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Sample records for chimerism analysis post

  1. Post-transplant monitoring of chimerism by lineage-specific analysis.

    PubMed

    Preuner, Sandra; Lion, Thomas

    2014-01-01

    Molecular surveillance of hematopoietic chimerism is an important part of the routine diagnostic program in patients after allogeneic stem cell transplantation. Chimerism testing permits early prediction and documentation of successful engraftment and facilitates early risk assessment of impending graft rejection. In patients transplanted for treatment of malignant hematologic disorders, monitoring of chimerism can provide an early indication of incipient disease relapse. The investigation of chimerism has therefore become an indispensable tool for the management of patients during the post-transplant period. Increasing use of reduced-intensity conditioning, which is associated with prolonged duration of mixed hematopoietic chimerism, has further increased the clinical importance of chimerism analysis. At present, the most commonly used technical approach to the investigation of chimerism is microsatellite analysis by polymerase chain reaction. The investigation of chimerism within specific leukocyte subsets isolated from peripheral blood or bone marrow samples by flow sorting- or magnetic bead-based techniques provides more specific information on processes underlying the dynamics of donor/recipient chimerism. Moreover, cell subset-specific analysis permits the assessment of impending complications at a significantly higher sensitivity, thus providing a basis for earlier treatment decisions.

  2. Chimerism analysis following nonmyeloablative stem cell transplantation.

    PubMed

    Lion, Thomas; Watzinger, Franz

    2006-01-01

    Molecular monitoring of hematopoietic chimerism has become a routine diagnostic approach in patients after allogeneic stem cell transplantation. Chimerism testing permits the documentation and surveillance of engraftment and facilitates early detection of impending graft rejection. In patients transplanted for treatment of malignant hematological disorders, monitoring of chimerism can provide an early indication of incipient disease relapse. The investigation of chimerism has therefore become an indispensable tool for the management of patients during the posttransplant period. Growing use of nonmyeloablative conditioning, which is associated with prolonged duration of mixed hematopoietic chimerism, has further increased the clinical importance of chimerism analysis. At present, the most commonly used technical approaches to the investigation of chimerism include microsatellite analysis by polymerase chain reaction and, in the gender-mismatched transplant setting, fluorescence in situ hybridization analysis of sex chromosomes. The investigation of chimerism within specific leukocyte subsets isolated from peripheral blood or bone marrow samples by flow-sorting or magnetic bead-based techniques provides more specific information on processes underlying the dynamics of donor/recipient chimerism. Moreover, cell subset-specific analysis permits the assessment of impending complications at a significantly higher sensitivity, thus providing a basis for earlier treatment decisions.

  3. Frontiers of stem cell transplantation monitoring: capturing graft dynamics through routine longitudinal chimerism analysis.

    PubMed

    Kristt, Don; Stein, Jerry; Klein, Tirza

    2007-03-01

    Quantitative chimerism testing has become an indispensable tool for following the course and success of allogeneic hematopoietic stem cell transplants. In this paper, we describe the current laboratory approach to quantitative chimerism testing based on an analysis of short tandem repeats, and explain why performing this analysis longitudinally is important and feasible. Longitudinal analysis focuses on relative changes appearing in the course of sequential samples, and as such exploits the ultimate potential of this intrinsically semi-quantitative platform. Such an analysis is more informative than single static values, less likely to be confused with platform artifacts, and is individualized to the particular patient. It is particularly useful with non-myeloablative conditioning, where mixed chimerism is common. When longitudinal chimerism analysis is performed on lineage-specific subpopulations, the sensitivity, specificity and mechanistic implications of the data are augmented. Importantly, longitudinal monitoring is a routinely feasible laboratory option because multiplex STR-PCR kits are available commercially, and modern software can be used to perform computation, reliability testing, and longitudinal tracking in a rapid, easy to use format. The ChimerTrack application, a shareware program developed in our laboratory for this purpose, produces a report that automatically summarizes and illustrates the quantitative temporal course of the patient's chimeric status. Such a longitudinal perspective enhances the value of quantitative chimerism monitoring for decisions regarding immunomodulatory post-transplant therapy. This information also provides unique insights into the biological dynamics of engraftment underlying the fluctuations in the temporal course of a patient's chimeric status.

  4. [Development of multiplex short tandem repeat (STR)-PCR for chimerism analysis in patients with hematological malignancies and comparison of chimerism in different sample sources].

    PubMed

    Taira, Chiaki; Matsuda, Kazuyuki; Takezawa, Yuka; Ito, Toshiro; Ishida, Fumihiro; Hidaka, Eiko; Kumagai, Toshiko; Honda, Takayuki

    2011-01-01

    Polymerase chain reaction analysis of short-tandem repeat (STR) markers (STR-PCR) has been used for chimerism testing to assess engraftment following hematopoietic stem cell transplantation (HSCT). We investigated the informativity of 7 STR loci (FGA, D5S818, SE33, TH01, VWF, PentaE, and D18S51) in 82 pre-HSCT DNA samples from 41 donor/recipient pairs and developed 2 multiplex STR-PCRs using VWF, SE33, and D18S51, D5S818 and FGA, respectively. The multiplex STR-PCRs could distinguish the recipients and donors in 92.7% of the cases. Dilution experiments using mixed DNA showed that the sensitivity of the multiplex STR-PCRs for detecting the minor population was 1-5%. To compare chimerism in different samples such as peripheral blood, mononuclear cells (MNC), and CD3-positive cells (CD3+), we investigated the relationship between the chimerisms at approximately day 30 post-HSCT and the interval from the day of HSCT to achievement of complete chimerism (CC) in 70 patients undergoing HSCT. CC was found in all samples of 54 patients at day 30 post-HSCT, and these samples showed CC thereafter. Eleven patients with mixed chimerism (MC) in all samples or in MNC and CD3+ showed CC at day 60-270 post-HSCT or persistent MC. The remaining 5 patients with MC in only CD3+ showed CC at day 30-60 post-HSCT. Taken together, MNC which can be separated easily may be a useful source for detecting patients who require longer time to achieve CC and those with high risk of graft failure.

  5. The EuroChimerism concept for a standardized approach to chimerism analysis after allogeneic stem cell transplantation.

    PubMed

    Lion, T; Watzinger, F; Preuner, S; Kreyenberg, H; Tilanus, M; de Weger, R; van Loon, J; de Vries, L; Cavé, H; Acquaviva, C; Lawler, M; Crampe, M; Serra, A; Saglio, B; Colnaghi, F; Biondi, A; van Dongen, J J M; van der Burg, M; Gonzalez, M; Alcoceba, M; Barbany, G; Hermanson, M; Roosnek, E; Steward, C; Harvey, J; Frommlet, F; Bader, P

    2012-08-01

    Hematopoietic stem cell transplantation is becoming an increasingly important approach to treatment of different malignant and non-malignant disorders. There is thus growing demand for diagnostic assays permitting the surveillance of donor/recipient chimerism posttransplant. Current techniques are heterogeneous, rendering uniform evaluation and comparison of diagnostic results between centers difficult. Leading laboratories from 10 European countries have therefore performed a collaborative study supported by a European grant, the EuroChimerism Concerted Action, with the aim to develop a standardized diagnostic methodology for the detection and monitoring of chimerism in patients undergoing allogeneic stem cell transplantation. Following extensive analysis of a large set of microsatellite/short tandem repeat (STR) loci, the EuroChimerism (EUC) panel comprising 13 STR markers was established with the aim to optimally meet the specific requirements of quantitative chimerism analysis. Based on highly stringent selection criteria, the EUC panel provides multiple informative markers in any transplant setting. The standardized STR-PCR tests permit detection of donor- or recipient-derived cells at a sensitivity ranging between 0.8 and 1.6%. Moreover, the EUC assay facilitates accurate and reproducible quantification of donor and recipient hematopoietic cells. Wide use of the European-harmonized protocol for chimerism analysis presented will provide a basis for optimal diagnostic support and timely treatment decisions.

  6. Detection of impending graft rejection and relapse by lineage-specific chimerism analysis.

    PubMed

    Lion, Thomas

    2007-01-01

    Molecular surveillance of hematopoietic chimerism has become part of the routine diagnostic program in patients after allogeneic stem cell transplantation. Chimerism testing permits early prediction and documentation of successful engraftment, and facilitates early detection of impending graft rejection. In patients transplanted for treatment of malignant hematological disorders, monitoring of chimerism can provide an early indication of incipient disease relapse. The investigation of chimerism has therefore become an indispensable tool for the management of patients during the posttransplant period. Growing use of nonmyeloablative conditioning, which is associated with prolonged duration of mixed hematopoietic chimerism, has further increased the clinical importance of chimerism analysis. At present, the most commonly used technical approach to the investigation of chimerism is microsatellite analysis by PCR. The investigation of chimerism within specific leukocyte subsets isolated from peripheral blood or bone marrow samples by flow-sorting or magnetic beads-based techniques provides more specific information on processes underlying the dynamics of donor/recipient chimerism. Moreover, cell subset-specific analysis permits the assessment of impending complications at a significantly higher sensitivity, thus providing a basis for earlier treatment decisions.

  7. Evaluation of immunomodulatory treatment based on conventional and lineage-specific chimerism analysis in patients with myeloid malignancies after myeloablative allogeneic hematopoietic cell transplantation.

