijing isolates (OR=4.47, p=0.018). There were 40 spoligotypes, with 118 patients (80.3%) being clustered. The biggest cluster, which consisted of 84 patients, was the Beijing spoligotype (57.1%). There were 16 novel spoligotypes from 16 patients compared to the Fourth International Spoligotyping Database, SpolDB4. Sixty-four percent of the patients were male, and the mean age of patients was 33.8 years. Beijing isolates from 2001 to 2005 were found in higher percentages than those from 1995 to 2000, but this difference was not significant (p=0.28).

  1. Electrostatic protein immobilization using charged polyacrylamide gels and cationic detergent microfluidic Western blotting. (United States)

    Kim, Dohyun; Karns, Kelly; Tia, Samuel Q; He, Mei; Herr, Amy E


    We report a novel protein immobilization matrix for fully integrated microfluidic Western blotting (WB). The electrostatic immobilization gel (EIG) enables immobilization of all proteins sized using cetyl trimethylammonium bromide polyacrylamide gel electrophoresis (CTAB-PAGE), for subsequent electrophoretic probing with detection affinity reagents (e.g., labeled antibodies). The "pan-analyte" capture strategy introduced here uses polyacrylamide gel grafted with concentrated point charges (zwitterionic macromolecules), in contrast to existing microfluidic WB strategies that rely on a sandwich immunoassay format for analyte immobilization and detection. Sandwich approaches limit analyte immobilization to capture of only a priori known targets. A charge interaction mechanism study supports the hypothesis that electrostatic interaction plays a major role in analyte immobilization on the EIG. We note that protein capture efficiency depends on both the concentration of copolymerized charges and ionic strength of the gel buffer. We demonstrate pan-analyte immobilization of sized CTAB-laden model proteins (protein G, ovalbumin, bovine serum albumin, β-galactosidase, lactoferrin) on the EIG with initial capture efficiencies ranging from 21 to 100%. Target proteins fixed on the EIG (protein G, lactoferrin) are detected using antibody probes with signal-to-noise ratios of 34 to 275. The approach advances protein immunoblotting performance through 200× reduction on sample consumption, 12× reduction in assay duration, and automated assay operation, compared to slab-gel WB. Using the microfluidic WB assay, assessment of lactoferrin in human tear fluid is demonstrated with a goal of advancing toward nonbiopsy-based diagnosis of Sjögren's Syndrome, an autoimmune disease.

  2. Evaluation of a Western Blot Test in an Outbreak of Acute Pulmonary Histoplasmosis (United States)

    Pizzini, Claudia V.; Zancopé-Oliveira, Rosely M.; Reiss, Errol; Hajjeh, Rana; Kaufman, Leo; Peralta, José Mauro


    A western blot (WB) test was evaluated for detection of antibodies against native glycosylated and chemically deglycosylated M and H antigens of Histoplasma capsulatum in serum obtained from patients during the acute phase of pulmonary histoplasmosis that occurred during an outbreak. Of 275 serum samples tested by immunodiffusion and complement fixation (CF) samples from 40 patients affected during this outbreak and from 37 negative controls were tested by WB test. A group of patients whose sera were negative for CF antibodies and precipitins early in the acute stage of histoplasmosis but who all seroconverted during convalescence 6 weeks later were tested with the WB test. Antibodies against untreated H and M antigens were detected at a 1:100 dilution by WB test in 45% of the 20 acute-phase serum samples and in all 20 of the convalescent-phase specimens. The WB test’s sensitivity for acute-phase specimens increased to 90% (18 of 20 specimens) when H and M antigens were treated by periodate oxidation to inactivate susceptible carbohydrate epitopes. When native glycosylated antigens were used in the WB test, positive reactions were observed in negative control serum specimens (3 of 37 specimens; 8%) and in serum specimens obtained from asymptomatic persons screened as part of the outbreak investigation (13 of 20 specimens; 65%). These positive reactions were also attributed to glycosidic epitopes since the specificity of the WB test increased from 78 to 100% when periodate-treated H and M antigens were used. WB test with deglycosylated H and M antigens of histoplasmin provides a rapid, sensitive, and specific test to diagnose acute pulmonary histoplasmosis before precipitins can be detected. PMID:9874658

  3. Evaluation of the Aspergillus Western blot IgG kit for diagnosis of chronic aspergillosis. (United States)

    Oliva, A; Flori, P; Hennequin, C; Dubus, J-C; Reynaud-Gaubert, M; Charpin, D; Vergnon, J M; Gay, P; Colly, A; Piarroux, R; Pelloux, H; Ranque, S


    Immunoprecipitin detection (IPD) is the current reference confirmatory technique for anti-Aspergillus antibody detection; however, the lack of standardization is a critical drawback of this assay. In this study, we evaluated the performance of the Aspergillus Western blot (Asp-WB) IgG kit (LDBio Diagnostics, Lyon, France), a recently commercialized immunoblot assay for the diagnosis of various clinical presentations of chronic aspergillosis. Three hundred eight serum samples from 158 patients with aspergillosis sensu lato (s.l.) were analyzed. More specifically, 267 serum samples were derived from patients with Aspergillus disease, including 89 cases of chronic pulmonary aspergillosis, 10 of aspergilloma, and 32 of allergic bronchopulmonary aspergillosis, while 41 samples were from patients with Aspergillus colonization, including 15 cystic fibrosis (CF) and 12 non-CF patients. For blood donor controls, the Asp-WB specificity was 94%, while the kit displayed a sensitivity for the aspergillosis s.l. diagnosis of 88.6%, with a diagnostic odds ratio (DOR) of 119 (95% confidence interval [CI], 57 to 251). The DOR values were 185.22 (95% CI,78.79 to 435.45) and 43.74 (95% CI, 15.65 to 122.20) for the diagnosis of Aspergillus disease and Aspergillus colonization, respectively. Among the patients, the sensitivities of the Asp-WB in the diagnosis of Aspergillus colonization were 100% and 41.7% in CF and non-CF patients, respectively. The Asp-WB yielded fewer false-negative results than did IPD. In conclusion, the Asp-WB kit performed well for the diagnosis of various clinical presentations of aspergillosis in nonimmunocompromised patients, with an enhanced standardization and a higher sensitivity than with IPD, which is the current reference method.

  4. Evaluation of immunodominant proteins of Mycobacterium avium paratuberculosis cell wall by Western blot analysis. (United States)

    Hashemi, Maryam; Madani, Rasool; Razmi, Nematollah


    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is a slow growing mycobactin, whose dependence on mycobacterial species is known to be the causative agent of Johne's disease (paratuberculosis) in all species of domestic ruminants worldwide. The organism is transmitted via close contact, ingestion, or transplacentally from mother to fetus and occurs commonly in grazing domestic animals. Johne's disease (JD) is characterized by gradual weight loss, decreased milk production, and diarrhea due to the chronic, progressive, granulomatous enteritis and lymphadenitis. The disease can cause serious economic damage to the dairy industry due to the loss of milk production and early culling of infected animals. In recent years, researchers have focused on the identification of a specific antigen of M. paratuberculosis to use in diagnosis test and preparation of effective vaccine. The goal of this study is evaluation of the immunodominant proteins of M. paratuberculosis cell wall. The amount of protein was determined with a Lowry assay (22.68 μg/100 μL). For production of polyclonal antibody against proteins of M. paratuberculosis cell wall, a New Zealand white rabbit was immunized with antigen and Freund's adjuvant. After immunization, the rabbit was bled to produce enriched serum. Antibodies were purified from serum with ion exchange chromatography. In the Ouchterlony test, the reactions between antigen and antibodies were seen in dilutions of one quarter for serum, one quarter for Ig, and one half for IgG by clear precipitation lines due to the well immunization of the rabbit. Electrophoresis and Western blot analysis were used and subsequently a sharp band appeared in nitrocellulose paper; these bands were about 25, 37, 50, 75, and 150 kDa molecular weight, which indicated immunodominant proteins.

  5. Detection of classical and atypical/Nor98 scrapie by the paraffin-embedded tissue blot method. (United States)

    Wemheuer, W M; Benestad, S L; Wrede, A; Wemheuer, W E; Brenig, B; Bratberg, B; Schulz-Schaeffer, W J


    The paraffin-embedded tissue (PET) blot method was used to investigate sections of the central nervous system and lymphatic tissues from 24 cases of classical scrapie and 25 cases of atypical/Nor98 scrapie in sheep and four healthy control sheep. The PET blot detected deposits of PrP(Sc) in the brain tissue of all 49 sheep with scrapie but no PrP(Sc) labelling could be detected in the control sheep. By contrast, not all the atypical/Nor98 scrapie cases were detectable by immunohistochemistry. The high sensitivity of the PET blot method made it possible to observe that in some atypical/Nor98 cases, deposits of PrP(Sc) may be restricted to supratentorial brain structures and that the diagnosis may be missed when only testing the obex area, where deposits are common in classical scrapie, and the cerebellar structures, where deposits are considered to be common in atypical/Nor98 cases.

  6. A slot blot immunoassay for quantitative detection of Plasmodium falciparum circumsporozoite protein in mosquito midgut oocyst.

    Directory of Open Access Journals (Sweden)

    Sanjai Kumar

    Full Text Available There is still a need for sensitive and reproducible immunoassays for quantitative detection of malarial antigens in preclinical and clinical phases of vaccine development and in epidemiology and surveillance studies, particularly in the vector host. Here we report the results of sensitivity and reproducibility studies for a research-grade, quantitative enhanced chemiluminescent-based slot blot assay (ECL-SB for detection of both recombinant Plasmodium falciparum circumsporozoite protein (rPfCSP and native PfCSP from Oocysts (Pf Oocyst developing in the midguts of Anopheles stephensi mosquitoes. The ECL-SB detects as little as 1.25 pg of rPfCSP (linear range of quantitation 2.5-20 pg; R2 = 0.9505. We also find the earliest detectable expression of native PfCSP in Pf Oocyst by ECL-SB occurs on day 7 post feeding with infected blood meal. The ECL-SB was able to detect approximately as few as 0.5 day 8 Pf Oocysts (linear quantitation range 1-4, R2 = 0.9795 and determined that one Pf Oocyst expressed approximately 2.0 pg (0.5-3 pg of native PfCSP, suggesting a similar range of detection for recombinant and native forms of Pf CSP. The ECL-SB is highly reproducible; the Coefficient of Variation (CV for inter-assay variability for rPf CSP and native PfCSP were 1.74% and 1.32%, respectively. The CVs for intra-assay variability performed on three days for rPf CSP were 2.41%, 0.82% and 2% and for native Pf CSP 1.52%, 0.57%, and 1.86%, respectively. In addition, the ECL-SB was comparable to microscopy in determining the P. falciparum prevalence in mosquito populations that distinctly contained either high and low midgut Pf Oocyst burden. In whole mosquito samples, estimations of positivity for P. falciparum in the high and low burden groups were 83.3% and 23.3% by ECL-SB and 85.7% and 27.6% by microscopy. Based on its performance characteristics, ECL-SB could be valuable in vaccine development and to measure the parasite prevalence in mosquitoes

  7. Assessment by Southern blot analysis of UV-induced damage and repair in human immunoglobulin genes. (United States)

    Bianchi, M S; Bianchi, N O; de la Chapelle, A


    Irradiation of DNA with UV light induces pyrimidine dimers and (6-4) photoproducts. The presence of one of these photolesions in the restriction site of a given endonuclease inhibits DNA cleavage and induces the formation of fragments by incomplete DNA digestion which appear as additional, facultative bands in Southern hybridization autoradiograms. The number and size of these fragments show a positive correlation with the UV dose. The response to UV light of immunoglobulin light-chain constant kappa and heavy-chain constant mu genes was analyzed with 2 specific probes. Constant kappa and mu genes when irradiated as part of the chromatin of living lymphocytes showed a UV sensitivity similar to that of naked DNA. The same genes from granulocytes had 50-60 times lower UV sensitivity. When cells were allowed to repair photolesions for 24 h the facultative bands from granulocytes disappeared indicating that these cells were able to remove photolesions from constant kappa and mu genes. Facultative bands from lymphocytes showed a smaller decrease of density after 24 h repair. This suggests that lymphocytes are less efficient than granulocytes in removing UV damage from constant kappa and mu genes.

  8. Hybrid intermediaries


    Cetorelli, Nicola


    I introduce the concept of hybrid intermediaries: financial conglomerates that control a multiplicity of entity types active in the "assembly line" process of modern financial intermediation, a system that has become known as shadow banking. The complex bank holding companies of today are the best example of hybrid intermediaries, but I argue that financial firms from the "nonbank" space can just as easily evolve into conglomerates with similar organizational structure, thus acquiring the cap...

  9. Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan

    Directory of Open Access Journals (Sweden)

    A Rasul


    Full Text Available Background: The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp. in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.Methods: Reverse line blot (RLB assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.Results: Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.Conclusion: Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001. Risk factor analysis revealed that male (P = 0.03, ani­mals infested by ticks (P = 0.03 and herd composed of sheep only (P = 0.001 were more infected by blood parasites.

  10. Highly Sensitive Fluorescent-labeled Probes and Glass Slide Hybridization for the Detection of Plant RNA Viruses and a Viroid

    Institute of Scientific and Technical Information of China (English)

    Zhiyou DU; Bo JIN; Wenhong LIU; Liang CHEN; Jishuang CHEN


    In this study, a modified method of the conventional RNA dot-blot hybridization was established, by replacing 32P labels with CY5 labels and replacing nylon membranes with positive-charged glass slides, for detecting plant RNA viruses and a viroid. The modified RNA dot-blot hybridization method was named glass slide hybridization. The optimum efficiency of RNA binding onto the surfaces of activated glass slide was achieved using aminosilane-coated glass slide as a solid matrix and 5×saline sodium citrate (SSC) as a spotting solution. Using a CY5-labeled DNA probe prepared through PCR amplification, the optimized glass slide hybridization could detect as little as 1.71 pg of tobacco mosaic virus (TMV) RNA.The sensitivity of the modified method was four times that of dot-blot hybridization on nylon membrane with a 32P-labeled probe. The absence of false positive within the genus Potyvirus [potato virus A, potato virus Y (PVY) and zucchini yellow mosaic virus] showed that this method was highly specific. Furthermore,potato spindle tuber viroid (PSTVd) was also detected specifically. A test of 40 field potato samples showed that this method was equivalent to the conventional dot-blot hybridization for detecting PVY and PSTVd. To our knowledge, this is the first report of using dot-blot hybridization on glass slides with fluorescent-labeled probes for detecting plant RNA viruses and a viroid.

  11. Comparison of Diagnostic Value of Antigen B and Protoscoleces Antigen in Diagnosis of Hydatid Cyst by Blotting Method

    Directory of Open Access Journals (Sweden)

    F. Oreizi


    Full Text Available Introduction & Objective : Hydatidosis, a disease caused by the cestod helminth echinococcus granulosus, is one of the most important parasitic zoonosis in man and a variety of animals. Sensitive and reliable serologic methods are necessary to confirm the diagnosis. In this study, Ag B and Psc Ag were purified as two specific parasitic antigens and evaluated by Dot blotting used on the serum of hydatidosis patients and control group in order to identify the most sensitive and specific subunits.Materials and Methods: In an analytic and comparative study, serum samples collected from 22 patients under operation of hydatid cyst. As a control group, 4 patients with acute toxoplasmosis, 4 patients with leishmaniasis, 4 patients infected by non-hydatid cestods(Tenia saginata and H.nana and 4 normal subjects were included in this investigation. Infected sheep’s liver and lung were used for the preparation of antigen. Cyst fluid containing protoscoleces was extracted and then partially purified with a protein A column. AgB and Psc Ags were interacted with hydatid and control sera, with Dot blot method and sensitivity and specificity of these antigens were evaluated. Results: Sensitivity and specificity were estimated 95.9% and 81% respectively, for AgB and 100% and 63% respectively, for Psc Ag in Dot blot Method. Conclusion: Evaluation of sensitivity and specificity of AgB and Psc Ag using Dot blotting revealed that AgB has high value for diagnosis of hydatidosis. and presumably can help physicians to diagnose hydatid cyst easier than other routine tests.

  12. Differentiation of Enterococcus faecium from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains by PCR and dot-blot hybridisation. (United States)

    Langa, S; Fernández, A; Martín, R; Reviriego, C; Marín, M L; Fernández, L; Rodríguez, J M


    Variations in length and sequence of the 16S/23S spacer region of Enterococcus faecium provided the basis for development of simple PCR and dot-blot hybridisation assays that enabled the differentiation of potentially probiotic Enterococcus faecium strains from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Such assays may be useful for differentiation of yoghurt starter cultures and enterococcal strains when they are simultaneously present in probiotic food products.

  13. Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids. (United States)

    Armua-Fernandez, Maria Teresa; Nonaka, Nariaki; Sakurai, Tatsuya; Nakamura, Seita; Gottstein, Bruno; Deplazes, Peter; Phiri, Isaac G K; Katakura, Ken; Oku, Yuzaburo


    We report the development of a colourimetric PCR/dot blot assay targeting the mitochondrial gene NADH dehydrogenase subunit 1 (nad1) for differential diagnosis of taeniid eggs. Partial sequences of the cestode nad1 gene were aligned and new primers were designed based on conserved regions. Species-specific oligonucleotide probes (S-SONP) for canine taeniid cestodes were then designed manually based on the variable region between the conserved primers. Specifically, S-SONP were designed for the Taenia crassiceps, T. hydatigena, T. multiceps, T. ovis, T. taeniaeformis, Echinococcus granulosus (genotype 1), E. multilocularis and E. vogeli. Each probe showed high specificity as no cross-hybridisation with any amplified nad1 fragment was observed. We evaluated the assay using 49 taeniid egg-positive samples collected from dogs in Zambia. DNA from 5 to 10 eggs was extracted in each sample. Using the PCR/dot blot assay, the probes successfully detected PCR products from T. hydatigena in 42 samples, T. multiceps in 3 samples, and both species (mixed infection) in the remaining 4 samples. The results indicate that the PCR/dot blot assay is a reliable alternative for differential diagnosis of taeniid eggs in faecal samples.

  14. Investigation of Anti-Toxocara Antibodies in Epileptic Patients and Comparison of Two Methods: ELISA and Western Blotting

    Directory of Open Access Journals (Sweden)

    Mohammad Zibaei


    Full Text Available The relationship between Toxocara infection and epilepsy was previously demonstrated by several case-control studies and case reports. These previous studies were often based on the enzyme-linked immunosorbent assay (ELISA using Toxocara excretory-secretory antigens, which are not specific due to cross-reactivity with other parasitic infections such as ascariasis, trichuriasis, and anisakiasis. An immunoblot analysis is highly specific and can detect low levels of Toxocara antibodies. Therefore, this assay may be useful in the identification of toxocariasis in epileptic patients. We examined patients who had epilepsy and healthy subjects for seropositivity for Toxocara infection by ELISA and Western blotting. Out of 85 epileptic patients, 10 (11.8% and 3 (3.5% persons exhibited Toxocara immunoglobulin G (IgG antibodies responses by ELISA and by both techniques, respectively. Moreover, in the healthy group (, 3 (3.5% persons were positive by ELISA, but none was detected by Western blotting. This study indicates that Toxocara infection is a risk factor for epilepsy in Iran. These findings strongly suggest the need to perform Western blotting immunodiagnosis, as well as the ELISA using Toxocara excretory-secretory antigens, to improve diagnosis of human toxocariasis in patients with epilepsy.

  15. Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

    DEFF Research Database (Denmark)

    Sahm, K.; MacGregor, BJ; Jørgensen, BB;


    , directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...

  16. Detection of Haemophilus influenzae by reverse dot-blot hybridization%反向斑点杂交法检测流感嗜血杆菌核酸

    Institute of Scientific and Technical Information of China (English)

    樊慧珍; 黄文杰; 梁昆; 方怡; 马立人; 刘耀清


    目的探讨早期、快速检测流感嗜血杆菌的方法.方法应用流感嗜血杆菌编码外膜蛋白特异的P6基因设计引物,通过聚合酶链反应合成特异探针,采用反向斑点杂交法检测生物素标记DNA,并应用于痰标本的检测.结果只有流感嗜血杆菌扩增出351 bp的DNA片段,该探针能检测出10 ng的细菌DNA,与其他细菌、病毒、真菌无交叉反应.该方法和培养法分别检测50份痰标本,两者的阳性率分别为30%和22%.结论该方法快速、特异,对流感嗜血杆菌感染有早期诊断价值.

  17. Detection of Sleeping Beauty transposition in the genome of host cells by non-radioactive Southern blot analysis. (United States)

    Aravalli, Rajagopal N; Park, Chang W; Steer, Clifford J


    The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type.

  18. Detection of Rickettsia in Rhipicephalus sanguineus ticks and Ctenocephalides felis fleas from southeastern Tunisia by reverse line blot assay. (United States)

    Khrouf, Fatma; M'Ghirbi, Youmna; Znazen, Abir; Ben Jemaa, Mounir; Hammami, Adnene; Bouattour, Ali


    Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia.

  19. How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot. (United States)

    Beyer, Mandy; Kiweler, Nicole; Mahboobi, Siavosh; Krämer, Oliver H


    Histone deacetylases (HDACs) catalyze the deacetylation of lysine residues in their target proteins. This biochemical modification can have profound effects on the functions of these proteins and a dysregulation of HDAC activity contributes to severe diseases, including neoplastic transformation. In the following chapter, we present a strategy that allows to distinguish between the inhibition of the class I HDACs HDAC1, 2, and 3 and of the class IIb HDAC HDAC6. This method is based on Western blot and relies on the detection of hyperacetylated substrates of class I or class IIb HDACs in lysates from cells that were treated with histone deacetylase inhibitors (HDACi).

  20. Hybrid microelectronic technology (United States)

    Moran, P.

    Various areas of hybrid microelectronic technology are discussed. The topics addressed include: basic thick film processing, thick film pastes and substrates, add-on components and attachment methods, thin film processing, and design of thick film hybrid circuits. Also considered are: packaging hybrid circuits, automating the production of hybrid circuits, application of hybrid techniques, customer's view of hybrid technology, and quality control and assurance in hybrid circuit production.

  1. 地高辛标记的Northern blot检测鼠疫菌sRNA%Detection of Yersinia pestis sRNA by digoxigenin-labeled Northern blot

    Institute of Scientific and Technical Information of China (English)

    邓仲良; 苏山春; 孟祥荣; 吴移谋; 杨瑞馥; 韩延平


    [目的]随着高通量测序方法的应用,越来越多的sRNA (small non-coding RNA,sRNA)需验证.本研究建立用地高辛标记Northern blot检测鼠疫菌sRNA的技术,为细菌sRNA验证提供一种灵敏、特异的方法.[方法]在低铁条件下,提取鼠疫菌总RNA,10% dPAGE分离后电转到尼龙膜上并用紫外线交联RNA.膜经地高辛标记RyhB1或RyhB2寡核苷酸RNA探针过夜杂交后洗脱、封闭和免疫检测,最后曝光显影.[结果]地高辛标记的Northern blot曝光时间为20 s-3 min,RyhB1或RyhB2检测灵敏度分别为0.005 μg和0.05 μg.RyhB1或RyhB2探针特异性好,相互间无交叉反应.带正电或中性的尼龙膜都适用于杂交反应.RNA探针在42℃-65℃内杂交均可,提高温度可减少非特异性反应;而DNA探针杂交温度需摸索.[结论]本研究成功构建一种地高辛标记Northern blot检测鼠疫菌sRNA技术,具有特异性好、灵敏度高、探针易保存、曝光时间短等优点,为细菌sRNA验证和功能研究提供有利工具.%[Objective] With the application of high-throughput sequencing methods, more and more sRNAs are required to be verified.In this study, we developed the digoxigenin-labeled Northern blot method for detection of Yersinia pestis sRNA.[Methods] Total RNAs extracted from Yersinia.pestis grown under low-iron conditions were loaded onto 10% denaturing urea polyacrylamide gel (dPAGE) , electrophoresed and transferred to nylon membranes.Northern blots were fixed to the membrane by UV cross-linking and subjected to hybridization with 3 ' -end digoxigenin-labeled oligonucleotides RNA probe for RyhBl and RyhB2 overnight and followed by washing, blocking, immunological detection and finally exposed to film.[Results] Exposure time of digoxigenin-labelled Northern blot was 20s-3min.The detection sensitivity of RyhBl and RyhB2 was 0.005μg and 0.05 μg, respectively.RyhBl and RyhB2 probe specificity was high and no cross reaction with each other was found

  2. Analysis of Sperm Membrane Protein Relevant to Antisperm Antibody by Two-Dimensional Gel Electrophoresis and Western Blotting

    Institute of Scientific and Technical Information of China (English)

    Hao-fei WANG; Zhu-qiong XIANG; Yi-xing WANG


    Objective To identify the sperm membrane proteins that are associated with antisperm antibodyMethods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensional gel electrophoresis followed by Western blot analysis to determine the molecular weights (MW) and isoelectric points (pI) of sperm membrane proteins that are associated with antisperm antibody.Results Eight kinds of MW with more than ten sperm membrane proteins can be recognized by antisperm antibody positive serum, of which the MWs and pI were 23 kD, 31 kD, 32 kD, 34 kD, 41 kD, 51 kD, 60 kD, 78 kD and 5.3, 5.5,5.7, 5.0, 5.3, 5.8, 6.0, 5.5~6.2, 4.6,5.1,5.5~5.8 respectively. The identification ratios of the sperm membrane proteins on 78 kD (60.7%), 60 kD (71.4%), 51 kD (14.9%) and 23 kD (14.29%) were higher.Conclusion The sperm membrane proteins with MW of 78 kD, 60 kD, 51 kD and 23 kD were associated with antisperm antibody and immunological infertility. Two-dimensional gel electrophoresis and Western blotting can precisely identify the sperm membrane proteins that are associated with antisperm antibody.

  3. Mere end blot pirringer

    DEFF Research Database (Denmark)

    Jantzen, Christian; Østergaard, Per


    Med udgangspunkt i en række cases på vellykkede oplevelsesøkonomiske forretningsmodeller argumenterer artiklen for, at oplevelsesprodukter skal bygge på et klart tema, som forbrugeren kan koble sig sanse- og følelsesmæssigt op på. Forbrugeren skal kunne omsætte produktets pirringer til egne erfar...

  4. Modification of T-cell antigenic properties of tetanus toxoid by SDS-PAGE separation. Implications for T-cell blotting

    DEFF Research Database (Denmark)

    Christensen, C B; Theander, T G


    Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20% of the ......Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20...... that SDS-PAGE alters the ability of TT to induce T-cell proliferation, possibly due to unpolymerized acrylamide binding to proteins during SDS-PAGE. The use of SDS-PAGE T-cell blotting in the screening for T-cell antigens must therefore be reconsidered. We suggest the use of SDS-Agarose Gel Electrophoresis...

  5. Hybrid Qualifications

    DEFF Research Database (Denmark)

    has turned out as a major focus of European education and training policies and certainly is a crucial principle underlying the European Qualifications Framework (EQF). In this context, «hybrid qualifications» (HQ) may be seen as an interesting approach to tackle these challenges as they serve «two...... masters», i.e. by producing skills for the labour market and enabling individuals to progress more or less directly to higher education. The specific focus of this book is placed on conditions, structures and processes which help to combine VET with qualifications leading into higher education...

  6. Hybrid Gear (United States)

    Handschuh, Robert F. (Inventor); Roberts, Gary D. (Inventor)


    A hybrid gear consisting of metallic outer rim with gear teeth and metallic hub in combination with a composite lay up between the shaft interface (hub) and gear tooth rim is described. The composite lay-up lightens the gear member while having similar torque carrying capability and it attenuates the impact loading driven noise/vibration that is typical in gear systems. The gear has the same operational capability with respect to shaft speed, torque, and temperature as an all-metallic gear as used in aerospace gear design.

  7. 用DNA探针检测沙眼衣原体%Detection of Chlamydia Trachomatis by DNA Probe Blot

    Institute of Scientific and Technical Information of China (English)

    王卫萍; 陈亚利; 武建国


    目的: 建立一种敏感而特异的沙眼衣原体分子生物学检测方法. 方法: 用PCR扩增517bp的沙眼衣原体种特异性基因片段并标记成探针,建立DNA探针杂交检测沙眼衣原体的方法.结果: 探针只与沙眼衣原体L2、TE55株DNA呈阳性杂交斑点,与其他两种衣原体、解脲支原体、淋病奈瑟菌、大肠埃希菌、金黄色葡萄球菌、流感嗜血杆菌及白色念珠菌DNA斑点膜无阳性杂交信号.从100例慢性宫颈炎和前列腺炎病人生殖道分泌物中检出阳性22例,阳性率22%. 结论: 建立的DNA探针检测沙眼衣原体方法具有较高的敏感性和特异性,可用于批量临床标本的检测.%Objectives: To establish a sensitive and specific molecular biologicalmethod for detecting Chlamydia trachomatis of genital tract infection patients. Methods: A DNA blot assay was developed by coating DNA of Chlamydia trachomatisand/or extract of clinical samples on nitrocellulose (NC) membrane, blotting witha DNA probe labeled with DIG. Results: There was no positive blotting in Chlamydiapneumoniae, Chlamydia psittaci, Ureaplasma urealyticum, Neisseria gonorrhoeae,Esherichia coli, Staphylococcus aureus, Haemophilus infuenzae and Candida albicansexcept Chlamydia trachomatis. The sensitivity could be improved to 1pg. The positivepercentage was 22% (22/100) in detection of swabs collected from 100 chroniccervicitis and prostatitis patients. Conclusions: This method was not only sensitive, rapid and specific but also could be applied to detect batch samples. Natl J Androl,2001,7(2):102~104

  8. Intuitionistic hybrid logic

    DEFF Research Database (Denmark)

    Braüner, Torben


    Intuitionistic hybrid logic is hybrid modal logic over an intuitionistic logic basis instead of a classical logical basis. In this short paper we introduce intuitionistic hybrid logic and we give a survey of work in the area.......Intuitionistic hybrid logic is hybrid modal logic over an intuitionistic logic basis instead of a classical logical basis. In this short paper we introduce intuitionistic hybrid logic and we give a survey of work in the area....

  9. Continuity Controlled Hybrid Automata


    Bergstra, J. A.; Middelburg, C.A.


    We investigate the connections between the process algebra for hybrid systems of Bergstra and Middelburg and the formalism of hybrid automata of Henzinger et al. We give interpretations of hybrid automata in the process algebra for hybrid systems and compare them with the standard interpretation of hybrid automata as timed transition systems. We also relate the synchronized product operator on hybrid automata to the parallel composition operator of the process algebra. It turns out that the f...

  10. Lectin staining and Western blot data showing differential sialylation of nutrient-deprived cancer cells to sialic acid supplementation. (United States)

    Badr, Haitham A; AlSadek, Dina M M; Mathew, Mohit P; Li, Chen-Zhong; Djansugurova, Leyla B; Yarema, Kevin J; Ahmed, Hafiz


    This report provides data that are specifically related to the differential sialylation of nutrient deprived breast cancer cells to sialic acid supplementation in support of the research article entitled, "Nutrient-deprived cancer cells preferentially use sialic acid to maintain cell surface glycosylation" [1]. Particularly, breast cancer cells, when supplemented with sialic acid under nutrient deprivation, display sialylated glycans at the cell surface, but non-malignant mammary cells show sialylated glycans intracellularly. The impact of sialic acid supplementation under nutrient deprivation was demonstrated by measuring levels of expression and sialylation of two markers, EGFR1 and MUC1. This Data in Brief article complements the main manuscript by providing detailed instructions and representative results for cell-level imaging and Western blot analyses of changes in sialylation during nutrient deprivation and sialic acid supplementation. These methods can be readily generalized for the study of many types of glycosylation and various glycoprotein markers through the appropriate selection of fluorescently-labeled lectins.

  11. Standardisation of Western blotting to detect HTLV-1 antibodies synthesised in the central nervous system of HAM/TSP patients

    Directory of Open Access Journals (Sweden)

    Luiz Claudio Pereira Ribeiro


    Full Text Available Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1 antibodies (Abs represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP patients. Western blotting (WB for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I and gag (p24 proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.

  12. MDR-TB Antibody Response (Western Blot) to Fractions of Isoniazid and Rifampicin Resistant Antigens of Mycobacterium tuberculosis. (United States)

    Hadizadeh Tasbiti, Alireza; Yari, Shamsi; Ghanei, Mostafa; Shokrgozar, Mohammad Ali; Bahrmand, Ahmadreza


    Drug-resistant TB poses a major threat to control of TB worldwide. Despite progress in the detection of Multidrug-resistant TB (MDR-TB) cases, a major diagnostic gap remains: 55% of reported TB patients estimated to have MDR-TB were not detected in 2013. MDR-TB antigens were conjugated to CNBr-activated Sepharose 4B. Specific polyclonal antibodies against MDR-TB Ags were prepared in rabbits using two boosted injections of the MDR-TB antigen. The antibodies were purified and treated with susceptible TB to remove any non-specific and cross-reactive antibodies. In the present study, comparative analysis of electrophoretic pattern of different antigens of INH/RIF-resistant TB were studied for identifying protein profiles. A RIF-resistant TB antigen was shown here to have different protein profiles from INH-resistant TB isolate. The results of Western blotting analysis showed that in the RIF- and INH-resistant antigenic fractions some bands of 14.4 and 45 kDa as immunogenic were common. Moreover, four bands of RIF-resistant TB antigen fractions (16, 19, 21, and 45 KDa) and one band of INH-resistant TB (about 26 KDa) were detected as diagnostic antigens. This study suggests that the Western blot is an accurate test to survey INH- and RIF-resistant TB antigens of M. tuberculosis infection. These findings indicate that MDR-TB diagnosis (based on Ag detection) could be useful in the identification of disease stages that precede symptomatic and microbiologically positive TB, such as subclinical and incipient TB.

  13. Hybridized tetraquarks

    Directory of Open Access Journals (Sweden)

    A. Esposito


    Full Text Available We propose a new interpretation of the neutral and charged X,Z exotic hadron resonances. Hybridized-tetraquarks are neither purely compact tetraquark states nor bound or loosely bound molecules but rather a manifestation of the interplay between the two. While meson molecules need a negative or zero binding energy, its counterpart for h-tetraquarks is required to be positive. The formation mechanism of this new class of hadrons is inspired by that of Feshbach metastable states in atomic physics. The recent claim of an exotic resonance in the Bs0π± channel by the D0 Collaboration and the negative result presented subsequently by the LHCb Collaboration are understood in this scheme, together with a considerable portion of available data on X,Z particles. Considerations on a state with the same quantum numbers as the X(5568 are also made.

  14. Hybridized Tetraquarks

    CERN Document Server

    Esposito, A.; Polosa, A.D.


    We propose a new interpretation of the neutral and charged X, Z exotic hadron resonances. Hybridized-tetraquarks are neither purely compact tetraquark states nor bound or loosely bound molecules. The latter would require a negative or zero binding energy whose counterpart in h-tetraquarks is a positive quantity. The formation mechanism of this new class of hadrons is inspired by that of Feshbach metastable states in atomic physics. The recent claim of an exotic resonance in the Bs pi+- channel by the D0 collaboration and the negative result presented subsequently by the LHCb collaboration are understood in this scheme, together with a considerable portion of available data on X, Z particles. Considerations on a state with the same quantum numbers as the X(5568) are also made.

  15. Fast hybridization solution for the detection of immobilized nucleic acids. (United States)

    Yang, T T; Kain, S R


    We have developed a fast hybridization solution, termed ExpressHyb, for the rapid and sensitive detection of nucleic acids immobilized on membrane supports. This solution reduces typical hybridization times of 12-24 h to as little as 1 h while simultaneously increasing the sensitivity of detection in many applications. Using ExpressHyb, human beta-actin mRNA was detected on a human multiple tissue Northern (MTN) blot following a 30-min hybridization, with optimal detection occurring with a 1-h hybridization interval. The moderately abundant human glyceraldehyde-3-phosphate dehydrogenase (G3PDH) mRNA was detected using similar hybridization conditions and yielded improved signal-to-background characteristics relative to overnight hybridizations in conventional solutions. ExpressHyb can be used with either 32P- or digoxigenin-labeled probes and works effectively with both cDNA and oligonucleotide probes. For non-isotopic detection in particular, ExpressHyb reduces the nonspecific background commonly encountered with this technique. In cDNA library screening, ExpressHyb was found to both reduce the time required for effective hybridizations and to increase the number of positive colonies obtained relative to conventional overnight procedures. Taken together, these results illustrate the broad capability of ExpressHyb Hybridization Solution to improve nucleic acid detection in a variety of important techniques.

  16. Continuity Controlled Hybrid Automata

    NARCIS (Netherlands)

    Bergstra, J.A.; Middelburg, C.A.


    We investigate the connections between the process algebra for hybrid systems of Bergstra and Middelburg and the formalism of hybrid automata of Henzinger et al. We give interpretations of hybrid automata in the process algebra for hybrid systems and compare them with the standard interpretation of

  17. Continuity controlled Hybrid Automata

    NARCIS (Netherlands)

    Bergstra, J.A.; Middelburg, C.A.


    We investigate the connections between the process algebra for hybrid systems of Bergstra and Middelburg and the formalism of hybrid automata of Henzinger et al. We give interpretations of hybrid automata in the process algebra for hybrid systems and compare them with the standard interpretation of

  18. Spectrophotometric quantitation of DNA on blots after ethanol-solubilization of the MTT-formazan from anti-digoxigenin-based detection of nucleic acids. (United States)

    Colgan, D J


    The tetrazolium salt, 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) has recently been established as a substitute for Nitro-blue tetrazolium (NBT) in stain mixtures using antibody-conjugated alkaline phosphatase for the location of proteins on Western blots (Heegaard, 1990). Experiments reported here show that MTT is as sensitive as NBT in digoxigenin-labeled probe localization (on nucleic acid blots) utilizing alkaline-phosphatase-labelled, anti-digoxigenin antibodies. Moreover, as the formazan from MTT is soluble in ethanol, it is shown that spectrophotometric quantitation can be used to estimate the amount of target DNA on dot and Southern blots. For dot blotting, pBR328 was used as the probe and pBR322 as target. For Southern blots, human rDNA was used as the probe and total genomic calf DNA as the target. Staining response was linear over at least six twofold DNA dilutions in both types of blot.

  19. [Optimization of the immunoelectrophoresis technic (western blot) for the confirmation of human immunodeficiency virus infection (HIV) in Panama]. (United States)

    Pascale, J M; de Austin, E; de Moreno, N O; Ledezma, C; de Márquez, E; Blanco, R; Quiroz, E; Calzada, J; Vincensini, A R; de Martin, M C


    The purpose of this study is to report the results of the authors' investigation to apply the western blot technique (WB UP-LCS) in the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection. To do this, the authors separated the proteins of the HIV-1 virus by electrophoresis, based on their molecular weight, in poliacilamide gel with SDS (SDS-PAGE) during 3 hours at 200 volts. Then they electrotransferred these proteins to nitrocellulose paper during four hours at 200 milliamperes, with the aid of external cooling. The nitrocellulose strips were evaluated considering the incubation time (1 and 16 hours), two conjugates (human anti IgG with Peroxidase and human anti IgG Biotin plus Streptatividine with Peroxidase) and two dilutions of the patients' sera (1/50 and 1/100). Based on their results the Authors conclude that, in the first place, the optimal conditions for the test include a dilution of 1/100 of the patients serum, incubation of the serum for 16 hours and the use of the conjugate of anti human IgG with Biotin and Streptavidine with Peroxidase; secondary, that the immunologic reactivity against proteins p24 and gp 160/120 is the most important diagnostic criterion for the confirmation of infection with HIV-1 and that they obtained a diagnostic correlation of 100% at a cost which was 5 to 7 times less than that of the commercial system.

  20. Development of rapid, sensitive and non-radioactive tissue-blot diagnostic method for the detection of citrus greening. (United States)

    Nageswara-Rao, Madhugiri; Miyata, Shin-Ichi; Ghosh, Dilip; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame


    Citrus huanglongbing (HLB or citrus greening) is one of the most devastating diseases of citrus worldwide. The disease is caused by Gram-negative, phloem-limited α-proteobacterium, 'Candidatus Liberibacter asiaticus', vectored by the psyllid, Diaphorina citri Kuwayama. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection and non-uniform distribution within the tree makes the detection of the pathogen very difficult. Efficient management of HLB disease requires rapid and sensitive detection early in the infection followed by eradication of the source of pathogen and the vector. The polymerase chain reaction (PCR) based method is most commonly employed for screening the infected/suspected HLB plants and psyllids. This is time consuming, cumbersome and not practical for screening large number of samples in the field. To overcome this, we developed a simple, sensitive, non-radioactive, tissue-blot diagnostic method for early detection and screening of HLB disease. Digoxigenin labeled molecular probes specific to 'Ca. L. asiaticus' nucleotide sequences have been developed and used for the detection of the pathogen of the HLB disease. The copy number of the target genes was also assessed using real-time PCR experiments and the optimized real-time PCR protocol allowed positive 'Ca. L. asiaticus' detection in citrus samples infected with 'Ca. L. asiaticus' bacterium.