    PubMed

    Zeiser, R; Spyridonidis, A; Wäsch, R; Ihorst, G; Grüllich, C; Bertz, H; Finke, J

    2005-05-01

    Both conventional chimerism analysis (CCA) and lineage-specific chimerism analysis (LCA) have potential pitfalls as diagnostic means for the detection of minimal residual disease after allogeneic hematopoietic cell transplantation (aHCT). Therefore, the present study examines the results of both methods in order to determine how predictive consecutive evaluations were, with respect to the risk that the patient would relapse during post-transplant follow-up and with respect to responsiveness to immunomodulatory treatment. A total of 168 individuals with acute myeloid leukemia (AML) (n = 137) and myelo dysplastic syndrome (n = 31) were investigated with CCA and LCA at mean intervals of 24 days (range: 11-116). The median follow-up after myeloablative aHCT was 22 months (range: 4-49). Of 168 patients, 65 experienced a clinical relapse after aHCT. CCA and LCA were comparatively sensitive and specific for relapse at the intervals of chimerism testing employed in this study. Of 32 patients, 10 who were offered donor lymphocyte infusions (DLI) treatment for increasing (n = 29) or stable (n = 3) mixed chimerism (MC) achieved at least transitory CC. The observation that all patients with increasing MC relapsed despite DLI treatment (54%) or withdrawal of immune suppression (24%) indicates that novel strategies to deal with rapidly evolving relapse in AML patients, such as shortening of chimerism monitoring intervals, need to be evaluated.

  8. Systematic analysis of stutters to enhance the accuracy of chimerism testing.

    PubMed

    Chen, Ding-Ping; Tseng, Ching-Ping; Tsai, Shu-Hui; Wu, Tsu-Lan; Chang, Pi-Yueh; Sun, Chien-Feng

    2008-01-01

    Post-transplantation chimerism testing is important to monitor the engraftment of donor stem cells and for the diagnosis of relapse. Detecting the presence of donor/recipient-specific short tandem repeats (STRs) is a frequently used method for engraftment study. Unfortunately, the interpretation of the STR-based chimerism tests is often subject to interference by the presence of a stutter peak, which is one 4-base repeat unit smaller than an authentic allele. The aim of this study was to systematically analyze and resolve the effect of stutter peaks on the interpretation of STR-based chimerism tests. The AmpFlSTR Identifiler Amplification kit (Applied Biosystems)was used to amplify 15 STR loci using genomic DNA from 30 randomly selected, healthy donors. We found that the stutter peaks had locus-specific characteristics. The stutter percentage was defined as the percentage of the stutter peak area/main STR peak area. Based on mean values for the 30 DNA samples, the stutter percentage varied from locus to locus and ranged from 3.12% to 10.71% for 15 STR loci. The locus-specific stutter effect can be eliminated through appropriately adjusted equations. The usefulness of these equations in the prediction of relapse was confirmed by the 5% sensitivity test. Hence, this report offers a valuable scheme to enhance the accuracy of chimerism testing.

  9. Platelet chimerism by polymerase chain reaction (PCR) utilizing variable number of tandem repeats (VNTR) in allogeneic stem cell transplant in children: a new novel approach to full chimerism analysis.

    PubMed

    Chou, P M; Olszewski, M; Huang, W; Silva, M; Kletzel, M

    2003-10-01

    Evaluation of chimerism following allogeneic transplantation has been performed traditionally focusing on two cellular compartments, namely lymphoid and myeloid. However, none has been described so far to evaluate platelet chimerism. In order to achieve full chimerism in all three cellular compartments, we prospectively obtained 138 samples of peripheral blood in 55 patients at different post transplant periods following allogeneic hematopoietic transplantation. Evaluation of chimerism was performed utilizing tests of variable number of tandem repeat (VNTR) and sex determination by quantitative polymerase chain reaction (PCR). Tests for platelet chimerism using platelet-rich plasma were simultaneously analyzed with samples for T-cell lymphoid and myeloid compartments. Complete donor chimerism was noted in 49 of 55 patients (89%), while the remaining six have split chimerism ranging from 34 to 98%. There is significant difference (P=0.0004) between the percentages of donor DNA in all three cellular compartments comparing the means+/-s.e.m. (myeloid 95.60+/-0.9, T-cell lymphocytes 87.6+/-1.9, and the platelets 90.8+/-1.5); however, comparison between the medians is not statistically significant. This study represents an additional step towards achieving full chimerism and the observation may help reduce the number of unnecessary platelet transfusions once chimerism is noted in that cellular compartment.

  10. Structure-Function Analysis of Peroxisomal ATP-binding Cassette Transporters Using Chimeric Dimers*

    PubMed Central

    Geillon, Flore; Gondcaille, Catherine; Charbonnier, Soëli; Van Roermund, Carlo W.; Lopez, Tatiana E.; Dias, Alexandre M. M.; Pais de Barros, Jean-Paul; Arnould, Christine; Wanders, Ronald J.; Trompier, Doriane; Savary, Stéphane

    2014-01-01

    ABCD1 and ABCD2 are two closely related ATP-binding cassette half-transporters predicted to homodimerize and form peroxisomal importers for fatty acyl-CoAs. Available evidence has shown that ABCD1 and ABCD2 display a distinct but overlapping substrate specificity, although much remains to be learned in this respect as well as in their capability to form functional heterodimers. Using a cell model expressing an ABCD2-EGFP fusion protein, we first demonstrated by proximity ligation assay and co-immunoprecipitation assay that ABCD1 interacts with ABCD2. Next, we tested in the pxa1/pxa2Δ yeast mutant the functionality of ABCD1/ABCD2 dimers by expressing chimeric proteins mimicking homo- or heterodimers. For further structure-function analysis of ABCD1/ABCD2 dimers, we expressed chimeric dimers fused to enhanced GFP in human skin fibroblasts of X-linked adrenoleukodystrophy patients. These cells are devoid of ABCD1 and accumulate very long-chain fatty acids (C26:0 and C26:1). We checked that the chimeric proteins were correctly expressed and targeted to the peroxisomes. Very long-chain fatty acid levels were partially restored in transfected X-linked adrenoleukodystrophy fibroblasts regardless of the chimeric construct used, thus demonstrating functionality of both homo- and heterodimers. Interestingly, the level of C24:6 n-3, the immediate precursor of docosahexaenoic acid, was decreased in cells expressing chimeric proteins containing at least one ABCD2 moiety. Our data demonstrate for the first time that both homo- and heterodimers of ABCD1 and ABCD2 are functionally active. Interestingly, the role of ABCD2 (in homo- and heterodimeric forms) in the metabolism of polyunsaturated fatty acids is clearly evidenced, and the chimeric dimers provide a novel tool to study substrate specificity of peroxisomal ATP-binding cassette transporters. PMID:25043761

  11. Assessment of the purity of isolated cell populations for lineage-specific chimerism monitoring post haematopoietic stem cell transplantation.

    PubMed

    Hanson, V; Adams, B; Lord, J; Barker, A; Poulton, K; Lee, H

    2013-10-01

    Following haematopoietic stem cell transplantation, monitoring the proportion of donor and recipient haematopoiesis in the patient (chimerism) is an influential tool in directing further treatment choices. Short tandem repeat (STR) analysis is a method of chimerism monitoring using DNA isolated from peripheral blood, bone marrow or specific isolated cell lineages such as CD3+ T cells. For lineage-specific STR analysis on cell populations isolated from peripheral blood, a qualitative estimation of the purity of each isolated population is essential for the correct interpretation of the test data. We describe a rapid, inexpensive method for the determination of purity using a simple flow cytometry method. The method described for assessing the purity of sorted CD3+ cells can be applied to any cell population isolated using the same technology. Data obtained were comparable to results from a commercial polymerase chain reaction (PCR)-based method for the assessment of purity (Non-T Genomic Detection Kit, Accumol, Calgary, AB, Canada) (P = 0.59). Of the 303 samples tested by flow cytometry, 290 (95.7%) exceeded 90% purity, and 215 (70.95%) were over 99% pure. There were some outlying samples, showing diversity between samples and the unpredictability of purity of isolated cell populations. This flow cytometry method can be easily assimilated into routine testing protocols, allowing purity assessment in multiple-sorted cell populations for lineage-specific chimerism monitoring using a single secondary antibody and giving results comparable to a PCR-based method. As purity of isolated cell lineages is affected by time after venepuncture and storage temperature, assessment of each sample is recommended to give a reliable indication of sample quality and confidence in the interpretation of the results.

  12. Standardisation of multiplex fluorescent short tandem repeat analysis for chimerism testing.

    PubMed

    Nollet, F; Billiet, J; Selleslag, D; Criel, A

    2001-09-01

    To evaluate the origin of cells after allogeneic haematopoietic stem cell transplantation we optimised and evaluated two commercially available systems (AmpFlSTR Profiler Plus and GenePrint Powerplex-16) which are based on multiplex fluorescent short tandem repeat (STR) analysis. A standard procedure for interpretation of electropherographs was found essential to obtain reproducible results. On the basis of the relative length of donor and recipient alleles, TYPE-I (no shared alleles are used to calculate chimerism), TYPE-II (one shared and one unshared allele is used to calculate chimerism) or TYPE-III (not informative) allelic distribution types were distinguished. Also, stutter peaks were recognised as an important criterion to exclude a marker for analysis. Intralaboratory and multicentre evaluation of the AmpFlSTR Profiler Plus system showed that mixed blood samples could be determined with an absolute deviation of less than 2%. A sensitivity threshold was set at 5% for TYPE-I and 10% for TYPE-II markers since relative imprecision increases at low chimerism values. No significant difference of calculated chimerism values was observed between STR markers shared between both systems. By monitoring 26 allogeneic peripheral blood stem cell transplants, the applicability of the proposed method was demonstrated.