  1. Cross-Reactions between Toxocara canis and Ascaris suum in the diagnosis of visceral larva migrans by western blotting technique

    Directory of Open Access Journals (Sweden)

    NUNES Cáris Maroni


    Full Text Available Visceral larva migrans (VLM is a clinical syndrome caused by infection of man by Toxocara spp, the common roundworm of dogs and cats. Tissue migration of larval stages causes illness specially in children. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. After the introduction of the enzyme-linked immunosorbent assay (ELISA using the larval excretory-secretory antigen of T. canis (TES, the diagnosis specificity was greatly improved although cross-reactivity with other helminths are still being reported. In Brazil, diagnosis is routinely made after absorption of serum samples with Ascaris suum antigens, a nematode antigenicaly related with Ascaris lumbricoides which is a common intestinal nematode of children. In order to identify T. canis antigens that cross react to A. suum antigens we analyzed TES antigen by SDS-PAGE and Western blotting techniques. When we used serum samples from patients suspected of VLM and positive result by ELISA as well as a reference serum sample numerous bands were seen (molecular weight of 210-200 kDa, 116-97 kDa, 55-50 kDa and 35-29 kDa. Among these there is at least one band with molecular weight around 55-66 kDa that seem to be responsible for the cross-reactivity between T. canis e A. suum once it disappears when previous absorption of serum samples with A. suum antigens is performed

  2. Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis

    Directory of Open Access Journals (Sweden)

    Luciana Pereira Silva


    Full Text Available The present study was conducted to evaluate the frequency of antigenic components recognized by serum IgG antibodies in Western blotting (WB using a Strongyloides ratti larval extract for the diagnosis of human strongyloidiasis. In addition, the WB results were compared to the enzyme-linked immunosorbent assay (ELISA and the indirect immunofluorescence antibody test (IFAT results. Serum samples of 180 individuals were analyzed (80 with strongyloidiasis, 60 with other intestinal parasitoses, and 40 healthy individuals. S. ratti was obtained from fecal culture of experimentally infected Rattus rattus. For IFAT, S. ratti larvae were used as antigen and S. ratti larval antigenic extracts were employed in WB and ELISA. Eleven S. ratti antigenic components were predominantly recognized by IgG antibodies in sera of patients with strongyloidiasis. There was a positive concordance for the three tests in 87.5% of the cases of strongyloidiasis. The negative concordance in the three tests was 94% and 97.5%, in patients with other intestinal parasitoses and healthy individuals, respectively. In cases of positive ELISA and negative IFAT results, diagnosis could be confirmed by WB. ELISA, IFAT, and WB using S. ratti antigens showed a high rate of sensitivity and specificity. In conclusion, WB using S. ratti larval extract was able to recognize 11 immunodominant antigenic components, showing to be a useful tool to define the diagnosis in cases of equivocal serology.

  3. "Enzyme-Linked Immunotransfer Blot Analysis of Somatic and Excretory- Secretory Antigens of Fasciola hepatica in Diagnosis of Human Fasciolosis"

    Directory of Open Access Journals (Sweden)

    MB Rokni


    Full Text Available The liver fluke Fasciola hepatica causes fascioliasis, a liver disease in most part of the world and particularly in north of Iran. Diagnosis of the diseases is anchored in coprological manner but serological methods are preferable due to some obscurities. In this study, sera obtained from human patients infected with Fasciola hepatica were tested by the enzymelinked immunotrotransfer blot (EITB technique with the parasite s somatic and excretory-secretory (ES antigens in order to evaluate the diagnostic potential of the assay. The study included sera from 40 patients infected with F. hepatica, 20 infected with hydatidosis, 6 with toxocariasis, 10 with strongyloidiasis, 10 with amoebiasis, 5 with malaria and 30 normal controls. By this assay, most pf the serum samples from humans with fascioliasis recognized two antigenic polypeptides of 27 and 29 kDa using both antigens. The sensitivity, specificity, positive and negative predictive values for somatic antigen were 91.0%, 96.2%, 95.2% and 92.7% respectively, while these parameters as for ES antigen were 95.2%, 98.0%, 97.5% and 96.2%, correspondingly. Totally, two cases of reactions for the first antigen and one for the latter were verified. The study suggests that the 27 and 29 kDa bands for two antigens in EITB test could be considered for the immunodiagnosis of human fascioliasis.

  4. A novel methylation PCR that offers standardized determination of FMR1 methylation and CGG repeat length without southern blot analysis. (United States)

    Grasso, Marina; Boon, Elles M J; Filipovic-Sadic, Stela; van Bunderen, Patrick A; Gennaro, Elena; Cao, Ru; Latham, Gary J; Hadd, Andrew G; Coviello, Domenico A


    Fragile X syndrome and associated disorders are characterized by the number of CGG repeats and methylation status of the FMR1 gene for which Southern blot (SB) historically has been required for analysis. This study describes a simple PCR-only workflow (mPCR) to replace SB analysis, that incorporates novel procedural controls, treatment of the DNA in separate control and methylation-sensitive restriction endonuclease reactions, amplification with labeled primers, and two-color amplicon sizing by capillary electrophoresis. mPCR was evaluated in two independent laboratories with 76 residual clinical samples that represented typical and challenging fragile X alleles in both males and females. mPCR enabled superior size resolution and analytical sensitivity for size and methylation mosaicism compared to SB. Full mutation mosaicism was detected down to 1% in a background of 99% normal allele with 50- to 100-fold less DNA than required for SB. A low level of full mutation mosaicism in one sample was detected using mPCR but not observed using SB. Overall, the sensitivity for detection of full mutation alleles was 100% (95% CI: 89%-100%) with an accuracy of 99% (95% CI: 93%-100%). mPCR analysis of DNA from individuals with Klinefelter and Turner syndromes, and DNA from sperm and blood, were consistent with SB. As such, mPCR enables accurate, sensitive, and standardized methods of FMR1 analysis that can harmonize results across different laboratories.

  5. Lectin staining and Western blot data showing differential sialylation of nutrient-deprived cancer cells to sialic acid supplementation

    Directory of Open Access Journals (Sweden)

    Haitham A. Badr


    Full Text Available This report provides data that are specifically related to the differential sialylation of nutrient deprived breast cancer cells to sialic acid supplementation in support of the research article entitled, “Nutrient-deprived cancer cells preferentially use sialic acid to maintain cell surface glycosylation" [1]. Particularly, breast cancer cells, when supplemented with sialic acid under nutrient deprivation, display sialylated glycans at the cell surface, but non-malignant mammary cells show sialylated glycans intracellularly. The impact of sialic acid supplementation under nutrient deprivation was demonstrated by measuring levels of expression and sialylation of two markers, EGFR1 and MUC1. This Data in Brief article complements the main manuscript by providing detailed instructions and representative results for cell-level imaging and Western blot analyses of changes in sialylation during nutrient deprivation and sialic acid supplementation. These methods can be readily generalized for the study of many types of glycosylation and various glycoprotein markers through the appropriate selection of fluorescently-labeled lectins.

  6. Evaluation of western blotting for the diagnosis of enzootic bovine leukemia Avaliação da técnica de western blot no diagnóstico da leucose enzoótica bovina

    Directory of Open Access Journals (Sweden)

    E.T. Gonzalez


    Full Text Available A western blotting (WB procedure has been developed for detecting antibodies to bovine leukosis virus (BLV in cattle sera. Two hundred and thirty three serum samples from naturally infected cattle with BLV virus and serial bleedings from experimentally BLV infected cows were used. An agar gel immunodiffusion test (AGID was used for comparing with the results obtained by WB. The AGID positive sera showed a different degree of reactivity by WB test against the two most important viral antigens (gp51 and p24, or against one of them. Other proteins (gp30, p15, p12 and p10 were not detected with any AGID positive sera, being observed occasionally three bands corresponding to the p24 protein. Using sera obtained by BLV experimental inoculation, the antibodies directed to p24 appeared early (between the 2nd and 4th week post inoculation and thereafter antibodies to gp51were detected in some animals. The analysis of field serum samples by AGID as compared to WB showed an agreement of 90.9%. Only 1.7% of sera were negative by AGID and positive by WB and 7.2% that were not conclusive by AGID and were defined by WB (4.2% as positive and 3.0% as negative.Um sistema de western blotting (WB foi desenvolvido para detecção de anticorpos contra o vírus da leucose em soros de bovinos. Foram utilizadas amostras de soros de 233 animais naturalmente infectados e soros de vacas experimentalmente infectadas. O teste de imunodifusão em ágar (AGID foi usado para comparação dos resultados. Graus diferentes de reatividade foram observados em soros positivos ao AGID, quando testados em WB frente a um ou aos dois antígenos mais importantes (gp51 e p24. Outras proteínas (gp30, p15, p12 e p10 não foram detectadas por nenhum soro positivo ao AGID, sendo que três bandas correspondentes à proteína p24 foram observadas ocasionalmente. Em soros obtidos por inoculação experimental, anticorpos contra a proteína p24 foram detectados entre a segunda e a quarta semanas

  7. Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa). (United States)

    Cuttell, Leigh; Gómez-Morales, Maria Angeles; Cookson, Beth; Adams, Peter J; Reid, Simon A; Vanderlinde, Paul B; Jackson, Louise A; Gray, C; Traub, Rebecca J


    Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory-secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value=0.66) that increased to very good (k-value=0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (PTrichinella DNA in any mainland animals, but did reveal the presence of a second larvae-positive wild boar on Gabba

  8. Detection of Babesia and Theileria species infection in cattle from Portugal using a reverse line blotting method. (United States)

    Silva, M G; Marques, P X; Oliva, A


    Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR-reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases.

  9. Estrogen receptors alpha and beta in rat placenta: detection by RT-PCR, real time PCR and Western blotting

    Directory of Open Access Journals (Sweden)

    Al-Bader Maie D


    Full Text Available Abstract Background High levels of estrogens during pregnancy not only retard placental and fetal growth but can lead to reproductive tract abnormalities in male progeny. Estrogens act through estrogen receptors (ER to modulate the transcription of target genes. These ER exist in two isoforms, ER alpha and ER beta and recently several variants of these isoforms have been identified. Methods The expressions of ER isoforms and variants have been studied in rat placenta at 16, 19 and 21 days gestation (dg. Gene expression was assessed using RT-PCR and real time PCR while protein expression was studied using Western blotting followed by immunodetection. Placental homogenates were probed with: a monoclonal antibody raised against the steroid binding domain of the ER alpha (ER alpha -S, a monoclonal antibody raised against the hinge region of ER alpha (ER alpha -H and a polyclonal antibody raised against the amino terminus of ER beta. Results ER alpha and ER beta mRNA and protein were detected from as early as 16 dg. Two PCR products were detected for ER alpha, one for the wild type ER alpha, and a smaller variant. Real time PCR results suggested the presence of a single product for ER beta. The antibodies used for detection of ER alpha protein both identified a single 67 kDa isoform; however a second 54 kDa band, which may be an ER alpha variant, was identified when using the ER alpha -H antibody. The abundance of both ER alpha bands decreased significantly between 16 and 19 dg. As for ER beta, four bands (76, 59, 54 and 41 kDa were detected. The abundance of the 59 and 54 kDa bands decreased significantly between 16 and 19 dg. Conclusion This study shows that both ER protein isoforms and their variants are present in rat placenta. The decrease in their expression near parturition suggests that the placenta may be relatively unresponsive to estrogens at this stage.

  10. 热带果树基因组DNA提取方法的改良及Southern blot分析%A Modified DNA Extraction Method for Tropical Fruit Trees and Southern Blot Analysis

    Institute of Scientific and Technical Information of China (English)

    彭军; 曾凡云; 龙海波; 黄俊生; 郭建荣


    Tissues of most tropical fruit trees contain polysaccharide and polyphenolics that restrict the extraction of high-quality genomie DNA for further molecular biology analysis. This experiment was conducted to develop a simple, universal and effective modified CTAB method for genomie DNA extraction from tissues of tropical fruit trees for southern blot analysis. This optimized procedure includes two steps. First, before the DNA was extrac- ted, the polysaccharide and polyphenolics were pre-removed through the STE lysis buffer by centrifugation. In addition, possible contamination by secondary metaboliates could be avoided by picking up the cotton-shaped ge- nomie DNA floccules resuspended in liquid at the DNA precipitation step with ethanol. With the modified CTAB method, high-quality DNAs were extracted from young leaves of the three representative tropical fruit trees, ba- nana (Musa spp. ) , papaya (Carica papaya L. ) and longan (Dimocarpus longan Lour. ). After digested with different restriction enzymes, fragments of banana ethylene receptor gene (GenBank accession No. AF113748), papaya phytoene desaturase gene (DQ779922) and longan flowering-related LEAFY homologous gene (DQ160214) were used as probes for further Southern blot analysis, respectively. The hybridization bands were clear with high background. The results indicate that the modified CTAB method, with pre-treated conjugation by DNA pick-up, is a simple and effective protocol for genomic DNA isolation from tropical fruit trees, which pro- duces high-quality genomic DNA available for further Southern blot analysis and other tests in molecular biology.%以香蕉、番木瓜、龙眼为代表材料,建立一套适合热带果树基因组DNA提取的方法——改良CTAB法。将提取各基因组DNA经限制性内切酶完全消化后,分别以香蕉内源乙烯受体基因(AF113748)、番木瓜八氢番茄红素脱氢酶基因PDS(DQ779922)、龙眼开花相关基因LEAFY(ADQ160214

  11. Molecular diagnosis of Theileria and Babesia species infecting cattle in Northern Spain using reverse line blot macroarrays

    Directory of Open Access Journals (Sweden)

    Juste Ramón A


    Full Text Available Abstract Background Piroplasmosis in cattle is caused by tick-borne haemoprotozoan parasites of the genera Theileria and Babesia. Molecular detection techniques offer higher sensitivity and specificity than microscopy examination methods and serological tests. A reverse line blot (RLB macroarray that included generic and species-specific probes for Theileria annulata, Theileria buffeli, Babesia bovis, Babesia bigemina, Babesia divergens and Babesia major was used to study the presence and identity of the piroplasm species infecting 263 bovine blood samples from 79 farms, most of them in Northern Spain. Microscopy examination of blood smears and haematology were also performed whenever possible to identify animals with parasitaemia. Results RLB hybridisation identified infection in 54.0% of the samples, whereas only 28.8% were positive by microscopy examination. The most frequently found species was T. buffeli, present in 42.6% of the samples. T. annulata was found in 22 samples (8.4% from 12 farms, including 9 farms (14 samples located in Northern Spain where presence of the vector is not very common. Babesia infections were less frequently detected: B. major was found in 3.0% of the samples, B. bigemina in 2.7%, B. bovis in 2.3% and B. divergens in 1.1%. Mixed infections were detected in 14 samples, accounting for six different combinations of species. Conclusion This is the first report in which B. major and B. divergens have been detected in Spain using molecular identification techniques and the first time that B. bovis has been detected in Northern Spain. The detection of T. annulata in Northern Spain suggests that the distribution of Mediterranean theileriosis might be changing. Samples with positive RLB hybridisation but negative microscopy had haematology values within the normal ranges suggesting that they corresponded to chronic carriers that may serve as reservoirs of the infection. In this sense, sensitive and specific laboratorial

  12. Evaluation of two sets of immunohistochemical and Western blot confirmatory methods in the detection of typical and atypical BSE cases

    Directory of Open Access Journals (Sweden)

    Greenlee Justin J


    Full Text Available Abstract Background Three distinct forms of bovine spongiform encephalopathy (BSE, defined as classical (C-, low (L- or high (H- type, have been detected through ongoing active and passive surveillance systems for the disease. The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC and Western blot (WB BSE confirmatory protocols to detect C- and atypical (L- and H-type BSE forms. Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. Results The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type. Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step. Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. Conclusions The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE

  13. Hybrid Management in Hospitals

    DEFF Research Database (Denmark)

    Byrkjeflot, Haldor; Jespersen, Peter Kragh


    Artiklen indeholder et litteraturbaseret studium af ledelsesformer i sygehuse, hvor sundhedsfaglig ledelse og generel ledelse mikses til hybride ledelsesformer......Artiklen indeholder et litteraturbaseret studium af ledelsesformer i sygehuse, hvor sundhedsfaglig ledelse og generel ledelse mikses til hybride ledelsesformer...

  14. From hybrid swarms to swarms of hybrids (United States)

    The introgression of modern humans (Homo sapiens) with Neanderthals 40,000 YBP after a half-million years of separation, may have led to the best example of a hybrid swarm on earth. Modern trade and transportation in support of the human hybrids has continued to introduce additional species, genotyp...

  15. The Hybrid Museum: Hybrid Economies of Meaning

    DEFF Research Database (Denmark)

    Vestergaard, Vitus


    this article shows that there are two different museum mindsets where the second mindset leans towards participatory practices. It is shown how a museum can support a hybrid economy of meaning that builds on both a user generated economy of meaning and an institutional economy of meaning and adds value to both....... Such a museum is referred to as a hybrid museum....

  16. Relative performance of Organon kit in comparison to Du Pont for confirmatory serological testing of HIV infection by western blot test in sera from blood donors. (United States)

    Aggarwal, R K; Chatterjee, R; Chattopadhya, D; Kumari, S


    A total of 32 specimens with different categories of reactivity by Du Pont Western Blot kit comprising of specimens showing full spectrum of HIV-I antigen specific bands, 19 specimens showing total absence of bands and four specimens showing non-specific bands (without any interpretative importance) were subjected to Western Blot testing by Organon test. Of the nine specimens showing full spectrum of bands by Du Pont the correlation with Organon kit was 100 per cent based on WHO criteria. Four specimens with non-specific indeterminate band pattern by Du Pont failed to show any band in Organon kit, indicating that latter to be more specific.

  17. Resin Catalyst Hybrids

    Institute of Scientific and Technical Information of China (English)

    S. Asaoka


    @@ 1Introduction: What are resin catalyst hybrids? There are typically two types of resin catalyst. One is acidic resin which representative is polystyrene sulfonic acid. The other is basic resin which is availed as metal complex support. The objective items of this study on resin catalyst are consisting of pellet hybrid, equilibrium hybrid and function hybrid of acid and base,as shown in Fig. 1[1-5].

  18. Mesoscale hybrid calibration artifact (United States)

    Tran, Hy D.; Claudet, Andre A.; Oliver, Andrew D.


    A mesoscale calibration artifact, also called a hybrid artifact, suitable for hybrid dimensional measurement and the method for make the artifact. The hybrid artifact has structural characteristics that make it suitable for dimensional measurement in both vision-based systems and touch-probe-based systems. The hybrid artifact employs the intersection of bulk-micromachined planes to fabricate edges that are sharp to the nanometer level and intersecting planes with crystal-lattice-defined angles.

  19. Intraply Hybrid Composite Design (United States)

    Chamis, C. C.; Sinclair, J. H.


    Several theoretical approaches combined in program. Intraply hybrid composites investigated theoretically and experimentally at Lewis Research Center. Theories developed during investigations and corroborated by attendant experiments used to develop computer program identified as INHYD (Intraply Hybrid Composite Design). INHYD includes several composites micromechanics theories, intraply hybrid composite theories, and integrated hygrothermomechanical theory. Equations from theories used by program as appropriate for user's specific applications.

  20. Homoploid hybrid expectations (United States)

    Homoploid hybrid speciation occurs when a stable, fertile, and reproductively isolated lineage results from hybridization between two distinct species without a change in ploidy level. Reproductive isolation between a homoploid hybrid species and its parents is generally attained via chromosomal re...

  1. Realizing the Hybrid Library. (United States)

    Pinfield, Stephen; Eaton, Jonathan; Edwards, Catherine; Russell, Rosemary; Wissenburg, Astrid; Wynne, Peter


    Outlines five projects currently funded by the United Kingdom's Electronic Libraries Program (eLib): HyLiFe (Hybrid Library of the Future), MALIBU (MAnaging the hybrid Library for the Benefit of Users), HeadLine (Hybrid Electronic Access and Delivery in the Library Networked Environment), ATHENS (authentication scheme), and BUILDER (Birmingham…

  2. Hybrid quantum information processing

    Energy Technology Data Exchange (ETDEWEB)

    Furusawa, Akira [Department of Applied Physics, School of Engineering, The University of Tokyo (Japan)


    I will briefly explain the definition and advantage of hybrid quantum information processing, which is hybridization of qubit and continuous-variable technologies. The final goal would be realization of universal gate sets both for qubit and continuous-variable quantum information processing with the hybrid technologies. For that purpose, qubit teleportation with a continuousvariable teleporter is one of the most important ingredients.

  3. Spoof Plasmon Hybridization

    CERN Document Server

    Zhang, Jingjing; Luo, Yu; Shen, Xiaopeng; Maier, Stefan A; Cui, Tie Jun


    Plasmon hybridization between closely spaced nanoparticles yields new hybrid modes not found in individual constituents, allowing for the engineering of resonance properties and field enhancement capabilities of metallic nanostructure. Experimental verifications of plasmon hybridization have been thus far mostly limited to optical frequencies, as metals cannot support surface plasmons at longer wavelengths. Here, we introduce the concept of 'spoof plasmon hybridization' in highly conductive metal structures and investigate experimentally the interaction of localized surface plasmon resonances (LSPR) in adjacent metal disks corrugated with subwavelength spiral patterns. We show that the hybridization results in the splitting of spoof plasmon modes into bonding and antibonding resonances analogous to molecular orbital rule and plasmonic hybridization in optical spectrum. These hybrid modes can be manipulated to produce enormous field enhancements (larger than 5000) by tuning the separation between disks or alte...

  4. Henkin and Hybrid Logic

    DEFF Research Database (Denmark)

    Blackburn, Patrick Rowan; Huertas, Antonia; Manzano, Maria;


    Leon Henkin was not a modal logician, but there is a branch of modal logic that has been deeply influenced by his work. That branch is hybrid logic, a family of logics that extend orthodox modal logic with special proposition symbols (called nominals) that name worlds. This paper explains why...... Henkin’s techniques are so important in hybrid logic. We do so by proving a completeness result for a hybrid type theory called HTT, probably the strongest hybrid logic that has yet been explored. Our completeness result builds on earlier work with a system called BHTT, or basic hybrid type theory...... is due to the first-order perspective, which lies at the heart of Henin’s best known work and hybrid logic....

  5. Nanoscale Organic Hybrid Electrolytes

    KAUST Repository

    Nugent, Jennifer L.


    Nanoscale organic hybrid electrolytes are composed of organic-inorganic hybrid nanostructures, each with a metal oxide or metallic nanoparticle core densely grafted with an ion-conducting polyethylene glycol corona - doped with lithium salt. These materials form novel solvent-free hybrid electrolytes that are particle-rich, soft glasses at room temperature; yet manifest high ionic conductivity and good electrochemical stability above 5V. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Hybrid Action Systems

    DEFF Research Database (Denmark)

    Ronkko, Mauno; Ravn, Anders P.


    a differential action, which allows differential equations as primitive actions. The extension allows us to model hybrid systems with both continuous and discrete behaviour. The main result of this paper is an extension of such a hybrid action system with parallel composition. The extension does not change...... the original meaning of the parallel composition, and therefore also the ordinary action systems can be composed in parallel with the hybrid action systems....

  7. BSA Hybrid Synthesized Polymer

    Institute of Scientific and Technical Information of China (English)

    Zong Bin LIU; Xiao Pei DENG; Chang Sheng ZHAO


    Bovine serum albumin (BSA), a naturally occurring biopolymer, was regarded as a polymeric material to graft to an acrylic acid (AA)-N-vinyl pyrrolidone (NVP) copolymer to form a biomacromolecular hybrid polymer. The hybrid polymer can be blended with polyethersulfone (PES) to increase the hydrophilicity of the PES membrane, which suggested that the hybrid polymer might have a wide application in the modification of biomaterials.


    Directory of Open Access Journals (Sweden)

    V. Dvadnenko


    Full Text Available The hybrid vehicle control system includes a start–stop system for an internal combustion engine. The system works in a hybrid mode and normal vehicle operation. To simplify the start–stop system, there were user new possibilities of a hybrid car, which appeared after the conversion. Results of the circuit design of the proposed system of basic blocks are analyzed.

  9. Hybrid radiator cooling system (United States)

    France, David M.; Smith, David S.; Yu, Wenhua; Routbort, Jules L.


    A method and hybrid radiator-cooling apparatus for implementing enhanced radiator-cooling are provided. The hybrid radiator-cooling apparatus includes an air-side finned surface for air cooling; an elongated vertically extending surface extending outwardly from the air-side finned surface on a downstream air-side of the hybrid radiator; and a water supply for selectively providing evaporative cooling with water flow by gravity on the elongated vertically extending surface.

  10. Hybrid Unifying Variable Supernetwork Model

    Institute of Scientific and Technical Information of China (English)

    LIU; Qiang; FANG; Jin-qing; LI; Yong


    In order to compare new phenomenon of topology change,evolution,hybrid ratio and network characteristics of unified hybrid network theoretical model with unified hybrid supernetwork model,this paper constructed unified hybrid variable supernetwork model(HUVSM).The first layer introduces a hybrid ratio dr,the

  11. Large Unifying Hybrid Supernetwork Model

    Institute of Scientific and Technical Information of China (English)

    LIU; Qiang; FANG; Jin-qing; LI; Yong


    For depicting multi-hybrid process,large unifying hybrid network model(so called LUHNM)has two sub-hybrid ratios except dr.They are deterministic hybrid ratio(so called fd)and random hybrid ratio(so called gr),respectively.

  12. Hybrid FOSS Project (United States)

    National Aeronautics and Space Administration — Armstrong researchers are continuing their efforts to further develop FOSS technologies. A hybrid FOSS technique (HyFOSS) employs conventional continuous grating...

  13. A simple explanation for a case of incompatibility with the reading frame theory in Duchenne muscular dystrophy: failure to detect an aberrant restriction fragment in Southern blot analysis. (United States)

    Patria, S Y; Takeshima, Y; Suminaga, R; Nakamura, H; Iwasaki, R; Minagawa, T; Matsuo, M


    According to the translational reading frame theory, Duchenne muscular dystrophy (DMD) patients harbor out-of-frame deletion mutations in the dystrophin gene. We identified a Japanese DMD case who appeared to have an in-frame deletion of exons 46-54 that was disclosed by Southern blot analysis using a dystrophin cDNA as a probe. Analysis of dystrophin mRNA in skeletal muscle revealed the presence of an out-of-frame deletion of exons 46-53. In agreement with this result, the region encompassing exon 54 could be amplified from genomic DNA by polymerase chain reaction (PCR). Furthermore, re-analysis by Southern blot using an exon specific probe disclosed that a HindIII fragment containing exon 54 was present at aberrant size, leading to the incorrect conclusion that exon 54 had been deleted. Thus, this particular DMD case does not constitute an exception to the reading frame theory.

  14. Characterization of Sm14 related components in different helminths by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis

    Directory of Open Access Journals (Sweden)

    Nilton Thaumaturgo


    Full Text Available Sm14 was the first fatty acid-binding protein homologue identified in helminths. Thereafter, members of the same family were identified in several helminth species, with high aminoacid sequence homology between them. In addition, immune crossprotection was also reported against Fasciola hepatica infection, in animals previously immunized with the Schistosoma mansoni vaccine candidate, r-Sm14. In the present study, data on preliminary sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis of nine different helminth extracts focusing the identification of Sm14 related proteins, is reported. Out of these, three extracts - Ascaris suum (males and females, Echinostoma paraensei, and Taenia saginata - presented components that comigrated with Sm14 in SDS-PAGE, and that were recognized by anti-rSm14 policlonal serum, in Western blotting tests.

  15. Comparison of linear array and line blot assay for detection of human papillomavirus and diagnosis of cervical precancer and cancer in the atypical squamous cell of undetermined significance and low-grade squamous intraepithelial lesion triage study. (United States)

    Castle, Philip E; Gravitt, Patti E; Solomon, Diane; Wheeler, Cosette M; Schiffman, Mark


    We evaluated Linear Array (LA), a newly commercialized PGMY09/11 L1 consensus primer PCR test that detects 37 human papillomavirus (HPV) genotypes by reverse line blot hybridization, for the detection of individual HPV genotypes and carcinogenic HPV and its clinical performance for detecting 2-year cumulative cervical precancer and cancer using archived specimens from the Atypical Squamous Cell of Undetermined Significance (ASCUS) and Low-Grade Squamous Intraepithelial Lesion Triage Study. LA testing was conducted on enrollment specimens from women referred because of an ASCUS Pap test. To gauge the performance of the new test, the results were compared to those of its prototype predecessor assay, Line Blot Assay (LBA), restricted to paired results (n = 3,335). LA testing was done masked to LBA results and clinical outcomes. The results of LA and LBA testing were compared for detection of carcinogenic HPV and clinical outcomes of cervical precancer and cancer. Overall, 50% and 55% of the women tested positive for carcinogenic HPV by LBA and LA, respectively (P < 0.0001). The percent agreement for carcinogenic HPV detection was 88%, percent positive agreement was 80%, and kappa was 0.76 for detection of carcinogenic HPV by the two assays. There was a significant increase in detection by LA for most of the 37 HPV genotypes targeted by both assays, including for 13 of 14 carcinogenic HPV genotypes. LA detected more multiple-genotype infections for all HPV genotypes among HPV-positive women (P < 0.0001) and for carcinogenic HPV genotypes among carcinogenic-HPV-positive women (P < 0.0001). LA was more sensitive (92.3% versus 87.1%; P = 0.003) and less specific (48.2% versus 54.0%; P < 0.0001) than LBA for 2-year cumulative cervical precancer and cancer as diagnosed by the Pathology Quality Control Group. In conclusion, we found LA to be a promising assay for the detection of HPV genotypes and carcinogenic HPV, and it may be clinically useful for the detection of

  16. Comparison of Linear Array and Line Blot Assay for Detection of Human Papillomavirus and Diagnosis of Cervical Precancer and Cancer in the Atypical Squamous Cell of Undetermined Significance and Low-Grade Squamous Intraepithelial Lesion Triage Study▿ (United States)

    Castle, Philip E.; Gravitt, Patti E.; Solomon, Diane; Wheeler, Cosette M.; Schiffman, Mark


    We evaluated Linear Array (LA), a newly commercialized PGMY09/11 L1 consensus primer PCR test that detects 37 human papillomavirus (HPV) genotypes by reverse line blot hybridization, for the detection of individual HPV genotypes and carcinogenic HPV and its clinical performance for detecting 2-year cumulative cervical precancer and cancer using archived specimens from the Atypical Squamous Cell of Undetermined Significance (ASCUS) and Low-Grade Squamous Intraepithelial Lesion Triage Study. LA testing was conducted on enrollment specimens from women referred because of an ASCUS Pap test. To gauge the performance of the new test, the results were compared to those of its prototype predecessor assay, Line Blot Assay (LBA), restricted to paired results (n = 3,335). LA testing was done masked to LBA results and clinical outcomes. The results of LA and LBA testing were compared for detection of carcinogenic HPV and clinical outcomes of cervical precancer and cancer. Overall, 50% and 55% of the women tested positive for carcinogenic HPV by LBA and LA, respectively (P < 0.0001). The percent agreement for carcinogenic HPV detection was 88%, percent positive agreement was 80%, and kappa was 0.76 for detection of carcinogenic HPV by the two assays. There was a significant increase in detection by LA for most of the 37 HPV genotypes targeted by both assays, including for 13 of 14 carcinogenic HPV genotypes. LA detected more multiple-genotype infections for all HPV genotypes among HPV-positive women (P < 0.0001) and for carcinogenic HPV genotypes among carcinogenic-HPV-positive women (P < 0.0001). LA was more sensitive (92.3% versus 87.1%; P = 0.003) and less specific (48.2% versus 54.0%; P < 0.0001) than LBA for 2-year cumulative cervical precancer and cancer as diagnosed by the Pathology Quality Control Group. In conclusion, we found LA to be a promising assay for the detection of HPV genotypes and carcinogenic HPV, and it may be clinically useful for the detection of

  17. Analysis of the Crude Antigen of Hymenolepis nana from Mice by SDS-PAGE and the Determination of Specific Antigens in Protein Structure by Western Blotting


    GÖNENÇ, Bahadır


    Protein bands of crude antigens of Hymenolepis nana were determined by SDS-PAGE and Western blotting. Thirty Swiss albino mice were allotted into two groups of 15 each as positive (infected with H. nana) and negative (non-infected with H. nana) groups. The natural infections of H. nana and other helminths were determined by centrifugal flotation of faeces. After bleeding, the mice were necropsied and their guts were examined for H. nana and other intestinal helminths. Sera from mice were test...

  18. Western blot analysis of a limited number of cells: a valuable adjunct to proteome analysis of paraffin wax-embedded, alcohol-fixed tissue after laser capture microdissection. (United States)

    Martinet, Wim; Abbeloos, Vanessa; Van Acker, Nathalie; De Meyer, Guido R Y; Herman, Arnold G; Kockx, Mark M


    In recent years, laser capture microdissection (LCM) has been used successfully to obtain distinct populations of cells for subsequent molecular analysis. Because of the limited sample availability and the absence of in vitro amplification steps for proteins, the use of LCM for proteome analysis largely depends on highly sensitive protein detection methods. In this study, a western blot protocol was developed and validated for the detection of beta-actin and the moderately expressed cell death protein caspase-3 in small numbers of cells. Initially, cultured human U937 monocytes and whole sections of paraffin wax-embedded, alcohol-fixed human tonsils were used to optimize protein electrophoresis and western blotting conditions. High-performance NuPAGE Bis-Tris gels in combination with high-quality transfer membranes, optimized antibody concentrations, and a sensitive chemiluminescent substrate provided a strong signal for beta-actin with approximately 500 U937 cells. In the same way, procaspase-3 could be identified with approximately 1000 cells. Similar results were obtained with germinal centre cells that were procured from paraffin wax-embedded, alcohol-fixed human tonsils by LCM. Treatment of U937 cells with etoposide rapidly induced cell death and allowed the detection of active caspase-3 with approximately 2500 cells (0.8 pg of protein). The findings of this study suggest that western blotting is a valuable adjunct to proteome analysis of LCM procured cells.

  19. Quantitative Expression Analysis in Brassica napus by Northern Blot Analysis and Reverse Transcription-Quantitative PCR in a Complex Experimental Setting. (United States)

    Rumlow, Annekathrin; Keunen, Els; Klein, Jan; Pallmann, Philip; Riemenschneider, Anja; Cuypers, Ann; Papenbrock, Jutta

    Analysis of gene expression is one of the major ways to better understand plant reactions to changes in environmental conditions. The comparison of many different factors influencing plant growth challenges the gene expression analysis for specific gene-targeted experiments, especially with regard to the choice of suitable reference genes. The aim of this study is to compare expression results obtained by Northern blot, semi-quantitative PCR and RT-qPCR, and to identify a reliable set of reference genes for oilseed rape (Brassica napus L.) suitable for comparing gene expression under complex experimental conditions. We investigated the influence of several factors such as sulfur deficiency, different time points during the day, varying light conditions, and their interaction on gene expression in oilseed rape plants. The expression of selected reference genes was indeed influenced under these conditions in different ways. Therefore, a recently developed algorithm, called GrayNorm, was applied to validate a set of reference genes for normalizing results obtained by Northern blot analysis. After careful comparison of the three methods mentioned above, Northern blot analysis seems to be a reliable and cost-effective alternative for gene expression analysis under a complex growth regime. For using this method in a quantitative way a number of references was validated revealing that for our experiment a set of three references provides an appropriate normalization. Semi-quantitative PCR was prone to many handling errors and difficult to control while RT-qPCR was very sensitive to expression fluctuations of the reference genes.

  20. From hybrid swarms to swarms of hybrids (United States)

    Stohlgren, Thomas J.; Szalanski, Allen L; Gaskin, John F.; Young, Nicholas E.; West, Amanda; Jarnevich, Catherine S.; Tripodi, Amber


    Science has shown that the introgression or hybridization of modern humans (Homo sapiens) with Neanderthals up to 40,000 YBP may have led to the swarm of modern humans on earth. However, there is little doubt that modern trade and transportation in support of the humans has continued to introduce additional species, genotypes, and hybrids to every country on the globe. We assessed the utility of species distributions modeling of genotypes to assess the risk of current and future invaders. We evaluated 93 locations of the genus Tamarix for which genetic data were available. Maxent models of habitat suitability showed that the hybrid, T. ramosissima x T. chinensis, was slightly greater than the parent taxa (AUCs > 0.83). General linear models of Africanized honey bees, a hybrid cross of Tanzanian Apis mellifera scutellata and a variety of European honey bee including A. m. ligustica, showed that the Africanized bees (AUC = 0.81) may be displacing European honey bees (AUC > 0.76) over large areas of the southwestern U.S. More important, Maxent modeling of sub-populations (A1 and A26 mitotypes based on mDNA) could be accurately modeled (AUC > 0.9), and they responded differently to environmental drivers. This suggests that rapid evolutionary change may be underway in the Africanized bees, allowing the bees to spread into new areas and extending their total range. Protecting native species and ecosystems may benefit from risk maps of harmful invasive species, hybrids, and genotypes.

  1. Hybrid intelligent engineering systems

    CERN Document Server

    Jain, L C; Adelaide, Australia University of


    This book on hybrid intelligent engineering systems is unique, in the sense that it presents the integration of expert systems, neural networks, fuzzy systems, genetic algorithms, and chaos engineering. It shows that these new techniques enhance the capabilities of one another. A number of hybrid systems for solving engineering problems are presented.

  2. Hybrid trajectory spaces

    NARCIS (Netherlands)

    Collins, P.J.


    In this paper, we present a general framework for describing and studying hybrid systems. We represent the trajectories of the system as functions on a hybrid time domain, and the system itself by its trajectory space, which is the set of all possible trajectories. The trajectory space is given a na

  3. Cardiac hybrid imaging

    Energy Technology Data Exchange (ETDEWEB)

    Gaemperli, Oliver [University Hospital Zurich, Cardiac Imaging, Zurich (Switzerland); University Hospital Zurich, Nuclear Cardiology, Cardiovascular Center, Zurich (Switzerland); Kaufmann, Philipp A. [University Hospital Zurich, Cardiac Imaging, Zurich (Switzerland); Alkadhi, Hatem [University Hospital Zurich, Institute of Diagnostic and Interventional Radiology, Zurich (Switzerland)


    Hybrid cardiac single photon emission computed tomography (SPECT)/CT imaging allows combined assessment of anatomical and functional aspects of cardiac disease. In coronary artery disease (CAD), hybrid SPECT/CT imaging allows detection of coronary artery stenosis and myocardial perfusion abnormalities. The clinical value of hybrid imaging has been documented in several subsets of patients. In selected groups of patients, hybrid imaging improves the diagnostic accuracy to detect CAD compared to the single imaging techniques. Additionally, this approach facilitates functional interrogation of coronary stenoses and guidance with regard to revascularization procedures. Moreover, the anatomical information obtained from CT coronary angiography or coronary artery calcium scores (CACS) adds prognostic information over perfusion data from SPECT. The use of cardiac hybrid imaging has been favoured by the dissemination of dedicated hybrid systems and the release of dedicated image fusion software, which allow simple patient throughput for hybrid SPECT/CT studies. Further technological improvements such as more efficient detector technology to allow for low-radiation protocols, ultra-fast image acquisition and improved low-noise image reconstruction algorithms will be instrumental to further promote hybrid SPECT/CT in research and clinical practice. (orig.)

  4. Editorial: Hybrid Systems

    DEFF Research Database (Denmark)

    Olderog, Ernst-Rüdiger; Ravn, Anders Peter


    An introduction to three papers in a special issue on Hybrid Systems. These paper were first presented at an IFIP WG 2.2 meeting in Skagen 2005.......An introduction to three papers in a special issue on Hybrid Systems. These paper were first presented at an IFIP WG 2.2 meeting in Skagen 2005....

  5. Hybrid propulsion technology program (United States)


    Technology was identified which will enable application of hybrid propulsion to manned and unmanned space launch vehicles. Two design concepts are proposed. The first is a hybrid propulsion system using the classical method of regression (classical hybrid) resulting from the flow of oxidizer across a fuel grain surface. The second system uses a self-sustaining gas generator (gas generator hybrid) to produce a fuel rich exhaust that was mixed with oxidizer in a separate combustor. Both systems offer cost and reliability improvement over the existing solid rocket booster and proposed liquid boosters. The designs were evaluated using life cycle cost and reliability. The program consisted of: (1) identification and evaluation of candidate oxidizers and fuels; (2) preliminary evaluation of booster design concepts; (3) preparation of a detailed point design including life cycle costs and reliability analyses; (4) identification of those hybrid specific technologies needing improvement; and (5) preperation of a technology acquisition plan and large scale demonstration plan.

  6. Hybrid reactors. [Fuel cycle

    Energy Technology Data Exchange (ETDEWEB)

    Moir, R.W.


    The rationale for hybrid fusion-fission reactors is the production of fissile fuel for fission reactors. A new class of reactor, the fission-suppressed hybrid promises unusually good safety features as well as the ability to support 25 light-water reactors of the same nuclear power rating, or even more high-conversion-ratio reactors such as the heavy-water type. One 4000-MW nuclear hybrid can produce 7200 kg of /sup 233/U per year. To obtain good economics, injector efficiency times plasma gain (eta/sub i/Q) should be greater than 2, the wall load should be greater than 1 MW.m/sup -2/, and the hybrid should cost less than 6 times the cost of a light-water reactor. Introduction rates for the fission-suppressed hybrid are usually rapid.

  7. Hybridization kinetics analysis of an oligonucleotide microarray for microRNA detection

    Institute of Scientific and Technical Information of China (English)

    Botao Zhao; Shuo Ding; Wei Li; Youxin Jin


    MicroRNA (miRNA) microarrays have been successfully used for profiling miRNA expression in many physiological processes such as development, differentiation, oncogenesis,and other disease processes. Detecting miRNA by miRNA microarray is actually based on nucleic acid hybridization between target molecules and their corresponding complementary probes. Due to the small size and high degree of similarity among miRNA sequences, the hybridization condition must be carefully optimized to get specific and reliable signals. Previously, we reported a microarray platform to detect miRNA expression. In this study, we evaluated the sensitivity and specificity of our microarray platform. After systematic analysis, we determined an optimized hybridization condition with high sensitivity and specificity for miRNA detection. Our results would be helpful for other hybridization-based miRNA detection methods, such as northern blot and nuclease protection assay.

  8. Modulation of tyrosine hydroxylase gene expression in the central nervous system visualized by in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Berod, A.; Biguet, N.F.; Dumas, S.; Bloch, B.; Mallet, J.


    cDNA probe was used for in situ hybridization studies on histological sections through the locus coeruleus, substantia nigra, and the ventral tegmental area of the rat brain. Experimental conditions were established that yielded no background and no signal when pBR322 was used as control probe. Using the tyrosine hydroxylase probe, the authors ascertained the specificity of the labeling over catecholaminergic cells by denervation experiments and comparison of the hybridization pattern with that of immunoreactivity. The use of /sup 35/S-labeled probe enabled the hybridization signal to be resolved at the cellular level. A single injection of reserpine into the rat led to an increase of the intensity of the autoradiographic signal over the locus coeruleus area, confirming an RNA gel blot analysis. The potential of in situ hybridization to analyze patterns of modulation of gene activity as a result of nervous activity is discussed.