  13. Pre-freeze and post-thaw characteristics on chimerism patterns in double-unit cord blood transplantation.

    PubMed

    Jaing, Tang-Her; Tsay, Pei-Kwei; Lin, Tung-Liang; Yang, Chao-Ping; Hung, Iou-Jih; Wen, Yu-Chuan

    2009-04-01

    We analyzed the pre-freeze and post-thaw characteristics on chimerism patterns in 20 cases of double-unit cord blood transplantation. The cord blood units (CBUs) were a 4/6 HLA match or better with recipients and achieved a minimum combined precryo-preservation cell dose of 3.7 x 10(7) total nucleated cell (TNC)/kg. The unit with a higher cell dose was infused first. All evaluable patients engrafted at a median of 18 days. By day 42, neutrophil engraftment was derived from both donors in 63% of cases and a single donor in 37% of patients. By day 100, one unit predominated in 80% of the patients. Higher pre-freeze TNC and CD34+ cell doses were associated with cord predominance in 67% of patients.

  14. Transcriptome analysis revealed chimeric RNAs, single nucleotide polymorphisms and allele-specific expression in porcine prenatal skeletal muscle

    PubMed Central

    Yang, Yalan; Tang, Zhonglin; Fan, Xinhao; Xu, Kui; Mu, Yulian; Zhou, Rong; Li, Kui

    2016-01-01

    Prenatal skeletal muscle development genetically determines postnatal muscle characteristics such as growth and meat quality in pigs. However, the molecular mechanisms underlying prenatal skeletal muscle development remain unclear. Here, we performed the first genome-wide analysis of chimeric RNAs, single nuclear polymorphisms (SNPs) and allele-specific expression (ASE) in prenatal skeletal muscle in pigs. We identified 14,810 protein coding genes and 163 high-confidence chimeric RNAs expressed in prenatal skeletal muscle. More than 94.5% of the chimeric RNAs obeyed the canonical GT/AG splice rule and were trans-splicing events. Ten and two RNAs were aligned to human and mouse chimeric transcripts, respectively. We detected 106,457 high-quality SNPs (6,955 novel), which were mostly (89.09%) located within QTLs for production traits. The high proportion of non-exonic SNPs revealed the incomplete annotation status of the current swine reference genome. ASE analysis revealed that 11,300 heterozygous SNPs showed allelic imbalance, whereas 131 ASE variants were located in the chimeric RNAs. Moreover, 4 ASE variants were associated with various economically relevant traits of pigs. Taken together, our data provide a source for studies of chimeric RNAs and biomarkers for pig breeding, while illuminating the complex transcriptional events underlying prenatal skeletal muscle development in mammals. PMID:27352850

  15. Transcriptome analysis revealed chimeric RNAs, single nucleotide polymorphisms and allele-specific expression in porcine prenatal skeletal muscle.

    PubMed

    Yang, Yalan; Tang, Zhonglin; Fan, Xinhao; Xu, Kui; Mu, Yulian; Zhou, Rong; Li, Kui

    2016-01-01

    Prenatal skeletal muscle development genetically determines postnatal muscle characteristics such as growth and meat quality in pigs. However, the molecular mechanisms underlying prenatal skeletal muscle development remain unclear. Here, we performed the first genome-wide analysis of chimeric RNAs, single nuclear polymorphisms (SNPs) and allele-specific expression (ASE) in prenatal skeletal muscle in pigs. We identified 14,810 protein coding genes and 163 high-confidence chimeric RNAs expressed in prenatal skeletal muscle. More than 94.5% of the chimeric RNAs obeyed the canonical GT/AG splice rule and were trans-splicing events. Ten and two RNAs were aligned to human and mouse chimeric transcripts, respectively. We detected 106,457 high-quality SNPs (6,955 novel), which were mostly (89.09%) located within QTLs for production traits. The high proportion of non-exonic SNPs revealed the incomplete annotation status of the current swine reference genome. ASE analysis revealed that 11,300 heterozygous SNPs showed allelic imbalance, whereas 131 ASE variants were located in the chimeric RNAs. Moreover, 4 ASE variants were associated with various economically relevant traits of pigs. Taken together, our data provide a source for studies of chimeric RNAs and biomarkers for pig breeding, while illuminating the complex transcriptional events underlying prenatal skeletal muscle development in mammals. PMID:27352850

  16. Evaluation of the sensitivity of two recently developed STR multiplexes for the analysis of chimerism after haematopoietic stem cell transplantation.

    PubMed

    Odriozola, A; Riancho, J A; Colorado, M; Zarrabeitia, M T

    2013-04-01

    Forensic-oriented kits analysing short tandem repeat (STR) polymorphisms are widely used to determine the proportions of donor and recipient cells after haematopoietic stem cell transplantation. The sensitivity of this technology is crucial for the early detection of relapse and, in consequence, the adjustment of the treatment to enhance donor-origin haematopoiesis in transplant recipients. The objective of this study was to compare the performance of two recently developed STR multiplex kits, AmpFℓSTR(®) Identifiler(®) Plus PCR Amplification Kit (Applied Biosystems) and Investigator™ IDplex(®) (Qiagen), in the analysis of chimerism. Fifteen STR loci were amplified with both kits in 26 peripheral blood samples of transplantated patients showing chimerism. Peak amplitude threshold, detection limit (%DL), per cent donor chimerism and efficacy of each multiplex and STR were determined, and the results with both kits were compared. The %DL and the estimated per cent donor chimerism were similar with both kits. On the other hand, Identifiler(®) Plus kit allowed chimerism identification only in 24 (92%) of the 26 cases with chimerism detected by using the Investigator™ IDplex(®) when only 'type 5' allelic constellations (i.e. without potential interference by stutter peaks) were taken into account. However, IDplex(®) efficacy was somewhat lower than that of Identifiler Plus when only the most informative loci (D2S1338, D21S11, D18S51 and FGA) were considered. Therefore, although each system had some particular advantages and disadvantages, overall both STR multiplexes showed similar performance in qualitative and quantitative chimerism analysis.

  17. Quantification of mixed chimerism allows early therapeutic interventions

    PubMed Central

    Merzoni, Jóice; Ewald, Gisele Menezes; Paz, Alessandra Aparecida; Daudt, Liane Esteves; Jobim, Luiz Fernando Job

    2014-01-01

    Hematopoietic stem cell transplantation is the curative option for patients with myelodysplastic syndrome; however, it requires a long post-transplantation follow-up. A 53-year-old woman with a diagnosis of myelodysplastic syndrome underwent related donor allogeneic hematopoietic stem cell transplantation in July 2006. Three months after transplantation, a comparative short tandem repeat analysis between donor and recipient revealed full chimerism, indicating complete, healthy bone marrow reconstitution. Three years and ten months after hematopoietic stem cell transplantation, the patient developed leukopenia and thrombocytopenia. Another short tandem repeat analysis was carried out which showed mixed chimerism (52.62%), indicating relapsed disease. A donor lymphocyte infusion was administered. The purpose of donor lymphocyte infusion is to induce a graft-versus-leukemia effect; in fact, this donor's lymphocyte infusion induced full chimerism. Successive short tandem repeat analyses were performed as part of post-transplantation follow-up, and in July 2010, one such analysis again showed mixed chimerism (64.25%). Based on this finding, a second donor lymphocyte infusion was administered, but failed to eradicate the disease. In September 2011, the patient presented with relapsed disease, and a second related donor allogeneic hematopoietic stem cell transplantation was performed. Subsequent short tandem repeat analyses revealed full chimerism, indicating complete bone marrow reconstitution. We conclude that quantitative detection of mixed chimerism is an important diagnostic tool that can guide early therapeutic intervention. PMID:25305171

  18. Clinical utility of chimerism status assessed by lineage-specific short tandem repeat analysis: experience from four cases of allogeneic stem cell transplantation.

    PubMed

    Goh, Ri-Young; Cho, Sung-Suk; Song, Yoo-Jeong; Heo, Kyeong; Oh, Sung-Yong; Kim, Sung-Hyun; Kwon, Hyeok-Chan; Kim, Hyo-Jin; Han, Jin-Yeong

    2009-08-01

    Chimerism testing permits early prediction and documentation of successful engraftment, and also facilitates detection of impending graft rejection. In this study, we serially monitored chimerism status by short tandem repeat-based PCR in nucleated cells (NC), T cells and natural killer (NK) cells after myeloablative allogeneic stem cell transplantation (SCT). Four patients with myeloid malignancies showed discrepant chimerism results among those three fractions. Three patients had mixed chimerism (MC) of donor/host T cells at a time point around the onset of chronic graft-versus-host disease (GVHD). In two patients with disease relapse, MC of NK cells preceded a morphological relapse or NK cells showed a higher percentage of patient cells compared to NC. Therefore, our study shows that chimerism analysis in lineage-specific cells might be useful in predicting clinical outcome after allogeneic SCT in certain patients.

  19. Functional analysis of aldehyde oxidase using expressed chimeric enzyme between monkey and rat.

    PubMed

    Itoh, Kunio; Asakawa, Tasuku; Hoshino, Kouichi; Adachi, Mayuko; Fukiya, Kensuke; Watanabe, Nobuaki; Tanaka, Yorihisa

    2009-01-01

    Aldehyde oxidase (AO) is a homodimer with a subunit molecular mass of approximately 150 kDa. Each subunit consists of about 20 kDa 2Fe-2S cluster domain storing reducing equivalents, about 40 kDa flavine adenine dinucleotide (FAD) domain and about 85 kDa molybdenum cofactor (MoCo) domain containing a substrate binding site. In order to clarify the properties of each domain, especially substrate binding domain, chimeric cDNAs were constructed by mutual exchange of 2Fe-2S/FAD and MoCo domains between monkey and rat. Chimeric monkey/rat AO was referred to one with monkey type 2Fe-2S/FAD domains and a rat type MoCo domain. Rat/monkey AO was vice versa. AO-catalyzed 2-oxidation activities of (S)-RS-8359 were measured using the expressed enzyme in Escherichia coli. Substrate inhibition was seen in rat AO and chimeric monkey/rat AO, but not in monkey AO and chimeric rat/monkey AO, suggesting that the phenomenon might be dependent on the natures of MoCo domain of rat. A biphasic Eadie-Hofstee profile was observed in monkey AO and chimeric rat/monkey AO, but not rat AO and chimeric monkey/rat AO, indicating that the biphasic profile might be related to the properties of MoCo domain of monkey. Two-fold greater V(max) values were observed in monkey AO than in chimeric rat/monkey AO, and in chimeric monkey/rat AO than in rat AO, suggesting that monkey has the more effective electron transfer system than rat. Thus, the use of chimeric enzymes revealed that 2Fe-2S/FAD and MoCo domains affect the velocity and the quantitative profiles of AO-catalyzed (S)-RS-8359 2-oxidation, respectively.

  20. IVGEN Post Flight Analysis

    NASA Technical Reports Server (NTRS)

    Mcquillen, John; Brown, Dan; Hussey, Sam; Zoldak, John

    2014-01-01

    The Intravenous Fluid Generation (IVGEN) Experiment was a technology demonstration experiment that purified ISS potable water, mixed it with salt, and transferred it through a sterilizing filter. On-orbit performance was verified as appropriate and two 1.5 l bags of normal saline solution were returned to earth for post-flight testing by a FDA certified laboratory for compliance with United States Pharmacopiea (USP) standards. Salt concentration deviated from required values and an analysis identified probable causes. Current efforts are focused on Total Organic Content (TOC) testing, and shelf life.The Intravenous Fluid Generation (IVGEN) Experiment demonstrated the purification of ISS potable water, the mixing of the purified water with sodium chloride, and sterilization of the solution via membrane filtration. On-orbit performance was monitored where feasible and two 1.5-liter bags of normal saline solution were returned to earth for post-flight testing by a FDA-registered laboratory for compliance with United States Pharmacopeia (USP)standards [1]. Current efforts have been focused on challenge testing with identified [2] impurities (total organic-carbon), and shelf life testing. The challenge testing flowed known concentrations of contaminants through the IVGEN deionizing cartridge and membrane filters to test their effectiveness. One finding was that the filters and DI-resin themselves contribute to the contaminant load during initial startup, suggesting that the first 100 ml of fluid be discarded. Shelf life testing is ongoing and involves periodic testing of stored DI cartridges and membrane filters that are capped and sealed in hermetic packages. The testing is conducted at six month intervals measuring conductivity and endotoxins in the effluent. Currently, the packaging technique has been successfully demonstrated for one year of storage testing. The USP standards specifies that the TOC be conducted at point of generation as opposed to point of

  1. Increasing chimerism after allogeneic stem cell transplantation is associated with longer survival time.

    PubMed

    Tang, Xiaowen; Alatrash, Gheath; Ning, Jing; Jakher, Haroon; Stafford, Patricia; Zope, Madhushree; Shpall, Elizabeth J; Jones, Roy B; Champlin, Richard E; Thall, Peter F; Andersson, Borje S

    2014-08-01

    Donor chimerism after allogeneic stem cell transplantation (allo-SCT) is commonly used to predict overall survival (OS) and disease-free survival (DFS). Because chimerism is observed at 1 or more times after allo-SCT and not at baseline, if chimerism is in fact associated with OS or DFS, then the occurrence of either disease progression or death informatively censors (terminates) the observed chimerism process. This violates the assumptions underlying standard statistical regression methods for survival analysis, which may lead to biased conclusions. To assess the association between the longitudinal post-allo-SCT donor chimerism process and OS or DFS, we analyzed data from 195 patients with acute myelogenous leukemia (n = 157) or myelodysplastic syndrome (n = 38) who achieved complete remission after allo-SCT following a reduced-toxicity conditioning regimen of fludarabine/intravenous busulfan. Median follow-up was 31 months (range, 1.1 to 105 months). Fitted joint longitudinal-survival time models showed that a binary indicator of complete (100%) donor chimerism and increasing percent of donor T cells were significantly associated with longer OS, whereas decreasing percent of donor T cells was highly significantly associated with shorter OS. Our analyses illustrate the usefulness of modeling repeated post-allo-SCT chimerism measurements as individual longitudinal processes jointly with OS and DFS to estimate their relationships.

  2. Antigenic and Cryo-Electron Microscopy Structure Analysis of a Chimeric Sapovirus Capsid

    PubMed Central

    Miyazaki, Naoyuki; Taylor, David W.

    2015-01-01

    ABSTRACT The capsid protein (VP1) of all caliciviruses forms an icosahedral particle with two principal domains, shell (S) and protruding (P) domains, which are connected via a flexible hinge region. The S domain forms a scaffold surrounding the nucleic acid, while the P domains form a homodimer that interacts with receptors. The P domain is further subdivided into two subdomains, termed P1 and P2. The P2 subdomain is likely an insertion in the P1 subdomain; consequently, the P domain is divided into the P1-1, P2, and P1-2 subdomains. In order to investigate capsid antigenicity, N-terminal (N-term)/S/P1-1 and P2/P1-2 were switched between two sapovirus genotypes GI.1 and GI.5. The chimeric VP1 constructs were expressed in insect cells and were shown to self-assemble into virus-like particles (VLPs) morphologically similar to the parental VLPs. Interestingly, the chimeric VLPs had higher levels of cross-reactivities to heterogeneous antisera than the parental VLPs. In order to better understand the antigenicity from a structural perspective, we determined an intermediate-resolution (8.5-Å) cryo-electron microscopy (cryo-EM) structure of a chimeric VLP and developed a VP1 homology model. The cryo-EM structure revealed that the P domain dimers were raised slightly (∼5 Å) above the S domain. The VP1 homology model allowed us predict the S domain (67–229) and P1-1 (229–280), P2 (281–447), and P1-2 (448–567) subdomains. Our results suggested that the raised P dimers might expose immunoreactive S/P1-1 subdomain epitopes. Consequently, the higher levels of cross-reactivities with the chimeric VLPs resulted from a combination of GI.1 and GI.5 epitopes. IMPORTANCE We developed sapovirus chimeric VP1 constructs and produced the chimeric VLPs in insect cells. We found that both chimeric VLPs had a higher level of cross-reactivity against heterogeneous VLP antisera than the parental VLPs. The cryo-EM structure of one chimeric VLP (Yokote/Mc114) was solved to 8.5-

  3. Lineage-specific chimerism analysis in nucleated cells, T cells and natural killer cells after myeloablative allogeneic hematopoietic stem cell transplantation

    PubMed Central

    Goh, Ri-Young; Kim, Sung-Hyun

    2011-01-01

    Background Chimerism analysis is an important tool for assessing the origin of hematopoietic cells after allogeneic stem cell transplantation (allo-SCT) and can be used to detect impending graft rejection and the recurrence of underlying malignant or nonmalignant diseases. Methods This study included 24 patients who underwent myeloablative allo-SCT. DNA was extracted from nucleated cells (NCs), T cells, and natural killer (NK) cells, and the chimerism status of these cell fractions was determined by STR-PCR performed using an automated fluorescent DNA analyzer. Results Twenty-three out of the 24 patients achieved engraftment. Mixed chimerism (MC) in NCs, but not in T cells and NK cells, was significantly correlated with disease relapse. MC in all cell fractions was correlated with mortality. Ten patients (41.6%) developed extensive chronic GVHD. Six patients had MC in T cells, and 3 of them had chronic GVHD. Four patients with MC and relapse received donor lymphocyte infusion (DLI), and among them, 3 had secondary relapse. Further, the chimerism status differed among different cell lineages in 6 patients with myeloid malignancies. Conclusion The implications of MC in lymphocyte subsets are an important area for future research. Chimerism analysis in lineage-specific cells permits detection of relapse and facilitates the monitoring of therapeutic interventions. These results can provide the basic data for chimerism analysis after myeloablative SCT. PMID:21461299

  4. Clinical significance of chimerism.

    PubMed

    Abuelo, Dianne

    2009-05-15

    Twins have been previously classified as either monozygotic or dizygotic. In recent years, fascinating, non-traditional mechanisms of twinning have been uncovered. We define chimerism versus mosaicism, touch on chimerism in the animal world, and explain timing of chimerism in humans. In addition, we discuss when to suspect chimerism in patients, and how to proceed with diagnostic evaluation and confirmation.

  5. Complex chimerism

    PubMed Central

    Ma, Kimberly K.; Petroff, Margaret G.; Coscia, Lisa A.; Armenti, Vincent T.; Adams Waldorf, Kristina M.

    2013-01-01

    Thousands of women with organ transplantation have undergone successful pregnancies, however little is known about how the profound immunologic changes associated with pregnancy might influence tolerance or rejection of the allograft. Pregnant women with a solid organ transplant are complex chimeras with multiple foreign cell populations from the donor organ, fetus, and mother of the pregnant woman. We consider the impact of complex chimerism and pregnancy-associated immunologic changes on tolerance of the allograft both during pregnancy and the postpartum period. Mechanisms of allograft tolerance are likely dynamic during pregnancy and affected by the influx of fetal microchimeric cells, HLA relationships (between the fetus, pregnant woman and/or donor), peripheral T cell tolerance to fetal cells, and fetal minor histocompatibility antigens. Further research is necessary to understand the complex immunology during pregnancy and the postpartum period of women with a solid organ transplant. PMID:23974274

  6. Comparing the Southern blot method and polymerase chain reaction product analysis for chimeric RCCX detection in CYP21A2 deficiency.