  9. Molecular cloning, expression and in situ hybridization of rat brain glutamic acid decarboxylase messenger RNA. (United States)

    Julien, J F; Legay, F; Dumas, S; Tappaz, M; Mallet, J


    A cDNA library was generated in the expression vector lambda GT11 from rat brain poly(A)+ RNAs and screened with a GAD antiserum. Two clones reacted positively. One of them was shown to express a GAD activity which was specifically trapped on anti-GAD immunogel and was inhibited by gamma-acetylenic-GABA. Blot hybridization analysis of RNAs from rat brain revealed a single 4 kilobases band. Preliminary in situ hybridizations showed numerous cells labelled by the GAD probe such as the Purkinje and stellate cells in the cerebellar cortex and the cells of the reticular thalamic nucleus.

  10. Glycophospholipid Formulation with NADH and CoQ10 Significantly Reduces Intractable Fatigue in Western Blot-Positive ‘Chronic Lyme Disease’ Patients: Preliminary Report

    Directory of Open Access Journals (Sweden)

    Garth L. Nicolson


    Full Text Available Background: An open label 8-week preliminary study was conducted in a small number of patients to determine if a combination oral supplement containing a mixture of phosphoglycolipids, coenzyme Q10 and microencapsulated NADH and other nutrients could affect fatigue levels in long-term, Western blot-positive, multi-symptom ‘chronic Lyme disease’ patients (also called ‘post-treatment Lyme disease’ or ‘post Lyme syndrome’ with intractable fatigue. Methods: The subjects in this study were 6 males (mean age = 45.1 ± 12.4 years and 10 females (mean age = 54.6 ± 7.4 years with ‘chronic Lyme disease’ (determined by multiple symptoms and positive Western blot analysis that had been symptomatic with chronic fatigue for an average of 12.7 ± 6.6 years. They had been seen by multiple physicians (13.3 ± 7.6 and had used many other remedies, supplements and drugs (14.4 ± 7.4 without fatigue relief. Fatigue was monitored at 0, 7, 30 and 60 days using a validated instrument, the Piper Fatigue Scale.Results: Patients in this preliminary study responded to the combination test supplement, showing a 26% reduction in overall fatigue by the end of the 8-week trial (p< 0.0003. Analysis of subcategories of fatigue indicated that there were significant improvements in the ability to complete tasks and activities as well as significant improvements in mood and cognitive abilities. Regression analysis of the data indicated that reductions in fatigue were consistent and occurred with a high degree of confidence (R2= 0.998. Functional Foods in Health and Disease 2012, 2(3:35-47 Conclusions: The combination supplement was a safe and effective method to significantly reduce intractable fatigue in long-term patients with Western blot-positive ‘chronic Lyme disease.’

  11. Study of a viral-dual infection in rainbow trout (Oncorhynchus mykiss) by seroneutralization, western blot and polymerase chain reaction assays. (United States)

    Rodríguez, S; Vilas, M P; Alonso, M; Pérez, S I


    Viral-dual infections in fish are of interest to aquaculture practices but they are rarely described and studied. Several methods were applied in this work to demonstrate a case of coinfection in a reared rainbow trout (Oncorhynchus mykiss) population. Inoculation in cell cultures and cross-neutralization tests were the standard procedures that made it possible to isolate and identify a birnavirus, the infectious pancreatic necrosis virus (IPNV), and suspect of a second virus. Western blotting with both polyclonal and monoclonal antibodies, and reverse transcriptional-polymerase chain reaction (RT-PCR) demonstrate coexistence of both, IPNV and a rhabdovirus.


    Directory of Open Access Journals (Sweden)

    Manoj G Tyagi


    Full Text Available Vasopressin, a posterior pituitary hormone is responsible for water reabsorption by the kidneys and maintenance of cardio-vascular homeostasis. Vasopressin receptors are characterized as VR 1 (V1a, VR2 (V2, and VR3 (V1b. VR1, which is abundant in vascular smooth muscles, causes vasoconstriction by increasing intracellular calcium via the phosphatidylinositol bisphosphonate pathway and a positive inotropic effect in cardiac muscle. VR2 has also been shown to be expressed in the heart. There is emerging role for vasopressin receptors in health and disease. This study describes the application of Western blotting to elucidate the importance of vasopressin receptors in the heart cells.

  13. Hybrid Bloch Brane

    CERN Document Server

    Bazeia, D; Losano, L


    This work reports on models described by two real scalar fields coupled with gravity in the five-dimensional spacetime, with a warped geometry involving one infinite extra dimension. Through a mechanism that smoothly changes a thick brane into a hybrid brane, one investigates the appearance of hybrid branes hosting internal structure, characterized by the splitting on the energy density and the volcano potential, induced by the parameter which controls interactions between the two scalar fields. In particular, we investigate distinct symmetric and asymmetric hybrid brane scenarios.

  14. Hybrid electric vehicles TOPTEC

    Energy Technology Data Exchange (ETDEWEB)



    This one-day TOPTEC session began with an overview of hybrid electric vehicle technology. Updates were given on alternative types of energy storage, APU control for low emissions, simulation programs, and industry and government activities. The keynote speech was about battery technology, a key element to the success of hybrids. The TOPEC concluded with a panel discussion on the mission of hybrid electric vehicles, with a perspective from industry and government experts from United States and Canada on their view of the role of this technology.

  15. Hybrid systems with constraints

    CERN Document Server

    Daafouz, Jamal; Sigalotti, Mario


    Control theory is the main subject of this title, in particular analysis and control design for hybrid dynamic systems.The notion of hybrid systems offers a strong theoretical and unified framework to cope with the modeling, analysis and control design of systems where both continuous and discrete dynamics interact. The theory of hybrid systems has been the subject of intensive research over the last decade and a large number of diverse and challenging problems have been investigated. Nevertheless, many important mathematical problems remain open.This book is dedicated mainly to

  16. Hybrid silicon evanescent devices

    Directory of Open Access Journals (Sweden)

    Alexander W. Fang


    Full Text Available Si photonics as an integration platform has recently been a focus of optoelectronics research because of the promise of low-cost manufacturing based on the ubiquitous electronics fabrication infrastructure. The key challenge for Si photonic systems is the realization of compact, electrically driven optical gain elements. We review our recent developments in hybrid Si evanescent devices. We have demonstrated electrically pumped lasers, amplifiers, and photodetectors that can provide a low-cost, scalable solution for hybrid integration on a Si platform by using a novel hybrid waveguide architecture, consisting of III-V quantum wells bonded to Si waveguides.

  17. Chaotic mixer improves microarray hybridization. (United States)

    McQuain, Mark K; Seale, Kevin; Peek, Joel; Fisher, Timothy S; Levy, Shawn; Stremler, Mark A; Haselton, Frederick R


    Hybridization is an important aspect of microarray experimental design which influences array signal levels and the repeatability of data within an array and across different arrays. Current methods typically require 24h and use target inefficiently. In these studies, we compare hybridization signals obtained in conventional static hybridization, which depends on diffusional target delivery, with signals obtained in a dynamic hybridization chamber, which employs a fluid mixer based on chaotic advection theory to deliver targets across a conventional glass slide array. Microarrays were printed with a pattern of 102 identical probe spots containing a 65-mer oligonucleotide capture probe. Hybridization of a 725-bp fluorescently labeled target was used to measure average target hybridization levels, local signal-to-noise ratios, and array hybridization uniformity. Dynamic hybridization for 1h with 1 or 10ng of target DNA increased hybridization signal intensities approximately threefold over a 24-h static hybridization. Similarly, a 10- or 60-min dynamic hybridization of 10ng of target DNA increased hybridization signal intensities fourfold over a 24h static hybridization. In time course studies, static hybridization reached a maximum within 8 to 12h using either 1 or 10ng of target. In time course studies using the dynamic hybridization chamber, hybridization using 1ng of target increased to a maximum at 4h and that using 10ng of target did not vary over the time points tested. In comparison to static hybridization, dynamic hybridization reduced the signal-to-noise ratios threefold and reduced spot-to-spot variation twofold. Therefore, we conclude that dynamic hybridization based on a chaotic mixer design improves both the speed of hybridization and the maximum level of hybridization while increasing signal-to-noise ratios and reducing spot-to-spot variation.

  18. Hybrid polymer microspheres (United States)

    Rembaum, A.


    Techniques have been successfully tested for bonding polymeric spheres, typically 0.1 micron in diameter, to spheres with diameter up to 100 microns. Hybrids are being developed as improved packing material for ion-exchange columns, filters, and separators.

  19. Hybrid adsorptive membrane reactor (United States)

    Tsotsis, Theodore T. (Inventor); Sahimi, Muhammad (Inventor); Fayyaz-Najafi, Babak (Inventor); Harale, Aadesh (Inventor); Park, Byoung-Gi (Inventor); Liu, Paul K. T. (Inventor)


    A hybrid adsorbent-membrane reactor in which the chemical reaction, membrane separation, and product adsorption are coupled. Also disclosed are a dual-reactor apparatus and a process using the reactor or the apparatus.

  20. Hybrid Rocket Technology

    Directory of Open Access Journals (Sweden)

    Sankaran Venugopal


    Full Text Available With their unique operational characteristics, hybrid rockets can potentially provide safer, lower-cost avenues for spacecraft and missiles than the current solid propellant and liquid propellant systems. Classical hybrids can be throttled for thrust tailoring, perform in-flight motor shutdown and restart. In classical hybrids, the fuel is stored in the form of a solid grain, requiring only half the feed system hardware of liquid bipropellant engines. The commonly used fuels are benign, nontoxic, and not hazardous to store and transport. Solid fuel grains are not highly susceptible to cracks, imperfections, and environmental temperature and are therefore safer to manufacture, store, transport, and use for launch. The status of development based on the experience of the last few decades indicating the maturity of the hybrid rocket technology is given in brief.Defence Science Journal, 2011, 61(3, pp.193-200, DOI:

  1. Nitrous Paraffin Hybrid Project (United States)

    National Aeronautics and Space Administration — The Nitrous Oxide Paraffin Hybrid engine (N2OP) is a proposed technology designed to provide small launch vehicles with high specific impulse, indefinitely storable...

  2. Hybrid photon detectors

    CERN Document Server

    D'Ambrosio, C


    Hybrid photon detectors detect light via vacuum photocathodes and accelerate the emitted photoelectrons by an electric field towards inversely polarized silicon anodes, where they are absorbed, thus producing electron-hole pairs. These, in turn, are collected and generate electronic signals on their ohmic contacts. This review first describes the characteristic properties of the main components of hybrid photon detectors: light entrance windows, photocathodes, and silicon anodes. Then, essential relations describing the trajectories of photoelectrons in electric and magnetic fields and their backscattering from the silicon anodes are derived. Depending on their anode configurations, three families of hybrid photon detectors are presented: hybrid photomultiplier tubes with single anodes for photon counting with high sensitivity and for gamma spectroscopy; multi-anode photon detector tubes with anodes subdivided into square or hexagonal pads for position-sensitive photon detection; imaging silicon pixel array t...

  3. Hybrid adsorptive membrane reactor (United States)

    Tsotsis, Theodore T.; Sahimi, Muhammad; Fayyaz-Najafi, Babak; Harale, Aadesh; Park, Byoung-Gi; Liu, Paul K. T.


    A hybrid adsorbent-membrane reactor in which the chemical reaction, membrane separation, and product adsorption are coupled. Also disclosed are a dual-reactor apparatus and a process using the reactor or the apparatus.

  4. Polymerase chain reaction and Southern blot-based analysis of the C9orf72 hexanucleotide repeat in different motor neuron diseases. (United States)

    Hübers, Annemarie; Marroquin, Nicolai; Schmoll, Birgit; Vielhaber, Stefan; Just, Marlies; Mayer, Benjamin; Högel, Josef; Dorst, Johannes; Mertens, Thomas; Just, Walter; Aulitzky, Anna; Wais, Verena; Ludolph, Albert C; Kubisch, Christian; Weishaupt, Jochen H; Volk, Alexander E


    The GGGGCC-hexanucleotide repeat expansion in C9orf72 is the most common genetic cause of familial amyotrophic lateral sclerosis and frontotemporal dementia. This study determined the frequency of C9orf72 repeat expansions in different motor neuron diseases (amyotrophic lateral sclerosis (ALS), motor neuron diseases affecting primarily the first or the second motor neuron and hereditary spastic paraplegia). Whereas most studies on C9orf72 repeat expansions published so far rely on a polymerase chain reaction-based screening, we applied both polymerase chain reaction-based techniques and Southern blotting. Furthermore, we determined the sensitivity and specificity of Southern blotting of the C9orf72 hexanucleotide repeat in DNA derived from lymphoblastoid cell lines. C9orf72 repeat expansions were found in 27.1% out of 166 familial ALS patients, only once in 68 sporadic ALS patients, and not in 61 hereditary spastic paraplegia patients or 52 patients with motor neuron diseases affecting clinically primarily either the first or the second motor neuron. We found hints for a correlation between C9orf72 repeat length and the age of onset. Somatic instability of the C9orf72 repeat was observed in lymphoblastoid cell lines compared with DNA derived from whole blood from the same patient and therefore caution is warranted for repeat length determination in immortalized cell lines.

  5. A Comparison of Antibacterial Activity of Selected Thyme (Thymus) Species by Means of the Dot Blot Test with Direct Bioautographic Detection. (United States)

    Orłowska, Marta; Kowalska, Teresa; Sajewicz, Mieczysław; Jesionek, Wioleta; Choma, Irena M; Majer-Dziedzic, Barbara; Szymczak, Grażyna; Waksmundzka-Hajnos, Monika


    Bioautography carried out with the aid of thin-layer chromatographic adsorbents can be used to assess antibacterial activity in samples of different origin. It can either be used as a simple and cost-effective detection method applied to a developed chromatogram, or to the dot blot test performed on a chromatographic plate, where total antibacterial activity of a sample is scrutinized. It was an aim of this study to compare antibacterial activity of 18 thyme (Thymus) specimens and species (originating from the same gardening plot and harvested in the same period of time) by means of a dot blot test with direct bioautography. A two-step extraction of herbal material was applied, and at step two the polar fraction of secondary metabolites was obtained under the earlier optimized extraction conditions [methanol-water (27+73, v/v), 130°C]. This fraction was then tested for its antibacterial activity against Bacillus subtilis bacteria. It was established that all investigated extracts exhibited antibacterial activity, yet distinct differences were perceived in the size of the bacterial growth inhibition zones among the compared thyme species. Based on the results obtained, T. citriodorus "golden dwarf" (sample No. 5) and T. marschallianus (sample No. 6) were selected as promising targets for further investigations and possible inclusion in a herbal pharmacopeia, which is an essential scientific novelty of this study.

  6. Detection of genetically modified microorganisms in soil using the most-probable-number method with multiplex PCR and DNA dot blot. (United States)

    Yeom, Jinki; Lee, Yunho; Noh, Jaemin; Jung, Jaejoon; Park, Jungsoon; Seo, Hyoju; Kim, Jisun; Han, Jiwon; Jeon, Che Ok; Kim, Taesung; Park, Woojun


    The principal objective of this study was to detect genetically modified microorganisms (GMMs) that might be accidentally released into the environment from laboratories. Two methods [plate counting and most-probable-number (MPN)] coupled with either multiplex PCR or DNA dot blots were compared using genetically modified Escherichia coli, Pseudomonas putida, and Acinetobacter oleivorans harboring an antibiotic-resistance gene with additional gfp and lacZ genes as markers. Alignments of sequences collected from databases using the Perl scripting language (Perl API) and from denaturing gradient gel electrophoresis analysis revealed that the gfp, lacZ and antibiotic-resistance genes (kanamycin, tetracycline, and ampicillin) in GMMs differed from the counterpart genes in many sequenced genomes and in soil DNA. Thus, specific multiplex PCR primer sets for detection of plasmid-based gfp and lacZ antibiotic-resistance genes could be generated. In the plate counting method, many antibiotic-resistant bacteria from a soil microcosm grew as colonies on antibiotic-containing agar plates. The multiplex PCR verification of randomly selected antibiotic-resistant colonies with specific primers proved ineffective. The MPN-multiplex PCR method and antibiotic-resistant phenotype could be successfully used to detect GMMs, although this method is quite laborious. The MPN-DNA dot blot method screened more cells at a time in a microtiter plate containing the corresponding antibiotics, and was shown to be a more efficient method for the detection of GMMs in soil using specific probes in terms of labor and accuracy.

  7. Requirements for Hybrid Cosimulation (United States)


    hybrid cosimulation version of the Functional Mockup Interface (FMI) standard. A cosimulation standard de nes interfaces that enable diverse simulation...cosimulation standards, and specifically provides guidance for development of a hybrid cosimulation version of the Functional Mockup Interface (FMI) standard...V. Peetz, and S. Wolf. The functional mockup interface for tool independent exchange of simulation models. In Proc. of the 8-th International

  8. Hybrid vertical cavity laser

    DEFF Research Database (Denmark)

    Chung, Il-Sug; Mørk, Jesper


    A new hybrid vertical cavity laser structure for silicon photonics is suggested and numerically investigated. It incorporates a silicon subwavelength grating as a mirror and a lateral output coupler to a silicon ridge waveguide.......A new hybrid vertical cavity laser structure for silicon photonics is suggested and numerically investigated. It incorporates a silicon subwavelength grating as a mirror and a lateral output coupler to a silicon ridge waveguide....

  9. Hybrid Baryon Signatures

    CERN Document Server

    Page, P R


    We discuss whether a low-lying hybrid baryon should be defined as a three quark - gluon bound state or as three quarks moving on an excited adiabatic potential. We show that the latter definition becomes exact, not only for very heavy quarks, but also for specific dynamics. We review the literature on the signatures of hybrid baryons, with specific reference to strong hadronic decays, electromagnetic couplings, diffractive production and production in psi decay.

  10. Hybridity in Disgrace

    Institute of Scientific and Technical Information of China (English)



    John Maxwell Coetzee's masterpiece-Disgrace is the representative work about post colonialism.The novel describes a series of disgraceful events happened between the white and the black in the post apartheid South Africa.The famous literature theory-hybridity of Homi K.Bhabha is the very key theory to analyze the work.In post apartheid South Africa,hybridity is the only way for the white and the black to coexist.

  11. 基于地高辛标记对小麦进行Southern杂交分析主要影响因素的优化和验证%Optimizing and Confirming of Digoxigenin Based Southern Blotting for Wheat Genome Analysis

    Institute of Scientific and Technical Information of China (English)

    刘禄; 牛焱焱; 雷昊; 林志珊; 赵翔宇; 叶兴国; 张宪省


    以转基因小麦和野生型小麦DNA为材料,对利用地高辛标记对小麦基因组DNA进行Southern杂交分析的影响因素进行了优化研究,包括探针制备与纯化、样品DNA量、酶切体系、真空转印条件、杂交条件、免疫检测方法等.结果表明,对随机引物标记的模板和标记后的探针进行纯化可明显提高探针的标记效率,10μg高质量的DNA样品在80μl的体系中,酶切8~12h可获得良好的效果;真空转膜时使用碱性液比中性液获得的转膜效果更干净;试剂纯度、杂交温度及杂交炉转速等均对杂交效果产生重要影响;配合改进的CSPD涂布方法,使用化学发光检测系统比单纯使用X光片显像更易操作,背景更干净;本研究所优化的地高辛标记的小麦Southern杂交分析显示出较高的灵敏度和信噪比,结果稳定,可克服同位素标记对实验条件、设备及实验人员身体状况等限制,在普通实验室推广应用.%The Southern blot analysis based on isotope labeling technique has some shortages such as facility limitation, environmental pollution, and poor stability. Therefore, establishing a safe, stable and efficient Southern hybridization protocol is very important to detect the integration of exogenous genes in transgenic wheat. In this study , by using the DNAs from wild-type wheat and transgenic wheat asmaterial, the DIG-labeled Southern blot analysis was improved through modifying several key steps, including probe preparation and purification, usage of DNA samples amount, digestion system, vacuum transfer conditions, stringent hybridization conditions, and immunoassay detect. The results showed that the purifications of DNA template and probe- labeled could significantly improve the efficiency of the probe labeling when random prime labeling method was used. Desirable digestion result could be a-chieved when 10|xg DNA samples with high quality was enzymed in 80|xl reaction system for 8 ~ 12

  12. GAPDH and β-actin protein decreases with aging, making Stain-Free technology a superior loading control in Western blotting of human skeletal muscle

    DEFF Research Database (Denmark)

    Vigelsø Hansen, Andreas; Dybboe, Rie; Hansen, Christina Neigaard;


    [β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and α-tubulin], as well as TP loaded measured by Stain-Free technology (SF) as normalization tool were tested. This was done using skeletal muscle samples from men subjected to physiological conditions often investigated in applied...... of β-actin and GAPDH was lower in older men compared with young men. In conclusion, β-actin, GAPDH, and α-tubulin may not be used for normalization in studies that include subjects with a large age difference. In contrast, the RPs may not be affected in studies that include muscle wasting...... and differences in muscle fiber type. The novel SF technology adds lower variation to the results compared with the existing methods for correcting for loading inaccuracy in Western blotting of human skeletal muscle in applied physiology....

  13. Serum detection of IgG antibodies against Demodex canis by western blot in healthy dogs and dogs with juvenile generalized demodicosis. (United States)

    Ravera, Ivan; Ferreira, Diana; Gallego, Laia Solano; Bardagí, Mar; Ferrer, Lluís


    The aim of this study was to investigate the presence of canine immunoglobulins (Ig) G against Demodex proteins in the sera of healthy dogs and of dogs with juvenile generalized demodicosis (CanJGD) with or without secondary pyoderma. Demodex mites were collected from dogs with CanJGD. Protein concentration was measured and a western blot technique was performed. Pooled sera from healthy dogs reacted mainly with antigen bands ranging from 55 to 72 kDa. Pooled sera from dogs with CanJGD without secondary pyoderma reacted either with 10 kDa antigen band or 55 to 72 kDa bands. Pooled sera from dogs with CanJGD with secondary pyoderma reacted only with a 10 kDa antigen band. The results of this study suggest that both healthy dogs and dogs with CanJGD develop a humoral response against different proteins of Demodex canis.

  14. A comparison of antigenic peptides in muscle larvae of several Trichinella species by two-dimensional western-blot analysis with monoclonal antibodies

    Directory of Open Access Journals (Sweden)

    Dea-Ayuela M.A.


    Full Text Available The antigens recognised by mAb US5 specific to 53 kDa glycoprotein (gp 53 in T. spiralis L-1 muscle larvae (TSL1 antigens, mAb US9 specific to gp 53 in TSL1 from all encapsulated species and mAb US4 specific to a tyvelose containing tetrasaccharide present in TSL1, were investigated in crude extracts from muscle larvae of T. spiralis, T. nativa and T. britovi by 2D-electrophoresis and western-blot. At least four proteins of different pI were recognised by mAb US5 on T. spiralis antigens. Recognition profile of mAb US9 on T. spiralis antigens exhibited some variation with regard to that of the US5. Polymorphism was apparent in gp 53. High reactivity was shown by the mAb US4 with the three species.

  15. Isolation and RNA gel blot analysis of genes that could serve as potential molecular markers for leaf senescence in Arabidopsis thaliana. (United States)

    Yoshida, S; Ito, M; Nishida, I; Watanabe, A


    Nine cDNAs, representing genes in which the transcripts accumulated in senescent leaves of Arabidopsis thaliana, were isolated by differential display reverse transcription polymerase chain reaction (DDRT-PCR) and the genes were designated yellow-leaf-specific gene 1 to 9 (YLS1-YLS9). Sequence analysis revealed that none of the YLS genes, except YLS6, had been reported as senescence-up-regulated genes. RNA gel blot analysis revealed that the transcripts of YLS3 accumulated at the highest level at an early senescence stage, whereas the transcripts from the other YLS genes reached their maximum levels in late senescence stages. Transcripts of YLS genes showed various accumulation patterns under natural senescence, and under artificial senescence induced by darkness, ethylene or ABA. These expression characteristics of YLS genes will be useful as potential molecular markers, which will enhance our understanding of natural and artificial senescence processes.

  16. Localization and Distribution of 'Candidatus Liberibacter asiaticus' in Citrus and Periwinkle by Direct Tissue Blot Immuno Assay with an Anti-OmpA Polyclonal Antibody. (United States)

    Ding, Fang; Duan, Yongping; Paul, Cristina; Brlansky, Ronald H; Hartung, John S


    'Candidatus Liberibacter asiaticus' (CaLas), a non-cultured member of the α-proteobacteria, is the causal agent of citrus Huanglongbing (HLB). Due to the difficulties of in vitro culture, antibodies against CaLas have not been widely used in studies of this pathogen. We have used an anti-OmpA polyclonal antibody based direct tissue blot immunoassay to localize CaLas in different citrus tissues and in periwinkle leaves. In citrus petioles, CaLas was unevenly distributed in the phloem sieve tubes, and tended to colonize in phloem sieve tubes on the underside of petioles in preference to the upper side of petioles. Both the leaf abscission zone and the junction of the petiole and leaf midrib had fewer CaLas bacteria compared to the main portions of the petiole and the midribs. Colonies of CaLas in phloem sieve tubes were more frequently found in stems with symptomatic leaves than in stems with asymptomatic leaves with an uneven distribution pattern. In serial sections taken from the receptacle to the peduncle, more CaLas were observed in the peduncle sections adjacent to the stem. In seed, CaLas was located in the seed coat. Many fewer CaLas were found in the roots, as compared to the seeds and petioles when samples were collected from trees with obvious foliar symptoms. The direct tissue blot immuno assay was adapted to whole periwinkle leaves infected by CaLas. The pathogen was distributed throughout the lateral veins and the results were correlated with results of qPCR. Our data provide direct spatial and anatomical information for CaLas in planta. This simple and scalable method may facilitate the future research on the interaction of CaLas and host plant.

  17. Localization and Distribution of 'Candidatus Liberibacter asiaticus' in Citrus and Periwinkle by Direct Tissue Blot Immuno Assay with an Anti-OmpA Polyclonal Antibody.

    Directory of Open Access Journals (Sweden)

    Fang Ding

    Full Text Available 'Candidatus Liberibacter asiaticus' (CaLas, a non-cultured member of the α-proteobacteria, is the causal agent of citrus Huanglongbing (HLB. Due to the difficulties of in vitro culture, antibodies against CaLas have not been widely used in studies of this pathogen. We have used an anti-OmpA polyclonal antibody based direct tissue blot immunoassay to localize CaLas in different citrus tissues and in periwinkle leaves. In citrus petioles, CaLas was unevenly distributed in the phloem sieve tubes, and tended to colonize in phloem sieve tubes on the underside of petioles in preference to the upper side of petioles. Both the leaf abscission zone and the junction of the petiole and leaf midrib had fewer CaLas bacteria compared to the main portions of the petiole and the midribs. Colonies of CaLas in phloem sieve tubes were more frequently found in stems with symptomatic leaves than in stems with asymptomatic leaves with an uneven distribution pattern. In serial sections taken from the receptacle to the peduncle, more CaLas were observed in the peduncle sections adjacent to the stem. In seed, CaLas was located in the seed coat. Many fewer CaLas were found in the roots, as compared to the seeds and petioles when samples were collected from trees with obvious foliar symptoms. The direct tissue blot immuno assay was adapted to whole periwinkle leaves infected by CaLas. The pathogen was distributed throughout the lateral veins and the results were correlated with results of qPCR. Our data provide direct spatial and anatomical information for CaLas in planta. This simple and scalable method may facilitate the future research on the interaction of CaLas and host plant.

  18. The Hybrids of Postmodernism

    Directory of Open Access Journals (Sweden)



    Full Text Available Hybridization is a fundamental characteristic of postmodernism, included by Ihab Hassan in his “catena” of features. This paper looks into the hybrids of postmodernism, which are the result of migration, displacement and uprooting, the re-visitation of myths, folklore and legends, or projections of their author’s imagination. The hybrids used as examples here are drawn from several novels written by Salman Rushdie, especially The Satanic Verses, two short stories, one by Márquez and the other by Donald Barthelme, Borges’s Book of Imaginary Beings, Cărtărescu’s Encyclopaedia of Dragons and Michelle Cliff’s No Telephone to Heaven. Diverse as they may be, these hybrids emphasize a defining characteristic of postmodernism, which is its pluralism. I conclude that the hybrids of postmodernism are aesthetically or politically subversive. Besides, what makes them difficult to grasp is their unfixed and protean nature. They ask for high leaps of the imagination, a total suspension of disbelief and a complete surrender to the powerful seduction of imagination on the reader’s part.

  19. 亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定%Screen and detection of specific antigens of Taenia asiatica cysticercus with two dimensional electrophoresis, western blotting and MS/MS spectrometry

    Institute of Scientific and Technical Information of China (English)

    方文; 肖靓靓; 包怀恩; 牟荣


    Objective To screen and identify specific antigens of Taenia asiatica cysticercus and to predict the function of target proteins using bioinformatics method. Methods Six three-way cross hybrid pigs were randomly divided into experimental group and control group and each experimental pig was infected with 84 000 eggs. Serum samples were collected 40 days after the infection. The total protein of Taenia asiatica cysticercus was separated by two dimensional electrophoresis and western blotting was performed to find out distinct antigens. Proteins from the two groups were identified by electrospray ion trap mass spectrometry(ESI-Trap MS). Results Totally 204 ± 11 spots were detected through Coomassie brilliant blue stained gels with relative molecular weight( Mr) of 14 400 -94 000 kDa and isoelectric point(pl) of 3. 0 - 10. 0. Western-blotting showed 6 specific antigen spots with pool sera of infected pigs. Two of the 6 antigens with known functions were ascribed to ATPase family gene 3-like protein 2 and ATP synthase beta subunit. Conclusion Two specific antigens of Taenia asiatica cysticercus have been identified.%目的 对亚洲带绦虫囊尾蚴的特异性抗原进行筛选、鉴定和生物信息学分析.方法 6头20d龄三元杂交乳猪随机分为实验组和对照组,实验组每头灌胃亚洲带绦虫虫卵悬液8.4万个;20 d后,分别制备实验组及对照组心脏血混合血清;收集、制备实验组亚洲带绦虫囊尾蚴蛋白并进行双向电泳分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜,用实验组及对照组混合血清(均为1∶10)为一抗,羊抗猪辣根过氧化物酶- IgG(1∶200)为二抗,进行蛋白质印迹分析和质谱鉴定.结果 双向电泳凝胶共检测到( 204±11)个蛋白质斑点,相对分子质量为14 400 ~94 000 kPa,等电点(pI)为3.0~10.0;筛选出6个特异性抗原,经质谱鉴定出其中2个蛋白,分别为ATP酶管家基因3-类蛋白2(AT-Pase family gene 3-like protein 2)

  20. Porosity in hybrid materials

    Energy Technology Data Exchange (ETDEWEB)

    Schaefer, D.W.; Beaucage, G.; Loy, D. [Sandia National Labs., Albuquerque, NM (United States)


    Multicomponent, or hybrid composites are emerging as precursors to porous materials. Sacrifice of an ephemeral phase can be used to generate porosity, the nature of which depends on precursor structure. Retention of an organic constituent, on the other hand, can add desirable toughness to an otherwise brittle ceramic. We use small-angle x-ray and neutron scattering to examine porosity in both simple and hybrid materials. We find that microphase separation controls porosity in almost all systems studied. Pore distributions are controlled by the detailed bonding within and between phases as well as the flexibility of polymeric constituents. Thus hybridization opens new regions of pore distributions not available in simple systems. We look at several sacrificial concepts and show that it is possible to generate multimodal pore size distributions due to the complicated phase structure in the precursor.

  1. Hybrid-secure MPC 

    DEFF Research Database (Denmark)

    Lucas, Christoph; Raub, Dominik; Maurer, Ueli


    of the adversary, without being aware of the actual adversarial setting. Thus, hybrid-secure MPC protocols allow for graceful degradation of security. We present a hybrid-secure MPC protocol that provides an optimal trade-off between IT robustness and computational privacy: For any robustness parameter ρ ...Most protocols for distributed, fault-tolerant computation, or multi-party computation (MPC), provide security guarantees in an all-or-nothing fashion. In contrast, a hybrid-secure protocol provides different security guarantees depending on the set of corrupted parties and the computational power...... obtain one MPC protocol that is simultaneously IT secure with robustness for up to t ≤ ρ actively corrupted parties, IT secure with fairness (no robustness) for up to t privacy and correctness only) for up to t

  2. Hybrid Action Systems

    DEFF Research Database (Denmark)

    Rönnkö, M.; Ravn, Anders Peter; Sere, K.


    In this paper we investigate the use of action systems with differential actions in the specifcation of hybrid systems. As the main contribution we generalize the definition of a differential action, allowing the use of arbitrary relations over model variables and their time-derivatives in modell......In this paper we investigate the use of action systems with differential actions in the specifcation of hybrid systems. As the main contribution we generalize the definition of a differential action, allowing the use of arbitrary relations over model variables and their time...... parallel composition. Moreover, as the strength of the action system formalism is the support for stepwise development by refinement, we investigate refinement involving a differential action. We show that, due to the predicate transformer semantics, standard action refinement techniques apply also...... to the differential action, thus, allowing stepwise development of hybrid systems Udgivelsesdato: JAN 1...

  3. for hybrid dynamical systems

    Directory of Open Access Journals (Sweden)

    Wassim M. Haddad


    Full Text Available In this paper we develop a unified dynamical systems framework for a general class of systems possessing left-continuous flows; that is, left-continuous dynamical systems. These systems are shown to generalize virtually all existing notions of dynamical systems and include hybrid, impulsive, and switching dynamical systems as special cases. Furthermore, we generalize dissipativity, passivity, and nonexpansivity theory to left-continuous dynamical systems. Specifically, the classical concepts of system storage functions and supply rates are extended to left-continuous dynamical systems providing a generalized hybrid system energy interpretation in terms of stored energy, dissipated energy over the continuous-time dynamics, and dissipated energy over the resetting events. Finally, the generalized dissipativity notions are used to develop general stability criteria for feedback interconnections of left-continuous dynamical systems. These results generalize the positivity and small gain theorems to the case of left-continuous, hybrid, and impulsive dynamical systems.

  4. Photoproduction of Hybrid Mesons

    CERN Document Server

    Barnes, T


    In this contribution I discuss prospects for photoproducing hybrid mesons at CEBAF, based on recent model results and experimental indications of possible hybrids. One excellent opportunity appears to be a search for the I=1, JPC=2+-, neutral "(b2)o" hybrid in (a2 pi)o through diffractive photoproduction. Other notable possibilities accessible through pi+ or pio exchange photoproduction are I=1, JPC=1-+, charged "pi1+" in f1 pi+, (b1 pi)+ and (rho pi)+; piJ(1770)+ in f2 pi+ and (b1 pi)+; pi(1800)+ in f0 pi+, f2 pi+, omega rho+ and (rho pi)+; a1 in f1 pi+ and f2 pi+; and omega in (rho pi)o, omega eta and (K1 K)o.

  5. Smart hybrid rotary damper (United States)

    Yang, C. S. Walter; DesRoches, Reginald


    This paper develops a smart hybrid rotary damper using a re-centering smart shape memory alloy (SMA) material as well as conventional energy-dissipating metallic plates that are easy to be replaced. The ends of the SMA and steel plates are inserted in the hinge. When the damper rotates, all the plates bend, providing energy dissipating and recentering characteristics. Such smart hybrid rotary dampers can be installed in structures to mitigate structural responses and to re-center automatically. The damaged energy-dissipating plates can be easily replaced promptly after an external excitation, reducing repair time and costs. An OpenSEES model of a smart hybrid rotary was established and calibrated to reproduce the realistic behavior measured from a full-scale experimental test. Furthermore, the seismic performance of a 3-story moment resisting model building with smart hybrid rotary dampers designed for downtown Los Angeles was also evaluated in the OpenSEES structural analysis software. Such a smart moment resisting frame exhibits perfect residual roof displacement, 0.006", extremely smaller than 18.04" for the conventional moment resisting frame subjected to a 2500 year return period ground motion for the downtown LA area (an amplified factor of 1.15 on Kobe earthquake). The smart hybrid rotary dampers are also applied into an eccentric braced steel frame, which combines a moment frame system and a bracing system. The results illustrate that adding smart hybrid rotaries in this braced system not only completely restores the building after an external excitation, but also significantly reduces peak interstory drifts.

  6. Analog and hybrid computing

    CERN Document Server

    Hyndman, D E


    Analog and Hybrid Computing focuses on the operations of analog and hybrid computers. The book first outlines the history of computing devices that influenced the creation of analog and digital computers. The types of problems to be solved on computers, computing systems, and digital computers are discussed. The text looks at the theory and operation of electronic analog computers, including linear and non-linear computing units and use of analog computers as operational amplifiers. The monograph examines the preparation of problems to be deciphered on computers. Flow diagrams, methods of ampl

  7. Hybrid Weyl semimetal (United States)

    Li, Fei-Ye; Luo, Xi; Dai, Xi; Yu, Yue; Zhang, Fan; Chen, Gang


    We construct a tight-binding model realizing one pair of Weyl nodes and three distinct Weyl semimetals. In the type-I (type-II) Weyl semimetal, both nodes belong to type-I (type-II) Weyl nodes. In addition, there exists a third type, previously undiscovered and dubbed "hybrid Weyl semimetal", in which one Weyl node is of type I while the other is of type II. For the hybrid Weyl semimetal, we further demonstrate the bulk Fermi surfaces and the topologically protected surface states, analyze the unique Landau-level structure and quantum oscillation, and discuss the conditions for possible material realization.

  8. The hybrid BCI

    Directory of Open Access Journals (Sweden)

    Gert Pfurtscheller


    Full Text Available Nowadays, everybody knows what a hybrid car is. A hybrid car normally has 2 engines, its main purpose being to enhance energy efficiency and reduce CO2 output. Similarly, a typical hybrid brain-computer interface (BCI is also composed of 2 BCIs or at least one BCI and another system. Such a hybrid BCI, like any BCI, must fulfil the following four criteria: (i the device must rely on signals recorded directly from the brain; (ii there must be at least one recordable brain signal that the user can intentionally modulate to effect goal-directed behaviour; (iii real time processing; and (iv the user must obtain feedback. This paper introduces some hybrid BCIs which have already been published or are currently in development or validation, and some concepts for future work. The BCIs described classify 2 EEG patterns: One is the event-related (desynchronisation (ERD, ERS of sensorimotor rhythms, and the other is the steady-state visual evoked potential (SSVEP. The hybrid BCI can either have more than one input whereby the inputs are typically processed simultaneously or operate 2 systems sequentially, whereby the first system can act as a “brain switch”. In the case of self-paced operation of a SSVEP-based hand orthosis control with an motor imagery-based switch it was possible to reduce the rate of false positives during resting periods by about 50% compared to the SSVEP BCI alone. It is shown that such a brain switch can also rely on hemodynamic changes measured through near-infrared spectroscopy (NIRS. Another interesting approach is a hybrid BCI with simultaneous operations of ERD- and SSVEP-based BCIs. Here it is important to prove the existing promising offline simulation results with online experiments. Hybrid BCIs can also use one brain signal and another input. Such an additional input can be a physiological signal like the heart rate but also a signal from an external device like, an eye gaze control system.

  9. Toyota hybrid synergy drive

    Energy Technology Data Exchange (ETDEWEB)

    Gautschi, H.


    This presentation made at the Swiss 2008 research conference on traffic by Hannes Gautschi, director of service and training at the Toyota company in Switzerland, takes a look at Toyota's hybrid drive vehicles. The construction of the vehicles and their combined combustion engines and electric generators and drives is presented and the combined operation of these components is described. Braking and energy recovery are discussed. Figures on the performance, fuel consumption and CO{sub 2} output of the hybrid vehicles are compared with those of conventional vehicles.

  10. Changes in rRNA levels during stress invalidates results from mRNA blotting: Fluorescence in situ rRNA hybridization permits renormalization for estimation of cellular mRNA levels

    DEFF Research Database (Denmark)

    Hansen, M.C.; Nielsen, A.K.; Molin, Søren;


    Regulation of gene expression can be analyzed by a number of different techniques. Some techniques monitor the level of specific mRNA directly, and others monitor indirectly by determining the level of enzymes encoded by the mRNA. Each method has its own inherent way of normalization. When result...

  11. A Mathematical Approach to Hybridization (United States)

    Matthews, P. S. C.; Thompson, J. J.


    Presents an approach to hybridization which exploits the similarities between the algebra of wave functions and vectors. This method will account satisfactorily for the number of orbitals formed when applied to hybrids involving the s and p orbitals. (GS)


    Institute of Scientific and Technical Information of China (English)

    LI Jin-feng; ZHANG Lei; SUN Su-lian


    Objective: The aim of this study was to detect micrometastases in bone marrow of primary breast cancer patients, and compare with other clinical parameters.Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization.Human breast cancer cell line T47D was mixed with bone marrow cells in different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and immunohistochemistry (IHC) methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples while the expression was not seen in 8 negative control samples. In all 54 patients 14 cases were CK-19 positive (25.9%) by RT-PCR, another positive signal was obtained in 5/54 (9.3%) of bone marrow samples by Southern blotting. The total positive cases are 19/54 (35.2%).CK-19 IHC+ cells were detected at a dilution of one T47D cell in 5×104 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1:5×105 and 1:1×106, respectively. This demonstrates that RT-PCR and Southern blotting was at least 20 times more sensitive than the IHC method. The micrometastases positive rate of the larger tumor size group (>5.0 cm) was significantly (P<0.05) greater than that of the smaller tumor size group (0-2.0 cm). Conclusion: detection of micrometastases in bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is highly sensitive and it is a method to be used for anticipating the prognosis of breast cancer patients.

  13. Hybrid architecture: object, landscape, infrastructure


    Pinto de Freitas, Rita


    The concept of "hybrid architecture" developed in this study considers hybrid all architecture that is at once object, landscape and infrastructure. Hybrid architecture, pushed by the fact that it concentrates in a single architectural intervention a triple object-, landscape- and infrastructure-related nature, generates architectural answers with very specific features, and its study achieves following goals: 1: Clarify the term hybrid related to architectural intervention; 2: Tran...