    PubMed

    Lee, Hsien-Hsiung; Lee, Yann-Jinn; Chao, Mei-Chyn

    2010-04-15

    The 3.2-kb TaqI-produced fragment of the CYP21A1P pseudogene and the 3.7-kb TaqI-produced fragment of the functional CYP21A2 gene exist on chromosome 6p21.3. We used the polymerase chain reaction (PCR) product and Southern blot method with TaqI endonuclease digestion to identify a chimeric RCCX module in two unrelated patients with congenital adrenal hyperplasia (CAH). After TaqI cleavage, the PCR product analysis revealed that patient 1 with the chimeric CYP21A1P/CYP21A2 gene in one allele and IVS2-12A/C>G in combination with the 707-714del mutation in the other allele produced a configuration of 3.2- and 2.4-kb fragments. Patient 2, who carried IVS2-12A/C>G in combination with the 707-714del mutation in one allele and the chimeric TNXA/TNXB gene in the other allele, presented with 3.2- and 2.3-kb fragments. However, Southern blot analysis showed that patients 1 and 2 produced 3.2-, 2.4-, and 2.5-kb fragments. We conclude that the chimeric CYP21A1P/CYP21A2 gene, IVS2-12A/C>G in combination with the 707-714del mutation, and the chimeric TNXA/TNXB gene cannot be distinguished by the Southern blot method. Conversely, the chimeric TNXA/TNXB gene was identified in the PCR product analysis due to the appearance of the 2.37-kb fragment, which indicates the occurrence of the chimeric TNXA/TNXB formation extending to the boundary of TNXA in the RCCX region. PMID:19961824

  7. Post Flight Dynamic Analysis Simulation

    NASA Technical Reports Server (NTRS)

    Gregory, B. R.

    1970-01-01

    Digital six-degrees-of-freedom, open loop Saturn 5 first stage flight evaluation simulation program obtains post flight simulation of the launch vehicle using actual flight data as input. Results are compared with measured data. For preflight analysis, the program uses predicted flight data as input.

  8. Possible formation of mitochondrial-RNA containing chimeric or trimeric RNA implies a post-transcriptional and post-splicing mechanism for RNA fusion.

    PubMed

    Yang, Wei; Wu, Jian-min; Bi, An-ding; Ou-Yang, Yong-chang; Shen, Hai-hong; Chirn, Gung-wei; Zhou, Jian-hua; Weiss, Emily; Holman, Emily Pauline; Liao, D Joshua

    2013-01-01

    Human cells are known to express many chimeric RNAs, i.e. RNAs containing two genes' sequences. Wondering whether there also is trimeric RNA, i.e. an RNA containing three genes' sequences, we wrote simple computer code to screen human expression sequence tags (ESTs) deposited in different public databases, and obtained hundreds of putative trimeric ESTs. We then used NCBI Blast and UCSC Blat browsers to further analyze their sequences, and identified 61 trimeric and two tetrameric ESTs (one EST containing four different sequences). We also identified 57 chimeric, trimeric or teterameric ESTs that contained both mitochondrial (mt) RNA and nuclear RNA (nRNA), i.e. were mtRNA-nRNA fusions. In some trimeric ESTs, the downstream partner was fused to the poly-A tail of the upstream partner, which, together with the mtRNA-nRNA fusions, suggests a possible new mechanism for RNA fusion that occurs after both transcription and splicing have been terminated, and possibly outside the nucleus, in contrast to the two current hypothetical mechanisms, trans-splicing and transcriptional-slippage, that occur in the nucleus. The mt-sequences in the mtRNA-nRNA fusions had pseudogenes in the nucleus but, surprisingly, localized mainly in chromosomes 1 and 5. In some mtRNA-nRNA fusions, as well as in some ESTs that were derived only from mtRNA, the mt-sequences might be cis- or trans-spliced. Actually, we cloned a new cis-spliced mtRNA, coined as 16SrRNA-s. Hence, mtDNA may not always be intron-less. Fusion of three or more RNAs to one, fusion of nRNA to mtRNA, and cis- or trans-splicing of mtRNA should all enlarge the cellular RNA repertoire, in turn enlarging the cellular functions. Therefore, future experimental verification of the existence of these novel classes of fusion RNAs and spliced mtRNAs in human cells should significantly advance our understanding of biology and medicine. PMID:24204722

  9. CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice

    PubMed Central

    Oji, Asami; Noda, Taichi; Fujihara, Yoshitaka; Miyata, Haruhiko; Kim, Yeon Joo; Muto, Masanaga; Nozawa, Kaori; Matsumura, Takafumi; Isotani, Ayako; Ikawa, Masahito

    2016-01-01

    Targeted gene disrupted mice can be efficiently generated by expressing a single guide RNA (sgRNA)/CAS9 complex in the zygote. However, the limited success of complicated genome editing, such as large deletions, point mutations, and knockins, remains to be improved. Further, the mosaicism in founder generations complicates the genotypic and phenotypic analyses in these animals. Here we show that large deletions with two sgRNAs as well as dsDNA-mediated point mutations are efficient in mouse embryonic stem cells (ESCs). The dsDNA-mediated gene knockins are also feasible in ESCs. Finally, we generated chimeric mice with biallelic mutant ESCs for a lethal gene, Dnajb13, and analyzed their phenotypes. Not only was the lethal phenotype of hydrocephalus suppressed, but we also found that Dnajb13 is required for sperm cilia formation. The combination of biallelic genome editing in ESCs and subsequent chimeric analysis provides a useful tool for rapid gene function analysis in the whole organism. PMID:27530713

  10. Chimerism Analysis of Cell-Free DNA in Patients Treated with Hematopoietic Stem Cell Transplantation May Predict Early Relapse in Patients with Hematologic Malignancies

    PubMed Central

    Aljurf, Mahmoud; Abalkhail, Hala; Alseraihy, Amal; Mohamed, Said Y.; Ayas, Mouhab; Alsharif, Fahad; Alzahrani, Hazza; Al-Jefri, Abdullah; Aldawsari, Ghuzayel; Al-Ahmari, Ali; Belgaumi, Asim F.; Walter, Claudia Ulrike; El-Solh, Hassan; Rasheed, Walid; Albitar, Maher

    2016-01-01

    Background. We studied DNA chimerism in cell-free DNA (cfDNA) in patients treated with HSCT. Methods. Chimerism analysis was performed on CD3+ cells, polymorphonuclear (PMN) cells, and cfDNA using 16 small tandem repeat loci. The resulting labeled PCR-products were size-fractionated and quantified. Results. Analyzing samples from 191 patients treated with HSCT for nonneoplastic hematologic disorders demonstrated that the cfDNA chimerism is comparable to that seen in PMN cells. Analyzing leukemia patients (N = 126) showed that, of 84 patients with 100% donor DNA in PMN, 16 (19%) had evidence of clinical relapse and >10% recipient DNA in the plasma. Additional 16 patients of the 84 (19%) showed >10% recipient DNA in plasma, but without evidence of relapse. Eight patients had mixed chimerism in granulocytes, lymphocytes, and plasma, but three of these patients had >10% recipient DNA in plasma compared to PMN cells and these three patients had clinical evidence of relapse. The remaining 34 patients showed 100% donor DNA in both PMN and lymphocytes, but cfDNA showed various levels of chimerism. Of these patients 14 (41%) showed laboratory or clinical evidence of relapse and all had >10% recipient DNA in cfDNA. Conclusion. Monitoring patients after HSCT using cfDNA might be more reliable than cellular DNA in predicting early relapse. PMID:27006832

  11. Evaluation of STR informativity for chimerism testing--comparative analysis of 27 STR systems in 203 matched related donor recipient pairs.

    PubMed

    Thiede, C; Bornhäuser, M; Ehninger, G

    2004-02-01

    Chimerism analysis has become a routine method to document engraftment and also for detection of residual disease. PCR-based procedures using STR analysis, especially commercially available multiplex assays, are frequently used. However, these assays have been optimized for forensic purposes and do not necessarily fulfil all needs for chimerism analysis. To improve these analyses, data on the level of informativity of STR systems in the context of chimerism analysis would be helpful. We evaluated 27 STR markers for their informativity in 203 patients and their HLA-matched related donors. These STRs included 18 from different multiplex kits, whereas nine were selected from the literature or STR databases. The STR profiles were ranked from Type 1 (not informative) to Type 5 (best suited for chimerism analysis). According to this ranking, the informativity of the STR systems was found highly variable, ranging from 4.4 to 49.0% Type 5 constellations. Among the most informative STRs were Penta E, SE33, D2S1338 and D18S51. Informativity of an STR was correlated with the degree of heterozygosity (r=0.86; P=0.0001), but not with the total number of alleles present. These data indicate that selection of suitable STR markers is important to improve diagnostics based on STR analysis.

  12. Assessing quantitative chimerism longitudinally: technical considerations, clinical applications and routine feasibility.

    PubMed

    Kristt, D; Stein, J; Yaniv, I; Klein, T

    2007-03-01

    In this review, we describe the current laboratory approach to quantitative chimerism testing based on short tandem repeats (STRs), focusing on a longitudinal analysis. The latter is based on relative changes appearing in the course of sequential samples, and as such exploits the ultimate potential of this intrinsically semiquantitative platform. Such an analysis is more informative than single static values, less likely to be confused with platform artifacts, and is individualized to the particular patient. It is particularly useful with non-myeloablative conditioning, where mixed chimerism is common. Importantly, longitudinal monitoring is a routinely feasible laboratory option because multiplex STR-polymerase chain reaction kits are available commercially, and modern software can be used to perform computation, reliability testing and longitudinal tracking in a rapid, easy to use format. The ChimerTrack application, a shareware, user friendly program developed for this purpose, produces a report that automatically summarizes and illustrates the quantitative temporal course of the patient's chimeric status. Such a longitudinal perspective enhances the value of quantitative chimerism monitoring for decisions regarding immunomodulatory post transplant therapy. This information also provides unique insights into the biological dynamics of engraftment underlying the fluctuations in the temporal course of a patient's chimeric status.

  13. Prenatal diagnosis and genetic analysis of a fetus with 47,XX,+21/46,XX mosaicism and XX/XY chimerism.

    PubMed

    Hwa, Hsiao-Lin; Ko, Tsang-Ming; Huang, Chien-Hao; Chang, Li-Shu

    2006-08-01

    Prenatal diagnosis of simultaneous occurrence of chimerism and autosomal mosaicism is extremely rare. We report the prenatal diagnosis and genetic analysis of a fetus in a twin pregnancy with mosaic 47,XX,+21/46,XX with chimeric XX/XY. A 36-year-old, para 1, woman was referred for genetic counseling at 20 weeks' gestation because of abnormal karyotype (47,XX,+21/46,XX) in one fetus in a twin pregnancy. Cordocentesis revealed 47,XX,+21[3]/46,XX[35]/46,XY[7] in this fetus. Postnatal cytogenetic analysis of cord blood confirmed three cell lines in this twin (A) and 46,XY in the co-twin (B). Postmortem pathologic findings of both fetuses were normal. Fluorescence in situ hybridization identified three cell lines in the cord blood of twin A. Molecular genetic analysis using polymorphic DNA markers revealed parental origin of fetal tissue, and confirmed the chimeric status. Molecular genetic analysis with polymorphic DNA markers help to differentiate chimerism from mosaicism and define the origin of cell lines, which may have importance in genetic counseling.

  14. Growth control of influenza A virus by M1 protein: analysis of transfectant viruses carrying the chimeric M gene.

    PubMed

    Yasuda, J; Bucher, D J; Ishihama, A

    1994-12-01

    Analysis of fast-growing reassortants (AWM viruses) of influenza A virus produced by mixed infection with a fast-growing WSN strain and a slowly growing Aichi strain indicated that the M gene plays a role in the regulation of virus growth rate at an early step of infection (J. Yasuda, T. Toyoda, M. Nakayama, and A. Ishihama, Arch. Virol. 133:283-294, 1993). To determine which of the two M gene products, M1 or M2, is responsible for the growth rate control, one recombinant WSN virus (CWA) clone possessing a chimeric M gene (WSN M1-Aichi M2) was generated by using an improved reverse genetics and transfection system. The recombinant CWA virus retained the phenotype of both large plaque formation and early onset of virus growth. This indicates that the WSN M1 protein is responsible for rapid virus growth. PMID:7966605

  15. Chimerism analysis in clinical practice and its relevance for the detection of graft rejection and malignant relapse in pediatric hematopoietic stem cell transplant patients.

    PubMed

    Mellgren, Karin; Arvidson, Johan; Toporski, Jacek; Winiarski, Jacek

    2015-11-01

    Chimerism and clinical outcome data from 244 hematopoietic stem cell transplants in 218 children were retrospectively analyzed to assess their relevance for the detection of graft rejection and malignant relapse. Patients transplanted for a non-malignant disease had significantly higher proportions of residual recipient T cells in peripheral blood at one, three, and six months compared with patients transplanted for malignant disease. Recipient T-cell levels were below 50% at one month after transplantation in most patients (129 of 152 transplants). Graft rejection occurred more frequently in the group of patients with high levels of recipient cells at one month (10 graft rejections in the 23 patients with recipient T cells >50% at one month as compared to seven graft rejections occurred in 129 patients with recipient T cells <50% (p < 0.001). Multilineage chimerism data in 87 children with leukemia at one, three, and six months after transplantation were not correlated with subsequent relapse of malignant disease. In conclusion, early analysis of lineage-specific chimerism in peripheral blood can be used to identify patients who are at high risk of graft rejection. However, the efficacy of early chimerism analysis for predicting leukemia relapse was limited.

  16. [Post-mortem microbiology analysis].

    PubMed

    Fernández-Rodríguez, Amparo; Alberola, Juan; Cohen, Marta Cecilia

    2013-12-01

    Post-mortem microbiology is useful in both clinical and forensic autopsies, and allows a suspected infection to be confirmed. Indeed, it is routinely applied to donor studies in the clinical setting, as well as in sudden and unexpected death in the forensic field. Implementation of specific sampling techniques in autopsy can minimize the possibility of contamination, making interpretation of the results easier. Specific interpretation criteria for post-mortem cultures, the use of molecular diagnosis, and its fusion with molecular biology and histopathology have led to post-mortem microbiology playing a major role in autopsy. Multidisciplinary work involving microbiologists, pathologists, and forensic physicians will help to improve the achievements of post-mortem microbiology, prevent infectious diseases, and contribute to a healthier population.

  17. [Post-mortem microbiology analysis].

    PubMed

    Fernández-Rodríguez, Amparo; Alberola, Juan; Cohen, Marta Cecilia

    2013-12-01

    Post-mortem microbiology is useful in both clinical and forensic autopsies, and allows a suspected infection to be confirmed. Indeed, it is routinely applied to donor studies in the clinical setting, as well as in sudden and unexpected death in the forensic field. Implementation of specific sampling techniques in autopsy can minimize the possibility of contamination, making interpretation of the results easier. Specific interpretation criteria for post-mortem cultures, the use of molecular diagnosis, and its fusion with molecular biology and histopathology have led to post-mortem microbiology playing a major role in autopsy. Multidisciplinary work involving microbiologists, pathologists, and forensic physicians will help to improve the achievements of post-mortem microbiology, prevent infectious diseases, and contribute to a healthier population. PMID:23195835

  18. Quo vadis chimerism?

    PubMed Central

    2011-01-01

    Although immunity in multicellular organisms is efficient in dealing with alien agents, it may fail for allogeneic chimerism. Natural chimerism is widely documented in nature, distributed in at least ten phyla of protists, invertebrates and plants, vertebrates and mammals, including humans; it is an important ecological/evolutionary tool manipulating metazoans' life history portraits. Instead of purging allogeneic nascent selfish cells, a ‘double edged sword’ chimerism emerges, displaying environmental dictated costs and benefits for the genotypes involved. Benefits include the development of synergistic complementation, the increase of genetic variability, the assurance of mate location, improved size-dependent ecological qualities (growth rates, reproduction, survivorship, competition, environmental tolerance) and more. Costs include the threat of somatic and germ cell parasitism, developmental instability, death, diseases, autoimmunity, sexual sterility and organ malformations, which develop as well in mammalian natural chimerism, including humans. Because of its importance, medical sciences should study and harness natural chimerism properties for clinical purposes. PMID:21547028

  19. Quo vadis chimerism?

    PubMed

    Rinkevich, Baruch

    2011-01-01

    Although immunity in multicellular organisms is efficient in dealing with alien agents, it may fail for allogeneic chimerism. Natural chimerism is widely documented in nature, distributed in at least ten phyla of protists, invertebrates and plants, vertebrates and mammals, including humans; it is an important ecological/evolutionary tool manipulating metazoans' life history portraits. Instead of purging allogeneic nascent selfish cells, a 'double edged sword' chimerism emerges, displaying environmental dictated costs and benefits for the genotypes involved. Benefits include the development of synergistic complementation, the increase of genetic variability, the assurance of mate location, improved size-dependent ecological qualities (growth rates, reproduction, survivorship, competition, environmental tolerance) and more. Costs include the threat of somatic and germ cell parasitism, developmental instability, death, diseases, autoimmunity, sexual sterility and organ malformations, which develop as well in mammalian natural chimerism, including humans. Because of its importance, medical sciences should study and harness natural chimerism properties for clinical purposes.

  20. Chimerism testing and detection of minimal residual disease after allogeneic hematopoietic transplantation using the bioView (Duet) combined morphological and cytogenetical analysis.

    PubMed

    Shimoni, A; Nagler, A; Kaplinsky, C; Reichart, M; Avigdor, A; Hardan, I; Yeshurun, M; Daniely, M; Zilberstein, Y; Amariglio, N; Brok-Simoni, F; Rechavi, G; Trakhtenbrot, L

    2002-08-01

    Recurrent disease remains a major obstacle to cure after allogeneic transplantation. Various methods have been developed to detect minimal residual disease (MRD) after transplantation to identify patients at risk for relapse. Chimerism tests differentiate recipient and donor cells and are used to identify MRD when there are no other disease-specific markers. The detection of MRD does not always correlate with relapse risk. Chimerism testing may also identify normal hematopoietic cells or other cells not contributing to relapse. In this study we report our initial experience with a novel system that provides combined morphological and cytogenetical analysis on the same cells. This system allows rapid automatic scanning of a large number of cells, thus increasing the sensitivity of detection of small recipient population. The clinical significance of MRD detection is improved by identifying the morphology of recipient cells. Identification of recipient characteristics within blasts predicts overt relapse in leukemia patients and precedes it by a few weeks to months. Identification within mature hematopoietic cells may not be closely associated with relapse. The system also allows chimerism testing after sex-mismatched transplants, within cellular subsets, with no need for sorting of cells. The system merits further study in larger scale trials.