  14. [Construction and Identification of the Bait Vector Containing Duck Circovirus Cap Gene for the Yeast Two-hybrid System]. (United States)

    Xu, Yu; Zhang, Zhilong; Lu, Yanyan; Zhang, Lei; Li, Pengfei; Jia, Renyong


    To construct a bait expression vector containing the duck circovirus Cap gene for use in the yeast two-hybrid system, the whole cap codon-optimized gene was inserted into pGBKT7 vector and confirmed by PCR, restriction enzyme digestion, and sequence analysis. After transformation into a Y2HGold yeast strain, the expression of Cap protein was analyzed by Western blotting. Toxicity and self-activation of the bait protein were detected using different dropout minimal base. PCR reaction, restriction enzyme digestion, and sequencing analyses indicated that the duck circovirus Cap gene was correctly inserted into pG- BKT7. Western blotting showed that the whole Cap protein was expressed. The recombinant bait protein had no toxicity and self-activation. Therefore, the bait vector with the Cap gene was constructed successfully, providing a foundation for future screening for interacting proteins in the yeast two-hybrid system.

  15. Hybrid Ventilation Air Flow Process

    DEFF Research Database (Denmark)

    Heiselberg, Per Kvols

    The scope of this annex is therefore to obtain better knowledge of the use of hybrid ventilation technologies. The annex focus on development of control strategies for hybrid ventilation, on development of methods to predict hybrid ventilation performance in office buildings and on implementation...

  16. (Hybrid) Baryons Symmetries and Masses

    CERN Document Server

    Page, P R


    We construct (hybrid) baryons in the flux-tube model of Isgur and Paton. In the limit of adiabatic quark motion, we build proper eigenstates of orbital angular momentum and construct the flavour, spin and J^P of hybrid baryons from the symmetries of the system. The lowest mass hybrid baryon is estimated at approximately 2 GeV.

  17. Workshop on hybrid rice

    Institute of Scientific and Technical Information of China (English)



    FAO, in collaboration with FEDEARROZ in Colombia and EMBRAPA / CNPAF in Brail, organized a workshop on the Establishment of a Coorperative Research Network on Hybrid Rice in Latin America and the Caribbean held from Mar 16 to 18, 1994 at EMBRAPA/CNPAF in Brazil. Dr MAO Changxiang,

  18. Improved hybrid rocket fuel (United States)

    Dean, David L.


    McDonnell Douglas Aerospace, as part of its Independent R&D, has initiated development of a clean burning, high performance hybrid fuel for consideration as an alternative to the solid rocket thrust augmentation currently utilized by American space launch systems including Atlas, Delta, Pegasus, Space Shuttle, and Titan. It could also be used in single stage to orbit or as the only propulsion system in a new launch vehicle. Compared to solid propellants based on aluminum and ammonium perchlorate, this fuel is more environmentally benign in that it totally eliminates hydrogen chloride and aluminum oxide by products, producing only water, hydrogen, nitrogen, carbon oxides, and trace amounts of nitrogen oxides. Compared to other hybrid fuel formulations under development, this fuel is cheaper, denser, and faster burning. The specific impulse of this fuel is comparable to other hybrid fuels and is between that of solids and liquids. The fuel also requires less oxygen than similar hybrid fuels to produce maximum specific impulse, thus reducing oxygen delivery system requirements.

  19. Organics go hybrid (United States)

    Lanzani, Guglielmo; Petrozza, Annamaria; Caironi, Mario


    From displays to solar cells, the field of organic optoelectronics has come a long way over the past 50 years, but the realization of an electrically pumped organic laser remains elusive. The answer may lie with hybrid organic-inorganic materials called perovskites.

  20. Teelt van hybride wintertarwerassen

    NARCIS (Netherlands)

    Timmer, R.D.; Paauw, J.G.M.


    Om de mogelijkheden van de teelt van hybride wintertarwerassen onder Nederlandse omstandigheden in beeld te brengen zijn er van 2000-2002 proeven uitgevoerd op het PPO-proefbedrijf te Lelystad. In deze proeven zijn een 4-tal hybriderassen (Hybnos, Hyno-braba, Hyno-esta, Mercury) vergeleken met een s

  1. Nuclear hybrid energy infrastructure

    Energy Technology Data Exchange (ETDEWEB)

    Agarwal, Vivek; Tawfik, Magdy S.


    The nuclear hybrid energy concept is becoming a reality for the US energy infrastructure where combinations of the various potential energy sources (nuclear, wind, solar, biomass, and so on) are integrated in a hybrid energy system. This paper focuses on challenges facing a hybrid system with a Small Modular Reactor at its core. The core of the paper will discuss efforts required to develop supervisory control center that collects data, supports decision-making, and serves as an information hub for supervisory control center. Such a center will also be a model for integrating future technologies and controls. In addition, advanced operations research, thermal cycle analysis, energy conversion analysis, control engineering, and human factors engineering will be part of the supervisory control center. Nuclear hybrid energy infrastructure would allow operators to optimize the cost of energy production by providing appropriate means of integrating different energy sources. The data needs to be stored, processed, analyzed, trended, and projected at right time to right operator to integrate different energy sources.

  2. Indexical Hybrid Tense Logic

    DEFF Research Database (Denmark)

    Blackburn, Patrick Rowan; Jørgensen, Klaus Frovin


    In this paper we explore the logic of now, yesterday, today and tomorrow by combining the semantic approach to indexicality pioneered by Hans Kamp [9] and refined by David Kaplan [10] with hybrid tense logic. We first introduce a special now nominal (our @now corresponds to Kamp’s original now...

  3. Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA

    Directory of Open Access Journals (Sweden)

    Yto André Y


    Full Text Available Abstract Background A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs at room temperature or -20°C after spotting on filter paper, for subsequent serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA. Methods Monoclonal antibodies (Mabs were spotted within a 0.5–1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20°C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20°C or at room temperature (RT in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS, class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS containing 0.2% gelatin. Results Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20°C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper. Conclusion The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG or M (IgM eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing

  4. Utility of slot-blot-ELISA as a new, fast, and sensitive immunoassay for detection of carcinoembryonic antigen in the urine samples of patients with various gastrointestinal malignancies. (United States)

    El-Masry, Samir; El-Sayed, Ibrahim H; Lotfy, Mahmoud; Mahmoud, Lamiaa; El-Naggar, Mohamed


    Carcinoembryonic antigen (CEA) is the most widely used clinical tumor marker. CEA immunoassay has found acceptance as a diagnostic adjunct in clinical diagnosis of gastrointestinal tumors (GIT). Several immunoassays have been established for detection of CEA in plasma, serum, tissue, feces, and urine of cancer patients using polyclonal or monoclonal antibodies raised against CEA. Some of these assays display both high sensitivity and specificity for the detection of CEA. However, these assays require special and highly expensive equipment and the procedures require long periods for their completion. In the present study, we established a Slot-Blot Enzyme Linked Immunosorbent Assay (SB-ELISA), based on anti-CEA monoclonal antibody (CEA-mAb), as a new, simple, fast, cheap, and non-invasive immunodiagnostic technique for detection of CEA in the urine of GIT patients. Urine and serum samples were collected from 248 GIT patients (58 with pancreatic cancer, 20 with hepatoma, 23 with ampullary carcinoma, 15 with hilar cholangiocarcinoma, 28 with gastric cancer, 14 with esophageal cancer, and 90 with colorectal cancer). Moreover, urine and serum samples were collected from 50 healthy individuals to serve as negative controls. The traditional ELISA technique was used for determination of CEA in the sera of GIT patients using anti-CEA monoclonal antibody. A comparison between the results of both techniques (ELISA and SB-ELISA) was carried out. The traditional ELISA detected CEA in the sera of 154 out of 248 GIT patients with a sensitivity of 59.8%, 51.7% positive predictive value (PPV) and 75.37% negative predictive value (NPV). In addition, it identified 15 false positive cases out of 50 healthy individuals with a specificity of 70%. The urinary CEA was identified by a Western blotting technique and CEA-mAb at a molecular mass of 180 Kda. The developed SB-ELISA showed higher sensitivity, specificity, PPV, and NPV (70.1%, 78%, 62.4%, and 82.13%, respectively) for detection

  5. Hybrid keyword search auctions

    KAUST Repository

    Goel, Ashish


    Search auctions have become a dominant source of revenue generation on the Internet. Such auctions have typically used per-click bidding and pricing. We propose the use of hybrid auctions where an advertiser can make a per-impression as well as a per-click bid, and the auctioneer then chooses one of the two as the pricing mechanism. We assume that the advertiser and the auctioneer both have separate beliefs (called priors) on the click-probability of an advertisement. We first prove that the hybrid auction is truthful, assuming that the advertisers are risk-neutral. We then show that this auction is superior to the existing per-click auction in multiple ways: 1. We show that risk-seeking advertisers will choose only a per-impression bid whereas risk-averse advertisers will choose only a per-click bid, and argue that both kind of advertisers arise naturally. Hence, the ability to bid in a hybrid fashion is important to account for the risk characteristics of the advertisers. 2. For obscure keywords, the auctioneer is unlikely to have a very sharp prior on the click-probabilities. In such situations, we show that having the extra information from the advertisers in the form of a per-impression bid can result in significantly higher revenue. 3. An advertiser who believes that its click-probability is much higher than the auctioneer\\'s estimate can use per-impression bids to correct the auctioneer\\'s prior without incurring any extra cost. 4. The hybrid auction can allow the advertiser and auctioneer to implement complex dynamic programming strategies to deal with the uncertainty in the click-probability using the same basic auction. The per-click and per-impression bidding schemes can only be used to implement two extreme cases of these strategies. As Internet commerce matures, we need more sophisticated pricing models to exploit all the information held by each of the participants. We believe that hybrid auctions could be an important step in this direction. The

  6. Simultaneous in situ hybridization for DNA and RNA reveals the presence of HPV in the majority of cervical cancer cells. (United States)

    D'Amato, L; Pilotti, S; Longoni, A; Donghi, R; Rilke, F


    Thirteen cases of invasive squamous cell carcinoma of the uterine cervix containing HPV types 16 or 18 DNA sequences, as detected by Southern blot analysis, were investigated by in situ hybridization on routine paraffin sections, using 35S nick-translated DNA probes. Simultaneous in situ hybridization for DNA and RNA showed that in ten out of 13 cases (77%) the percentage of tumor cells containing HPV 16 or 18 varied from 75 to 100%. In one case, harboring both in situ and invasive carcinoma, the same type of HPV DNA was detected in both components. This finding suggests that neoplastic cells retained the viral genome during progression to invasiveness.

  7. Direct detection of expanded trinucleotide repeats using DNA hybridization techniques

    Energy Technology Data Exchange (ETDEWEB)

    Petronis, A.; Tatuch, Y.; Kennedy, J.L. [Univ. of Toronto (Canada)] [and others


    Recently, unstable trinucleotide repeats have been shown to be the etiologic factor in several neuropsychiatric diseases, and they may play a similar role in other disorders. To our knowledge, a method that detects expanded trinucleotide sequences with the opportunity for direct localization and cloning has not been achieved. We have developed a set of hybridization-based methods for direct detection of unstable DNA expansion. Our analysis of myotonic dystrophy patients that possess different degrees of (CTG){sub n} expansion, versus unaffected controls, has demonstrated the identification of the trinucleotide instability site without any prior information regarding genetic map location. High stringency modified Southern blot hybridization with a PCR-generated trinucleotide repeat probe allowed us to detect the DNA fragment containing the expansion in myotonic dystrophy patients. The same probe was used for fluorescent in situ hybridization and several regions of (CTG){sub n}/(CAG){sub n} repeats in the human genome were detected, including the myotonic dystrophy locus on chromosome 19q. These strategies can be applied to directly clone genes involved in disorders caused by unstable DNA.

  8. Ants exhibit asymmetric hybridization in a mosaic hybrid zone. (United States)

    Purcell, Jessica; Zahnd, Sacha; Athanasiades, Anouk; Türler, Rebecca; Chapuisat, Michel; Brelsford, Alan


    Research on hybridization between species provides unparalleled insights into the pre- and postzygotic isolating mechanisms that drive speciation. In social organisms, colony-level incompatibilities may provide additional reproductive barriers not present in solitary species, and hybrid zones offer an opportunity to identify these barriers. Here, we use genotyping-by-sequencing to sequence hundreds of markers in a hybrid zone between two socially polymorphic ant species, Formica selysi and Formica cinerea. We characterize the zone, determine the frequency of hybrid workers, infer whether hybrid queens or males are produced and investigate whether hybridization is influenced by colony social organization. We also compare cuticular hydrocarbon profiles and aggression levels between the two species. The hybrid zone exhibits a mosaic structure. The asymmetric distribution of hybrids skewed towards F. cinerea suggests a pattern of unidirectional nuclear gene flow from F. selysi into F. cinerea. The occurrence of backcrossed individuals indicates that hybrid queens and/or males are fertile, and the presence of the F. cinerea mitochondrial haplotype in 97% of hybrids shows that successful F1 hybrids will generally have F. cinerea mothers and F. selysi fathers. We found no evidence that social organization contributes to speciation, because hybrids occur in both single-queen and multiple-queen colonies. Strongly differentiated cuticular hydrocarbon profiles and heightened interspecific aggression further reveal that species recognition cues are both present and perceived. The discovery of fertile hybrids and asymmetrical gene flow is unusual in ants, and this hybrid zone will therefore provide an ideal system with which to investigate speciation in social insects.

  9. Whole genomic sequencing of RT98 mitochondria derived from Oryza rufipogon and northern blot analysis to uncover a cytoplasmic male sterility-associated gene. (United States)

    Igarashi, Keisuke; Kazama, Tomohiko; Motomura, Keiji; Toriyama, Kinya


    Cytoplasmic male sterility (CMS) is a maternally inherited trait resulting in the failure to produce functional pollen and is often observed when an alien cytoplasm is transferred into a cultivated species. An RT98A CMS line and an RT98C fertility restorer line were obtained by successive backcrossing between Oryza rufipogon W1109 and Oryza sativa cultivar Taichung 65. To uncover the CMS-associated mitochondrial genes, we determined the complete sequence of the RT98-CMS mitochondrial genome using next-generation pyrosequencing, and searched new open reading frames (orfs) absent in a reported mitochondrial genome of O. sativa Nipponbare. Then, six candidates were selected for the CMS-associated genes based on the criteria in which they were chimeric in structure or encoded a peptide with transmembrane domains. One of the candidates, orf113, showed different transcript sizes between RT98A and RT98C on Northern blot analysis. The orf113 gene was shown to be co-transcribed with atp4 and cox3 encoding ATP synthase F0 subunit 4 and Cyt c oxidase subunit 3, respectively, and their transcripts were distinctly processed in the presence of a fertility restorer gene. Our results indicate that orf113 is a CMS-associated gene of RT98-CMS.

  10. Western blot seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 in Fortaleza (Brazil: a serological and molecular diagnostic and epidemiological approach

    Directory of Open Access Journals (Sweden)

    Terezinha de Jesus Teixeira Santos


    Full Text Available How to handle Western blot (WB seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 constitutes a challenge for blood banks and fam ilies. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA reactive individuals from the hematological center (HEMOCE of Fortaleza (Brazil, examining their serological (WB and molecular (PCR diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22% were positive and 32 (78% were negative for HTLV-1/2. The demographic analysis indicated a trend towards a predominance of males among the seroindeterminate individuals and females in the seropositive ones. The seroindeterminate individuals were younger than the seropositive ones. We did not find any association of these conditions with syphilis, Chagas disease or HIV or hepatitis, and with risk factors such as breast-feeding, blood transfusion, STD (syphilis and IDU.

  11. Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants. (United States)

    Stefano, Biricolti; Patrizia, Bogani; Matteo, Cerboneschi; Massimo, Gori


    One of the major unanswered questions with respect to the commercial use of genetic transformation in woody plants is the stability of the transgene expression over several decades within the same individual. Gene expression is strongly affected by the copy number which has been integrated into the plant genome and by the local DNA features close to the integration sites. Because woody plants cannot be subjected to selfing or backcrossing to modify the transgenic allelic structure without affecting the valuable traits of the cultivar, molecular characterization of the transformation event is therefore crucial. After assessing the transgene copy number of a set of apple transgenic clones with Southern blotting, we describe two alternative methods: the first is based on inverse PCR (i-PCR) and the second on the quantitative PCR (q-PCR). The methods produced comparable results with the exception of the data regarding a high copy number clone, but while the q-PCR-based system is rapid and easily adaptable to high throughput systems, the i-PCR-based method can provide information regarding the transformation event and the characteristics of the sequences flanking the transgenic construct.

  12. Utility of Schistosoma bovis Adult Worm Antigens for Diagnosis of Human Schistosomiasis by Enzyme-Linked Immunosorbent Assay and Electroimmunotransfer Blot Techniques (United States)

    Pardo, J.; Carranza, C.; Turrientes, M. C.; Arellano, J. L. Pérez; Vélez, R. López; Ramajo, V.; Muro, A.


    Immunodiagnostic methods based on the detection of antibodies continue to be the most effective and practical methods for the diagnosis of imported schistosomiasis. Schistosoma bovis is a species whose final natural hosts are bovines, ovines, caprines, and small wild ruminants. Different studies have demonstrated the analogies existing between S. bovis and other Schistosoma species which affect humans. The objective of this work was to evaluate the utility of S. bovis adult worm antigens (AWA) for the diagnosis of imported human schistosomiasis by enzyme-linked immunosorbent assay (ELISA) and electroimmunotransfer blotting (EITB) techniques. By detecting eggs, the ELISA for S. bovis AWA was able to definitively detect imported cases with a sensitivity of 94%. The specificity of the ELISA for S. bovis AWA was 97%. There were no differences between the results of the S. bovis AWA ELISA for patients infected with Schistosoma mansoni and those infected with Schistosoma haematobium. The EITB technique showed bands of 85, 37, and 20 kDa, which are characteristic of infections with Schistosoma spp. Specific bands to indicate infection by different species of Schistosoma have not been detected. The combined use of the ELISA for S. bovis AWA and EITB increased the global sensitivity of the study to 97%. Our findings suggest that the ELISA for S. bovis AWA is a useful test for the immunodiagnosis of imported schistosomiasis and that EITB for detecting S. bovis AWA permits the confirmation of diagnosis when the ELISA for S. bovis AWA is positive. PMID:15539523

  13. Diversity of Borrelia genospecies in Ixodes ricinus ticks in a Lyme borreliosis endemic area in Switzerland identified by using new probes for reverse line blotting. (United States)

    Gern, Lise; Douet, Véronique; López, Zully; Rais, Olivier; Cadenas, Francisca Morán


    In Europe, 7 Borrelia species belonging to the Borrelia burgdorferi sensu lato complex have been reported in Ixodes ricinus ticks. In addition, another Borrelia, related to the relapsing fever spirochaetes, has also been described. In the present study, we designed probes for reverse line blotting allowing detection and identification of all these Borrelia species after amplification of the variable spacer region between the 23S and 5S ribosomal genes. These new probes allowed us investigate the diversity of Borrelia in 915 I. ricinus collected on the south-facing slope of Chaumont (Switzerland). Among the 159 infected ticks, 7 Borrelia species were identified, and B. spielmanii and relapsing fever-like (RFL) spirochaetes were identified in this area for the first time. B. valaisiana and B. spielmanii were significantly less present in male than in female or nymphal ticks. Mixed infection with RFL spirochaetes and Lyme borreliosis spirochaetes were detected in 4 ticks. In addition, the set of probes could identify the recently described species, B. bavariensis.

  14. A Western blot-based investigation of the yeast secretory pathway designed for an intermediate-level undergraduate cell biology laboratory. (United States)

    Hood-Degrenier, Jennifer K


    The movement of newly synthesized proteins through the endomembrane system of eukaryotic cells, often referred to generally as the secretory pathway, is a topic covered in most intermediate-level undergraduate cell biology courses. An article previously published in this journal described a laboratory exercise in which yeast mutants defective in two distinct steps of protein secretion were differentiated using a genetic reporter designed specifically to identify defects in the first step of the pathway, the insertion of proteins into the endoplasmic reticulum (Vallen, 2002). We have developed two versions of a Western blotting assay that serves as a second way of distinguishing the two secretory mutants, which we pair with the genetic assay in a 3-wk laboratory module. A quiz administered before and after students participated in the lab activities revealed significant postlab gains in their understanding of the secretory pathway and experimental techniques used to study it. A second survey administered at the end of the lab module assessed student perceptions of the efficacy of the lab activities; the results of this survey indicated that the experiments were successful in meeting a set of educational goals defined by the instructor.

  15. Identification of liver protein targets modified by tienilic acid metabolites using a two-dimensional Western blot-mass spectrometry approach (United States)

    Methogo, Ruth Menque; Dansette, Patrick M.; Klarskov, Klaus


    A combined approach based on two-dimensional electrophoresis-immuno-blotting and nanoliquid chromatography coupled on-line with electrospray ionization mass spectrometry (nLC-MS/MS) was used to identify proteins modified by a reactive intermediate of tienilic acid (TA). Liver homogenates from rats exposed to TA were fractionated using ultra centrifugation; four fractions were obtained and subjected to 2D electrophoresis. Following transfer to PVDF membranes, modified proteins were visualized after India ink staining, using an anti-serum raised against TA and ECL detection. Immuno-reactive spots were localized on the PVDF membrane by superposition of the ECL image, protein spots of interest were excised, digested on the membrane with trypsin followed by nLC-MS/MS analysis and protein identification. A total of 15 proteins were identified as likely targets modified by a TA reactive metabolite. These include selenium binding protein 2, senescence marker protein SMP-30, adenosine kinase, Acy1 protein, adenosylhomocysteinase, capping protein (actin filament), protein disulfide isomerase, fumarylacetoacetase, arginase chain A, ketohexokinase, proteasome endopeptidase complex, triosephosphate isomerase, superoxide dismutase, dna-type molecular chaperone hsc73 and malate dehydrogenase.

  16. Immunohistochemical and Western blot analysis of two protein tyrosine phosphatase receptors, R and Z1, in colorectal carcinoma, colon adenoma and normal colon tissues. (United States)

    Woźniak, Marta; Gamian, Elżbieta; Łaczmańska, Izabela; Sąsiadek, Maria M; Duś-Szachniewicz, Kamila; Ziółkowski, Piotr


    Two classes of proteins, namely tyrosine kinases (PTK) and phosphatases (PTP), play an important role in cell proliferation and differentiation, thus leading to an acceleration or inhibition of tumour growth. The role of the above proteins in colorectal carcinoma (CRC) growth is a well-known event. In this study we carried out immunohistochemical and Western blot analysis of colorectal carcinoma, adenoma and normal colon tissue in relation to two protein tyrosine phosphatase receptors, R and Z1. Twenty-five cases of CRC were analyzed and the results were compared with similar data obtained in non-malignant tissues. High expression of both PTP receptors was observed in all examined cases of CRC, adenoma and normal colon tissue in this study. These results are not in line with recently published data, showing that genetic coding for PTPRR and PTPRZ1 were hypermethylated in CRC's. We presume that the protein tyrosine phosphatase overexpression in colorectal carcinoma is not enough to protect from the progression of disease.

  17. Direct measurement of tubulin and bulk message distributions on polysomes of growing, starved and deciliated Tetrahymena using RNA gel blots of sucrose gradients containing acrylamide. (United States)

    Calzone, F J; Callahan, R; Gorovsky, M A


    A method was developed using sucrose gradients containing acrylamide which greatly simplifies the measurement of the polysomal distribution of messages. After centrifugation, the acrylamide was polymerized, forming a "polysome gel". RNA gel blots of polysome gels were used to determine the polysomal distributions of alpha-tubulin and total polyadenylated mRNA in growing, starved (nongrowing) and starved-deciliated Tetrahymena and the number of messages loaded onto polysomes was calculated. These measurements indicated that the translational efficiencies of alpha-tubulin mRNA and total polyadenylated mRNA are largely unaffected when the rates of tubulin and total protein synthesis vary dramatically. Thus, differential regulation of alpha-tubulin mRNA translation initiation does not contribute to the greater than 100-fold induction of tubulin synthesis observed during cilia regeneration and in growing cells. The major translation-level process regulating tubulin synthesis in Tetrahymena appears to be a change in message loading mediated by a non-specific message recruitment or unmasking factor.

  18. Identification of differentially expressed radiation-induced genes in cervix carcinoma cells using suppression subtractive hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jun Sang; Lee, Young Sook; Lee, Jeung Hoon; Lee, Woong Hee; Seo, Eun Young; Cho, Moon June [Chungnam National University, Daejeon (Korea, Republic of)


    A number of genes and their products are induced early or late following exposure of cells to ionizing radiation. These radiation-induced genes have various effects of irradiated cells and tissues. Suppression subtractive hybridization (SSH) based on PCR was used to identify the differentially expressed genes by radiation in cervix carcinoma cells. Total RNA and poly (A){sup +} mRNA were isolated from irradiated and non-irradiated HeLa cells. Forward-and reverse-subtracted cDNA libraries were constructed using SSH. Eighty-eight clones of each were used to randomly select differentially expressed genes using reverse Northern blotting (dot blot analysis). Northern blotting was used to verify the screened genes. Of the 176 clones, 10 genes in the forward-subtracted library and 9 genes in the reverse-subtracted library were identified as differentially expressed radiation-induced genes by PCR-select differential screening. Three clones from the forward-subtracted library were confirmed by Northern blotting, and showed increased expression in a dose-dependent manner, including a telomerase catalytic subunit and sodium channel-like protein gene, and an ESTs (expressed sequence tags) gene. We identified differentially expressed radiation-induced genes with low-abundance genes with SSH, but further characterization of theses genes are necessary to clarify the biological functions of them.

  19. Homologous genes for mouse 4.5S hybRNA are found in all eukaryotes and their low molecular weight RNA transcripts intermolecularly hybridize with eukaryotic 18S ribosomal RNAs. (United States)

    Trinh-Rohlik, Q; Maxwell, E S


    Previous work has reported the isolation and sequencing of a mouse low molecular weight RNA species designated 4.5S hybridizing RNA or hybRNA because of its ability to intermolecularly hybridize with mouse mRNA and 18S rRNA sequences. Using synthetic DNA oligonucleotide probes we have examined the conservation of this gene sequence and its expression as a lmwRNA transcript across evolution. Southern blot analysis has shown that homologous genes of single or low copy number are found in all eukaryotes examined as well as in E. coli. Northern blot analysis has demonstrated 4.5S hybRNA transcription in all mouse tissues as well as expression in yeast and Xenopus laevis as lmwRNAs of approximately 130 and 100 nucleotides, respectively, as compared with mouse/rat/hamster species of approximately 87 nucleotides. Yeast and X. laevis 4.5S hybRNA homologs, isolated by hybrid-selection, were shown by Northern blot analysis to intermolecularly hybridize with homologous as well as heterologous 18S rRNA sequences. The conservation of 4.5S hybRNA homologous genes and their expression as lmwRNA transcripts with common intermolecular RNA:RNA hybridization capabilities in fungi, amphibians, and mammals argues for a common, conserved and required biological function for this lmwRNA in all eukaryotes and potential utilization of its intermolecular RNA:RNA hybridization capabilities to carry out this function.

  20. Energy Efficiency Comparison between Hydraulic Hybrid and Hybrid Electric Vehicles

    Directory of Open Access Journals (Sweden)

    Jia-Shiun Chen


    Full Text Available Conventional vehicles tend to consume considerable amounts of fuel, which generates exhaust gases and environmental pollution during intermittent driving cycles. Therefore, prospective vehicle designs favor improved exhaust emissions and energy consumption without compromising vehicle performance. Although pure electric vehicles feature high performance and low pollution characteristics, their limitations are their short driving range and high battery costs. Hybrid electric vehicles (HEVs are comparatively environmentally friendly and energy efficient, but cost substantially more compared with conventional vehicles. Hydraulic hybrid vehicles (HHVs are mainly operated using engines, or using alternate combinations of engine and hydraulic power sources while vehicles accelerate. When the hydraulic system accumulator is depleted, the conventional engine reengages; concurrently, brake-regenerated power is recycled and reused by employing hydraulic motor–pump modules in circulation patterns to conserve fuel and recycle brake energy. This study adopted MATLAB Simulink to construct complete HHV and HEV models for backward simulations. New European Driving Cycles were used to determine the changes in fuel economy. The output of power components and the state-of-charge of energy could be retrieved. Varying power component models, energy storage component models, and series or parallel configurations were combined into seven different vehicle configurations: the conventional manual transmission vehicle, series hybrid electric vehicle, series hydraulic hybrid vehicle, parallel hybrid electric vehicle, parallel hydraulic hybrid vehicle, purely electric vehicle, and hydraulic-electric hybrid vehicle. The simulation results show that fuel consumption was 21.80% lower in the series hydraulic hybrid vehicle compared to the series hybrid electric vehicle; additionally, fuel consumption was 3.80% lower in the parallel hybrid electric vehicle compared to the

  1. Phoxonic Hybrid Superlattice. (United States)

    Alonso-Redondo, Elena; Huesmann, Hannah; El Boudouti, El-Houssaine; Tremel, Wolfgang; Djafari-Rouhani, Bahram; Butt, Hans-Juergen; Fytas, George


    We studied experimentally and theoretically the direction-dependent elastic and electromagnetic wave propagation in a supported film of hybrid PMMA (poly[methyl-methacrylate])-TiO2 superlattice (SL). In the direction normal to the layers, this one-dimensional periodic structure opens propagation band gaps for both hypersonic (GHz) phonons and near-UV photons. The high mismatch of elastic and optical impedance results in a large dual phoxonic band gap. The presence of defects inherent to the spin-coating fabrication technique is sensitively manifested in the band gap region. Utilizing Brillouin light scattering, phonon propagation along the layers was observed to be distinctly different from propagation normal to them and can, under certain conditions (SL thickness and substrate elasticity), reveal the nanomechanical properties of the constituent layers. Besides the first realization of unidirectional phoxonic behavior, hybrid (soft-hard) periodic materials are a promising simple platform for opto-acoustic interactions and applications such as filters and Bragg mirrors.

  2. Hybrid plasma modeling.

    Energy Technology Data Exchange (ETDEWEB)

    Hopkins, Matthew Morgan; DeChant, Lawrence Justin.; Piekos, Edward Stanley; Pointon, Timothy David


    This report summarizes the work completed during FY2007 and FY2008 for the LDRD project ''Hybrid Plasma Modeling''. The goal of this project was to develop hybrid methods to model plasmas across the non-continuum-to-continuum collisionality spectrum. The primary methodology to span these regimes was to couple a kinetic method (e.g., Particle-In-Cell) in the non-continuum regions to a continuum PDE-based method (e.g., finite differences) in continuum regions. The interface between the two would be adjusted dynamically ased on statistical sampling of the kinetic results. Although originally a three-year project, it became clear during the second year (FY2008) that there were not sufficient resources to complete the project and it was terminated mid-year.

  3. The Power of Hybridization

    CERN Document Server

    CERN. Geneva


    Programming languages always seem to do some things well but not others: Python punts when it comes to user interfaces, Java’s artificial complexity prevents rapid development and produces tangles, and it will be awhile before we see benefits from C++ concurrency work. The cognitive load of languages and their blind spots increases the cost of experimentation, impeding your ability to fail fast and iterate. If you use a single language to solve your problem, you are binding yourself to the worldview limitations and the mistakes made by the creator of that language. Consider increasing your wiggle room by crossing language boundaries, complementing a language that is powerful in one area with a different language powerful in another. Language hybridization can speed development to quickly discover your real problems, giving you more time to fix them. After making a case for hybridizing your thinking in general, I will present a number of simple examples; first showing the benefits of using other languages...

  4. Hybrid undulator numerical optimization

    Energy Technology Data Exchange (ETDEWEB)

    Hairetdinov, A.H. [Kurchatov Institute, Moscow (Russian Federation); Zukov, A.A. [Solid State Physics Institute, Chernogolovka (Russian Federation)


    3D properties of the hybrid undulator scheme arc studied numerically using PANDIRA code. It is shown that there exist two well defined sets of undulator parameters which provide either maximum on-axis field amplitude or minimal higher harmonics amplitude of the basic undulator field. Thus the alternative between higher field amplitude or pure sinusoidal field exists. The behavior of the undulator field amplitude and harmonics structure for a large set of (undulator gap)/(undulator wavelength) values is demonstrated.

  5. A Pseudoscalar Hybrid Meson?

    CERN Document Server

    Page, P R


    New experimental information on the non--exotic J^{PC} = 0^{-+} isovector seen at 1.8 GeV by VES yields convincing evidence of its excited gluonic (hybrid) nature when a critical study of alternative quarkonium assignments is made in the context of ^3 P_0 decay by flux--tube breaking. Production of this gluonic excitation via meson exchange is promising, although its two photon production vanishes.

  6. Fibonacci-Pell Hybridities (United States)

    Koshy, Thomas; Gao, Zhenguang


    We develop a recurrence satisfied by the Fibonacci and Pell families. We then use it to find explicit formulae and generating functions for the hybrids "F[subscript n]P[subscript n]", "L[subscript n]P[subscript n]", "F[subscript n]Q[subscript n]" and "L[subscript n]Q[subscript n]", where "F[subscript n]", "L[subscript n]", "P[subscript n]" and…

  7. Military Hybrid Vehicle Survey (United States)


    Furthermore, a standard duty cycle that is accepted for measuring fuel economy does not exist nor does a focus towards a particular technology. This...expanded into mild hybrid with the addition of a clutch connecting the generator to the transmission and additional energy storage [16-17...speed control and one for engine/generator torque [35]. Urban, Highway, Composite 33%, 27.9%, 49% General vehicle simulation [30]. Urban 19.0

  8. Hybrid electroluminescent devices (United States)

    Shiang, Joseph John; Duggal, Anil Raj; Michael, Joseph Darryl


    A hybrid electroluminescent (EL) device comprises at least one inorganic diode element and at least one organic EL element that are electrically connected in series. The absolute value of the breakdown voltage of the inorganic diode element is greater than the absolute value of the maximum reverse bias voltage across the series. The inorganic diode element can be a power diode, a Schottky barrier diode, or a light-emitting diode.

  9. Printed hybrid systems (United States)

    Karioja, Pentti; Mäkinen, Jukka-Tapani; Keränen, Kimmo; Aikio, Janne; Alajoki, Teemu; Jaakola, Tuomo; Koponen, Matti; Keränen, Antti; Heikkinen, Mikko; Tuomikoski, Markus; Suhonen, Riikka; Hakalahti, Leena; Kopola, Pälvi; Hast, Jukka; Liedert, Ralf; Hiltunen, Jussi; Masuda, Noriyuki; Kemppainen, Antti; Rönkä, Kari; Korhonen, Raimo


    This paper presents research activities carried out at VTT Technical Research Centre of Finland in the field of hybrid integration of optics, electronics and mechanics. Main focus area in our research is the manufacturing of electronic modules and product structures with printed electronics, film-over-molding and polymer sheet lamination technologies and the goal is in the next generation of smart systems utilizing monolithic polymer packages. The combination of manufacturing technologies such as roll-to-roll -printing, injection molding and traditional component assembly is called Printed Hybrid Systems (PHS). Several demonstrator structures have been made, which show the potential of polymer packaging technology. One demonstrator example is a laminated structure with embedded LED chips. Element thickness is only 0.3mm and the flexible stack of foils can be bent in two directions after assembly process and was shaped curved using heat and pressure. The combination of printed flexible circuit boards and injection molding has also been demonstrated with several functional modules. The demonstrators illustrate the potential of origami electronics, which can be cut and folded to 3D shapes. It shows that several manufacturing process steps can be eliminated by Printed Hybrid Systems technology. The main benefits of this combination are small size, ruggedness and conformality. The devices are ideally suited for medical applications as the sensitive electronic components are well protected inside the plastic and the structures can be cleaned easily due to the fact that they have no joints or seams that can accumulate dirt or bacteria.

  10. Hybrid Keyword Search Auctions

    CERN Document Server

    Goel, Ashish


    Search auctions have become a dominant source of revenue generation on the Internet. Such auctions have typically used per-click bidding and pricing. We propose the use of hybrid auctions where an advertiser can make a per-impression as well as a per-click bid, and the auctioneer then chooses one of the two as the pricing mechanism. We assume that the advertiser and the auctioneer both have separate beliefs (called priors) on the click-probability of an advertisement. We first prove that the hybrid auction is truthful, assuming that the advertisers are risk-neutral. We then show that this auction is superior to the existing per-click auction in multiple ways: 1) It takes into account the risk characteristics of the advertisers. 2) For obscure keywords, the auctioneer is unlikely to have a very sharp prior on the click-probabilities. In such situations, the hybrid auction can result in significantly higher revenue. 3) An advertiser who believes that its click-probability is much higher than the auctioneer's es...

  11. Asymmetric Hybrid Nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Chumanov, George [Clemson Univ., SC (United States)


    Hybrid Nanoparticles (AHNs) are rationally-designed multifunctional nanostructures and novel building blocks for the next generation of advanced materials and devices. Nanoscale materials attract considerable interest because of their unusual properties and potential for practical applications. Most of the activity in this field is focused on the synthesis of homogeneous nanoparticles from metals, metal oxides, semiconductors, and polymers. It is well recognized that properties of nanoparticles can be further enhanced if they are made as hybrid structures. This program is concerned with the synthesis, characterization, and application of such hybrid structures termed AHNs. AHNs are composed of a homogeneous core and several caps of different materials deposited on its surface (Fig. 1). Combined properties of the core and the caps as well as new properties that arise from core-cap and cap-cap interactions render AHNs multifunctional. In addition, specific chemical reactivity of the caps enables directional self-assembly of AHNs into complex architectures that are not possible with only spherical nanoparticles.

  12. A comparative study of Toxoplasma gondii seroprevalence in mink using a modified agglutination test, a Western blot, and enzyme-linked immunosorbent assays. (United States)

    Gu, Yi; Wang, Zedong; Cai, Yufeng; Li, Xiaoxing; Wei, Feng; Shang, Limin; Li, Jiping; Liu, Quan


    Toxoplasma gondii can infect almost all warm-blooded animals, and many serological methods have been developed to detect T. gondii infection in a variety of animal species. In the present study, the seroprevalence of T. gondii infection in farmed mink in northeast China was determined using the modified agglutination test (MAT), a Western blot (WB), and 3 enzyme-linked immunosorbent assays (ELISAs) with protein A/G conjugate, using either of 2 recombinant dense granule antigens, GRA1 and GRA7, or Toxoplasma soluble antigens (TSA). There was no significant difference between the detection results of the GRA1-, GRA7-, and TSA-ELISAs and WB (McNemar chi-square, P > 0.05), but a significant difference was observed between MAT and WB (P < 0.05). A near perfect agreement (97.0%) was found between the GRA7-ELISA and WB (κ = 0.83), and a substantial agreement (92.4-93.1%) was observed in the TSA- and GRA1-ELISAs (κ = 0.68-0.73). The GRA7-ELISA showed the highest sensitivity and specificity, and the lowest false-positive and negative rates, while the MAT gave both a low sensitivity and frequent false positives in comparison to the WB. Receiver operating characteristic analysis revealed the largest area under curve of 0.85 (95% confidence interval: 0.74-0.96), and the highest relative sensitivity (72.7%) and specificity (99.0%) for a cutoff value of 0.19 in the GRA7-ELISA. These results indicate that the GRA7-ELISA is suitable for detection of T. gondii infection in mink and that MAT should be used with caution.

  13. Western blot data using two distinct anti-O-GlcNAc monoclonal antibodies showing unique glycosylation status on cellular proteins under 2-deoxy-d-glucose treatment

    Directory of Open Access Journals (Sweden)

    Tetsuya Okuda


    Full Text Available Protein modification by O-linked N-acetylglucosamine (O-GlcNAcylation is one of the post transcriptional modifications occurring on cellular proteins. This paper provides a data set relating to the O-GlcNAcylation of cellular proteins detected by RL2 and CTD110.6 antibodies, which are commonly used for detection of protein O-GlcNAcylation, in 2-deoxy-d-glucose (2DG-treated human teratocarcinoma NCCIT cells in support of the research article entitled “A novel, promoter-based, target-specific assay identifies 2-deoxy-d-glucose as an inhibitor of globotriaosylceramide biosynthesis” (Okuda et al., 2009 [1]. The main article described a suppressive effect of 2DG on an Sp1 target gene in NCCIT cells and discussed the relationship between the effect of 2DG and O-GlcNAcylation status of Sp1. The data in this paper complements this relationship by Western blotting and clearly showed that the 2DG treatment increased O-GlcNAcylation of cellular proteins in NCCIT cells, whereas the RL2 and CTD110.6 epitopes were detected in a different manner. The RL2 epitope was detected on Sp1 during 2DG treatment, and the level was transiently increased at 24 h. In contrast, the CTD110.6 epitope became detectable on Sp1 over 72 h after 2DG treatment, and then the other proteins containing CTD110.6 epitopes also appeared in the cell lysates and the anti-Sp1 antibody precipitates.

  14. HIV-1/2 indeterminate Western blot results: follow-up of asymptomatic blood donors in Belo Horizonte, Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)



    Full Text Available The clinical and public health importance of indeterminate results in HIV-1/2 testing is still difficult to evaluate in volunteer blood donors. At Fundação Hemominas, HIV-1/2 ELISA is used as the screening test and, if reactive, is followed by Western blot (WB. We have evaluated 84 blood donors who had repeatedly reactive ELISA tests for HIV-1/2, but indeterminate WB results. Sixteen of the 84 donors (19.0% had history of sexually transmitted diseases; 18/84 (21.4% informed receiving or paying for sex; 3/84 (3.6% had homosexual contact; 2/26 women (7.6% had past history of multiple illegal abortions and 3/84 (3.6% had been previously transfused. Four out of 62 donors (6.5% had positive anti-nuclear factor (Hep2, with titles up to 1:640. Parasitological examination of the stool revealed eggs of S. mansoni in 4/62 (6.4% donors and other parasites in 8/62 (12.9%. Five (5.9% of the subjects presented overt seroconversion for HIV-1/2, 43/84 (51.2% had negative results on the last visit, while 36/84 (42.9% remained WB indeterminate. Although some conditions could be found associated with the HIV-1/2 indeterminate WB results and many donors had past of risky behavior, the significance of the majority of the results remains to be determined.

  15. Identification of reference proteins for Western blot analyses in mouse model systems of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD toxicity.

    Directory of Open Access Journals (Sweden)

    Stephenie D Prokopec

    Full Text Available Western blotting is a well-established, inexpensive and accurate way of measuring protein content. Because of technical variation between wells, normalization is required for valid interpretation of results across multiple samples. Typically this involves the use of one or more endogenous controls to adjust the measured levels of experimental molecules. Although some endogenous controls are widely used, validation is required for each experimental system. This is critical when studying transcriptional-modulators, such as toxicants like 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD.To address this issue, we examined hepatic tissue from 192 mice representing 47 unique combinations of strain, sex, Ahr-genotype, TCDD dose and treatment time. We examined 7 candidate reference proteins in each animal and assessed consistency of protein abundance through: 1 TCDD-induced fold-difference in protein content from basal levels, 2 inter- and intra- animal stability, and 3 the ability of each candidate to reduce instability of the other candidates. Univariate analyses identified HPRT as the most stable protein. Multivariate analysis indicated that stability generally increased with the number of proteins used, but gains from using >3 proteins were small. Lastly, by comparing these new data to our previous studies of mRNA controls on the same animals, we were able to show that the ideal mRNA and protein control-genes are distinct, and use of only 2-3 proteins provides strong stability, unlike in mRNA studies in the same cohort, where larger control-gene batteries were needed.

  16. Hybrid2 - The hybrid power system simulation model

    Energy Technology Data Exchange (ETDEWEB)

    Baring-Gould, E.I.; Green, H.J.; Dijk, V.A.P. van [National Renewable Energy Lab., Golden, CO (United States); Manwell, J.F. [Univ. of Massachusetts, Amherst, MA (United States)


    There is a large-scale need and desire for energy in remote communities, especially in the developing world; however the lack of a user friendly, flexible performance prediction model for hybrid power systems incorporating renewables hindered the analysis of hybrids as options to conventional solutions. A user friendly model was needed with the versatility to simulate the many system locations, widely varying hardware configurations, and differing control options for potential hybrid power systems. To meet these ends, researchers from the National Renewable Energy Laboratory (NREL) and the University of Massachusetts (UMass) developed the Hybrid2 software. This paper provides an overview of the capabilities, features, and functionality of the Hybrid2 code, discusses its validation and future plans. Model availability and technical support provided to Hybrid2 users are also discussed. 12 refs., 3 figs., 4 tabs.

  17. Atypical haemolytic uraemic syndrome associated with a hybrid complement gene.

    Directory of Open Access Journals (Sweden)

    Julian P Venables


    Full Text Available BACKGROUND: Sequence analysis of the regulators of complement activation (RCA cluster of genes at chromosome position 1q32 shows evidence of several large genomic duplications. These duplications have resulted in a high degree of sequence identity between the gene for factor H (CFH and the genes for the five factor H-related proteins (CFHL1-5; aliases CFHR1-5. CFH mutations have been described in association with atypical haemolytic uraemic syndrome (aHUS. The majority of the mutations are missense changes that cluster in the C-terminal region and impair the ability of factor H to regulate surface-bound C3b. Some have arisen as a result of gene conversion between CFH and CFHL1. In this study we tested the hypothesis that nonallelic homologous recombination between low-copy repeats in the RCA cluster could result in the formation of a hybrid CFH/CFHL1 gene that predisposes to the development of aHUS. METHODS AND FINDINGS: In a family with many cases of aHUS that segregate with the RCA cluster we used cDNA analysis, gene sequencing, and Southern blotting to show that affected individuals carry a heterozygous CFH/CFHL1 hybrid gene in which exons 1-21 are derived from CFH and exons 22/23 from CFHL1. This hybrid encodes a protein product identical to a functionally significant CFH mutant (c.3572C>T, S1191L and c.3590T>C, V1197A that has been previously described in association with aHUS. CONCLUSIONS: CFH mutation screening is recommended in all aHUS patients prior to renal transplantation because of the high risk of disease recurrence post-transplant in those known to have a CFH mutation. Because of our finding it will be necessary to implement additional screening strategies that will detect a hybrid CFH/CFHL1 gene.

  18. Effect of ploidy increase on transgene expression: example from Citrus diploid cybrid and allotetraploid somatic hybrid expressing the EGFP gene. (United States)

    Xu, Shi-Xiao; Cai, Xiao-Dong; Tan, Bin; Li, Ding-Li; Guo, Wen-Wu


    Polyploidization is an important speciation mechanism for all eukaryotes, and it has profound impacts on biodiversity dynamics and ecosystem functioning. Green fluorescent protein (GFP) has been used as an effective marker to visually screen somatic hybrids at an early stage in protoplast fusion. We have previously reported that the intensity of GFP fluorescence of regenerated embryoids was also an early indicator of ploidy level. However, little is known concerning the effects of ploidy increase on the GFP expression in citrus somatic hybrids at the plant level. Herein, allotetraploid and diploid cybrid plants with enhanced GFP (EGFP) expression were regenerated from the fusion of embryogenic callus protoplasts from 'Murcott' tangor (Citrus reticulata Blanco × Citrus sinensis (L.) Osbeck) and mesophyll protoplasts from transgenic 'Valencia' orange (C. sinensis (L.) Osbeck) expressing the EGFP gene, via electrofusion. Subsequent simple sequence repeat (SSR), chloroplast simple sequence repeat and cleaved amplified polymorphic sequence analysis revealed that the two regenerated tetraploid plants were true allotetraploid somatic hybrids possessing nuclear genomic DNA of both parents and cytoplasmic DNA from the callus parent, while the five regenerated diploid plants were cybrids containing nuclear DNA of the leaf parent and with complex segregation of cytoplasmic DNA. Furthermore, EGFP expression was compared in cells and protoplasts from mature leaves of these diploid cybrids and allotetraploid somatic hybrids. Results showed that the intensity of GFP fluorescence per cell or protoplast in diploid was generally brighter than in allotetraploid. Moreover, same hybridization signal was detected on allotetraploid and diploid plants by Southern blot analysis. By real-time RT-PCR and Western blot analysis, GFP expression level of the diploid cybrid was revealed significantly higher than that of the allotetraploid somatic hybrid. These results suggest that ploidy

  19. First-Order Hybrid Logic

    DEFF Research Database (Denmark)

    Braüner, Torben


    Hybrid logic is an extension of modal logic which allows us to refer explicitly to points of the model in the syntax of formulas. It is easy to justify interest in hybrid logic on applied grounds, with the usefulness of the additional expressive power. For example, when reasoning about time one...... often wants to build up a series of assertions about what happens at a particular instant, and standard modal formalisms do not allow this. What is less obvious is that the route hybrid logic takes to overcome this problem often actually improves the behaviour of the underlying modal formalism....... For example, it becomes far simpler to formulate proof-systems for hybrid logic, and completeness results can be proved of a generality that is simply not available in modal logic. That is, hybridization is a systematic way of remedying a number of known deficiencies of modal logic. First-order hybrid logic...

  20. Hybridization in geese: a review


    Ottenburghs, Jente; Van Hooft, Pim; van Wieren, Sipke E.; Ydenberg, Ronald C; Herbert H. T. Prins


    The high incidence of hybridization in waterfowl (ducks, geese and swans) makes this bird group an excellent study system to answer questions related to the evolution and maintenance of species boundaries. However, knowledge on waterfowl hybridization is biased towards ducks, with a large knowledge gap in geese. In this review, we assemble the available information on hybrid geese by focusing on three main themes: (1) incidence and frequency, (2) behavioural mechanisms leading to hybridizatio...

  1. Hybrid solar lighting distribution systems and components (United States)

    Muhs, Jeffrey D.; Earl, Dennis D.; Beshears, David L.; Maxey, Lonnie C.; Jordan, John K.; Lind, Randall F.


    A hybrid solar lighting distribution system and components having at least one hybrid solar concentrator, at least one fiber receiver, at least one hybrid luminaire, and a light distribution system operably connected to each hybrid solar concentrator and each hybrid luminaire. A controller operates all components.

  2. Hybrid solar lighting systems and components (United States)

    Muhs, Jeffrey D.; Earl, Dennis D.; Beshears, David L.; Maxey, Lonnie C.; Jordan, John K.; Lind, Randall F.


    A hybrid solar lighting system and components having at least one hybrid solar concentrator, at least one fiber receiver, at least one hybrid luminaire, and a light distribution system operably connected to each hybrid solar concentrator and each hybrid luminaire. A controller operates each component.

  3. Evaluación de la técnica Western blot para la detección de antígenos de Hymenolepis nana

    Directory of Open Access Journals (Sweden)

    Flora Chávez-Salas


    Full Text Available Este trabajo tuvo como objetivo evaluar la técnica de inmunoelectrotransferencia (Western Blot para detectar los antígenos específicos de excreción/secreción de Hymenolepis nana en sueros de pacientes con himenolepiosis y con otras helmintiosis confirmadas. Se utilizó a Mesocricetus auratus “hamster” para obtener ejemplares adultos de H. nana. Los antígenos de excreción/secreción fueron obtenidos en el medio MEM (Minimum Essential Medium Eagle, y enfrentados con un grupo de sueros de pacientes con himenolepiosis confirmada para evaluar su calidad inmunológica y con sueros individuales de pacientes con himenolepiosis y con otras helmintiosis confirmadas para detectar mediante la técnica de “Western Blot”, los antígenos específicos de este cestode. El grupo de sueros de pacientes con himenolepiosis confirmada reconoció las bandas antigénicas de 50,1; 42,6; 38,9; 32,9; 26,3; 22,4 y 18,6 kDa; sin embargo, los sueros individuales reconocieron diferente número de bandas, siendo la de 50,1 KDa la que fue reconocida por todos ellos. Los sueros de pacientes con helmintiosis confirmadas no reconocieron la banda de 50,1 kDa; sin embargo, dieron reacción cruzada con algunas de las demás bandas, a excepción de los sueros de pacientes con cisticercosis que no reconocieron a ninguna de las bandas de estos antígenos. Se concluye que el antígeno de excreción/secreción de H. nana de 50,1 kDa es específico de este cestode por ser reconocido por todos los sueros de pacientes con himenolepiosis confirmada y no por sueros de pacientes con otras helmintiosis utilizando la técnica de “Western Blot”.

  4. The governance of hybrid organisations

    DEFF Research Database (Denmark)

    Spear, Roger; Cornforth, Chris


    The focus of this chapter is on the governance of third sector organizations (TSOs) and the challenges that are raised by hybridity. In particular it will focus on the question how does hybridity affect governance structures and processes and the challenges that governing bodies face?......The focus of this chapter is on the governance of third sector organizations (TSOs) and the challenges that are raised by hybridity. In particular it will focus on the question how does hybridity affect governance structures and processes and the challenges that governing bodies face?...

  5. Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization

    Institute of Scientific and Technical Information of China (English)

    Christine Fallsehr; Christina Zapletal; Michael Kremer; Resit Demir; Magnus von Knebel Doeberitz; Ernst Klar


    AIM: To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs.METHODS: We used two strategies (SSH suppression subtractive hybridization and hybridization of cDNA arrays)to determine early changes in gene expression profiles in a rat model of partial WI/R, comparing postischemic and adjacent nonischemic liver lobes. Differential gene expression was verified (WT/R; 1 h/2 h) and analyzed in more detail after warm ischemia (1 h) in a reperfusion time kinetics (0, 1, 2 and 6 h) and compared to untreated livers by Northern blot hybridizations. Protein expression was examined on Western blots and by immunohistochemistry for four differentially expressed target genes (Hsp70,Hsp27, Gadd45a and IL-1rl).RESULTS: Thirty-two individual WI/R target genes showing altered RNA levels after confirmation by Northern blot analyzes were identified. Among them, six functionally uncharacteristic expressed sequences and 26 known genes (12 induced in postischemic liver lobes, 14 with higher transcriptional expression in adjacent nonischemic liver lobes). Functional categories of the verified marker genes indicate on the one hand cellular stress and tissue damage but otherwise activation of protective cellular reactions (AP-1 transcription factors, apoptosis related genes, heat shock genes). In order to assign the transcriptional status to the biological relevant protein level we demonstrated that Hsp70, Hsp27, Gadd45a and IL-1rI were clearly up-regulated comparing postischemic and untreated rat livers, suggesting their involvement in the WI/R context.CONCLUSION: This study unveils a WI/R response gene set that will help to explore molecular pathways involved in the tissue damage after WI/R. In addition, these genes especially Hsp70and Gadd45a might represent promising new candidates indicating WI/R liver damage.

  6. A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques Comparação dos parâmetros imunológicos de cães infectados com leishmaniose visceral usando as técnicas de Western blot e neutralização

    Directory of Open Access Journals (Sweden)

    Yeda L. Nogueira


    Full Text Available The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.A técnica de Western blot foi utilizada para demonstrar a presença de anticorpos do soro de cães, que apresentavam leishmaniose visceral canina, contra algumas moléculas específicas no lisado da forma promastigota de Leshmania chagasi.Através da associa

  7. Hybrid Keyword Search Auctions


    Goel, Ashish; Munagala, Kamesh


    Search auctions have become a dominant source of revenue generation on the Internet. Such auctions have typically used per-click bidding and pricing. We propose the use of hybrid auctions where an advertiser can make a per-impression as well as a per-click bid, and the auctioneer then chooses one of the two as the pricing mechanism. We assume that the advertiser and the auctioneer both have separate beliefs (called priors) on the click-probability of an advertisement. We first prove that the ...

  8. Reflections on Intellectual Hybridity

    Directory of Open Access Journals (Sweden)

    Kimala Price


    Full Text Available Drawing from the growing literature on interdisciplinarity and my own experiences as an intellectual hybrid, I discuss the personal and institutional challenges inherent in crossing disciplinary boundaries in the academy. I argue that boundary crossing is a natural occurrence and that the issue of (interdisciplinarity is a matter of degree and of determining who gets to define the boundaries. Defining boundaries is not merely an intellectual enterprise, but also a political act that delineates what is, or is not, legitimate scholarship. This issue is especially salient to women's and gender studies during times of economic distress and educational budget cuts.

  9. Hybrid Random Fields

    CERN Document Server

    Freno, Antonino


    This book presents an exciting new synthesis of directed and undirected, discrete and continuous graphical models. Combining elements of Bayesian networks and Markov random fields, the newly introduced hybrid random fields are an interesting approach to get the best of both these worlds, with an added promise of modularity and scalability. The authors have written an enjoyable book---rigorous in the treatment of the mathematical background, but also enlivened by interesting and original historical and philosophical perspectives. -- Manfred Jaeger, Aalborg Universitet The book not only marks an

  10. Hybrid power semiconductor (United States)

    Chen, D. Y.


    The voltage rating of a bipolar transistor may be greatly extended while at the same time reducing its switching time by operating it in conjunction with FETs in a hybrid circuit. One FET is used to drive the bipolar transistor while the other FET is connected in series with the transistor and an inductive load. Both FETs are turned on or off by a single drive signal of load power, the second FET upon ceasing conductions, rendering one power electrode of the bipolar transistor open. Means are provided to dissipate currents which flow after the bipolar transistor is rendered nonconducting.

  11. Cryogenic Hybrid Magnetic Bearing (United States)

    Meeks, Crawford R.; Dirusso, Eliseo; Brown, Gerald V.


    Cryogenic hybrid magnetic bearing is example of class of magnetic bearings in which permanent magnets and electromagnets used to suspend shafts. Electromagnets provide active control of position of shaft. Bearing operates at temperatures from -320 degrees F (-196 degrees C) to 650 degrees F (343 degrees C); designed for possible use in rocket-engine turbopumps, where effects of cryogenic environment and fluid severely limit lubrication of conventional ball bearings. This and similar bearings also suitable for terrestrial rotating machinery; for example, gas-turbine engines, high-vacuum pumps, canned pumps, precise gimbals that suspend sensors, and pumps that handle corrosive or gritty fluids.

  12. Molecular cytogenetics of Alstroemeria: identification of parental genomes in interspecific hybrids and characterization of repetitive DNA families in constitutive heterochromatin. (United States)

    Kuipers, A G; van Os, D P; de Jong, J H; Ramanna, M S


    The genus Alstroemeria consists of diploid (2n = 2x = 16) species originating mainly from Chile and Brazil. Most cultivars are triploid or tetraploid interspecific hybrids. C-banding of eight species revealed obvious differentiation of constitutive heterochromatin within the genus. The present study focused on the molecular (cyto)genetic background of this differentiation. Genomic slot-blot analysis demonstrated strong conservation of major parts of the genomes among six species. The chromosomes of A. aurea and A. ligtu, species with pronounced interstitial C-bands, were found to contain large amounts of highly repetitive and species-specific DNA. The variation in size, number and intensity of strongly probed bands of major repetitive DNA families observed in genomic Southern blots of Sau3A, HaeIII, and MseI digests indicated a strong correlation between variation in genomic DNA composition and different C-banding patterns among Alstroemeria species. Genomic in situ hybridization (GISH) revealed a clear distinction between parental chromosomes in the hybrids between Chilean and Brazilian species and also between Chilean species, as long as at least one of the parental species possessed prominent C-banding. Regarding the latter, discriminative hybridization resulted from highly repetitive species specific DNA in the heterochromatic chromosome regions of A. aurea and A. ligtu, and caused GISH banding patterns that coincided with the C-banding patterns.

  13. Hybrid vehicle potential assessment. Volume 7. Hybrid vehicle review

    Energy Technology Data Exchange (ETDEWEB)

    Leschly, K.O.


    Review of hybrid vehicles (HVs) built during the past ten years or planned to be built in the near future is presented. An attempt is made to classify and analyze these vehicles to get an overall picture of their key characteristics. The review includes on-road hybrid passenger cars, trucks, vans, and buses.

  14. The Hybrid Advantage: Graduate Student Perspectives of Hybrid Education Courses (United States)

    Hall, Sarah; Villareal, Donna


    Hybrid courses combine online and face-to-face learning environments. To organize and teach hybrid courses, instructors must understand the uses of multiple online learning tools and face-toface classroom activities to promote and monitor the progress of students. The purpose of this phenomenological study was to explore the perspectives of…

  15. Analysis of hybrid viscous damper by real time hybrid simulations

    DEFF Research Database (Denmark)

    Brodersen, Mark Laier; Ou, Ge; Høgsberg, Jan Becker


    Results from real time hybrid simulations are compared to full numerical simulations for a hybrid viscous damper, composed of a viscous dashpot in series with an active actuator and a load cell. By controlling the actuator displacement via filtered integral force feedback the damping performance...... of the hybrid viscous damper is improved, while for pure integral force feedback the damper stroke is instead increased. In the real time hybrid simulations viscous damping is emulated by a bang-bang controlled Magneto-Rheological (MR) damper. The controller activates high-frequency modes and generates drift...... in the actuator displacement, and only a fraction of the measured damper force can therefore be used as input to the investigated integral force feedback in the real time hybrid simulations....

  16. Unified Hybrid Network Theoretical Model Trilogy

    Institute of Scientific and Technical Information of China (English)


    The first of the unified hybrid network theoretical model trilogy (UHNTF) is the harmonious unification hybrid preferential model (HUHPM), seen in the inner loop of Fig. 1, the unified hybrid ratio is defined.

  17. Overview on hybrid propulsion (United States)

    Calabro, M.


    Aside of research works, this historical survey shows propulsion units used by students for small satellites and for gas generation, or those for the Space Ship One, even if LOx/HTPB was studied and tested in large motors for its potential very low cost; however, this combination highlights a series of technical problems without any performance advantage over the existing LOx/Kerosene family and never been operational for ETO applications. The particularity of hybrid propulsion is to use the state-of-the-art of both liquids and solids; the only show stopper is the propellant itself. The past work focused on LOx/HTPB (selected for its low cost) appears to be a dead-end (combustion problems and global low performances resulting from a high level of residuals). The solution that appears through the past experience is the addition of hydrides to a binder (HTPB or other) or to a binder and a homogeneous fuel or a mixture of both, with or without others additives; within these solutions some will not present any manufacturing problem and some may have a low cost. Nevertheless, the studies of the following phases have to demonstrate the compatibility of the potential regression rate range with a high-performance global design of a hybrid Motor and the manufacturing at a reasonable cost of a hydride giving a high level of performances.

  18. Landmarks in Hybrid Planning

    Directory of Open Access Journals (Sweden)

    Mohamed Elkawkagy


    Full Text Available Although planning techniques achieved a significant progress during recent years, solving many planning problem still difficult even for modern planners. In this paper, we will adopt landmark concept to hybrid planning setting - a method that combines reasoning about procedural knowledge and causalities. Land-marks are a well-known concept in the realm of classical planning. Recently, they have been adapted to hierarchical approaches. Such landmarks can be extracted in a pre-processing step from a declarative hierarchical planning domain and problem description. It was shown how this technique allows for a considerable reduction of the search space by eliminating futile plan development options before the actual planning. Therefore, we will present a new approach to in¬tegrate landmark pre-processing technique in the context of hierarchical planning with landmark technique in the classical planning. This integration allows to incorporate the ability of using extracted landmark tasks from hierarchical domain knowledge in the form of HTN and using landmark literals from classical planning. To this end, we will construct a transformation technique to transform the hybrid planning domain into a classical domain model. The method¬ologies in this paper have been implemented successfully, and we will present some experimental results that give evidence for the consid-erable performance increase gained through planning system.

  19. Suppression subtractive hybridization. (United States)

    Ghorbel, Mohamed T; Murphy, David


    Comparing two RNA populations that differ from the effects of a single independent variable, such as a drug treatment or a specific genetic defect, can establish differences in the abundance of specific transcripts that vary in a population dependent manner. There are different methods for identifying differentially expressed genes. These methods include microarray, Serial Analysis of Gene Expression (SAGE), and quantitative Reverse-Transcriptase Polymerase Chain Reaction (qRT-PCR). Herein, the protocol describes an easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under examination. It is specifically relevant when low levels of RNA starting material are available. This protocol describes the use of Switching Mechanism At RNA Termini Polymerase Chain Reaction (SMART-PCR) to amplify cDNA from small amounts of RNA. The amplified cDNA populations under comparison are then subjected to Suppression Subtractive Hybridization (SSH-PCR). SSH-PCR is a technique that couples subtractive hybridization with suppression PCR to selectively amplify fragments of differentially expressed genes. The resulting products are cDNA populations enriched for significantly overrepresented transcripts in either of the two input RNAs. These cDNA populations can then be cloned to generate subtracted cDNA library. Microarrays made with clones from the subtracted forward and reverse cDNA libraries are then screened for differentially expressed genes using targets generated from tester and driver total RNAs.

  20. Arabidopsis hybrid speciation processes. (United States)

    Schmickl, Roswitha; Koch, Marcus A


    The genus Arabidopsis provides a unique opportunity to study fundamental biological questions in plant sciences using the diploid model species Arabidopsis thaliana and Arabidopsis lyrata. However, only a few studies have focused on introgression and hybrid speciation in Arabidopsis, although polyploidy is a common phenomenon within this genus. More recently, there is growing evidence of significant gene flow between the various Arabidopsis species. So far, we know Arabidopsis suecica and Arabidopsis kamchatica as fully stabilized allopolyploid species. Both species evolved during Pleistocene glaciation and deglaciation cycles in Fennoscandinavia and the amphi-Beringian region, respectively. These hybrid studies were conducted either on a phylogeographic scale or reconstructed experimentally in the laboratory. In our study we focus at a regional and population level. Our research area is located in the foothills of the eastern Austrian Alps, where two Arabidopsis species, Arabidopsis arenosa and A. lyrata ssp. petraea, are sympatrically distributed. Our hypothesis of genetic introgression, migration, and adaptation to the changing environment during the Pleistocene has been confirmed: We observed significant, mainly unidirectional gene flow between the two species, which has given rise to the tetraploid A. lyrata. This cytotype was able to escape from the narrow ecological niche occupied by diploid A. lyrata ssp. petraea on limestone outcrops by migrating northward into siliceous areas, leaving behind a trail of genetic differentiation.

  1. Hybridity in Embedded Computing Systems

    Institute of Scientific and Technical Information of China (English)

    虞慧群; 孙永强


    An embedded system is a system that computer is used as a component in a larger device.In this paper,we study hybridity in embedded systems and present an interval based temporal logic to express and reason about hybrid properties of such kind of systems.

  2. Design Principles for Hybrid Ventilation

    DEFF Research Database (Denmark)

    Heiselberg, Per

    For many years mechanical and natural ventilation systems have developed separately. Naturally, the next step in this development is the development of ventilation concepts that utilize and combine the best features from each system to create a new type of ventilation system -Hybrid Ventilation....... The hybrid ventilation concepts, design challenges and - principles are discussed and illustrated by four building examples....

  3. Expanding Discourse Repertoires with Hybridity (United States)

    Kelly, Gregory J.


    In "Hybrid discourse practice and science learning" Kamberelis and Wehunt present a theoretically rich argument about the potential of hybrid discourses for science learning. These discourses draw from different forms of "talk, social practice, and material practices" to create interactions that are "intertextually complex" and "interactionally…

  4. Hybrid Charmonium from Lattice QCD

    CERN Document Server

    Luo, X Q


    We review our recent results on the JPC = 0¡¡ exotic hybrid charmonium mass and JPC = 0¡+, 1¡¡ and 1++ nonexotic hybrid charmonium spectrum from anisotropic improved lattice QCD and discuss the relevance to the recent discovery of the Y(4260) state and future experimental search for other states.

  5. Hybrid spread spectrum radio system (United States)

    Smith, Stephen F [London, TN; Dress, William B [Camas, WA


    Systems and methods are described for hybrid spread spectrum radio systems. A method, includes receiving a hybrid spread spectrum signal including: fast frequency hopping demodulating and direct sequence demodulating a direct sequence spread spectrum signal, wherein multiple frequency hops occur within a single data-bit time and each bit is represented by chip transmissions at multiple frequencies.

  6. Detection of isoform-specific fibroblast growth factor receptors by whole-mount in situ hybridization in early chick embryos. (United States)

    Nishita, Junko; Ohta, Sho; Bleyl, Steven B; Schoenwolf, Gary C


    We have developed "b" and "c" isoform-specific chicken fibroblast growth factor (FGF) receptor 1-3 probes for in situ hybridization. We rigorously demonstrate the specificity of these probes by using both dot blot hybridization and whole-mount in situ hybridization during neurulation and early postneurulation stages, and we compare expression patterns of each of the three isoform-specific probes to one another and to generic probes to each of the three (non-isoform-specific) FGF receptors. We show that the expression pattern of each receptor is represented by the collective expression of each of its two isoforms, with the expression of each FGF receptor being most similar to that of its "c" isoform at two of the three stages studied, and that tissue and stage differences exist in the patterns of expression of the six isoforms. We demonstrate the usefulness of these probes for defining the differential tissue expression of FGF receptor 1-3 isoforms.

  7. Model Reduction of Hybrid Systems

    DEFF Research Database (Denmark)

    Shaker, Hamid Reza

    High-Technological solutions of today are characterized by complex dynamical models. A lot of these models have inherent hybrid/switching structure. Hybrid/switched systems are powerful models for distributed embedded systems design where discrete controls are applied to continuous processes...... of hybrid systems, designing controllers and implementations is very high so that the use of these models is limited in applications where the size of the state space is large. To cope with complexity, model reduction is a powerful technique. This thesis presents methods for model reduction and stability....... Hybrid systems are also an important modeling class for nonlinear systems because a wide variety of nonlinearities are either piecewise-affine (e.g., a saturated linear actuator characteristic) or can be approximated as hybrid systems. The complexity of verifying and assessing general properties...

  8. Detecting hybridization using ancient DNA. (United States)

    Schaefer, Nathan K; Shapiro, Beth; Green, Richard E


    It is well established that related species hybridize and that this can have varied but significant effects on speciation and environmental adaptation. It should therefore come as no surprise that hybridization is not limited to species that are alive today. In the last several decades, advances in technologies for recovering and sequencing DNA from fossil remains have enabled the assembly of high-coverage genome sequences for a growing diversity of organisms, including many that are extinct. Thanks to the development of new statistical approaches for detecting and quantifying admixture from genomic data, genomes from extinct populations have proven useful both in revealing previously unknown hybridization events and informing the study of hybridization between living organisms. Here, we review some of the key recent statistical innovations for detecting ancient hybridization using genomewide sequence data and discuss how these innovations have revised our understanding of human evolutionary history.

  9. Hybrid laser-arc welding

    DEFF Research Database (Denmark)

    Hybrid laser-arc welding (HLAW) is a combination of laser welding with arc welding that overcomes many of the shortfalls of both processes. This important book gives a comprehensive account of hybrid laser-arc welding technology and applications. The first part of the book reviews...... the characteristics of the process, including the properties of joints produced by hybrid laser-arc welding and ways of assessing weld quality. Part II discusses applications of the process to such metals as magnesium alloys, aluminium and steel as well as the use of hybrid laser-arc welding in such sectors as ship...... building and the automotive industry. With its distinguished editor and international team of contributors, Hybrid laser-arc welding, will be a valuable source of reference for all those using this important welding technology. Professor Flemming Ove Olsen works in the Department of Manufacturing...

  10. Small RNA Detection by in Situ Hybridization Methods

    Directory of Open Access Journals (Sweden)

    Martyna O. Urbanek


    Full Text Available Small noncoding RNAs perform multiple regulatory functions in cells, and their exogenous mimics are widely used in research and experimental therapies to interfere with target gene expression. MicroRNAs (miRNAs are the most thoroughly investigated representatives of the small RNA family, which includes short interfering RNAs (siRNAs, PIWI-associated RNA (piRNAs, and others. Numerous methods have been adopted for the detection and characterization of small RNAs, which is challenging due to their short length and low level of expression. These include molecular biology methods such as real-time RT-PCR, northern blotting, hybridization to microarrays, cloning and sequencing, as well as single cell miRNA detection by microscopy with in situ hybridization (ISH. In this review, we focus on the ISH method, including its fluorescent version (FISH, and we present recent methodological advances that facilitated its successful adaptation for small RNA detection. We discuss relevant technical aspects as well as the advantages and limitations of ISH. We also refer to numerous applications of small RNA ISH in basic research and molecular diagnostics.

  11. Evaluación de las pruebas dot blot y aglutinación de látex para el diagnóstico de cisticercosis en Perú

    Directory of Open Access Journals (Sweden)

    Eduardo Miranda-Ulloa


    Full Text Available Con el objetivo de evaluar las pruebas dot blot y aglutinación de látex para la detección de cisticercosis humana con antígeno de líquido de cisticerco de Taenia solium, se usaron 125 sueros humanos, de los cuales 60 procedían de personas con cisticercosis confirmada por Western Blot, 45 de personas con otras enfermedades parasitarias y 20 de personas aparentemente sanas. La concentración óptima del antígeno para impregnar las tiras dot blot fue de 0,01 ug/uL, y para impregnar las partículas de látex fue de 0,092 ug/uL. Para la prueba dot blot se encontró una sensibilidad del 100% y especificidad del 87,7%; para la aglutinación de látex una sensibilidad del 93,3% y especificidad del 89,2%. Ambas pruebas podrían ser de utilidad y factibles de implementar como alternativas de diagnóstico serológico en laboratorios de áreas endémicas del Perú

  12. Evaluación de las pruebas dot blot y aglutinación de látex para el diagnóstico de cisticercosis en Perú

    Directory of Open Access Journals (Sweden)

    Eduardo Miranda-Ulloa

    Full Text Available Con el objetivo de evaluar las pruebas dot blot y aglutinación de látex para la detección de cisticercosis humana con antígeno de líquido de cisticerco de Taenia solium, se usaron 125 sueros humanos, de los cuales 60 procedían de personas con cisticercosis confirmada por Western Blot, 45 de personas con otras enfermedades parasitarias y 20 de personas aparentemente sanas. La concentración óptima del antígeno para impregnar las tiras dot blot fue de 0,01 ug/uL, y para impregnar las partículas de látex fue de 0,092 ug/uL. Para la prueba dot blot se encontró una sensibilidad del 100% y especificidad del 87,7%; para la aglutinación de látex una sensibilidad del 93,3% y especificidad del 89,2%. Ambas pruebas podrían ser de utilidad y factibles de implementar como alternativas de diagnóstico serológico en laboratorios de áreas endémicas del Perú

  13. Chromosome engineering: generation of mono- and dicentric isochromosomes in a somatic cell hybrid system. (United States)

    Higgins, A W; Schueler, M G; Willard, H F


    The most common isochromosome found in humans involves the long arm of the X, i(Xq), and is associated with a subset of Turner syndrome cases. To study the formation and behavior of isochromosomes in a more tractable experimental system, we have developed a somatic cell hybrid model system that allows for the selection of mono- or dicentric isochromosomes involving the short arm of the X, i(Xp). Simultaneous positive and negative counterselection of a mouse/human somatic cell hybrid containing a human X chromosome, selecting for retention of the UBE1 locus in Xp but against the HPRT locus in Xq, results in a variety of abnormalities of the X chromosome involving deletions of Xq. We have generated 70 such "Pushmi-Pullyu" hybrids derived from seven independent X chromosomes. Cytogenetic analysis of these hybrids using fluorescence in situ hybridization showed i(Xp) chromosomes in approximately 19% of the hybrids. Southern blot and polymerase chain reaction analyses of the Pushmi-Pullyu hybrids revealed a distribution of breakpoints along Xq. The distance between the centromeres of the dicentric i(Xp)s generated ranged from approximately 2 Mb to approximately 20 Mb. To examine centromeric activity in these dicentric i(Xp)s, we used indirect immunofluorescence with antibodies to centromere protein E (CENP-E). CENP-E was detected at only one of the centromeres of a dicentric i(Xp) with approximately 2-3 Mb of Xq DNA. In contrast, CENP-E was detected at both centromeres of a dicentric i(Xp) with approximately 14 Mb of Xq DNA. Two other dicentric i(Xp) chromosomes were heterogeneous with respect to centromeric activity, suggesting that centromeric activity and chromosome stability of dicentric chromosomes may be more complicated than previously thought. The Pushmi-Pullyu model system presented in this study may provide a tool for examining the structure and function of mammalian centromeres.

  14. Molecular evidence for hybridization in Colias (Lepidoptera: Pieridae): are Colias hybrids really hybrids? (United States)

    Dwyer, Heather E; Jasieniuk, Marie; Okada, Miki; Shapiro, Arthur M


    Gene flow and hybridization among species dramatically affect our understanding of the species as a biological unit, species relationships, and species adaptations. In North American Colias eurytheme and Colias eriphyle, there has been historical debate over the extent of hybridization occurring and the identity of phenotypically intermediate individuals as genetic hybrids. This study assesses the population structure of these two species to measure the extent of hybridization and the genetic identity of phenotypic intermediates as hybrids. Amplified fragment length polymorphism (AFLP) marker analysis was performed on 378 specimens collected from northern California and Nevada. Population structure was inferred using a Bayesian/Markov chain Monte Carlo method, which probabilistically assigns individuals to genetic clusters. Three genetic clusters provided the best fit for the data. C. eurytheme individuals were primarily assigned to two closely related clusters, and C. eriphyle individuals were mostly assigned to a third, more distantly related cluster. There appeared to be significant hybridization between the two species. Individuals of intermediate phenotype (putative hybrids) were found to be genetically indistinguishable from C. eriphyle, indicating that previous work based on the assumption that these intermediate forms are hybrids may warrant reconsideration. PMID:26306172

  15. Identification and analysis of differentially expressed genes involved in dark-induced photoperiod response and senescence of soybean leaves by suppression subtractive hybridization

    Institute of Scientific and Technical Information of China (English)


    A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putat...

  16. Hybrid vehicle control

    Energy Technology Data Exchange (ETDEWEB)

    Shallvari, Iva; Velnati, Sashidhar; DeGroot, Kenneth P.


    A method and apparatus for heating a catalytic converter's catalyst to an efficient operating temperature in a hybrid electric vehicle when the vehicle is in a charge limited mode such as e.g., the charge depleting mode or when the vehicle's high voltage battery is otherwise charge limited. The method and apparatus determine whether a high voltage battery of the vehicle is incapable of accepting a first amount of charge associated with a first procedure to warm-up the catalyst. If it is determined that the high voltage battery is incapable of accepting the first amount of charge, a second procedure with an acceptable amount of charge is performed to warm-up the catalyst.

  17. Hybrid Heat Exchangers (United States)

    Tu, Jianping Gene; Shih, Wei


    A hybrid light-weight heat exchanger concept has been developed that uses high-conductivity carbon-carbon (C-C) composites as the heat-transfer fins and uses conventional high-temperature metals, such as Inconel, nickel, and titanium as the parting sheets to meet leakage and structural requirements. In order to maximize thermal conductivity, the majority of carbon fiber is aligned in the fin direction resulting in 300 W/m.K or higher conductivity in the fin directions. As a result of this fiber orientation, the coefficient of thermal expansion (CTE) of the C-C composite in both non-fiber directions matches well with the CTE of various high-temperature metal alloys. This allows the joining of fins and parting sheets by using high-temperature braze alloys.

  18. Hybrid Batch Bayesian Optimization

    CERN Document Server

    Azimi, Javad; Fern, Xiaoli


    Bayesian Optimization aims at optimizing an unknown non-convex/concave function that is costly to evaluate. We are interested in application scenarios where concurrent function evaluations are possible. Under such a setting, BO could choose to either sequentially evaluate the function, one input at a time and wait for the output of the function before making the next selection, or evaluate the function at a batch of multiple inputs at once. These two different settings are commonly referred to as the sequential and batch settings of Bayesian Optimization. In general, the sequential setting leads to better optimization performance as each function evaluation is selected with more information, whereas the batch setting has an advantage in terms of the total experimental time (the number of iterations). In this work, our goal is to combine the strength of both settings. Specifically, we systematically analyze Bayesian optimization using Gaussian process as the posterior estimator and provide a hybrid algorithm t...

  19. Advanced Photonic Hybrid Materials (United States)


    distribution is unlimited.   TECHNICAL DESCRIPTION OF THE RESULTS:    1/  Synthesis , characterization and modelling of anisotropic  gold   nanoparticles ...preparations  and  purification  of  the AuNPs  are  described  elsewhere.4,6,9,14,15   Synthesis  of the  gold  spherical  nanoparticles   The spherical...presentations.  This report is based on 3 articles (joined to the report):  ‐ Dispersion  and  self‐orientation  of  gold   nanoparticles   in  sol‐gel  hybrid

  20. Nanoporous hybrid electrolytes

    KAUST Repository

    Schaefer, Jennifer L.


    Oligomer-suspended SiO2-polyethylene glycol nanoparticles are studied as porous media electrolytes. At SiO2 volume fractions, , bracketing a critical value y ≈ 0.29, the suspensions jam and their mechanical modulus increase by more than seven orders. For >y, the mean pore diameter is close to the anion size, yet the ionic conductivity remains surprisingly high and can be understood, at all , using a simple effective medium model proposed by Maxwell. SiO 2-polyethylene glycol hybrid electrolytes are also reported to manifest attractive electrochemical stability windows (0.3-6.3 V) and to reach a steady-state interfacial impedance when in contact with metallic lithium. © 2010 The Royal Society of Chemistry.

  1. Photochromic mesoporous hybrid coatings (United States)

    Raboin, L.; Matheron, M.; Gacoin, T.; Boilot, J.-P.


    Spirooxazine (SO) photochromic molecules were trapped in sol-gel matrices. In order to increase the colourability and improve mechanical properties of sol-gel photochromic films, we present an original strategy in which SO photochromic molecules were dispersed in mesoporous organized films using the impregnation technique. Well-ordered organosilicate mesoporous coatings with the 3D-hexagonal symmetry were prepared by the sol-gel technique. These robust mesoporous films, which contain high amounts of hydrophobic methyl groups at the pore surface, offer optimized environments for photochromic dyes dispersed by impregnation technique. After impregnation by a spirooxazine solution, the photochromic response is only slightly slower when compared with mesostructured or soft sol-gel matrices, showing that mesoporous organized hybrid matrix are good host for photochromic dyes. Moreover, the molecular loading in films is easily adjustable in a large range using multi-impregnation procedure and increasing the film thickness leading to coatings for optical switching devices.

  2. Auditing Hybrid IT Environments

    Directory of Open Access Journals (Sweden)

    Georgiana Mateescu


    Full Text Available This paper presents a personal approach of auditing the hybrid IT environments consisting in both on premise and on demand services and systems. The analysis is performed from both safety and profitability perspectives and it aims to offer to strategy, technical and business teams a representation of the value added by the cloud programme within the company’s portfolio. Starting from the importance of the IT Governance in the actual business environments, we presented in the first section the main principles that drive the technology strategy in order to maximize the value added by IT assets in the business products. Section two summarizes the frameworks leveraged by our approach in order to implement the safety and profitability computation algorithms described in the third section. The paper concludes with benefits of our personal frameworks and presents the future developments.

  3. Hybrid Natural Inflation (United States)

    Ross, Graham G.; Germán, Gabriel; Vázquez, J. Alberto


    We construct two simple effective field theory versions of Hybrid Natural Inflation (HNI) that illustrate the range of its phenomenological implications. The resulting inflationary sector potential, V = Δ4(1 + acos( ϕ/f)), arises naturally, with the inflaton field a pseudo-Nambu-Goldstone boson. The end of inflation is triggered by a waterfall field and the conditions for this to happen are determined. Also of interest is the fact that the slow-roll parameter ɛ (and hence the tensor r) is a non-monotonic function of the field with a maximum where observables take universal values that determines the maximum possible tensor to scalar ratio r. In one of the models the inflationary scale can be as low as the electroweak scale. We explore in detail the associated HNI phenomenology, taking account of the constraints from Black Hole production, and perform a detailed fit to the Planck 2015 temperature and polarisation data.

  4. A Hybrid Imagination

    DEFF Research Database (Denmark)

    Jamison, Andrew; Christensen, Steen Hyldgaard; Botin, Lars

    This book presents a cultural perspective on scientific and technological development. As opposed to the “story-lines” of economic innovation and social construction that tend to dominate the both the popular and scholarly literature on science, technology and society (or STS, the authors offer...... an alternative approach, devoting special attention to the role played by social and cultural movements in the making of science and technology. They show how social and cultural movements, from the Renaissance of the late 15th century to the environmental and global justice movements of our time, have provided......” on the part of many a scientist and engineer and neglect the consequences - and a hybrid imagination, connecting scientific “facts” and technological “artifacts” with cultural understanding. The book concludes with chapters on the recent transformations in the modes of scientific and technological production...