  1. Chimeric analysis of Notch2 function: a role for Notch2 in the development of the roof plate of the mouse brain.

    PubMed

    Kadokawa, Yuzo; Marunouchi, Tohru

    2002-10-01

    Notch proteins are transmembrane receptors involved in cell-fate determination throughout development. Targeted disruption of either the Notch1 or Notch2 gene in mice results in embryonic lethality around embryonic day (E) 10.5 with widespread cell death. Although Notch1-deficient mice show disorganized somitogenesis, Notch2 mutants did not show definitive abnormalities in any tissue expressing high levels of the Notch2 gene, including the central nervous system. To study Notch2 function in development beyond the embryonic lethal stage, we performed chimeric analysis between Notch2 mutant and wild-type mouse embryos. Chimeric embryos developed normally and homozygous Notch2 mutant-specific cell death was not observed. Although chimeric embryos showed normal mosaicism until E9.5 in all tissues studied to date, Notch2 homozygous mutant cells failed to contribute to formation of the roof plate of the diencephalon and mesencephalon at later developmental stages, when Notch2 is normally expressed at high levels at there. Furthermore, Notch2 heterozygous mutant cells were also excluded from the roof plate of the chimera, however, Notch2 heterozygous mutant mice developed normally. We also showed that Wnt-1 and Mash1 expression patterns at the roof plate were disorganized in Notch2 homozygous mutant embryos. These results indicate that Notch2 plays an important role in development of the roof plate of the diencephalon and mesencephalon, and suggest that cellular rearrangement is involved in this process.

  2. Detection and quantification of chimerism by droplet digital PCR.

    PubMed

    George, David; Czech, Juliann; John, Bobby; Yu, Min; Jennings, Lawrence J

    2013-01-01

    Accurate quantification of chimerism and microchimerism is proving to be increasingly valuable for hematopoietic cell transplantation as well as non-transplant conditions. However, methods that are available to quantify low-level chimerism lack accuracy. Therefore, we developed and validated a method for quantifying chimerism based on digital PCR technology. We demonstrate accurate quantification that far exceeds what is possible with analog qPCR down to 0.01% with the potential to go even lower. Also, this method is inherently more informative than qPCR. We expect the advantages of digital PCR will make it the preferred method for chimerism analysis.

  3. Crystal Structure of PG16 and Chimeric Dissection with Somatically Related PG9: Structure-Function Analysis of Two Quaternary-Specific Antibodies That Effectively Neutralize HIV-1

    SciTech Connect

    Pancera, Marie; McLellan, Jason S.; Wu, Xueling; Zhu, Jiang; Changela, Anita; Schmidt, Stephen D.; Yang, Yongping; Zhou, Tongqing; Phogat, Sanjay; Mascola, John R.; Kwong, Peter D.

    2010-11-03

    HIV-1 resists neutralization by most antibodies. Two somatically related human antibodies, PG9 and PG16, however, each neutralize 70 to 80% of circulating HIV-1 isolates. Here we present the structure of the antigen-binding fragment of PG16 in monoclinic and orthorhombic lattices at 2.4 and 4.0 {angstrom}, respectively, and use a combination of structural analysis, paratope dissection, and neutralization assessment to determine the functional relevance of three unusual PG9/PG16 features: N-linked glycosylation, extensive affinity maturation, and a heavy chain-third complementarity-determining region (CDR H3) that is one of the longest observed in human antibodies. Glycosylation extended off the side of the light chain variable domain and was not required for neutralization. The CDR H3 formed an axe-shaped subdomain, which comprised 42% of the CDR surface, with the axe head looming {approx}20 {angstrom} above the other combining loops. Comprehensive sets of chimeric swaps between PG9 and PG16 of light chain, heavy chain, and CDR H3 were employed to decipher structure-function relationships. Chimeric swaps generally complemented functionally, with differences in PG9/PG16 neutralization related primarily to residue differences in CDR H3. Meanwhile, chimeric reversions to genomic V genes showed isolate-dependent effects, with affinity maturation playing a significant role in augmenting neutralization breadth (P = 0.036) and potency (P < 0.0001). The structural and functional details of extraordinary CDR H3 and extensive affinity maturation provide insights into the neutralization mechanism of and the elicitation pathway for broadly neutralizing antibodies like PG9 and PG16.

  4. Chimerism testing by quantitative PCR using Indel markers.

    PubMed

    Gendzekhadze, Ketevan; Gaidulis, Laima; Senitzer, David

    2013-01-01

    Engraftment monitoring is critical for patients after Hematopoietic Stem Cell Transplantation (HSCT). Complete donor chimerism is the goal; therefore, early detection of rejection and relapse is crucial for guiding the patient post HSCT treatment. Quantitative PCR for chimerism testing has been reported to be highly sensitive. In this chapter we discuss the quantitative PCR (qPCR) method using 34 Indel (Insertion and Deletion) genetic markers spread over 20 different chromosomes.

  5. Long-Term Stable Mixed Chimerism after Hematopoietic Stem Cell Transplantation in Patients with Non-Malignant Disease, Shall We Be Tolerant?

    PubMed Central

    Stikvoort, Arwen; Sundin, Mikael; Uzunel, Mehmet; Gertow, Jens; Sundberg, Berit; Schaffer, Marie; Mattsson, Jonas; Uhlin, Michael

    2016-01-01

    Long-term stable mixed chimerism is a rare and poorly understood phenomenon post hematopoietic stem cell transplantation. This study aims to shed light on whether the two hematopoietic systems in patients with mixed chimerism remain functional. Additionally, we investigate possible immunologic differences in these individuals compared to patients with only donor derived immune cells. Patients with donor and mixed chimerism, at median 10 (5–16) years post-HSCT for non-malignant diseases, were assessed regarding clinical situation and immune system (phenotypical and functional). No difference in long-term outcome was seen in terms of general wellbeing, central phenotypic immune system features (e.g., differentiation status, CD4/CD8 ratio, B and NK-cell frequency) and antibody responses to immunizations. At a median of 10 years post transplantation, patients with mixed chimerism had significantly higher IgG3 and platelet levels. Additionally, these patients had higher NKT-cell levels (CD94+CD8+ and CD56+CD8+) than patients with donor chimerism. In depth phenotypic analysis of patients with mixed chimerism demonstrated recipient-derived fractions in most immune cell lineages (e.g., T-cell, B-cell and NK-cell subsets). Recipient cells were also capable of responding to mitogenic stimulation with production of several cytokines. In conclusion, long-term mixed chimerism did not negatively affect patient wellbeing and long-term outcome. Moreover, recipient-derived immunity may still be functional in these patients, suggesting an active state of tolerance and immunologic dependence on both hematopoietic systems. PMID:27152621

  6. Long-Term Stable Mixed Chimerism after Hematopoietic Stem Cell Transplantation in Patients with Non-Malignant Disease, Shall We Be Tolerant?

    PubMed

    Stikvoort, Arwen; Sundin, Mikael; Uzunel, Mehmet; Gertow, Jens; Sundberg, Berit; Schaffer, Marie; Mattsson, Jonas; Uhlin, Michael

    2016-01-01

    Long-term stable mixed chimerism is a rare and poorly understood phenomenon post hematopoietic stem cell transplantation. This study aims to shed light on whether the two hematopoietic systems in patients with mixed chimerism remain functional. Additionally, we investigate possible immunologic differences in these individuals compared to patients with only donor derived immune cells. Patients with donor and mixed chimerism, at median 10 (5-16) years post-HSCT for non-malignant diseases, were assessed regarding clinical situation and immune system (phenotypical and functional). No difference in long-term outcome was seen in terms of general wellbeing, central phenotypic immune system features (e.g., differentiation status, CD4/CD8 ratio, B and NK-cell frequency) and antibody responses to immunizations. At a median of 10 years post transplantation, patients with mixed chimerism had significantly higher IgG3 and platelet levels. Additionally, these patients had higher NKT-cell levels (CD94+CD8+ and CD56+CD8+) than patients with donor chimerism. In depth phenotypic analysis of patients with mixed chimerism demonstrated recipient-derived fractions in most immune cell lineages (e.g., T-cell, B-cell and NK-cell subsets). Recipient cells were also capable of responding to mitogenic stimulation with production of several cytokines. In conclusion, long-term mixed chimerism did not negatively affect patient wellbeing and long-term outcome. Moreover, recipient-derived immunity may still be functional in these patients, suggesting an active state of tolerance and immunologic dependence on both hematopoietic systems.

  7. Long-Term Stable Mixed Chimerism after Hematopoietic Stem Cell Transplantation in Patients with Non-Malignant Disease, Shall We Be Tolerant?

    PubMed

    Stikvoort, Arwen; Sundin, Mikael; Uzunel, Mehmet; Gertow, Jens; Sundberg, Berit; Schaffer, Marie; Mattsson, Jonas; Uhlin, Michael

    2016-01-01

    Long-term stable mixed chimerism is a rare and poorly understood phenomenon post hematopoietic stem cell transplantation. This study aims to shed light on whether the two hematopoietic systems in patients with mixed chimerism remain functional. Additionally, we investigate possible immunologic differences in these individuals compared to patients with only donor derived immune cells. Patients with donor and mixed chimerism, at median 10 (5-16) years post-HSCT for non-malignant diseases, were assessed regarding clinical situation and immune system (phenotypical and functional). No difference in long-term outcome was seen in terms of general wellbeing, central phenotypic immune system features (e.g., differentiation status, CD4/CD8 ratio, B and NK-cell frequency) and antibody responses to immunizations. At a median of 10 years post transplantation, patients with mixed chimerism had significantly higher IgG3 and platelet levels. Additionally, these patients had higher NKT-cell levels (CD94+CD8+ and CD56+CD8+) than patients with donor chimerism. In depth phenotypic analysis of patients with mixed chimerism demonstrated recipient-derived fractions in most immune cell lineages (e.g., T-cell, B-cell and NK-cell subsets). Recipient cells were also capable of responding to mitogenic stimulation with production of several cytokines. In conclusion, long-term mixed chimerism did not negatively affect patient wellbeing and long-term outcome. Moreover, recipient-derived immunity may still be functional in these patients, suggesting an active state of tolerance and immunologic dependence on both hematopoietic systems. PMID:27152621

  8. Chimeric Pestivirus Experimental Vaccines.

    PubMed

    Reimann, Ilona; Blome, Sandra; Beer, Martin

    2016-01-01

    Chimeric pestiviruses have shown great potential as marker vaccine candidates against pestiviral infections. Exemplarily, we describe here the construction and testing of the most promising classical swine fever vaccine candidate "CP7_E2alf" in detail. The description is focused on classical cloning technologies in combination with reverse genetics. PMID:26458840

  9. Achievement of early complete donor chimerism in CD25+-activated leukocytes is a strong predictor of the development of graft-versus-host-disease after stem cell transplantation.