  5. Hybrid Filter Membrane (United States)

    Laicer, Castro; Rasimick, Brian; Green, Zachary


    Cabin environmental control is an important issue for a successful Moon mission. Due to the unique environment of the Moon, lunar dust control is one of the main problems that significantly diminishes the air quality inside spacecraft cabins. Therefore, this innovation was motivated by NASA s need to minimize the negative health impact that air-suspended lunar dust particles have on astronauts in spacecraft cabins. It is based on fabrication of a hybrid filter comprising nanofiber nonwoven layers coated on porous polymer membranes with uniform cylindrical pores. This design results in a high-efficiency gas particulate filter with low pressure drop and the ability to be easily regenerated to restore filtration performance. A hybrid filter was developed consisting of a porous membrane with uniform, micron-sized, cylindrical pore channels coated with a thin nanofiber layer. Compared to conventional filter media such as a high-efficiency particulate air (HEPA) filter, this filter is designed to provide high particle efficiency, low pressure drop, and the ability to be regenerated. These membranes have well-defined micron-sized pores and can be used independently as air filters with discreet particle size cut-off, or coated with nanofiber layers for filtration of ultrafine nanoscale particles. The filter consists of a thin design intended to facilitate filter regeneration by localized air pulsing. The two main features of this invention are the concept of combining a micro-engineered straight-pore membrane with nanofibers. The micro-engineered straight pore membrane can be prepared with extremely high precision. Because the resulting membrane pores are straight and not tortuous like those found in conventional filters, the pressure drop across the filter is significantly reduced. The nanofiber layer is applied as a very thin coating to enhance filtration efficiency for fine nanoscale particles. Additionally, the thin nanofiber coating is designed to promote capture of

  6. Wankel engine for hybrid powertrain

    Energy Technology Data Exchange (ETDEWEB)

    Butti, A. [Univ. of Florence (Italy); Site, V.D.


    The Wankel engine is suited to be used to drive hybrid propulsion systems. The main disadvantage of hybrid propulsion systems is the complexity that causes a high weight and large dimensions. For these reason hybrid systems are more suitable for large size vehicle (buses, vans) rather than for small passenger cars. A considerable reduction of hybrid systems weight and dimensions can be obtained using a Wankel rotary engine instead of a conventional engine. The Wankel engine is light, compact, simple, and produces low noise and low vibrations. Therefore a Wankel engine powered hybrid system is suited to be used on small cars. In this paper a 1,000 kg parallel hybrid car with continuously variable transmission and a 6,000 kg series hybrid minibus both equipped with Wankel engines are considered. The Wankel engine works at steady state to minimize fuel consumption and exhaust emissions. The simulation of the behavior of these two vehicles during a ECE + EUDC test cycle is presented in order to evaluate the performances of the systems.

  7. Fabrication of Hybrid Petroelectric Vehicle

    Directory of Open Access Journals (Sweden)

    G. Adinarayana


    Full Text Available In automobile sector, the need for alternative fuel as a replacement of conventional fossil fuel, due to its depletion and amount of emission has given way for new technologies like Fuel cells vehicles, Electric vehicles. Still a lot of advancement has to take place in these technologies for commercialization. The gap between the current fossil fuel technology and zero emission vehicles can be bridged by hybrid technology. Hybrid vehicles are those which can run on two or more powering sources/fuels. Feasibility of this technology is been proved in four wheelers and automobile giants like Toyota, Honda, and Hyundai have launched successful vehicles like Toyota prius, Honda insight etc. This technology maximizes the advantages of the two fuels and minimizes the disadvantages of the same. The best preferred hybrid pair is electric and fossil fuel. This increases the mileage of the vehicle twice the existing and also reduces the emission to half. At present, we like to explore the hybrid technology in the two wheeler sector and its feasibility on road. This paper deals with an attempt to make a hybrid with electric start and petrol run. Further a design of basic hybrid elements like motor, battery, and engine. As on today, hybrid products are one of the best solutions for all pollution hazards at a fairly nominal price. An investment within the means of a common man that guarantees a better environment to live in.

  8. Hybrid codes: Methods and applications

    Energy Technology Data Exchange (ETDEWEB)

    Winske, D. (Los Alamos National Lab., NM (USA)); Omidi, N. (California Univ., San Diego, La Jolla, CA (USA))


    In this chapter we discuss hybrid'' algorithms used in the study of low frequency electromagnetic phenomena, where one or more ion species are treated kinetically via standard PIC methods used in particle codes and the electrons are treated as a single charge neutralizing massless fluid. Other types of hybrid models are possible, as discussed in Winske and Quest, but hybrid codes with particle ions and massless fluid electrons have become the most common for simulating space plasma physics phenomena in the last decade, as we discuss in this paper.

  9. Completeness in Hybrid Type Theory

    DEFF Research Database (Denmark)

    Areces, Carlos; Blackburn, Patrick Rowan; Huertas, Antonia;


    We show that basic hybridization (adding nominals and @ operators) makes it possible to give straightforward Henkin-style completeness proofs even when the modal logic being hybridized is higher-order. The key ideas are to add nominals as expressions of type t, and to extend to arbitrary types...... the way we interpret @i in propositional and first-order hybrid logic. This means: interpret @iαa , where αa is an expression of any type a , as an expression of type a that rigidly returns the value that αa receives at the i-world. The axiomatization and completeness proofs are generalizations of those...

  10. Pseudovector mesons, hybrids and glueballs

    CERN Document Server

    Burakovsky, L; Burakovsky, Leonid; Page, Philip R.


    We consider glueball- (hybrid) meson mixing for the low-lying four pseudovector states. The h_1'(1380) decays dominantly to K*K with some presence in rho pi and omega eta. The newly observed h_1(1600) has a D- to S-wave width ratio to omega eta which does not enable differentiation between a conventional and hybrid meson interpretation. We predict the decay pattern of the isopartner conventional or hybrid meson b_1(1650). A notably narrow s sbar partner h_1'(1810) is predicted.

  11. Hybrid Vehicle Program. Final report

    Energy Technology Data Exchange (ETDEWEB)



    This report summarizes the activities on the Hybrid Vehicle Program. The program objectives and the vehicle specifications are reviewed. The Hybrid Vehicle has been designed so that maximum use can be made of existing production components with a minimum compromise to program goals. The program status as of the February 9-10 Hardware Test Review is presented, and discussions of the vehicle subsystem, the hybrid propulsion subsystem, the battery subsystem, and the test mule programs are included. Other program aspects included are quality assurance and support equipment. 16 references, 132 figures, 47 tables.

  12. How common is homoploid hybrid speciation? (United States)

    Schumer, Molly; Rosenthal, Gil G; Andolfatto, Peter


    Hybridization has long been considered a process that prevents divergence between species. In contrast to this historical view, an increasing number of empirical studies claim to show evidence for hybrid speciation without a ploidy change. However, the importance of hybridization as a route to speciation is poorly understood, and many claims have been made with insufficient evidence that hybridization played a role in the speciation process. We propose criteria to determine the strength of evidence for homoploid hybrid speciation. Based on an evaluation of the literature using this framework, we conclude that although hybridization appears to be common, evidence for an important role of hybridization in homoploid speciation is more circumscribed.

  13. Towards Modelling of Hybrid Systems

    DEFF Research Database (Denmark)

    Wisniewski, Rafal


    The article is an attempt to use methods of category theory and topology for analysis of hybrid systems. We use the notion of a directed topological space; it is a topological space together with a set of privileged paths. Dynamical systems are examples of directed topological spaces. A hybrid...... system consists of a number of dynamical systems that are glued together according to information encoded in the discrete part of the system. We develop a definition of a hybrid system as a functor from the category generated by a transition system to the category of directed topological spaces. Its...... directed homotopy colimit (geometric realization) is a single directed topological space. The behavior of hybrid systems can be then understood in terms of the behavior of dynamical systems through the directed homotopy colimit....

  14. Annular Hybrid Rocket Motor Project (United States)

    National Aeronautics and Space Administration — Engineers at SpaceDev have conducted a preliminary design and analysis of a proprietary annular design concept for a hybrid motor. A U.S. Patent application has been...

  15. Hybrid models for complex fluids

    CERN Document Server

    Tronci, Cesare


    This paper formulates a new approach to complex fluid dynamics, which accounts for microscopic statistical effects in the micromotion. While the ordinary fluid variables (mass density and momentum) undergo usual dynamics, the order parameter field is replaced by a statistical distribution on the order parameter space. This distribution depends also on the point in physical space and its dynamics retains the usual fluid transport features while containing the statistical information on the order parameter space. This approach is based on a hybrid moment closure for Yang-Mills Vlasov plasmas, which replaces the usual cold-plasma assumption. After presenting the basic properties of the hybrid closure, such as momentum map features, singular solutions and Casimir invariants, the effect of Yang-Mills fields is considered and a direct application to ferromagnetic fluids is presented. Hybrid models are also formulated for complex fluids with symmetry breaking. For the special case of liquid crystals, a hybrid formul...

  16. Design Procedure for Hybrid Ventilation

    DEFF Research Database (Denmark)

    Heiselberg, Per; Tjelflaat, Per Olaf

    Mechanical and natural ventilation systems have developed separately during many years. The natural next step in this development is development of ventilation concepts that utilises and combines the best features from each system into a new type of ventilation system - Hybrid Ventilation....... Buildings with hybrid ventilation often include other sustainable technologies and an energy optimisation requires an integrated approach in the design of the building and its mechanical systems. Therefore, the hybrid ventilation design procedure differs from the design procedure for conventional HVAC....... The first ideas on a design procedure for hybrid ventilation is presented and the different types of design methods, that is needed in different phases of the design process, is discussed....

  17. Real and Hybrid Atomic Orbitals. (United States)

    Cook, D. B.; Fowler, P. W.


    Demonstrates that the Schrodinger equation for the hydrogenlike atom separates in both spheroconal and prolate spheroidal coordinates and that these separations provide a sound theoretical basis for the real and hybrid atomic orbitals. (Author/SK)

  18. Hybrid-Vehicle Transmission System (United States)

    Lupo, G.; Dotti, G.


    Continuously-variable transmission system for hybrid vehicles couples internal-combustion engine and electric motor section, either individually or in parallel, to power vehicle wheels during steering and braking.

  19. Hybrid Fuel Cell Technology Overview

    Energy Technology Data Exchange (ETDEWEB)

    None available


    For the purpose of this STI product and unless otherwise stated, hybrid fuel cell systems are power generation systems in which a high temperature fuel cell is combined with another power generating technology. The resulting system exhibits a synergism in which the combination performs with an efficiency far greater than can be provided by either system alone. Hybrid fuel cell designs under development include fuel cell with gas turbine, fuel cell with reciprocating (piston) engine, and designs that combine different fuel cell technologies. Hybrid systems have been extensively analyzed and studied over the past five years by the Department of Energy (DOE), industry, and others. These efforts have revealed that this combination is capable of providing remarkably high efficiencies. This attribute, combined with an inherent low level of pollutant emission, suggests that hybrid systems are likely to serve as the next generation of advanced power generation systems.

  20. Information component of hybrid war


    Bohdanov, Aleksander


    A hybrid warfare in the three-dimensional coordinate system «Matter-Information-measure» is considering. In particular, the information component is highlighted and analyzed. The factors of preparation of information operations is defined, which are disclosed as an example of experience of volunteer group information warfare of Institute of special communication and information security of NTUU «KPI».Keywords: coordinate system «Matter-Information-Measure» hybrid warfare, information componen...

  1. Formal Description of Hybrid Systems

    DEFF Research Database (Denmark)

    Zhou, Chaochen; Ji, Wang; Ravn, Anders P.


    A language to describe hybrid systems, i.e. networks of communicating discrete and continuous processes, is proposed. A semantics of the language is given in Extended Duration Calculus, a real-time interval logic with a proof system that allows reasoning in mathematical analysis about continuous ...... processes to be embedded into the logic. The semantics thus provides a secure link to hybrid system models based on a general theory of dynamical systems....

  2. Ion-atom hybrid systems

    CERN Document Server

    Willitsch, Stefan


    The study of interactions between simultaneously trapped cold ions and atoms has emerged as a new research direction in recent years. The development of ion-atom hybrid experiments has paved the way for investigating elastic, inelastic and reactive collisions between these species at very low temperatures, for exploring new cooling mechanisms of ions by atoms and for implementing new hybrid quantum systems. The present lecture reviews experimental methods, recent results and upcoming developments in this emerging field.

  3. A hybrid base isolation system

    Energy Technology Data Exchange (ETDEWEB)

    Hart, G.C. [Univ. of California, Los Angeles, CA (United States); Lobo, R.F.; Srinivasan, M. [Hart Consultant Group, Santa Monica, CA (United States); Asher, J.W. [kpff Engineers, Santa Monica, CA (United States)


    This paper proposes a new analysis procedure for hybrid base isolation buildings when considering the displacement response of a base isolated building to wind loads. The system is considered hybrid because of the presence of viscous dampers in the building above the isolator level. The proposed analysis approach incorporates a detailed site specific wind study combined with a dynamic nonlinear analysis of the building response.

  4. Hybrid particles and associated methods (United States)

    Fox, Robert V; Rodriguez, Rene; Pak, Joshua J; Sun, Chivin


    Hybrid particles that comprise a coating surrounding a chalcopyrite material, the coating comprising a metal, a semiconductive material, or a polymer; a core comprising a chalcopyrite material and a shell comprising a functionalized chalcopyrite material, the shell enveloping the core; or a reaction product of a chalcopyrite material and at least one of a reagent, heat, and radiation. Methods of forming the hybrid particles are also disclosed.

  5. Hybrid superconductor magnet bearings (United States)

    Chu, Wei-Kan


    Hybrid superconductor magnet bearings (HSMB's) utilize high temperature superconductors (HTS's) together with permanent magnets to form a frictionless interface between relatively rotating parts. They are low mass, stable, and do not incur expenditure of energy during normal operation. There is no direct physical contact between rotor and stator, and hence there is no wear and tear. However, just as any other applications of HTS's, it requires a very cold temperature to function. Whereas this might be perceived as a disadvantage on earth, it is of no great concern in space or on the moon. To astronomers, the moon is an excellent site for an observatory, but the cold and dusty vacuum environment on the moon precludes the use of mechanical bearings on the telescope mounts. Furthermore, drive mechanisms with very fine steps, and hence bearings with extremely low friction are needed to track a star from the moon, because the moon rotates very slowly. All aspects considered, the HSMB is about the only candidate that fits in naturally. Here, we present a design for one such bearing, capable of supporting a telescope that weighs about 3 lbs on Earth.

  6. Hybrid Power Management (HPM) (United States)

    Eichenberg, Dennis J.


    The NASA Glenn Research Center s Avionics, Power and Communications Branch of the Engineering and Systems Division initiated the Hybrid Power Management (HPM) Program for the GRC Technology Transfer and Partnership Office. HPM is the innovative integration of diverse, state-of-the-art power devices in an optimal configuration for space and terrestrial applications. The appropriate application and control of the various power devices significantly improves overall system performance and efficiency. The advanced power devices include ultracapacitors and fuel cells. HPM has extremely wide potential. Applications include power generation, transportation systems, biotechnology systems, and space power systems. HPM has the potential to significantly alleviate global energy concerns, improve the environment, and stimulate the economy. One of the unique power devices being utilized by HPM for energy storage is the ultracapacitor. An ultracapacitor is an electrochemical energy storage device, which has extremely high volumetric capacitance energy due to high surface area electrodes, and very small electrode separation. Ultracapacitors are a reliable, long life, maintenance free, energy storage system. This flexible operating system can be applied to all power systems to significantly improve system efficiency, reliability, and performance. There are many existing and conceptual applications of HPM.

  7. Hybrid manifold embedding. (United States)

    Liu, Yang; Liu, Yan; Chan, Keith C C; Hua, Kien A


    In this brief, we present a novel supervised manifold learning framework dubbed hybrid manifold embedding (HyME). Unlike most of the existing supervised manifold learning algorithms that give linear explicit mapping functions, the HyME aims to provide a more general nonlinear explicit mapping function by performing a two-layer learning procedure. In the first layer, a new clustering strategy called geodesic clustering is proposed to divide the original data set into several subsets with minimum nonlinearity. In the second layer, a supervised dimensionality reduction scheme called locally conjugate discriminant projection is performed on each subset for maximizing the discriminant information and minimizing the dimension redundancy simultaneously in the reduced low-dimensional space. By integrating these two layers in a unified mapping function, a supervised manifold embedding framework is established to describe both global and local manifold structure as well as to preserve the discriminative ability in the learned subspace. Experiments on various data sets validate the effectiveness of the proposed method.

  8. Hybrid cluster identification (United States)

    Martín-Herrero, J.


    I present a hybrid method for the labelling of clusters in two-dimensional lattices, which combines the recursive approach with iterative scanning to reduce the stack size required by the pure recursive technique, while keeping its benefits: single pass and straightforward cluster characterization and percolation detection parallel to the labelling. While the capacity to hold the entire lattice in memory is usually regarded as the major constraint for the applicability of the recursive technique, the required stack size is the real limiting factor. Resorting to recursion only for the transverse direction greatly reduces the recursion depth and therefore the required stack. It also enhances the overall performance of the recursive technique, as is shown by results on a set of uniform random binary lattices and on a set of samples of the Ising model. I also show how this technique may replace the recursive technique in Wolff's cluster algorithm, decreasing the risk of stack overflow and increasing its speed, and the Hoshen-Kopelman algorithm in the Swendsen-Wang cluster algorithm, allowing effortless characterization during generation of the samples and increasing its speed.

  9. Hybrid power source (United States)

    Singh, Harmohan N.


    A hybrid power system is comprised of a high energy density element such as a fuel-cell and high power density elements such as a supercapacitor banks. A DC/DC converter electrically connected to the fuel cell and converting the energy level of the energy supplied by the fuel cell. A first switch is electrically connected to the DC/DC converter. First and second supercapacitors are electrically connected to the first switch and a second switch. A controller is connected to the first switch and the second switch, monitoring charge levels of the supercapacitors and controls the switching in response to the charge levels. A load is electrically connected to the second switch. The first switch connects the DC/DC converter to the first supercapacitor when the second switch connects the second supercapacitor to the load. The first switch connects the DC/DC converter to the second supercapacitor when the second switch connects the first supercapacitor to the load.

  10. Risk assessment of transgenic apomictic tetraploid bahiagrass, cytogenetics, breeding behavior and performance of intra-specific hybrids. (United States)

    Sandhu, Sukhpreet; James, Victoria A; Quesenberry, Kenneth H; Altpeter, Fredy


    Pollen-mediated gene transfer from stress tolerant or herbicide-resistant transgenic plants may cause environmental or agronomic problems. Apomictic seed production found in some bahiagrass cultivars may serve as a natural transgene containment system. Under greenhouse conditions, the average gene transfer frequency from an herbicide-resistant apomictic tetraploid to a population of sexual diploid bahiagrass genotypes or apomictic tetraploid bahiagrass was 0.16% when the transgenic pollen donor was placed at 0.5-1.5 m distance from the non-transgenic pollen receptors. The herbicide-resistant hybrids were characterized for transgene integration, expression and ploidy, by Southern blot analysis, immuno-chromatography and flow cytometry, respectively. Hybrids resulting from open pollination of non-transgenic diploid female plants with transgenic tetraploid male plants were triploids or near-triploids, with 2n = 26-34. These hybrids displayed a wide range of phenotypic variability, including some non-persistent or non-flowering dwarf-type hybrids with good vigor, or hybrids with vegetative growth similar to non-transgenic plants, but with significantly reduced seed set. Non-flowering aneu-triploids with good vigor/field performance will provide the highest level of transgene containment. Embryo sac analysis of pollinated spikelets confirmed a high proportion of aborted ovules. An apospory-linked RFLP marker was detected in 13 of the 15 near-triploid hybrids. All flowering aneuploid hybrids displayed significantly reduced seed set, and none of the sexual near-triploid hybrids produced any seeds. All tetraploid gene transfer events carried the apospory-linked RFLP marker, suggesting that despite the presence of the aposporus locus, a low degree of sexuality co-exists in apomictic tetraploid cultivars. Thus, tetraploid apomictic bahiagrass does not provide complete transgene containment, although intra-specific gene transfer is drastically reduced compared to sexually

  11. Hybridization in a warmer world. (United States)

    Chunco, Amanda J


    Climate change is profoundly affecting the evolutionary trajectory of individual species and ecological communities, in part through the creation of novel species assemblages. How climate change will influence competitive interactions has been an active area of research. Far less attention, however, has been given to altered reproductive interactions. Yet, reproductive interactions between formerly isolated species are inevitable as populations shift geographically and temporally as a result of climate change, potentially resulting in introgression, speciation, or even extinction. The susceptibility of hybridization rates to anthropogenic disturbance was first recognized in the 1930s. To date, work on anthropogenically mediated hybridization has focused primarily on either physical habitat disturbance or species invasion. Here, I review recent literature on hybridization to identify how ecological responses to climate change will increase the likelihood of hybridization via the dissolution of species barriers maintained by habitat, time, or behavior. Using this literature, I identify several cases where novel hybrid zones have recently formed, likely as a result of changing climate. Future research should focus on identifying areas and taxonomic groups where reproductive species interactions are most likely to be influenced by climate change. Furthermore, a better understanding of the evolutionary consequences of climate-mediated secondary contact is urgently needed. Paradoxically, hybridization is both a major conservation concern and an important source of novel genetic and phenotypic variation. Hybridization may therefore both contribute to increasing rates of extinction and stimulate the creation of novel phenotypes that will speed adaptation to novel climates. Predicting which result will occur following secondary contact will be an important contribution to conservation for many species.

  12. Combined Detection of Breast Cancer Micrometastases in the Lymph Nodes and Bone Marrow Using Reverse-transcriptase chain Reaction and Southern Hybridization

    Institute of Scientific and Technical Information of China (English)


    The presence of lymph nodes and bone marrow micrometastases of patients with breast carcinoma by immunohistochemistry (IHC) methods has been strongly correlated to early recurrence and shorter overall survival. The aim of this study was to detect micrometastases in matched sample pairs of lymph nodes and the bone marrow of primary breast cancer patients using a more sensitive method, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells at different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and IHC methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples, while the expression wasn't seen in 18 negative control samples. CK-19 IHC positive cells were detected at a dilution of one T47D cell in 5×105 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1:5′ 104 and 1:106, respectively. In the samples from the 35 patients, we found CK-19 positive cells in 2 cases (5.7%) by IHC. CK-19 gene expression signal was detected in 14/35 (40%) by RT-PCR,and 17/35 (48.6%) by southern blotting. Four cases were micrometastases positive both in lymph node and bone marrow (11.4%). There was no correlation between CK-1 9 detection and other clinical parameters. Conclusion: combined detection of micrometastases in lymph node and bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is a highly sensitive method for breast cancer.

  13. Somatic hybridization in higher plants. (United States)

    Constabel, F


    Somatic hybridization in higher plants has come into focus since methods have been established for protoplast fusion and uptake of foreign DNA and organelles by protoplasts. Polyethylene glycol (PEG) was an effective agent for inducing fusion. Treatment of protoplasts with PEG resulted in 5 to 30% heterospecific fusion products. Protoplasts of different species, genera and even families were compatible when fused. A number of protoplast combinations (soybean + corn, soybean + pea, soybean + tobacco, carrot + barley, etc.) provided fusion products which underwent cell division and callus formation. Fusion products initially were heterokaryocytes. In dividing heterokaryocytes, random distribution of mitotic nuclei was observed to be accompanied by multiple wall formation and to result in chimeral callus. Juxtaposition of mitotic nuclei suggested nuclear fusion and hybrid formation. Fusion of heterospecific interphase nuclei was demonstrated in soybean + pea and carrot + barley heterokaryons. Provided parental protoplasts carry suitable markers, the fusion products can be recognized. For the isolation and cloning of hybrid cells, fusion experiments must be supplemented with a selective system. Complementation of two non-allelic genes that prevent or inhibit growth under special culture conditions appears as the principle on which to base the selection of somatic hybrids. As protoplasts of some species have been induced to regenerate entire plants, the development of hybrid plants from protoplast fusion products is feasible and has already been demonstrated for tobacco.

  14. Hybrid origins of cultivated potatoes. (United States)

    Rodríguez, Flor; Ghislain, Marc; Clausen, Andrea M; Jansky, Shelley H; Spooner, David M


    Solanum section Petota is taxonomically difficult, partly because of interspecific hybridization at both the diploid and polyploid levels. The taxonomy of cultivated potatoes is particularly controversial. Using DNA sequence data of the waxy gene, we here infer relationships among the four species of cultivated potatoes accepted in the latest taxonomic treatment (S. ajanhuiri, S. curtilobum, S. juzepczukii and S. tuberosum, the latter divided into the Andigenum and Chilotanum Cultivar Groups). The data support prior ideas of hybrid origins of S. ajanhuiri from the S. tuberosum Andigenum Group (2x = S. stenotomum) × S. megistacrolobum; S. juzepczukii from the S. tuberosum Andigenum Group (2x = S. stenotomum) × S. acaule; and S. curtilobum from the S. tuberosum Andigenum Group (4x = S. tuberosum subsp. andigenum) × S. juzepczukii. For the tetraploid cultivar-groups of S. tuberosum, hybrid origins are suggested entirely within much more closely related species, except for two of three examined accessions of the S. tuberosum Chilotanum Group that appear to have hybridized with the wild species S. maglia. Hybrid origins of the crop/weed species S. sucrense are more difficult to support and S. vernei is not supported as a wild species progenitor of the S. tuberosum Andigenum Group.

  15. Symbolic Algorithmic Analysis of Rectangular Hybrid Systems

    Institute of Scientific and Technical Information of China (English)

    Hai-Bin Zhang; Zhen-Hua Duan


    This paper investigates symbolic algorithmic analysis of rectangular hybrid systems. To deal with the symbolic reachability problem, a restricted constraint system called hybrid zone is formalized for the representation and manipulation of rectangular automata state-spaces. Hybrid zones are proved to be closed over symbolic reachability operations of rectangular hybrid systems. They are also applied to model-checking procedures for verifying some important classes of timed computation tree logic formulas. To represent hybrid zones, a data structure called difference constraint matrix is defined.These enable us to deal with the symbolic algorithmic analysis of rectangular hybrid systems in an efficient way.

  16. Intercalated hybrid graphite fiber composite (United States)

    Gaier, James R. (Inventor)


    The invention is directed to a highly conductive lightweight hybrid material and methods of producing the same. The hybrid composite is obtained by weaving strands of a high strength carbon or graphite fiber into a fabric-like structure, depositing a layer of carbon onto the structure, heat treating the structure to graphitize the carbon layer, and intercalating the graphitic carbon layer structure. A laminate composite material useful for protection against lightning strikes comprises at least one layer of the hybrid material over at least one layer of high strength carbon or graphite fibers. The composite material of the present invention is compatible with matrix compounds, has a coefficient of thermal expansion which is the same as underlying fiber layers, and is resistant to galvanic corrosion in addition to being highly conductive. These materials are useful in the aerospace industry, in particular as lightning strike protection for airplanes.

  17. Hybrid Simulation of Composite Structures

    DEFF Research Database (Denmark)

    Høgh, Jacob Herold

    of freedom. In this dissertation the main focus is to develop hybrid simulation for composite structures e.g. wind turbine blades where the boundary between the numerical model and the physical experiment is continues i.e. in principal infinite amount of degrees of freedom. This highly complicates...... at the shared boundary. The hybrid simulation programs have been tested on different simple composite structures and they have proven able to increase the accuracy in tests with a complex transfer system.......Hybrid simulation is a substructural method combining a numerical simulation with a physical experiment. A structure is thereby simulated under the assumption that a substructure’s response is well known and easily modelled while a given substructure is studied more accurately in a physical...

  18. Optimizing Hybrid Spreading in Metapopulations

    CERN Document Server

    Zhang, Changwang; Cox, Ingemar J; Chain, Benjamin M


    Epidemic spreading phenomena are ubiquitous in nature and society. Examples include the spreading of diseases, information, and computer viruses. Epidemics can spread by \\textit{local spreading}, where infected nodes can only infect a limited set of direct target nodes and \\textit{global spreading}, where an infected node can infect every other node. In reality, many epidemics spread using a hybrid mixture of both types of spreading. In this study we develop a theoretical framework for studying hybrid epidemics, and examine the optimum balance between spreading mechanisms in terms of achieving the maximum outbreak size. In a metapopulation, made up of many weakly connected subpopulations, we show that one can calculate an optimal tradeoff between local and global spreading which will maximise the extent of the epidemic. As an example we analyse the 2008 outbreak of the Internet worm Conficker, which uses hybrid spreading to propagate through the internet. Our results suggests that the worm would have been eve...

  19. New Supercapacitors of Hybrid Configurations

    Directory of Open Access Journals (Sweden)

    Wako Naoi


    Full Text Available In recent years, the improvement of the energydensity of nano-composite battery materials hasbeen object of great study. Hybridizing battery andcapacitor materials overcome the energy densitylimitation of existing generation-I capacitors withoutmuch sacrificing the cycling performances. Normalbattery-capacitor hybrids employ high-energy &sluggish redox electrode and low-energy & fastdouble-layer electrodes, possibly producing a largerworking voltage and higher over-all capacitance. Inorder to smoothly operate such asymmetric systems,however, the rates of the two different electrodes mustbe highly balanced. Especially, the redox rates of thebattery electrodes must be substantially increasedto the levels of double-layer process. In this report,we attempt to identify the essential issues for therealizable hybrids and suggest ways to overcomethe rate enhancement by exemplifying ultrafastperformance of the Li4Ti5O12 nanocrystal preparedvia a unique in-situ material processing technologyunder ultra-centrifuging.

  20. Additive Manufacturing of Hybrid Circuits (United States)

    Sarobol, Pylin; Cook, Adam; Clem, Paul G.; Keicher, David; Hirschfeld, Deidre; Hall, Aaron C.; Bell, Nelson S.


    There is a rising interest in developing functional electronics using additively manufactured components. Considerations in materials selection and pathways to forming hybrid circuits and devices must demonstrate useful electronic function; must enable integration; and must complement the complex shape, low cost, high volume, and high functionality of structural but generally electronically passive additively manufactured components. This article reviews several emerging technologies being used in industry and research/development to provide integration advantages of fabricating multilayer hybrid circuits or devices. First, we review a maskless, noncontact, direct write (DW) technology that excels in the deposition of metallic colloid inks for electrical interconnects. Second, we review a complementary technology, aerosol deposition (AD), which excels in the deposition of metallic and ceramic powder as consolidated, thick conformal coatings and is additionally patternable through masking. Finally, we show examples of hybrid circuits/devices integrated beyond 2-D planes, using combinations of DW or AD processes and conventional, established processes.

  1. DNA-based hybrid catalysis. (United States)

    Rioz-Martínez, Ana; Roelfes, Gerard


    In the past decade, DNA-based hybrid catalysis has merged as a promising novel approach to homogeneous (asymmetric) catalysis. A DNA hybrid catalysts comprises a transition metal complex that is covalently or supramolecularly bound to DNA. The chiral microenvironment and the second coordination sphere interactions provided by the DNA are key to achieve high enantioselectivities and, often, additional rate accelerations in catalysis. Nowadays, current efforts are focused on improved designs, understanding the origin of the enantioselectivity and DNA-induced rate accelerations, expanding the catalytic scope of the concept and further increasing the practicality of the method for applications in synthesis. Herein, the recent developments will be reviewed and the perspectives for the emerging field of DNA-based hybrid catalysis will be discussed.

  2. Genomic networks of hybrid sterility. (United States)

    Turner, Leslie M; White, Michael A; Tautz, Diethard; Payseur, Bret A


    Hybrid dysfunction, a common feature of reproductive barriers between species, is often caused by negative epistasis between loci ("Dobzhansky-Muller incompatibilities"). The nature and complexity of hybrid incompatibilities remain poorly understood because identifying interacting loci that affect complex phenotypes is difficult. With subspecies in the early stages of speciation, an array of genetic tools, and detailed knowledge of reproductive biology, house mice (Mus musculus) provide a model system for dissecting hybrid incompatibilities. Male hybrids between M. musculus subspecies often show reduced fertility. Previous studies identified loci and several X chromosome-autosome interactions that contribute to sterility. To characterize the genetic basis of hybrid sterility in detail, we used a systems genetics approach, integrating mapping of gene expression traits with sterility phenotypes and QTL. We measured genome-wide testis expression in 305 male F2s from a cross between wild-derived inbred strains of M. musculus musculus and M. m. domesticus. We identified several thousand cis- and trans-acting QTL contributing to expression variation (eQTL). Many trans eQTL cluster into eleven 'hotspots,' seven of which co-localize with QTL for sterility phenotypes identified in the cross. The number and clustering of trans eQTL-but not cis eQTL-were substantially lower when mapping was restricted to a 'fertile' subset of mice, providing evidence that trans eQTL hotspots are related to sterility. Functional annotation of transcripts with eQTL provides insights into the biological processes disrupted by sterility loci and guides prioritization of candidate genes. Using a conditional mapping approach, we identified eQTL dependent on interactions between loci, revealing a complex system of epistasis. Our results illuminate established patterns, including the role of the X chromosome in hybrid sterility. The integrated mapping approach we employed is applicable in a broad

  3. Genomic networks of hybrid sterility.

    Directory of Open Access Journals (Sweden)

    Leslie M Turner


    Full Text Available Hybrid dysfunction, a common feature of reproductive barriers between species, is often caused by negative epistasis between loci ("Dobzhansky-Muller incompatibilities". The nature and complexity of hybrid incompatibilities remain poorly understood because identifying interacting loci that affect complex phenotypes is difficult. With subspecies in the early stages of speciation, an array of genetic tools, and detailed knowledge of reproductive biology, house mice (Mus musculus provide a model system for dissecting hybrid incompatibilities. Male hybrids between M. musculus subspecies often show reduced fertility. Previous studies identified loci and several X chromosome-autosome interactions that contribute to sterility. To characterize the genetic basis of hybrid sterility in detail, we used a systems genetics approach, integrating mapping of gene expression traits with sterility phenotypes and QTL. We measured genome-wide testis expression in 305 male F2s from a cross between wild-derived inbred strains of M. musculus musculus and M. m. domesticus. We identified several thousand cis- and trans-acting QTL contributing to expression variation (eQTL. Many trans eQTL cluster into eleven 'hotspots,' seven of which co-localize with QTL for sterility phenotypes identified in the cross. The number and clustering of trans eQTL-but not cis eQTL-were substantially lower when mapping was restricted to a 'fertile' subset of mice, providing evidence that trans eQTL hotspots are related to sterility. Functional annotation of transcripts with eQTL provides insights into the biological processes disrupted by sterility loci and guides prioritization of candidate genes. Using a conditional mapping approach, we identified eQTL dependent on interactions between loci, revealing a complex system of epistasis. Our results illuminate established patterns, including the role of the X chromosome in hybrid sterility. The integrated mapping approach we employed is

  4. Charge Qubit-Atom Hybrid

    CERN Document Server

    Yu, Deshui; Hufnagel, C; Kwek, L C; Amico, Luigi; Dumke, R


    We investigate a novel hybrid system of a superconducting charge qubit interacting directly with a single neutral atom via electric dipole coupling. Interfacing of the macroscopic superconducting circuit with the microscopic atomic system is accomplished by varying the gate capacitance of the charge qubit. To achieve strong interaction, we employ two Rydberg states with an electric-dipole-allowed transition, which alters the polarizability of the dielectric medium of the gate capacitor. Sweeping the gate voltage with different rates leads to a precise control of hybrid quantum states. Furthermore, we show a possible implementation of a universal two-qubit gate.

  5. Towards an expansive hybrid psychology

    DEFF Research Database (Denmark)

    Brinkmann, Svend


    This article develops an integrative theory of the mind by examining how the mind, understood as a set of skills and dispositions, depends upon four sources of mediators. Harré’s hybrid psychology is taken as a meta-theoretical starting point, but is expanded significantly by including the four...... sources of mediators that are the brain, the body, social practices and technological artefacts. It is argued that the mind is normative in the sense that mental processes do not simply happen, but can be done more or less well, and thus are subject to normative appraisal. The expanded hybrid psychology...

  6. Interspecific Hybridization within Ornamental Plants

    DEFF Research Database (Denmark)

    Kuligowska, Katarzyna

    The economic importance of the ornamental plant industry requires constant development of novel and high quality varieties. Traits attractive for production of new ornamental plants may not be available within the commercial cultivars, but broad genetic variation is present within the plant genera...... commercially important genera of ornamental plants: Kalanchoë and Hibiscus. The nature of hybridization barriers hampering hybrid production was investigated during pre- and post-fertilization stages. For each genus the interspecific crosses of Kalanchoë species and Hibiscus species, abnormal germination...

  7. Hybrid winding concept for toroids

    DEFF Research Database (Denmark)

    Schneider, Henrik; Andersen, Thomas; Knott, Arnold;


    This paper proposes a hybrid winding concept for toroids using the traces in a printed circuit board to make connection to bended copper foil cutouts. In a final product a number of strips with a certain thickness would be held by a former and the whole assembly could be placed by pick...... and placement machinery. This opens up the possibility for both an automated manufacturing process and an automated production process of toroidal magnetics such as power inductors, filtering inductors, air core inductors, transformers etc. Both the proposed hybrid and the common wire wound winding...

  8. A "Hybrid" Approach for Synthesizing Optimal Controllers of Hybrid Systems

    DEFF Research Database (Denmark)

    Zhao, Hengjun; Zhan, Naijun; Kapur, Deepak;


    We propose an approach to reduce the optimal controller synthesis problem of hybrid systems to quantifier elimination; furthermore, we also show how to combine quantifier elimination with numerical computation in order to make it more scalable but at the same time, keep arising errors due to disc...

  9. Modeling and analysis using hybrid Petri nets

    CERN Document Server

    Ghomri, Latéfa


    This paper is devoted to the use of hybrid Petri nets (PNs) for modeling and control of hybrid dynamic systems (HDS). Modeling, analysis and control of HDS attract ever more of researchers' attention and several works have been devoted to these topics. We consider in this paper the extensions of the PN formalism (initially conceived for modeling and analysis of discrete event systems) in the direction of hybrid modeling. We present, first, the continuous PN models. These models are obtained from discrete PNs by the fluidification of the markings. They constitute the first steps in the extension of PNs toward hybrid modeling. Then, we present two hybrid PN models, which differ in the class of HDS they can deal with. The first one is used for deterministic HDS modeling, whereas the second one can deal with HDS with nondeterministic behavior. Keywords: Hybrid dynamic systems; D-elementary hybrid Petri nets; Hybrid automata; Controller synthesis

  10. Field errors in hybrid insertion devices

    Energy Technology Data Exchange (ETDEWEB)

    Schlueter, R.D. [Lawrence Berkeley Lab., CA (United States)


    Hybrid magnet theory as applied to the error analyses used in the design of Advanced Light Source (ALS) insertion devices is reviewed. Sources of field errors in hybrid insertion devices are discussed.

  11. Hybrid Power Management System and Method (United States)

    Eichenberg, Dennis J. (Inventor)


    A system and method for hybrid power management. The system includes photovoltaic cells, ultracapacitors, and pulse generators. In one embodiment, the hybrid power management system is used to provide power for a highway safety flasher.

  12. How to distinguish Hybrids from Radial Quarkonia

    CERN Document Server

    Close, Francis Edwin; Close, Frank E; Page, Philip R.


    We present arguments that reinforce the hybrid interpretation of pi(1800) and we establish that the rho(1450) and the omega(1420) can be interpreted as radial-hybrid mixtures. Some questions for future experiments are raised.


    Energy Technology Data Exchange (ETDEWEB)

    Ye Zhuang; Stanley J. Miller; Michelle R. Olderbak; Rich Gebert


    A new concept in particulate control, called an advanced hybrid particulate collector (AHPC), is being developed under funding from the U.S. Department of Energy. The AHPC combines the best features of electrostatic precipitators (ESPs) and baghouses in an entirely novel manner. The AHPC concept combines fabric filtration and electrostatic precipitation in the same housing, providing major synergism between the two methods, both in the particulate collection step and in transfer of dust to the hopper. The AHPC provides ultrahigh collection efficiency, overcoming the problem of excessive fine-particle emissions with conventional ESPs, and solves the problem of reentrainment and re-collection of dust in conventional baghouses. Phase I of the development effort consisted of design, construction, and testing of a 5.7-m{sup 3}/min (200-acfm) working AHPC model. Results from both 8-hr parametric tests and 100-hr proof-of-concept tests with two different coals demonstrated excellent operability and greater than 99.99% fine-particle collection efficiency. Since all of the developmental goals of Phase I were met, the approach was scaled up in Phase II to a size of 255 m{sup 3}/min (9000 acfm) (equivalent in size to 2.5 MW) and was installed on a slipstream at the Big Stone Power Plant. For Phase II, the AHPC at Big Stone Power Plant was operated continuously from late July 1999 until mid-December 1999. The Phase II results were highly successful in that ultrahigh particle collection efficiency was achieved, pressure drop was well controlled, and system operability was excellent. For Phase III, the AHPC was modified into a more compact configuration, and components were installed that were closer to what would be used in a full-scale commercial design. The modified AHPC was operated from April to July 2000. While operational results were acceptable during this time, inspection of bags in the summer of 2000 revealed some membrane damage to the fabric that appeared to be

  14. Distant hybridization leads to different ploidy fishes

    Institute of Scientific and Technical Information of China (English)


    Distant hybridization makes it possible to transfer the genome of one species to another, which results in changes in phenotypes and genotypes of the progenies. This study shows that distant hybridization or the combination of this method with gynogenesis or androgenesis lead to different ploidy fishes with genetic variation, including fertile tetraploid hybrids, sterile triploid hybrids, fertile diploid hybrids, fertile diploid gynogenetic fish, and their derived progenies. The formations of the different ploidy fishes depend on the genetic relationship between the parents. In this study, several types of distant hybridization, including red crucian carp (Carassius auratus red var.) (2n=100, abbreviated as RCC) (♀)×common carp (Cyprinus carpio L.) (2n=100, abbreviated as CC) (♂), and RCC (2n=100) (♀)×blunt snout bream (Megalobrama amblycephala) (2n=48, abbreviated as BSB) (♂) are described. In the distant hybridization of RCC (♀)×CC (♂), bisexual fertile F3–F18 allotetraploid hybrids (4n=200, abbreviated as 4nAT) were formed. The diploid hybrid eggs and diploid sperm generated by the females and males of 4nAT developed into diploid gynogenetic hybrids and diploid androgenetic hybrids, respectively, by gynogenesis and androgenesis, without treatment for doubling the chromosome. Improved tetraploid hybrids and improved diploid fishes with genetic variation were derived from the gynogenetic hybrid line. The improved diploid fishes included the high-body RCC and high-body goldfish. The formation of the tetraploid hybrids was related to the occurrence of unreduced gametes generated from the diploid hybrids, which involved in premeiotic endoreduplication, endomitosis, or fusion of germ cells. The sterile triploid hybrids (3n=150) were produced on a large scale by crossing the males of tetraploid hybrids with females of diploid fish (2n=100). In another distant hybridization of RCC (♀)×BSB (♂), different ploidy fishes were obtained, including

  15. Distant hybridization leads to different ploidy fishes. (United States)

    Liu, ShaoJun


    Distant hybridization makes it possible to transfer the genome of one species to another, which results in changes in phenotypes and genotypes of the progenies. This study shows that distant hybridization or the combination of this method with gynogenesis or androgenesis lead to different ploidy fishes with genetic variation, including fertile tetraploid hybrids, sterile triploid hybrids, fertile diploid hybrids, fertile diploid gynogenetic fish, and their derived progenies. The formations of the different ploidy fishes depend on the genetic relationship between the parents. In this study, several types of distant hybridization, including red crucian carp (Carassius auratus red var.) (2n=100, abbreviated as RCC) (female) x common carp (Cyprinus carpio L.) (2n=100, abbreviated as CC) (male), and RCC (2n=100) (female) x blunt snout bream (Megalobrama amblycephala) (2n=48, abbreviated as BSB) (male) are described. In the distant hybridization of RCC (female) x CC (male), bisexual fertile F(3)-F(18) allotetraploid hybrids (4n=200, abbreviated as 4nAT) were formed. The diploid hybrid eggs and diploid sperm generated by the females and males of 4nAT developed into diploid gynogenetic hybrids and diploid androgenetic hybrids, respectively, by gynogenesis and androgenesis, without treatment for doubling the chromosome. Improved tetraploid hybrids and improved diploid fishes with genetic variation were derived from the gynogenetic hybrid line. The improved diploid fishes included the high-body RCC and high-body goldfish. The formation of the tetraploid hybrids was related to the occurrence of unreduced gametes generated from the diploid hybrids, which involved in premeiotic endoreduplication, endomitosis, or fusion of germ cells. The sterile triploid hybrids (3n=150) were produced on a large scale by crossing the males of tetraploid hybrids with females of diploid fish (2n=100). In another distant hybridization of RCC (female) x BSB (male), different ploidy fishes were

  16. Advanced Digital Signal Processing for Hybrid Lidar (United States)


    Technical 4. TITLE AND SUBTITLE Advance Digital Signal Processing for Hybrid Lidar 6. AUTHOR(S) William D. Jemison 7. PERFORMING ORGANIZATION NAME(S...development of signed processing algorithms for hybrid lidar - radar designed to improve detection performance. 15. SUBJECT TERMS Hybrid Lidar - Radar 16...Award Number N000141110371 Title of Research Advanced Digital Signal Processing for Hybrid Lidar Principal Investigator William D. Jemison

  17. Contribution of interspecific hybridization to sunflower breeding


    Christov M.


    This investigation is directed at improving sunflower using hybrid forms resulted from interspecific hybridization. The aim is to create new B/A and R lines from interspecific hybrid forms that are resistant to diseases, the parasite broomrape, herbicides, and other stress factors and are characterized with high combining ability and to obtain on this basis highly productive oilseed sunflower hybrids with varied fatty acid composition of oil. The investigat...

  18. Dynamical investigation of macromolecular hybridization bioassays

    CERN Document Server

    Bittner, R; Wixforth, A


    A novel sensoric technique for the dynamical in situ investigation of a hybridization bio assay is presented, which utilizes a metal bead labeling method. Therein, hybridization results in an increased metal coverage on parts of a substrate, where it takes place. Our sensing principle relies on the measurement of the radio frequency impedance of the hybridization spots. We propose several examples for sensor devices.

  19. Hybrid Logical Analyses of the Ambient Calculus

    DEFF Research Database (Denmark)

    Bolander, Thomas; Hansen, Rene Rydhof


    In this paper, hybrid logic is used to formulate three control flow analyses for Mobile Ambients, a process calculus designed for modelling mobility. We show that hybrid logic is very well-suited to express the semantic structure of the ambient calculus and how features of hybrid logic can...

  20. The Hybrid Automobile and the Atkinson Cycle (United States)

    Feldman, Bernard J.


    The hybrid automobile is a strikingly new automobile technology with a number of new technological features that dramatically improve energy efficiency. This paper will briefly describe how hybrid automobiles work; what are these new technological features; why the Toyota Prius hybrid internal combustion engine operates on the Atkinson cycle…

  1. Hybrid Doctoral Program: Innovative Practices and Partnerships (United States)

    Alvich, Dori; Manning, JoAnn; McCormick, Kathy; Campbell, Robert


    This paper reflects on how one mid-Atlantic University innovatively incorporated technology into the development of a hybrid doctoral program in educational leadership. The paper describes a hybrid doctoral degree program using a rigorous design; challenges of reworking a traditional syllabus of record to a hybrid doctoral program; the perceptions…

  2. Review of laser hybrid welding

    DEFF Research Database (Denmark)

    Bagger, Claus


    In this artucle an overview og the hybrid welding process is given. After a short historic overview, a review of the fundamental phenomenon taking place when a laser (CO2 or Nd:YAG) interacts in the same molten pool as a more conventional source of energy, e.g. tungsten in-active gas, plasma...

  3. Hybrid winding concept for toroids

    DEFF Research Database (Denmark)

    Schneider, Henrik; Andersen, Thomas; Knott, Arnold;


    This paper proposes a hybrid winding concept for toroids using the traces in a printed circuit board to make connection to bended copper foil cutouts. In a final product a number of strips with a certain thickness would be held by a former and the whole assembly could be placed by pick and placem...

  4. Organic and hybrid solar cells

    CERN Document Server

    Huang, Hui


    This book delivers a comprehensive evaluation of organic and hybrid solar cells and identifies their fundamental principles and numerous applications. Great attention is given to the charge transport mechanism, donor and acceptor materials, interfacial materials, alternative electrodes, device engineering and physics, and device stability. The authors provide an industrial perspective on the future of photovoltaic technologies.

  5. Multifunctional hybrids for electromagnetic absorption

    Energy Technology Data Exchange (ETDEWEB)

    Huynen, I. [Research Center in Micro and Nanoscopic Materials and Electronic Devices, CeRMiN, Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Information and Communications Technologies, Electronics and Applied Mathematics (ICTEAM), Microwave Laboratory, Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Quievy, N. [Institute of Condensed Matter and Nanosciences (IMCN), Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Bailly, C. [Research Center in Micro and Nanoscopic Materials and Electronic Devices, CeRMiN, Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Institute of Condensed Matter and Nanosciences (IMCN), Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Institute of Mechanics, Materials and Civil Engineering (iMMC), Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Bollen, P. [Information and Communications Technologies, Electronics and Applied Mathematics (ICTEAM), Microwave Laboratory, Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Institute of Condensed Matter and Nanosciences (IMCN), Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Institute of Mechanics, Materials and Civil Engineering (iMMC), Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Detrembleur, C. [Center for Education and Research on Macromolecules (CERM), University of Liege, Sart-Tilman B6a, 4000 Liege (Belgium); Eggermont, S.; Molenberg, I. [Information and Communications Technologies, Electronics and Applied Mathematics (ICTEAM), Microwave Laboratory, Universite catholique de Louvain, B-1348 Louvain-la-Neuve (Belgium); Thomassin, J.M.; Urbanczyk, L. [Center for Education and Research on Macromolecules (CERM), University of Liege, Sart-Tilman B6a, 4000 Liege (Belgium)


    Highlights: > EM absorption requires low dielectric constant and {approx}1 S/m electrical conductivity. > New hybrids were processed with CNT-filled polymer foam inserted in Al honeycomb. > The EM absorption in the GHz range is superior to any known material. > A closed form model is used to guide the design of the hybrid. > The architectured material is light with potential for thermal management. - Abstract: Electromagnetic (EM) interferences are ubiquitous in modern technologies and impact on the reliability of electronic devices and on living cells. Shielding by EM absorption, which is preferable over reflection in certain instances, requires combining a low dielectric constant with high electrical conductivity, which are antagonist properties in the world of materials. A novel class of hybrid materials for EM absorption in the gigahertz range has been developed based on a hierarchical architecture involving a metallic honeycomb filled with a carbon nanotube-reinforced polymer foam. The waveguide characteristics of the honeycomb combined with the performance of the foam lead to unexpectedly large EM power absorption over a wide frequency range, superior to any known material. The peak absorption frequency can be tuned by varying the shape of the honeycomb unit cell. A closed form model of the EM reflection and absorption provides a tool for the optimization of the hybrid. This designed material sets the stage for a new class of sandwich panels combining high EM absorption with mass efficiency, stiffness and thermal management.


    Institute of Scientific and Technical Information of China (English)

    Yi Tong; Wu Fangjun


    This letter proposes a hybrid method for computing dynamic program slicing. The key element is to construct a Coverage-Testing-based Dynamic Dependence Graph (CTDDG),which makes use of both dynamic and static information to get execution status. The approach overcomes the limitations of previous dynamic slicing methods, which have to redo slicing if slice criterion changes.

  7. Inference in hybrid Bayesian networks

    DEFF Research Database (Denmark)

    Lanseth, Helge; Nielsen, Thomas Dyhre; Rumí, Rafael


    Since the 1980s, Bayesian Networks (BNs) have become increasingly popular for building statistical models of complex systems. This is particularly true for boolean systems, where BNs often prove to be a more efficient modelling framework than traditional reliability-techniques (like fault trees...... decade's research on inference in hybrid Bayesian networks. The discussions are linked to an example model for estimating human reliability....

  8. Hybrid Rice for Dry Regions

    Institute of Scientific and Technical Information of China (English)


    Luo Lijun, director of the Shanghai Agrobiological Gene Center, recently announced that Hanyou No. 3 - a genetically modified rice capable of producing high yields in dry areas - has been developed at the center.The hybrid yields 7,571 kilograms per hectare, about the same as ordinary rice.

  9. 1997 hybrid electric vehicle specifications

    Energy Technology Data Exchange (ETDEWEB)

    Sluder, S.; Larsen, R.; Duoba, M.


    The US DOE sponsors Advanced Vehicle Technology competitions to help educate the public and advance new vehicle technologies. For several years, DOE has provided financial and technical support for the American Tour de Sol. This event showcases electric and hybrid electric vehicles in a road rally across portions of the northeastern United States. The specifications contained in this technical memorandum apply to vehicles that will be entered in the 1997 American Tour de Sol. However, the specifications were prepared to be general enough for use by other teams and individuals interested in developing hybrid electric vehicles. The purpose of the specifications is to ensure that the vehicles developed do not present a safety hazard to the teams that build and drive them or to the judges, sponsors, or public who attend the competitions. The specifications are by no means the definitive sources of information on constructing hybrid electric vehicles - as electric and hybrid vehicles technologies advance, so will the standards and practices for their construction. In some cases, the new standards and practices will make portions of these specifications obsolete.

  10. Yeast two-hybrid screen. (United States)

    Makuch, Lauren


    Yeast two-hybrid is a method for screening large numbers of gene products (encoded by cDNA libraries) for their ability to interact with a protein of interest. This system can also be used for characterizing and manipulating candidate protein: protein interactions. Interactions between proteins are monitored by the growth of yeast plated on selective media.

  11. Hybrid Model of Content Extraction

    DEFF Research Database (Denmark)

    Qureshi, Pir Abdul Rasool; Memon, Nasrullah


    We present a hybrid model for content extraction from HTML documents. The model operates on Document Object Model (DOM) tree of the corresponding HTML document. It evaluates each tree node and associated statistical features like link density and text distribution across the node to predict signi...

  12. Comparative genomic hybridization: practical guidelines.

    NARCIS (Netherlands)

    Jeuken, J.W.M.; Sprenger, S.H.; Wesseling, P.


    Comparative genomic hybridization (CGH) is a technique used to identify copy number changes throughout a genome. Until now, hundreds of CGH studies have been published reporting chromosomal imbalances in a large variety of human neoplasms. Additionally, technical improvements of specific steps in a

  13. Shape Memory Composite Hybrid Hinge (United States)

    Fang, Houfei; Im, Eastwood; Lin, John; Scarborough, Stephen


    There are two conventional types of hinges for in-space deployment applications. The first type is mechanically deploying hinges. A typical mechanically deploying hinge is usually composed of several tens of components. It is complicated, heavy, and bulky. More components imply higher deployment failure probability. Due to the existence of relatively moving components among a mechanically deploying hinge, it unavoidably has microdynamic problems. The second type of conventional hinge relies on strain energy for deployment. A tape-spring hinge is a typical strain energy hinge. A fundamental problem of a strain energy hinge is that its deployment dynamic is uncontrollable. Usually, its deployment is associated with a large impact, which is unacceptable for many space applications. Some damping technologies have been experimented with to reduce the impact, but they increased the risks of an unsuccessful deployment. Coalescing strain energy components with shape memory composite (SMC) components to form a hybrid hinge is the solution. SMCs are well suited for deployable structures. A SMC is created from a high-performance fiber and a shape memory polymer resin. When the resin is heated to above its glass transition temperature, the composite becomes flexible and can be folded or packed. Once cooled to below the glass transition temperature, the composite remains in the packed state. When the structure is ready to be deployed, the SMC component is reheated to above the glass transition temperature, and it returns to its as-fabricated shape. A hybrid hinge is composed of two strain energy flanges (also called tape-springs) and one SMC tube. Two folding lines are placed on the SMC tube to avoid excessive strain on the SMC during folding. Two adapters are used to connect the hybrid hinge to its adjacent structural components. While the SMC tube is heated to above its glass transition temperature, a hybrid hinge can be folded and stays at folded status after the temperature

  14. The Hybrid Mindset and Operationalizing Innovation: Toward a Theory of Hybrid (United States)


    explored the existence of a hybrid mindset from a Middle-Eastern context. Military professionals, defense think tanks, and academics regard this...Approved for Public Release; Distribution is Unlimited THE HYBRID MINDSET AND OPERATIONALIZING INNOVATION: TOWARD A THEORY OF HYBRID A Monograph...CONTRACT NUMBER The Hybrid Mindset and Operationalizing Innovation: Toward a Theory of Hybrid 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER

  15. SCAN-based hybrid and double-hybrid density functionals from models without fitted parameters


    Hui, Kerwin; Chai, Jeng-Da


    By incorporating the nonempirical SCAN semilocal density functional [Sun, Ruzsinszky, and Perdew, Phys. Rev. Lett. 115, 036402 (2015)] in the underlying expression of four existing hybrid and double-hybrid models, we propose one hybrid (SCAN0) and three double-hybrid (SCAN0-DH, SCAN-QIDH, and SCAN0-2) density functionals, which are free from any fitted parameters. The SCAN-based double-hybrid functionals consistently outperform their parent SCAN semilocal functional for self-interaction probl...

  16. Reproduction and hybrid load in all-hybrid populations of Rana esculenta water frogs in Denmark

    DEFF Research Database (Denmark)

    Christiansen, Ditte Guldager; Fog, Kåre; Pedersen, Bo Vest;


    All-hybrid populations of the water frog, Rana esculenta, are exceptional in consisting of independently and to some extent sexually reproducing interspecific hybrids. In most of its range R. esculenta reproduces hemiclonally with one of the parental species, R. lessonae or R. ridibunda, but viable...... gametogenesis and mating between frogs with incompatible gametes induce a significant hybrid load in all-hybrid populations of R. esculenta, and we discuss compensating advantages and potential evolutionary trajectories to reduce this hybrid load....

  17. Direct detection of expanded trinucleotide repeats using PCR and DNA hybridization techniques

    Energy Technology Data Exchange (ETDEWEB)

    Petronis, A.; Tatuch, Y.; Klempan, T.A.; Kennedy, J.L. [Hospital for Sick Children, Toronto (Canada)] [and others


    Recently, unstable trinucleotide repeats have been shown to be the etiologic factor in seven neuropsychiatric diseases, and they may play a similar role in other genetic disorders which exhibit genetic anticipation. We have tested one polymerase chain reaction (PCR)-based and two hybridization-based methods for direct detection of unstable DNA expansion in genomic DNA. This technique employs a single primer (asymmetric) PCR using total genomic DNA as a template to efficiently screen for the presence of large trinucleotide repeat expansions. High-stringency Southern blot hybridization with a PCR-generated trinucleotide repeat probe allowed detection of the DNA fragment containing the expansion. Analysis of myotonic dystrophy patients containing different degrees of (CTG){sub n} expansion demonstrated the identification of the site of trinucleotide instability in some affected individuals without any prior information regarding genetic map location. The same probe was used for fluorescent in situ hybridization and several regions of (CTG){sub n}/(CAG){sub n} repeats in the human genome were detected, including the myotonic dystrophy locus on chromosome 19q. Although limited at present to large trinucleotide repeat expansions, these strategies can be applied to directly clone genes involved in disorders caused by large expansions of unstable DNA. 33 refs., 4 figs.

  18. Fusion of CCL21 non-migratory active breast epithelial and breast cancer cells give rise to CCL21 migratory active tumor hybrid cell lines.

    Directory of Open Access Journals (Sweden)

    Benjamin Berndt

    Full Text Available The biological phenomenon of cell fusion has been linked to tumor progression because several data provided evidence that fusion of tumor cells and normal cells gave rise to hybrid cell lines exhibiting novel properties, such as increased metastatogenic capacity and an enhanced drug resistance. Here we investigated M13HS hybrid cell lines, derived from spontaneous fusion events between M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics and HS578T-Hyg breast cancer cells, concerning CCL21/CCR7 signaling. Western Blot analysis showed that all cell lines varied in their CCR7 expression levels as well as differed in the induction and kinetics of CCR7 specific signal transduction cascades. Flow cytometry-based calcium measurements revealed that a CCL21 induced calcium influx was solely detected in M13HS hybrid cell lines. Cell migration demonstrated that only M13HS hybrid cell lines, but not parental derivatives, responded to CCL21 stimulation with an increased migratory activity. Knockdown of CCR7 expression by siRNA completely abrogated the CCL21 induced migration of hybrid cell lines indicating the necessity of CCL21/CCR7 signaling. Because the CCL21/CCR7 axis has been linked to metastatic spreading of breast cancer to lymph nodes we conclude from our data that cell fusion could be a mechanism explaining the origin of metastatic cancer (hybrid cells.

  19. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures. (United States)

    Lamer, S; Jungblut, P R


    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

  20. Collective Identity Formation in Hybrid Organizations

    DEFF Research Database (Denmark)

    Boulongne, Romain; Boxenbaum, Eva

    The present article examines the process of collective identity formation in the context of hybrid organizing. Empirically, we investigate hybrid organizing in a collaborative structure at the interface of two heterogeneous organizations in the domain of new renewable energies. We draw...... organizations. Our findings point to two factors that limit knowledge sharing and hence to new collective identity formation in a hybrid space: 1) ambiguous or multiple organizational roles and 2) strong identities of the collaborating organizations. These findings contribute to illuminating the initial...... formation of a new collective identity in hybrid organizing, and hence how new hybrid organizational forms may emerge non-intentionally....

  1. Detection of MYCN Gene Amplification in Neuroblastoma by Fluorescence In Situ Hybridization: A Pediatric Oncology Group Study

    Directory of Open Access Journals (Sweden)

    Prasad Mathew


    Full Text Available To assess the utility of fluorescence in situ hybridization (FISH for analysis of MYCN gene amplification in neuroblastoma, we compared this assay with Southern blot analysis using tumor specimens collected from 232 patients with presenting characteristics typical of this disease. The FISH technique identified MYCN amplification in 47 cases, compared with 39 by Southern blotting, thus increasing the total number of positive cases by 21%. The major cause of discordancy was a low fraction of tumor cells (≤30% replacement in clinical specimens, which prevented an accurate estimate of MYCN copy number by Southern blotting. With FISH, by contrast, it was possible to analyze multiple interphase nuclei of tumor cells, regardless of the proportion of normal peripheral blood, bone marrow, or stromal cells in clinical samples. Thus, FISH could be performed accurately with very small numbers of tumor cells from touch preparations of needle biopsies. Moreover, this procedure allowed us to discern the heterogeneous pattern of MYCN amplification that is characteristic of neuroblastoma. We conclude that FISH improves the detection of MYCN gene amplification in childhood neuroblastomas in a clinical setting, thus facilitating therapeutic decisions based on the presence or absence of this prognostically important biologic marker.

  2. Gasoline hybrid pneumatic engine for efficient vehicle powertrain hybridization


    Dimitrova, Zlatina; Maréchal, François


    The largest applied convertors in passenger cars are the internal combustion engines – gasoline, diesel, adapted also for operating on alternative fuels and hybrid modes. The number of components that are necessary to realize modern future propulsion system is inexorably increasing. The need for efficiency improvement of the vehicle energy system induces the search for an innovative methodology during the design process. In this article the compressed air is investigated as an innovative solu...

  3. Towards stable silicon nanoarray hybrid solar cells. (United States)

    He, W W; Wu, K J; Wang, K; Shi, T F; Wu, L; Li, S X; Teng, D Y; Ye, C H


    Silicon nanoarray hybrid solar cells benefit from the ease of fabrication and the cost-effectiveness of the hybrid structure, and represent a new research focus towards the utilization of solar energy. However, hybrid solar cells composed of both inorganic and organic components suffer from the notorious stability issue, which has to be tackled before the hybrid solar cells could become a viable alternative for harvesting solar energy. Here we show that Si nanoarray/PEDOT:PSS hybrid solar cells with improved stability can be fabricated via eliminating the water inclusion in the initial formation of the heterojunction between Si nanoarray and PEDOT:PSS. The Si nanoarray hybrid solar cells are stable against rapid degradation in the atmosphere environment for several months without encapsulation. This finding paves the way towards the real-world applications of Si nanoarray hybrid solar cells.

  4. Hybrid methods for cybersecurity analysis :

    Energy Technology Data Exchange (ETDEWEB)

    Davis, Warren Leon,; Dunlavy, Daniel M.


    Early 2010 saw a signi cant change in adversarial techniques aimed at network intrusion: a shift from malware delivered via email attachments toward the use of hidden, embedded hyperlinks to initiate sequences of downloads and interactions with web sites and network servers containing malicious software. Enterprise security groups were well poised and experienced in defending the former attacks, but the new types of attacks were larger in number, more challenging to detect, dynamic in nature, and required the development of new technologies and analytic capabilities. The Hybrid LDRD project was aimed at delivering new capabilities in large-scale data modeling and analysis to enterprise security operators and analysts and understanding the challenges of detection and prevention of emerging cybersecurity threats. Leveraging previous LDRD research e orts and capabilities in large-scale relational data analysis, large-scale discrete data analysis and visualization, and streaming data analysis, new modeling and analysis capabilities were quickly brought to bear on the problems in email phishing and spear phishing attacks in the Sandia enterprise security operational groups at the onset of the Hybrid project. As part of this project, a software development and deployment framework was created within the security analyst work ow tool sets to facilitate the delivery and testing of new capabilities as they became available, and machine learning algorithms were developed to address the challenge of dynamic threats. Furthermore, researchers from the Hybrid project were embedded in the security analyst groups for almost a full year, engaged in daily operational activities and routines, creating an atmosphere of trust and collaboration between the researchers and security personnel. The Hybrid project has altered the way that research ideas can be incorporated into the production environments of Sandias enterprise security groups, reducing time to deployment from months and

  5. Developing a Hybrid Graduate Program

    Directory of Open Access Journals (Sweden)

    Ronda Sturgill


    Full Text Available There is a continuing need for flexibility and adaptability in the dynamic world of program development in higher education. Students today have more responsibilities and obligations outside of the classroom. Therefore, educational programs that offer alternative class meeting times and other flexible options are attractive to the nontraditional student. The purpose of this paper is to describe and demonstrate a model for a graduate program delivered by a hybrid, or blended, format. The model will be a master's degree program in exercise and nutrition science where the program is delivered through blending both face to face classroom learning and e-learning teaching methodologies. Challenges of development, lessons learned, and future recommendations will also be presented. This hybrid model is interdisciplinary and can be adapted and utilized across a variety of disciplines.

  6. Segmented Hybrid Gasostatic Bearing Optimization

    Directory of Open Access Journals (Sweden)

    Prodan Nikolay Vasilevich


    Full Text Available The purpose of research-development of methods of numerical optimization rotatable support pads gasostatic hybrid bearing. In the world‘s aerospace engineering the gas-dynamic bearings are currently most common. They are characterized by the supporting layer of different designs, which ensures the workability of the rotors during starts and stops. The main problem of this bearing type, apart from the construction complexity is the wear of this supporting layer. Gas-static bearing has no such defect, since there is no physical contact between solid surfaces. This study presents the results of the hybrid bearing’s calculation, combining both technologies. The slotted nozzle of non-conventional shape that mirrors the solution of Reynolds equation’s isoline is studied. The dependences of the main parameters on the speed of the shaft’s rotation are discussed. The aerodynamic resistance of pads for different regimes of operation is investigated.

  7. Clinical Role of Hybrid Imaging. (United States)

    Hsiao, Edward M; Ali, Bilal; Dorbala, Sharmila


    Recent technological advances have fueled the growth in hybrid radionuclide and CT imaging of the heart. Noninvasive imaging studies are reliable means to diagnose coronary artery disease (CAD), stratify risk, and guide clinical management. Myocardial perfusion scintigraphy is a robust, widely available noninvasive modality for the evaluation of ischemia from known or suspected CAD. Cardiac CT (coronary artery calcium score and coronary CT angiography) has emerged as a clinically robust noninvasive anatomic imaging test, capable of rapidly diagnosing or excluding obstructive CAD. Both anatomic and functional modalities have strengths and weaknesses, and can complement each other by offering integrated structural and physiologic information. As we discuss below, in selected patients, hybrid imaging may facilitate more accurate diagnosis, risk stratification, and management in a "one-stop shop" setting.

  8. Photoelastic tomography as hybrid mechanics

    Directory of Open Access Journals (Sweden)

    Errapart A.


    Full Text Available Photoelastic tomography is a non-destructive method of 3D stress analysis. It permits determination of normal stress distribution in an arbitrary section of a 3D test object. In case of axial symmetry also the shear stress distribution can be determined directly from the measurement data. To determine also the other stress components one can use equations of the theory of elasticity. Such a combined application of experimental measurements and numerical handling of the equations of the theory of elasticity is named hybrid mechanics. It is shown that if stresses are due to external loads, the hybrid mechanics algorithm is based on the equations of equilibrium and compatibility. In the case of the measurement of the residual stress in glass the compatibility equation can not be applied. In this case a new relationship of axisymmetric thermoelasticity, the generalized sum rule can be applied.

  9. Photoelastic tomography as hybrid mechanics (United States)

    Aben, H.; Ainola, L.; Errapart, A.


    Photoelastic tomography is a non-destructive method of 3D stress analysis. It permits determination of normal stress distribution in an arbitrary section of a 3D test object. In case of axial symmetry also the shear stress distribution can be determined directly from the measurement data. To determine also the other stress components one can use equations of the theory of elasticity. Such a combined application of experimental measurements and numerical handling of the equations of the theory of elasticity is named hybrid mechanics. It is shown that if stresses are due to external loads, the hybrid mechanics algorithm is based on the equations of equilibrium and compatibility. In the case of the measurement of the residual stress in glass the compatibility equation can not be applied. In this case a new relationship of axisymmetric thermoelasticity, the generalized sum rule can be applied.

  10. Hybrid power plants; Thermische Hybridkraftwerke

    Energy Technology Data Exchange (ETDEWEB)

    Tuschy, I. [ALSTOM Power Support, Mannheim (Germany). Bereiche Forschung und Entwicklung/Ingenieurdienstleistungen; Franke, U. [Fachhochschule Flensburg (Germany). Thermodynamik


    New German laws concerning the use of biomass and other renewable energy sources have improved the conditions for power generation with renewable energies. A promising alternative to common small-scale (combined-heat-and-) power plants could be found in so-called hybrid power plants which use renewable energies and fossil fuels in combination. A thermodynamic process analysis shows how to determine the renewable energies' portion of a hybrid power plant's total output. [German] Mit den veraenderten gesetzlichen Rahmenbedingungen erhoeht sich die Motivation, regenerative Energiequellen zur Stromerzeugung zu nutzen. Neben dem exklusiven Einsatz in kleineren (Heiz-) Kraftwerken bietet sich die gemeinsame Nutzung mit fossilen Energietraegern in sogenannten Hybridkraftwerken an. Hier stellt sich die Frage, wie der Anteil der regenerativen Energien an der Gesamt-Stromproduktion des Hybridkraftwerkes zu ermitteln ist. Eine thermodynamische Prozessanalyse kann darueber Aufschluss geben.

  11. Hybrid optical acoustic seafloor mapping (United States)

    Inglis, Gabrielle

    The oceanographic research and industrial communities have a persistent demand for detailed three dimensional sea floor maps which convey both shape and texture. Such data products are used for archeology, geology, ship inspection, biology, and habitat classification. There are a variety of sensing modalities and processing techniques available to produce these maps and each have their own potential benefits and related challenges. Multibeam sonar and stereo vision are such two sensors with complementary strengths making them ideally suited for data fusion. Data fusion approaches however, have seen only limited application to underwater mapping and there are no established methods for creating hybrid, 3D reconstructions from two underwater sensing modalities. This thesis develops a processing pipeline to synthesize hybrid maps from multi-modal survey data. It is helpful to think of this processing pipeline as having two distinct phases: Navigation Refinement and Map Construction. This thesis extends existing work in underwater navigation refinement by incorporating methods which increase measurement consistency between both multibeam and camera. The result is a self consistent 3D point cloud comprised of camera and multibeam measurements. In map construction phase, a subset of the multi-modal point cloud retaining the best characteristics of each sensor is selected to be part of the final map. To quantify the desired traits of a map several characteristics of a useful map are distilled into specific criteria. The different ways that hybrid maps can address these criteria provides justification for producing them as an alternative to current methodologies. The processing pipeline implements multi-modal data fusion and outlier rejection with emphasis on different aspects of map fidelity. The resulting point cloud is evaluated in terms of how well it addresses the map criteria. The final hybrid maps retain the strengths of both sensors and show significant improvement

  12. Complex hybrid inflation and baryogenesis. (United States)

    Delepine, David; Martínez, Carlos; Ureña-López, L Arturo


    We propose a hybrid inflation model with a complex waterfall field which contains an interaction term that breaks the U(1) global symmetry associated with the waterfall field charge. We show that the asymmetric evolution of the real and imaginary parts of the complex field during the phase transition at the end of inflation translates into a charge asymmetry. The latter strongly depends on the vacuum expectation value of the waterfall field, which is well constrained by diverse cosmological observations.

  13. Performance of hybrid photovoltaic collector


    Garbisu Eugui, Josu


    The aim of the present project is the study of the performance of a combined photovoltaic-thermal plant, called also hybrid system, located in south Italy, evaluating the efficiency of the photovoltaic and thermal systems and the advantage respect to the two single plants (photovoltaic and thermal ). This research project has two objectives fundamentals of efficiency improvement energy from solar photovoltaic panels. On one hand, increase photovoltaic efficiency, at the same time an...

  14. Hybrid Systems: Computation and Control. (United States)


    elbow) and a pinned first joint (shoul- der) (see Figure 2); it is termed an underactuated system since it is a mechanical system with fewer...Montreal, PQ, Canada, 1998. [10] M. W. Spong. Partial feedback linearization of underactuated mechanical systems . In Proceedings, IROS󈨢, pages 314-321...control mechanism and search for optimal combinations of control variables. Besides the nonlinear and hybrid nature of powertrain systems , hardware

  15. Hybrid Maritime Satellite Communication Antenna

    DEFF Research Database (Denmark)

    Smith, Thomas Gunst

    Hybrid antennas for a maritime satellite communication terminal with simultaneous operation at L- and Ka-band have been investigated. The frequency bands of interest are 1; 525:0 1; 660:5 MHz (RX+TX, RHCP), 19:7 20:2 (RX, LHCP) and 29:5 30:0 GHz (TX, RHCP), which are all part of the Inmarsat BGAN...

  16. CERN manufactured hybrid photon detectors

    CERN Multimedia

    Maximilien Brice


    These hybrid photon detectors (HPDs) produce an electric signal from a single photon. An electron is liberated from a photocathode and accelerated to a silicon pixel array allowing the location of the photon on the cathode to be recorded. The electronics and optics for these devices have been developed in close collaboration with industry. HPDs have potential for further use in astrophysics and medical imaging.

  17. Hybrid Tower, Designing Soft Structures

    DEFF Research Database (Denmark)

    Ramsgaard Thomsen, Mette; Tamke, Martin; Holden Deleuran, Anders;


    predictions about the inherent interdependency and material dependent performance of the hybrid structure and (2) the inter-scalar design strategies for specification and fabrication. The first investigation focuses on the design pipelines developed between the implementation of realtime physics...... (membrane reinforcement strategy) and micro scale (design of bespoke textile membrane). The paper concludes with a post construction analysis. Comparing structural and environmental data, the predicted and the actual performance of tower are evaluated and discussed....

  18. Hybrid metric-Palatini stars

    CERN Document Server

    Danila, Bogdan; Lobo, Francisco S N; Mak, M K


    We consider the internal structure and the physical properties of specific classes of neutron, quark and Bose-Einstein Condensate stars in the hybrid metric-Palatini gravity theory, which is a combination of the metric and Palatini $f(R)$ formalisms. The theory is very successful in accounting for the observed phenomenology, since it unifies local constraints at the Solar System level and the late-time cosmic acceleration, even if the scalar field is very light. We derive the equilibrium equations for a spherically symmetric configuration (mass continuity and Tolman-Oppenheimer-Volkoff) in the framework of hybrid metric-Palatini theory, and we investigate their solutions numerically for different equations of state of neutron and quark matter, by adopting for the scalar field potential a Higgs-type form. Stellar models, described by the stiff fluid, radiation-like, the bag model and the Bose-Einstein Condensate equations of state are explicitly constructed in both General Relativity and hybrid metric-Palatini...

  19. Gravity localization on hybrid branes

    Energy Technology Data Exchange (ETDEWEB)

    Veras, D.F.S., E-mail: [Universidade Federal do Ceará (UFC), Departamento de Física, Campus do Pici, Caixa Postal 6030, 60455-760, Fortaleza, Ceará (Brazil); Cruz, W.T., E-mail: [Instituto Federal de Educação, Ciência e Tecnologia do Ceará (IFCE), Campus Juazeiro do Norte, 63040-540 Juazeiro do Norte, Ceará (Brazil); Maluf, R.V., E-mail: [Universidade Federal do Ceará (UFC), Departamento de Física, Campus do Pici, Caixa Postal 6030, 60455-760, Fortaleza, Ceará (Brazil); Almeida, C.A.S., E-mail: [Universidade Federal do Ceará (UFC), Departamento de Física, Campus do Pici, Caixa Postal 6030, 60455-760, Fortaleza, Ceará (Brazil)


    This work deals with gravity localization on codimension-1 brane worlds engendered by compacton-like kinks, the so-called hybrid branes. In such scenarios, the thin brane behavior is manifested when the extra dimension is outside the compact domain, where the energy density is non-trivial, instead of asymptotically as in the usual thick brane models. The zero mode is trapped in the brane, as required. The massive modes, although not localized in the brane, have important phenomenological implications such as corrections to the Newton's law. We study such corrections in the usual thick domain wall and in the hybrid brane scenarios. By means of suitable numerical methods, we attain the mass spectrum for the graviton and the corresponding wavefunctions. The spectra possess the usual linearly increasing behavior from the Kaluza–Klein theories. Further, we show that the 4D gravitational force is slightly increased at short distances. The first eigenstate contributes highly for the correction to the Newton's law. The subsequent normalized solutions have diminishing contributions. Moreover, we find out that the phenomenology of the hybrid brane is not different from the usual thick domain wall. The use of numerical techniques for solving the equations of the massive modes is useful for matching possible phenomenological measurements in the gravitational law as a probe to warped extra dimensions.

  20. Electroactive functional hybrid layered nanocomposites

    Energy Technology Data Exchange (ETDEWEB)

    Destri, Giovanni Li; Torrisi, Vanna; Marletta, Giovanni [Laboratory for Molecular Surfaces and Nanotechnology (LAMSUN) - University of Catania and CSGI - Catania (Italy)


    Two methodologies to build new nanostructured hybrid layered nanocomposites are presented. The first one involves the preparation of hybrid metal/polymer nanolayers (NLs) by combining two monolayer preparation techniques: Horizontal Precipitation Langmuir Blodgett method (HP-ML), for copolymer monolayers and sputter deposition technique, for Au NLs deposition. The second methodology is aimed to prepare regular arrays of nanopores, with diameter ranging between 40-100 nm, in ultra-thin films of electroactive polymers, to obtain embedded regular arrays of nanopores filled by a further electroactive organic component. The produced hybrid MLs have been characterized by means of X-ray Photoelectron Spectroscopy (XPS), Atomic Force Microscopy (AFM) and Grazing Incidence X-ray Diffraction (GI-XRD). In the first case, current-voltage (I-V) measurements demonstrate that the multilayers exhibit a bipolar conduction behaviour (electrons and holes carriers), with a peculiar transition in the nature of the majority carriers (from holes to electrons) above a threshold number of bilayers. In the second case, it is found that the degree of pore filling, as well as the polymer crystallinity can be easily modulated, prompting the tuning of the photoresponse of the nanocomposites.

  1. Gravity localization on hybrid branes (United States)

    Veras, D. F. S.; Cruz, W. T.; Maluf, R. V.; Almeida, C. A. S.


    This work deals with gravity localization on codimension-1 brane worlds engendered by compacton-like kinks, the so-called hybrid branes. In such scenarios, the thin brane behavior is manifested when the extra dimension is outside the compact domain, where the energy density is non-trivial, instead of asymptotically as in the usual thick brane models. The zero mode is trapped in the brane, as required. The massive modes, although not localized in the brane, have important phenomenological implications such as corrections to the Newton's law. We study such corrections in the usual thick domain wall and in the hybrid brane scenarios. By means of suitable numerical methods, we attain the mass spectrum for the graviton and the corresponding wavefunctions. The spectra possess the usual linearly increasing behavior from the Kaluza-Klein theories. Further, we show that the 4D gravitational force is slightly increased at short distances. The first eigenstate contributes highly for the correction to the Newton's law. The subsequent normalized solutions have diminishing contributions. Moreover, we find out that the phenomenology of the hybrid brane is not different from the usual thick domain wall. The use of numerical techniques for solving the equations of the massive modes is useful for matching possible phenomenological measurements in the gravitational law as a probe to warped extra dimensions.

  2. The plus-hybrid effect on the grain yield of two ZP maize hybrids

    Directory of Open Access Journals (Sweden)

    Božinović Sofija


    Full Text Available The combined effect of cytoplasmic male sterility and xenia on maize hybrid traits is referred to as the plus-hybrid effect. Two studied ZP hybrids differently responded to this effect for grain yield. All plus-hybrid combinations of the firstly observed hybrid had a higher yield than their fertile counterparts, but not significantly, while only one combination of the second hybrid positively responded, also without statistical significance. It seems that the observed effect mostly depended on the genotype of the female component.

  3. Comparison analysis between Western blot method and Magnetic particle Chemiluminescence method detection SLE autoantibody%免疫印迹法和磁微粒化学发光法检测SLE自身抗体的比对分析

    Institute of Scientific and Technical Information of China (English)



    目的:比较免疫印迹法(Western blot)和磁微粒化学发光法(Magnetic particle Chemiluminescence method)检测系统性红斑狼疮(SLE)自身抗体的一致性。方法:用免疫印迹法和磁微粒化学发光法对123份血清进行检测,其中系统性红斑狼疮阳性患者63例,健康体检者60例,用卡方检验对两种方法结果进行比较分析。结果:免疫印迹法和磁微粒化学发光法中抗Sm抗体阳性率分别为27.0%和14.3%、抗RNP/Sm阳性率分别为54.0%和42.9%、dsDNA阳性率分别为41.3%和30.2%、抗核小体抗体阳性率分别为31.7%和20.6%、抗核糖体P蛋白抗体阳性率分别为28.6%和11.1%。两种方法的准确性无显著性差异(>0.05)总符合率为80.6%,Kappa值为0.669。结论:免疫印迹法和磁微粒化学发光法检测系统性红斑狼疮自身抗体的检测结果具有可靠性,较好的符合率和准确性,磁微粒化学发光法可以作为简便、快速、经济的检测方法应用于系统性红斑狼疮自身抗体的检测。%Objective: The present study aims to compare Western blot and Magnetic particle Chemiluminescence method consistency on autoimmune antibody detection of systemic lupus erythematosus. Methods: Western blot assay and Magnetic particle Chemiluminescence method were used to detect autoimmune antibody from 123 serum samples, of which 63 patients with systemic lupus erythematosus, the others from 60 healthy persons. The results of the two methods were compared by X2 test. Results: Anti-Sm antibody positive rate of Western blot and Magnetic particle Chemiluminescence method were 27.0 and 14.3 percent respectively. Anti- RNP/Sm antibody positive rates were 39.7% and 42.9% respectively. Anti- ds- DNA antibody positive rates were 41.3% and 30.2% respectively. Anti- nucleosome antibody positive rates were 31.7% and 20.6% respectively. Anti- ribosomal protein antibody positive rates were 28.6% and 11.1% respectively. The accuracy


    Institute of Scientific and Technical Information of China (English)

    Wang Caifeng; Li Xu; Zhang Yunjing


    Objective To explore if strand breaks of DNA in human early chorionic villus cells in uterus were induced by diagnostic ultrasound and to evaluate the method used for detection of single-stranded breaks and doublestranded breaks in human DNA. Methods 60 normal pregnant women aged 20-30, who underwent artificial abortion during 6-8 weeks of gestation, were randomly divided into 2 experimental groups: All 30 cases were exposed to diagnostic ultrasound in uterus for 10 minutes, and 24 hours later chorionic villi were extracted; the other 30 cases were taken as the control group. Single-stranded DNA and double-stranded DNA in villus cells in all cases were isolated by the alkaline unwinding combined with hydroxylapatite chromatography, and were quantitatively detected using32 P-labeled Alu probe for dot-blotting hybridization. Results There was no significant difference in quantity and percentage in single-stranded DNA and double-stranded DNA between 2 groups (P>0.05). 32 P-Alu probe could only hybridize with human DNA, and could detect DNA isolated from as few as 2.5 × 103 chorionic villus cells and 0.45 ng DNA in human leukocytes. Conclusion The results suggested that there were no DNA strand damages in human chorionic villus cells when the uterus was exposed to diagnostic ultrasound for 10 minutes. The method, 32P-Alu probe for dot-blotting hybridization, was even more specific, sensitive and accurate than conventional approaches.

  5. Identification of a deletion in the mismatch repair gene, MSH2, using mouse-human cell hybrids monosomal for chromosome 2. (United States)

    Pyatt, R E; Nakagawa, H; Hampel, H; Sedra, M; Fuchik, M B; Comeras, I; de la Chapelle, A; Prior, T W


    Hereditary non-polyposis colorectal cancer is characterized by mutations in one of the DNA mismatch repair genes, primarily MLH1, MSH2, or MSH6. We report here the identification of a genomic deletion of approximately 11.4 kb encompassing the first two exons of the MSH2 gene in two generations of an Ohio family. By Southern blot analysis, using a cDNA probe spanning the first seven exons of MSH2, an alteration in each of three different enzyme digests (including a unique 13-kb band on HindIII digests) was observed, which suggested the presence of a large alteration in the 5' region of this gene. Mouse-human cell hybrids from a mutation carrier were then generated which contained a single copy each of human chromosome 2 on which the MSH2 gene resides. Southern blots on DNA from the cell hybrids demonstrated the same, unique 13-kb band from one MSH2 allele, as seen in the diploid DNA. DNA from this same monosomal cell hybrid failed to amplify in polymerase chain reactions (PCRs) using primers to exons 1 and 2, demonstrating the deletion of these sequences in one MSH2 allele, and the breakpoints involving Alu repeats were identified by PCR amplification and sequence analysis.

  6. Identification of differentially expressed genes in uveal melanoma using suppressive subtractive hybridization (United States)

    Landreville, Solange; Lupien, Caroline B.; Vigneault, Francois; Gaudreault, Manon; Mathieu, Mélissa; Rousseau, Alain P.; Guérin, Sylvain L.


    Purpose Uveal melanoma (UM) is the most common primary cancer of the eye, resulting not only in vision loss, but also in metastatic death. This study attempts to identify changes in the patterns of gene expression that lead to malignant transformation and proliferation of normal uveal melanocytes (UVM) using the Suppressive Subtractive Hybridization (SSH) technique. Methods The SSH technique was used to isolate genes that are differentially expressed in the TP31 cell line derived from a primary UM compared to UVM. The expression level of selected genes was further validated by microarray, semi-quantitative RT–PCR and western blot analyses. Results Analysis of the subtracted libraries revealed that 37 and 36 genes were, respectively, up- and downregulated in TP31 cells compared to UVM. Differential expression of the majority of these genes was confirmed by comparing UM cells with UVM by microarray. The expression pattern of selected genes was analyzed by semi-quantitative RT–PCR and western blot, and was found to be consistent with the SSH findings. Conclusions We demonstrated that the SSH technique is efficient to detect differentially expressed genes in UM. The genes identified in this study represent valuable candidates for further functional analysis in UM and should be informative in studying the biology of this tumor. PMID:21647268

  7. Enhancing disease resistances of Super Hybrid Rice with four antifungal genes

    Institute of Scientific and Technical Information of China (English)


    A plant expression vector harboring four antifungal genes was delivered into the embryogenic calli of ‘9311’, an indica restorer line of Super Hybrid Rice, via modified biolistic particle bombardment. Southern blot analysis indicated that in the regenerated hygromycin-resistant plants, all the four anti-fungal genes, including RCH10, RAC22, β-Glu and B-RIP, were integrated into the genome of ‘9311’, co-transmitted altogether with the marker gene hpt in a Mendelian pattern. Some transgenic R1 and R2 progenies, with all transgenes displaying a normal expression level in the Northern blot analysis, showed high resistance to Magnaporthe grisea when tested in the typical blast nurseries located in Yanxi and Sanya respectively. Furthermore, transgenic F1 plants, resulting from a cross of R2 homo-zygous lines with high resistance to rice blast with the non-transgenic male sterile line Peiai 64S, showed not only high resistance to M. grisea but also enhanced resistance to rice false smut (a disease caused by Ustilaginoidea virens) and rice kernel smut (another disease caused by Tilletia barclayana).

  8. Hybrid Airy Plasmons with Dynamically Steerable Trajectories

    CERN Document Server

    Li, Rujiang; Lin, Xiao; Wang, Huaping; Xu, Zhiwei; Chen, Hongsheng


    With the intriguing properties of diffraction-free, self-accelerating, and self-healing, Airy plasmons are promising to be used in the trapping, transporting, and sorting of micro-objects, imaging, and chip scale signal processing. However, the high dissipative loss and the lack of dynamical steerability restrict the implementation of Airy plasmons in these applications. Here we reveal the hybrid Airy plasmons for the first time by taking a hybrid graphene-based plasmonic waveguide in the terahertz (THz) domain as an example. Due to the coupling between an optical mode and a plasmonic mode, the hybrid Airy plasmons can have large propagation lengths and effective transverse deflections, where the transverse waveguide confinements are governed by the hybrid modes with moderate quality factors. Meanwhile, the propagation trajectories of hybrid Airy plasmons are dynamically steerable by changing the chemical potential of graphene. These hybrid Airy plasmons may promote the further discovery of non-diffracting be...

  9. Hybrid soft computing approaches research and applications

    CERN Document Server

    Dutta, Paramartha; Chakraborty, Susanta


    The book provides a platform for dealing with the flaws and failings of the soft computing paradigm through different manifestations. The different chapters highlight the necessity of the hybrid soft computing methodology in general with emphasis on several application perspectives in particular. Typical examples include (a) Study of Economic Load Dispatch by Various Hybrid Optimization Techniques, (b) An Application of Color Magnetic Resonance Brain Image Segmentation by ParaOptiMUSIG activation Function, (c) Hybrid Rough-PSO Approach in Remote Sensing Imagery Analysis,  (d) A Study and Analysis of Hybrid Intelligent Techniques for Breast Cancer Detection using Breast Thermograms, and (e) Hybridization of 2D-3D Images for Human Face Recognition. The elaborate findings of the chapters enhance the exhibition of the hybrid soft computing paradigm in the field of intelligent computing.

  10. A Hybrid Architecture Approach for Quantum Algorithms

    Directory of Open Access Journals (Sweden)

    Mohammad R.S. Aghaei


    Full Text Available Problem statement: In this study, a general plan of hybrid architecture for quantum algorithms is proposed. Approach: Analysis of the quantum algorithms shows that these algorithms were hybrid with two parts. First, the relationship of classical and quantum parts of the hybrid algorithms was extracted. Then a general plan of hybrid structure was designed. Results: This plan was illustrated the hybrid architecture and the relationship of classical and quantum parts of the algorithms. This general plan was used to increase implementation performance of quantum algorithms. Conclusion/Recommendations: Moreover, simulation results of quantum algorithms on the hybrid architecture proved that quantum algorithms can be implemented on the general plan as well.

  11. Hybrid Airy plasmons with dynamically steerable trajectories. (United States)

    Li, Rujiang; Imran, Muhammad; Lin, Xiao; Wang, Huaping; Xu, Zhiwei; Chen, Hongsheng


    With their intriguing diffraction-free, self-accelerating, and self-healing properties, Airy plasmons show promise for use in the trapping, transporting, and sorting of micro-objects, imaging, and chip scale signal processing. However, high dissipative loss and lack of dynamical steerability restrict the implementation of Airy plasmons in these applications. Here we reveal hybrid Airy plasmons for the first time by taking a hybrid graphene-based plasmonic waveguide in the terahertz (THz) domain as an example. Due to coupling between optical modes and plasmonic modes, the hybrid Airy plasmons can have large propagation lengths and effective transverse deflections, where the transverse waveguide confinements are governed by the hybrid modes with moderate quality factors. Meanwhile, the propagation trajectories of the hybrid Airy plasmons are dynamically steerable by changing the chemical potential of graphene. These hybrid Airy plasmons may promote the further discovery of non-diffracting beams along with the emerging developments of optical tweezers and tractor beams.

  12. Infection of great apes and a zoo keeper with the same Mycobacterium tuberculosis spoligotype

    NARCIS (Netherlands)

    Akkerman, O.W.; Werf, van de T.S.; Rietkerk, F.; Eger, A.; Soolingen, D.; Loo, v.d. K.; Zanden, v.d. A.G.M.


    An animal keeper was diagnosed with pulmonary tuberculosis (TB) after bi-annual screening for latent TB infection in zoo employees. In the same period, several bonobos of the zoo were suffering from TB as well. The Mycobacterium tuberculosis strains from both the animal keeper and the bonobos appear

  13. [Hybridization of crucian carp, Carassius carassius (Linnaeus, 1758), in Ukrainian reservoirs and genetic structure of hybrids]. (United States)

    Mezhzheryn, S V; Kokodyĭĭ, S V; Kulysh, A V; Verlat'iĭĭ, D B; Fedorenko, L V


    Hybridization of crucian carps Carassius carassius in polyspecific crucian populations of reservoirs of Ukraine and genetic structure of the hybrids were investigated using biochemical gene marking and cytometric procedure. The fact of wide hybridization between C. auratus and C. carassius was proved to be true by large number of hybrids which can form populations consisting only from hybrid individuals. Hybrids C. auratus x C. carassius were diploid, tryploid and in exceptional cases tetraploid; females and males which most likely breed by hybridogenesis. Besides, some clonal hybrids C. carassius x C. gibelio-1 appearing as tetraploid females, and one triploid female C. carassius x Tinca tinca were revealed. It is supported that hybridization of alien C. auratus with endemic C. carassius became one of mechanisms of replacement and depressions of populations of the last.

  14. Hybrid and Plug-in Electric Vehicles

    Energy Technology Data Exchange (ETDEWEB)



    Hybrid and plug-in electric vehicles use electricity either as their primary fuel or to improve the efficiency of conventional vehicle designs. This new generation of vehicles, often called electric drive vehicles, can be divided into three categories: hybrid electric vehicles (HEVs), plug-in hybrid electric vehicles(PHEVs), and all-electric vehicles (EVs). Together, they have great potential to reduce U.S. petroleum use.

  15. Hybrid synchronization of hyperchaotic Lu system

    Indian Academy of Sciences (India)

    K Sebastian Sudheer; M Sabir


    In this paper, we study the hybrid synchronization between two identical hyperchaotic Lu systems. Hybrid synchronization of hyperchaotic Lu system is achieved through synchronization of two pairs of states and anti-synchronization of the other two pairs of states. Active controls are designed to achieve hybrid synchronization between drive and response systems using the sum and difference of relevant variables of the chaotic systems. Numerical simulations are presented to evaluate the analysis and effectiveness of the controllers.

  16. Parallel Hybrid Vehicle Optimal Storage System (United States)

    Bloomfield, Aaron P.


    A paper reports the results of a Hybrid Diesel Vehicle Project focused on a parallel hybrid configuration suitable for diesel-powered, medium-sized, commercial vehicles commonly used for parcel delivery and shuttle buses, as the missions of these types of vehicles require frequent stops. During these stops, electric hybridization can effectively recover the vehicle's kinetic energy during the deceleration, store it onboard, and then use that energy to assist in the subsequent acceleration.

  17. On Hybrid and mixed finite element methods (United States)

    Pian, T. H. H.


    Three versions of the assumed stress hybrid model in finite element methods and the corresponding variational principles for the formulation are presented. Examples of rank deficiency for stiffness matrices by the hybrid stress model are given and their corresponding kinematic deformation modes are identified. A discussion of the derivation of general semi-Loof elements for plates and shells by the hybrid stress method is given. It is shown that the equilibrium model by Fraeijs de Veubeke can be derived by the approach of the hybrid stress model as a special case of semi-Loof elements.

  18. Consensus of Hybrid Multi-Agent Systems. (United States)

    Zheng, Yuanshi; Ma, Jingying; Wang, Long


    In this brief, we consider the consensus problem of hybrid multiagent systems. First, the hybrid multiagent system is proposed, which is composed of continuous-time and discrete-time dynamic agents. Then, three kinds of consensus protocols are presented for the hybrid multiagent system. The analysis tool developed in this brief is based on the matrix theory and graph theory. With different restrictions of the sampling period, some necessary and sufficient conditions are established for solving the consensus of the hybrid multiagent system. The consensus states are also obtained under different protocols. Finally, simulation examples are provided to demonstrate the effectiveness of our theoretical results.

  19. Hybrid slab-microchannel gel electrophoresis system (United States)

    Balch, Joseph W.; Carrano, Anthony V.; Davidson, James C.; Koo, Jackson C.


    A hybrid slab-microchannel gel electrophoresis system. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate.

  20. Nanostructured organic and hybrid solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Weickert, Jonas; Dunbar, Ricky B.; Hesse, Holger C.; Wiedemann, Wolfgang; Schmidt-Mende, Lukas [Department of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians University (LMU) Munich, Amalienstr. 54, 80799 Munich (Germany)


    This progress report highlights recent developments in nanostructured organic and hybrid solar cells. The authors discuss novel approaches to control the film morphology in fully organic solar cells and the design of nanostructured hybrid solar cells. The motivation and recent results concerning fabrication and effects on device physics are emphasized. The aim of this review is not to give a summary of all recent results in organic and hybrid solar cells, but rather to focus on the fabrication, device physics, and light trapping properties of nanostructured organic and hybrid devices. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  1. Development of a Hybrid Photopolymer for Stereolithography

    Institute of Scientific and Technical Information of China (English)


    New liquid free radical-cationic hybrid photopolymer, consisting of acrylate-based photocurable resin and epoxy-based photosensitive resin for stereolithography by UV laser was developed.The experiment results indicated that the hybrid photopolymer exhibits advantages of both the acrylate- based photosensitive resin and the epoxy- based photosensitive polymer contained in the hybrid system with relatively high photospeed and low linear shrinkage.Stereolithography parts without obvious distortion were built on the stereolithography apparatus HRPLAI from this hybrid resin successfully and efficiently.

  2. Cluster Tree Based Hybrid Document Similarity Measure

    Directory of Open Access Journals (Sweden)

    M. Varshana Devi


    Full Text Available hybrid similarity measure is established to measure the hybrid similarity. In cluster tree, the hybrid similarity measure can be calculated for the random data even it may not be the co-occurred and generate different views. Different views of tree can be combined and choose the one which is significant in cost. A method is proposed to combine the multiple views. Multiple views are represented by different distance measures into a single cluster. Comparing the cluster tree based hybrid similarity with the traditional statistical methods it gives the better feasibility for intelligent based search. It helps in improving the dimensionality reduction and semantic analysis.

  3. Hybrid Dynamical Systems Modeling, Stability, and Robustness

    CERN Document Server

    Goebel, Rafal; Teel, Andrew R


    Hybrid dynamical systems exhibit continuous and instantaneous changes, having features of continuous-time and discrete-time dynamical systems. Filled with a wealth of examples to illustrate concepts, this book presents a complete theory of robust asymptotic stability for hybrid dynamical systems that is applicable to the design of hybrid control algorithms--algorithms that feature logic, timers, or combinations of digital and analog components. With the tools of modern mathematical analysis, Hybrid Dynamical Systems unifies and generalizes earlier developments in continuous-time and discret

  4. Organic/inorganic hybrid coatings for anticorrosion (United States)

    He, Zhouying

    Compared to organic coatings, organic-inorganic hybrid coatings can potentially improve the anticorrosion performance. The organic phase provides the excellent mechaincal and barrier properties while the inorganic phase acts as an adhesion promoter and corrosion inhibitor. Despite that many studies on alkoxylsilane-based hybrid coatings have been developed and studied, their weatherability and anticorrosion performance has been rarely evaluated. On the other hand, organic-inorganic hybrid coatings based on mixed sol-gel precursors have received much less attention compared to alkoxylsilane-based hybrid coatings. In the first part, polyurethane hybrid coatings with a unique hybrid crosslinked structure as an improved unicoat were successfully prepared. The effect of polyesters on physical properties of the hybrid coatings was studied. Polyurethane coatings derived from cycloaliphatic polyester show comparable properties than those derived from the commercially viable aromatic polyester. Introducing the polysiloxane part into the polyurethane coatings enhanced the crosslinking density, Tg, mechanical properties, and general coating properties. The increased adhesion between the hybrid coating and the substrate make the hybrid coating a good candidate for anticorrosion application, which is shown by electrochemical impedance spectroscopy (EIS). The degradation mechanism of the polyurethane/polysiloxane hybrid coatings under various weathering conditions was shown to be the scission of the urethane and ester groups in the organic phase along with reorganizing and rearranging of the inorganic phase. The anticorrosion performance of the cycloaliphatic hybrid was much better than that of aromatic based hybrid under outdoor weathering based on visual observation and EIS analysis. Acid undercutting is an issue for TEOS based hybrid coating. In the second part, design of experiments (DOEs) was used to statistically investigate on the effect of sol-gel precursors. The

  5. Techno-economic comparison of series hybrid, plug-in hybrid, fuel cell and regular cars

    NARCIS (Netherlands)

    van Vliet, O.P.R.; Kruithof, T.; Turkenburg, W.C.; Faaij, A.P.C.


    We examine the competitiveness of series hybrid compared to fuel cell, parallel hybrid, and regular cars. We use public domain data to determine efficiency, fuel consumption, total costs of ownership and greenhouse gas emissions resulting from drivetrain choices. The series hybrid drivetrain can be


    Directory of Open Access Journals (Sweden)

    A. A. Lobaty


    Full Text Available This paper extends the recently introduced approach for modeling and solving the optimal control problem of fixedswitched mode DC-DC power converter. DCDC converters are a class of electric power circuits that used extensively in regulated DC power supplies, DC motor drives of different types, in Photovoltaic Station energy conversion and other applications due to its advantageous features in terms of size, weight and reliable performance. The main problem in controlling this type converters is in their hybrid nature as the switched circuit topology entails different modes of operation, each of it with its own associated linear continuous-time dynamics.This paper analyses the modeling and controller synthesis of the fixed-frequency buck DC-DC converter, in which the transistor switch is operated by a pulse sequence with constant frequency. In this case the regulation of the DC component of the output voltage is via the duty cycle. The optimization of the control system is based on the formation of the control signal at the output.It is proposed to solve the problem of optimal control of a hybrid system based on the formation of the control signal at the output of the controller, which minimizes a given functional integral quality, which is regarded as a linear quadratic Letov-Kalman functional. Search method of optimal control depends on the type of mathematical model of control object. In this case, we consider a linear deterministic model of the control system, which is common for the majority of hybrid electrical systems. For this formulation of the optimal control problem of search is a problem of analytical design of optimal controller, which has the analytical solution.As an example of the hybrid system is considered a step-down switching DC-DC converter, which is widely used in various electrical systems: as an uninterruptible power supply, battery charger for electric vehicles, the inverter in solar photovoltaic power plants.. A

  7. Hybrid-Source Impedance Networks

    DEFF Research Database (Denmark)

    Li, Ding; Gao, Feng; Loh, Poh Chiang;


    the cascaded networks would have a higher output voltage gain and other unique advantages that currently have not been investigated yet. It is anticipated that these advantages would help the formed inverters find applications in photovoltaic and other renewable systems, where a high voltage gain is usually......Hybrid-source impedance networks have attracted attention among researchers because of their flexibility in performing buck-boost energy conversion. To date, three distinct types of impedance networks can be summarized for implementing voltage-type inverters with another three types summarized...

  8. Essentialism, hybridism and cultural critique

    DEFF Research Database (Denmark)

    Frello, Birgitta


    Transgression concepts - such as ‘hybridity', ‘diaspora', ‘creolization', ‘transculturalization' and ‘syncretism' - have to an increasing extent become key concepts in various attempts at escaping the problems of suppression and exclusion involved in notions of purity - be it the purity of race......, culture or whatever. The purpose of focusing on concepts of transgression is usually to try to develop conceptual spaces within which it is possible to grasp and to study cultural identity without resorting to cultural essentialism. These attempts open up new possibilities but also new problems - e...

  9. Hydrostatic and hybrid bearing design

    CERN Document Server

    Rowe, W B


    Hydrostatic and Hybrid Bearing Design is a 15-chapter book that focuses on the bearing design and testing. This book first describes the application of hydrostatic bearings, as well as the device pressure, flow, force, power, and temperature. Subsequent chapters discuss the load and flow rate of thrust pads; circuit design, flow control, load, and stiffness; and the basis of the design procedures and selection of tolerances. The specific types of bearings, their design, dynamics, and experimental methods and testing are also shown. This book will be very valuable to students of engineering des

  10. Coordinated hybrid automatic repeat request

    KAUST Repository

    Makki, Behrooz


    We develop a coordinated hybrid automatic repeat request (HARQ) approach. With the proposed scheme, if a user message is correctly decoded in the first HARQ rounds, its spectrum is allocated to other users, to improve the network outage probability and the users\\' fairness. The results, which are obtained for single- and multiple-antenna setups, demonstrate the efficiency of the proposed approach in different conditions. For instance, with a maximum of M retransmissions and single transmit/receive antennas, the diversity gain of a user increases from M to (J+1)(M-1)+1 where J is the number of users helping that user.

  11. Development and functional analysis of novel genetic promoters using DNA shuffling, hybridization and a combination thereof.

    Directory of Open Access Journals (Sweden)

    Rajiv Ranjan

    Full Text Available BACKGROUND: Development of novel synthetic promoters with enhanced regulatory activity is of great value for a diverse range of plant biotechnology applications. METHODOLOGY: Using the Figwort mosaic virus full-length transcript promoter (F and the sub-genomic transcript promoter (FS sequences, we generated two single shuffled promoter libraries (LssF and LssFS, two multiple shuffled promoter libraries (LmsFS-F and LmsF-FS, two hybrid promoters (FuasFScp and FSuasFcp and two hybrid-shuffled promoter libraries (LhsFuasFScp and LhsFSuasFcp. Transient expression activities of approximately 50 shuffled promoter clones from each of these libraries were assayed in tobacco (Nicotiana tabacum cv. Xanthi protoplasts. It was observed that most of the shuffled promoters showed reduced activity compared to the two parent promoters (F and FS and the CaMV35S promoter. In silico studies (computer simulated analyses revealed that the reduced promoter activities of the shuffled promoters could be due to their higher helical stability. On the contrary, the hybrid promoters FuasFScp and FSuasFcp showed enhanced activities compared to F, FS and CaMV 35S in both transient and transgenic Nicotiana tabacum and Arabidopsis plants. Northern-blot and qRT-PCR data revealed a positive correlation between transcription and enzymatic activity in transgenic tobacco plants expressing hybrid promoters. Histochemical/X-gluc staining of whole transgenic seedlings/tissue-sections and fluorescence images of ImaGene Green™ treated roots and stems expressing the GUS reporter gene under the control of the FuasFScp and FSuasFcp promoters also support the above findings. Furthermore, protein extracts made from protoplasts expressing the human defensin (HNP-1 gene driven by hybrid promoters showed enhanced antibacterial activity compared to the CaMV35S promoter. SIGNIFICANCE/CONCLUSION: Both shuffled and hybrid promoters developed in the present study can be used as molecular tools to

  12. Hybrid polypeptides having cellobiohydrolase activity and polynucleotides encoding same (United States)

    Liu, Ye; Shaghasi, Tarana


    The present invention provides hybrid polypeptides having cellobiohydrolase activity. The present invention also provides polynucleotides encoding the hybrid polypeptides; nucleic acid constructs, vectors and host cells comprising the polynucleotides; and processes of using the hybrid polypeptides.

  13. Quantum technologies with hybrid systems. (United States)

    Kurizki, Gershon; Bertet, Patrice; Kubo, Yuimaru; Mølmer, Klaus; Petrosyan, David; Rabl, Peter; Schmiedmayer, Jörg


    An extensively pursued current direction of research in physics aims at the development of practical technologies that exploit the effects of quantum mechanics. As part of this ongoing effort, devices for quantum information processing, secure communication, and high-precision sensing are being implemented with diverse systems, ranging from photons, atoms, and spins to mesoscopic superconducting and nanomechanical structures. Their physical properties make some of these systems better suited than others for specific tasks; thus, photons are well suited for transmitting quantum information, weakly interacting spins can serve as long-lived quantum memories, and superconducting elements can rapidly process information encoded in their quantum states. A central goal of the envisaged quantum technologies is to develop devices that can simultaneously perform several of these tasks, namely, reliably store, process, and transmit quantum information. Hybrid quantum systems composed of different physical components with complementary functionalities may provide precisely such multitasking capabilities. This article reviews some of the driving theoretical ideas and first experimental realizations of hybrid quantum systems and the opportunities and challenges they present and offers a glance at the near- and long-term perspectives of this fascinating and rapidly expanding field.

  14. Nanofabrication of Hybrid Optoelectronic Devices (United States)

    Dibos, Alan Michael

    The material requirements for optoelectronic devices can vary dramatically depending on the application. Often disparate material systems need to be combined to allow for full device functionality. At the nanometer scale, this can often be challenging because of the inherent chemical and structural incompatibilities of nanofabrication. This dissertation concerns the integration of seemingly dissimilar materials into hybrid optoelectronic devices for photovoltaic, plasmonic, and photonic applications. First, we show that combining a single strip of conjugated polymer and inorganic nanowire can yield a nanoscale solar cell, and modeling of optical absorption and exciton diffusion in this device can provide insight into the efficiency of charge separation. Second, we use an on-chip nanowire light emitting diode to pump a colloidal quantum dot coupled to a silver waveguide. The resulting device is an electro-optic single plasmon source. Finally, we transfer diamond waveguides onto near-field avalanche photodiodes fabricated from GaAs. Embedded in the diamond waveguides are nitrogen vacancy color centers, and the mapping of emission from these single-photon sources is demonstrated using our on-chip detectors, eliminating the need for external photodetectors on an optical table. These studies show the promise of hybrid optoelectronic devices at the nanoscale with applications in alternative energy, optical communication, and quantum optics.

  15. Evaluating the Pedagogical Potential of Hybrid Models (United States)

    Levin, Tzur; Levin, Ilya


    The paper examines how the use of hybrid models--that consist of the interacting continuous and discrete processes--may assist in teaching system thinking. We report an experiment in which undergraduate students were asked to choose between a hybrid and a continuous solution for a number of control problems. A correlation has been found between…

  16. Hybrid Composite of Polyaniline Containing Carbon Nanotube

    Institute of Scientific and Technical Information of China (English)


    Carbon nanotube-polyaniline hybrid material was synthesized by emulsion polymerization in-situ. The morphology of hybrid material was studied by TEM and X-ray diffraction. The conductivity of nanocomposite increases with the increasing of carbon nanotube content because of the new conductivity passageways formed by carbon nanotubes.

  17. Computer code for intraply hybrid composite design (United States)

    Chamis, C. C.; Sinclair, J. H.


    A computer program has been developed and is described herein for intraply hybrid composite design (INHYD). The program includes several composite micromechanics theories, intraply hybrid composite theories and a hygrothermomechanical theory. These theories provide INHYD with considerable flexibility and capability which the user can exercise through several available options. Key features and capabilities of INHYD are illustrated through selected samples.

  18. Symmetry reduction for stochastic hybrid systems

    NARCIS (Netherlands)

    Bujorianu, L.M.; Katoen, J.P.


    This paper is focused on adapting symmetry reduction, a technique that is highly successful in traditional model checking, to stochastic hybrid systems. We first show that performability analysis of stochastic hybrid systems can be reduced to a stochastic reachability analysis (SRA). Then, we genera

  19. Symmetry Reduction For Stochastic Hybrid Systems

    NARCIS (Netherlands)

    Bujorianu, L.M.; Katoen, J.P.


    This paper is focused on adapting symmetry reduction, a technique that is highly successful in traditional model checking, to stochastic hybrid systems. To that end, we first show that performability analysis of stochastic hybrid systems can be reduced to a stochastic reachability analysis (SRA). Th

  20. Turbo Codes with Hybrid Interleaving Mode

    Institute of Scientific and Technical Information of China (English)


    In the investigation of turbo codes, either random interleavers or structured interleavers are used. Combined two kinds of interleavers in one turbo encoder, a hybrid interleaving mode is proposed in this paper. Computer simulations show that the performance of turbo codes with the hybrid interleaving mode is better than that with the typical interleaving mode.

  1. Hybrid reactors: Nuclear breeding or energy production?

    Energy Technology Data Exchange (ETDEWEB)

    Piera, Mireia [UNED, ETSII-Dp Ingenieria Energetica, c/Juan del Rosal 12, 28040 Madrid (Spain); Lafuente, Antonio; Abanades, Alberto; Martinez-Val, J.M. [ETSII-UPM, c/Jose Gutierrez Abascal 2, 28006 Madrid (Spain)


    After reviewing the long-standing tradition on hybrid research, an assessment model is presented in order to characterize the hybrid performance under different objectives. In hybrids, neutron multiplication in the subcritical blanket plays a major role, not only for energy production and nuclear breeding, but also for tritium breeding, which is fundamental requirement in fusion-fission hybrids. All three objectives are better achieved with high values of the neutron multiplication factor (k-eff) with the obvious and fundamental limitation that it cannot reach criticality under any event, particularly, in the case of a loss of coolant accident. This limitation will be very important in the selection of the coolant. Some general considerations will be proposed, as guidelines for assessing the hybrid potential in a given scenario. Those guidelines point out that hybrids can be of great interest for the future of nuclear energy in a framework of Sustainable Development, because they can contribute to the efficient exploitation of nuclear fuels, with very high safety features. Additionally, a proposal is presented on a blanket specially suited for fusion-fission hybrids, although this reactor concept is still under review, and new work is needed for identifying the most suitable blanket composition, which can vary depending on the main objective of the hybrid. (author)

  2. DSP Control of Line Hybrid Active Filter

    DEFF Research Database (Denmark)

    Dan, Stan George; Benjamin, Doniga Daniel; Magureanu, R.;


    Active Power Filters have been intensively explored in the past decade. Hybrid active filters inherit the efficiency of passive filters and the improved performance of active filters, and thus constitute a viable improved approach for harmonic compensation. In this paper a parallel hybrid filter ...

  3. Hybrid Therapy Regimen for Helicobacter Pylori Eradication

    Directory of Open Access Journals (Sweden)

    Zhi-Qiang Song


    Conclusions: Hybrid therapy showed wide differences in the efficacy but consistently good compliance and safety across different regions. Dual clarithromycin and metronidazole resistance were the key factor to efficacy. Hybrid therapy was similar to sequential or concomitant therapy in the efficacy, safety, and compliance.

  4. Hybrid Compounding in New Zealand English (United States)

    Degani, Marta; Onysko, Alexander


    This study investigates hybrid compound formation of Maori and English terms in present day New Zealand English (NZE). On the background of Maori and English language contact, the phenomenon of hybrid compounding emerges as a process that, on the one hand, symbolizes the vitality of the Maori element in NZE and, on the other hand, marks the…

  5. (Hybrid) Baryons Quantum Numbers and Adiabatic Potentials

    CERN Document Server

    Page, P R


    We construct (hybrid) baryons in the flux-tube model of Isgur and Paton. In the limit of adiabatic quark motion, we build proper eigenstates of orbital angular momentum and indicate the flavour, spin, chirality and J^P of (hybrid) baryons. The adiabatic potential is calculated as a function of the quark positions.

  6. Design Principles for Natural and Hybrid Ventilation

    DEFF Research Database (Denmark)

    Heiselberg, Per

    For many years mechanical and natural ventilation systems have developed separately. Naturally, the next step in this development is the development of ventilation concepts that utilize and combine the best features from each system to create a new type of ventilation system- Hybrid Ventilation....... The hybrid ventilation concepts, design challenges and principles are discussed and illustrated by four building examples....

  7. Development of hybrid rice in China

    Institute of Scientific and Technical Information of China (English)

    Yuan Longping; Wu Xiaojin


    @@ Introduction The success achieved in development of hybrid rice isa great breakthrough in rice breeding which provides aneffective way to markedly enhance rice yield on a largescale. China is the first country in the world to exploit riceheterosis commercially. Research on hybrid rice was initiatedin 1964.

  8. Hybrid logics with infinitary proof systems

    NARCIS (Netherlands)

    Kooi, Barteld; Renardel de Lavalette, Gerard; Verbrugge, Rineke


    We provide a strongly complete infinitary proof system for hybrid logic. This proof system can be extended with countably many sequents. Thus, although these logics may be non-compact, strong completeness proofs are provided for infinitary hybrid versions of non-compact logics like ancestral logic a

  9. On Fractional Order Hybrid Differential Equations

    Directory of Open Access Journals (Sweden)

    Mohamed A. E. Herzallah


    Full Text Available We develop the theory of fractional hybrid differential equations with linear and nonlinear perturbations involving the Caputo fractional derivative of order 0<α<1. Using some fixed point theorems we prove the existence of mild solutions for two types of hybrid equations. Examples are given to illustrate the obtained results.

  10. Statistical Model Checking for Stochastic Hybrid Systems

    DEFF Research Database (Denmark)

    David, Alexandre; Du, Dehui; Larsen, Kim Guldstrand


    This paper presents novel extensions and applications of the UPPAAL-SMC model checker. The extensions allow for statistical model checking of stochastic hybrid systems. We show how our race-based stochastic semantics extends to networks of hybrid systems, and indicate the integration technique ap...

  11. Harmonious Unifying Hybrid Preferential Supernetwork Model

    Institute of Scientific and Technical Information of China (English)

    LIU; Qiang; FANG; Jin-qing; LI; Yong


    The basic concepts and methods for harmonious unifying hybrid preferential model(HUHPM)are based on random preferential attachment(RPA)mixed with deterministic preferential attachment(DPA),so there is only one unified hybrid ratio dr,which is defined as:

  12. Hybrid2: The hybrid system simulation model, Version 1.0, user manual

    Energy Technology Data Exchange (ETDEWEB)

    Baring-Gould, E.I.


    In light of the large scale desire for energy in remote communities, especially in the developing world, the need for a detailed long term performance prediction model for hybrid power systems was seen. To meet these ends, engineers from the National Renewable Energy Laboratory (NREL) and the University of Massachusetts (UMass) have spent the last three years developing the Hybrid2 software. The Hybrid2 code provides a means to conduct long term, detailed simulations of the performance of a large array of hybrid power systems. This work acts as an introduction and users manual to the Hybrid2 software. The manual describes the Hybrid2 code, what is included with the software and instructs the user on the structure of the code. The manual also describes some of the major features of the Hybrid2 code as well as how to create projects and run hybrid system simulations. The Hybrid2 code test program is also discussed. Although every attempt has been made to make the Hybrid2 code easy to understand and use, this manual will allow many organizations to consider the long term advantages of using hybrid power systems instead of conventional petroleum based systems for remote power generation.

  13. SCAN-based hybrid and double-hybrid density functionals from parameter-free models

    CERN Document Server

    Hui, Kerwin


    By incorporating the nonempirical SCAN semilocal density functional [Sun, Ruzsinszky, and Perdew, Phys. Rev. Lett. 115, 036402 (2015)] in the underlying expression, we propose one hybrid (SCAN0) and three double-hybrid (SCAN0-DH, SCAN-QIDH, and SCAN0-2) density functionals, which are free of any empirical parameter. The SCAN-based hybrid and double-hybrid functionals consistently outperform their parent SCAN semilocal functional for a wide range of applications. The SCAN-based semilocal, hybrid, and double-hybrid functionals generally perform better than the corresponding PBE-based functionals. In addition, the SCAN0-2 and SCAN-QIDH double-hybrid functionals significantly reduce the qualitative failures of the SCAN semilocal functional, such as the self-interaction error and noncovalent interaction error, extending the applicability of the SCAN-based functionals to a very diverse range of systems.

  14. A comprehensive overview of hybrid construction machinery

    Directory of Open Access Journals (Sweden)

    Jixin Wang


    Full Text Available With the increasing attention of energy saving and emission reduction technology, the recent application of hybrid powertrain technology affects the development of construction machinery industry. This article reviews these publications and provides comprehensive references. This article reviews the state-of-art for the hybrid wheel loader and excavator, which focuses on powertrain configuration, energy storage devices, and energy management strategies. The basis of classification and characteristic of each powertrain configuration are described. Advantages and disadvantages of batteries, supercapacitors, hydraulic accumulators, and flywheel used in hybrid construction machinery are summarized. The existing energy management strategies for hybrid construction machinery are also elaborated. The technological challenges and developing trends in the near future for hybrid construction machinery are discussed.

  15. Mechanical property characterization of intraply hybrid composites (United States)

    Chamis, C. C.; Lark, R. F.; Sinclair, J. H.


    An investigation of the mechanical properties of intraply hybrids made from graphite fiber/epoxy matrix hybridized with secondary S-glass or Kevlar 49 fiber composites is presented. The specimen stress-strain behavior was determined, showing that mechanical properties of intraply hybrid composites can be measured with available methods such as the ten-degree off-axis test for intralaminar shear, and conventional tests for tensile, flexure, and Izod impact properties. The results also showed that combinations of high modulus graphite/S-glass/epoxy matrix composites exist which yield intraply hybrid laminates with the best 'balanced' properties, and that the translation efficiency of mechanical properties from the constituent composites to intraply hybrids may be assessed with a simple equation.

  16. Programmed cell death and hybrid incompatibility. (United States)

    Frank, S A; Barr, C M


    We propose a new theory to explain developmental aberrations in plant hybrids. In our theory, hybrid incompatibilities arise from imbalances in the mechanisms that cause male sterility in hermaphroditic plants. Mitochondria often cause male sterility by killing the tapetal tissue that nurtures pollen mother cells. Recent evidence suggests that mitochondria destroy the tapetum by triggering standard pathways of programmed cell death. Some nuclear genotypes repress mitochondrial male sterility and restore pollen fertility. Normal regulation of tapetal development therefore arises from a delicate balance between the disruptive effects of mitochondria and the defensive countermeasures of the nuclear genes. In hybrids, incompatibilities between male-sterile mitochondria and nuclear restorers may frequently upset the regulatory control of programmed cell death, causing tapetal abnormalities and male sterility. We propose that hybrid misregulation of programmed cell death may also spill over into other tissues, explaining various developmental aberrations observed in hybrids.

  17. Hybrid sterility in plant: stories from rice. (United States)

    Ouyang, Yidan; Liu, Yao-Guang; Zhang, Qifa


    Hybrid sterility is the most common form of postzygotic reproductive isolation in plants. The best-known example is perhaps the hybrid sterility between indica and japonica subspecies of Asian cultivated rice (Oryza sativa L.). Major progress has been reported recently in rice in identifying and cloning hybrid sterility genes at two loci regulating female and male fertility, respectively. Genetic analyses and molecular characterization of these genes, together with the results from other model organisms especially Drosophila, have advanced the understanding of the processes underlying reproductive isolation and speciation. These findings also have significant implications for crop genetic improvement, by providing the feasibility and strategies for overcoming intersubspecific hybrid sterility thus allowing the development of intersubspecific hybrids.

  18. Hybrid percolation transition in complex networks (United States)

    Kahng, Byungnam

    Percolation has been one of the most applied statistical models. Percolation transition is one of the most robust continuous transitions known thus far. However, recent extensive researches reveal that it exhibits diverse types of phase transitions such as discontinuous and hybrid phase transitions. Here hybrid phase transition means the phase transition exhibiting natures of both continuous and discontinuous phase transitions simultaneously. Examples include k-core percolation, cascading failures in interdependent networks, synchronization, etc. Thus far, it is not manifest if the critical behavior of hybrid percolation transitions conforms to the conventional scaling laws of second-order phase transition. Here, we investigate the critical behaviors of hybrid percolation transitions in the cascading failure model in inter-dependent networks and the restricted Erdos-Renyi model. We find that the critical behaviors of the hybrid percolation transitions contain some features that cannot be described by the conventional theory of second-order percolation transitions.

  19. Stochastic Reachability Analysis of Hybrid Systems

    CERN Document Server

    Bujorianu, Luminita Manuela


    Stochastic reachability analysis (SRA) is a method of analyzing the behavior of control systems which mix discrete and continuous dynamics. For probabilistic discrete systems it has been shown to be a practical verification method but for stochastic hybrid systems it can be rather more. As a verification technique SRA can assess the safety and performance of, for example, autonomous systems, robot and aircraft path planning and multi-agent coordination but it can also be used for the adaptive control of such systems. Stochastic Reachability Analysis of Hybrid Systems is a self-contained and accessible introduction to this novel topic in the analysis and development of stochastic hybrid systems. Beginning with the relevant aspects of Markov models and introducing stochastic hybrid systems, the book then moves on to coverage of reachability analysis for stochastic hybrid systems. Following this build up, the core of the text first formally defines the concept of reachability in the stochastic framework and then...

  20. The role of epigenetics in hybrid vigour. (United States)

    Groszmann, Michael; Greaves, Ian K; Fujimoto, Ryo; Peacock, W James; Dennis, Elizabeth S


    Hybrid vigour, or heterosis, refers to the increased yield and biomass of hybrid offspring relative to the parents. Although this has been exploited in plants for agriculture and horticulture, the molecular and cellular mechanisms underlying hybrid vigour are largely unknown. Genetic analyses show that there are a large number of quantitative trait loci (QTLs) that contribute to the heterotic phenotype, indicating that it is a complex phenomenon. Gene expression in hybrids is regulated by the interactions of the two parental epigenetic systems and the underlying genomes. Increasing understanding of the interplay of small RNA (sRNA) molecules, DNA methylation, and histone marks provides new opportunities to define the basis of hybrid vigour and to understand why F1 heterosis is not passed on to subsequent generations. We discuss recent findings that suggest the existence of several pathways that alter DNA methylation patterns, which may lead to transcriptional changes resulting in the heterotic phenotype.