    PubMed

    Martínez-Laperche, Carolina; Noriega, Víctor; Kwon, Mi; Balsalobre, Pascual; González-Rivera, Milagros; Serrano, David; Anguita, Javier; Gayoso, Jorge; Díez-Martín, José Luis; Buño, Ismael

    2015-01-01

    Chimerism dynamics in bone marrow, peripheral blood (PB), and T lymphocytes (TL) has been associated with the development of various complications after allogeneic stem-cell transplantation (allo-SCT). In the present study, the usefulness of chimerism monitoring in CD25(+)-activated leukocytes (AL), together with that in bone marrow, PB, and TL, for the anticipation of complications after allo-SCT, has been analyzed in 68 patients. In AL, we observed a slower dynamics toward complete chimerism (CC) than in PB (p = 0.042), while no significant differences were found between TL and PB (p = 0.12). Complete chimerism achievement in AL at day +30 has shown to be an independent risk factor for the development of grade II-IV acute graft-versus-host disease (aGvHD; hazard ratio [95% confidence interval]: 11.9 [1.5-91.7]; p = 0.017). Moreover, among patients achieving CC in TL and AL at different time-points after SCT (n = 17/68), the incidence of grade II-IV aGvHD was significantly higher in patients who achieved CC earlier in AL (5/5) than in those who achieved CC earlier in TL (1/11; p = 0.001). Therefore, achievement of early complete donor chimerism in CD25(+) AL is a strong predictor for the development of aGvHD. Prospective analysis of chimerism in AL could improve the post-SCT management of immunosuppressive therapy in transplanted patients.

  10. Allosteric regulation and substrate channeling in multifunctional pyrimidine biosynthetic complexes: analysis of isolated domains and yeast-mammalian chimeric proteins.

    PubMed

    Serre, V; Guy, H; Liu, X; Penverne, B; Hervé, G; Evans, D

    1998-08-14

    The initial steps of pyrimidine biosynthesis in yeast and mammals are catalyzed by large multifunctional proteins of similar size, sequence and domain structure, but appreciable functional differences. The mammalian protein, CAD, has carbamyl phosphate synthetase (CPSase), aspartate transcarbamylase (ATCase) and dihydroorotase (DHOase) activities. The yeast protein, ura2, catalyzes the first two reactions and has a domain, called pDHO, which is homologous to mammalian DHOase, but is inactive. In CAD, only CPSase is regulated, whereas both CPSase and ATCase in the yeast protein are inhibited by UTP. These functional differences were explored by constructing a series of mammalian yeast chimeras. The isolated ATCase domain is catalytically active, but is not regulated. The inclusion of the yeast sequences homologous to the mammalian regulatory domain (B3) and the intervening pDHO domain did not confer regulation. Chimeric proteins in which the homologous regions of the mammalian protein were replaced by the corresponding domains of ura2 exhibited full catalytic activity, as well regulation of the CPSase, but not the ATCase, activities. The yeast B3 subdomain confers UTP sensitivity on the mammalian CPSase, suggesting that it is the locus of CPSase regulation in ura2. Taken together, these results indicate that there are regulatory site(s) in ura2. Channeling is impaired in all the chimeric complexes and completely abolished in the chimera in which the pDHO domain of yeast is replaced by the mammalian DHO domain. PMID:9698553

  11. In silico analysis and modeling of ACP-MIP–PilQ chimeric antigen from Neisseria meningitidis serogroup B

    PubMed Central

    Gholami, Mehrdad; Salimi Chirani, Alireza; Moshiri, Mona; Sedighi, Mansour; Pournajaf, Abazar; Tohidfar, Masoud; Irajian, Gholamreza

    2015-01-01

    Background: Neisseria meningitidis, a life-threatening human pathogen with the potential to cause large epidemics, can be isolated from the nasopharynx of 5–15% of adults. The aim of the current study was to evaluate biophysical and biochemical properties and immunological aspects of chimeric acyl-carrier protein-macrophage infectivity potentiator protein-type IV pilus biogenesis protein antigen (ACP-MIP-PilQ) from N. meningitidis serogroup B strain. Methods: Biochemical properties and multiple alignments were predicted by appropriate web servers. Secondary molecular structures were predicted based on Chou and Fasman, Garnier-Osguthorpe-Robson, and Neural Network methods. Tertiary modeling elucidated conformational properties of the chimeric protein. Proteasome cleavage and transporter associated with antigen processing (TAP) binding sites, and T- and B-cell antigenic epitopes, were predicted using bioinformatic web servers. Results: Based on our in silico and immunoinformatics analyses, the ACP-MIP-PilQ protein (AMP) can induce high-level cross-strain bactericidal activity. In addition, several immune proteasomal cleavage sites were detected. The 22 epitopes associated with MHC class I and class II (DR) alleles were confirmed in the AMP. Thirty linear B-cell epitopes as antigenic regions were predicted from the full-length protein. Conclusion: All predicted properties of the AMP indicate it could be a good candidate for further immunological in vitro and in vivo studies. PMID:26989750

  12. Acute GVHD is a strong predictor of full donor CD3+ T cell chimerism after reduced intensity conditioning allogeneic stem cell transplantation.

    PubMed

    El-Cheikh, Jean; Vazquez, Alberto; Crocchiolo, Roberto; Furst, Sabine; Calmels, Boris; Castagna, Luca; Lemarie, Claude; Granata, Angela; Ladaique, Patrick; Oudin, Claire; Faucher, Catherine; Chabannon, Christian; Blaise, Didier

    2012-12-01

    The monitoring of chimerism is a standard procedure to assess engraftment and achievement of full donor lymphoid cells after reduced intensity conditioning (RIC) stem cell transplantation (Allo-SCT). However, there is no consensus on when and how often to monitor post-transplant chimerism. We retrospectively analyzed our experience regarding the impact of acute graft versus host disease (GVHD) for the prediction of allograft chimerism. One-hundred-and-fifteen patients transplanted between 2001 and 2010 were identified. This group included 57 females and 58 males with a median age of 50 years (range: 26-68). Patients evaluated in this study were adult patients with hematologic malignancies, who received transplants from an HLA-matched sibling donor or matched unrelated donor (MUD) at allele level so-called 10/10, and received the RIC regimen including fludarabine/busulfan and anti-thymoglobulin (ATG). Mixed T-cell chimerism was defined as between 5 and 94% recipient cells, and full chimerism was defined as the presence of more than 95% donor T-cell chimerism (TCC). Full donor TCC was achieved in 93 patients (81%) at a median of 77 days (range: 30-120) post-transplant. The cumulative incidence of Grade 2-4 GVHD in our population was 25% (95% CI 17-34). The analysis of the population of patients with acute GVHD grade ≥2 showed that at day 120 after Allo-SCT they all had a total full donor TCC. On the other hand, 78 (68%) patients without acute GVHD grade ≥2 presented with mixed chimerism (p = 0.002) on day 120 post-transplant. Interestingly, patients who received ATG 5 mg/kg obtained a higher probability of complete chimerism compared with those receiving 2.5 mg/kg (p = 0.03). In conclusion, our study demonstrates that acute GVHD was predictive of full donor TCC after RIC Allo-SCT. Therefore, our data may challenge the concept of the frequent or close monitoring of donor chimerism in some patients with ongoing acute GVHD. However, chimerism testing could represent

  13. Comparison of chimerism and minimal residual disease monitoring for relapse prediction after allogeneic stem cell transplantation for adult acute lymphoblastic leukemia.

    PubMed

    Terwey, Theis Helge; Hemmati, Philipp Georg; Nagy, Marion; Pfeifer, Heike; Gökbuget, Nicola; Brüggemann, Monika; Le Duc, Tanja Melinh; le Coutre, Philipp; Dörken, Bernd; Arnold, Renate

    2014-10-01

    Little data are available on the relative merits of chimerism and minimal residual disease (MRD) monitoring for relapse prediction after allogeneic hematopoietic stem cell transplantation (HCT). We performed a retrospective analysis of serial chimerism assessments in 101 adult HCT recipients with acute lymphoblastic leukemia (ALL) and of serial MRD assessments in a subgroup of 22 patients. All patients had received myeloablative conditioning. The cumulative incidence of relapse was significantly higher in the patients with increasing mixed chimerism (in-MC) compared with those with complete chimerism, low-level MC, and decreasing MC, but the sensitivity of in-MC detection with regard to relapse prediction was only modest. In contrast, MRD assessment was highly sensitive and specific. Patients with MRD positivity after HCT had the highest incidence of relapse among all prognostic groups analyzed. The median time from MRD positivity to relapse was longer than the median time from detection of in-MC, but in some cases in-MC preceded MRD positivity. We conclude that MRD assessment is a powerful prognostic tool that should be included in the routine post-transplantation monitoring of patients with ALL, but chimerism analysis may provide additional information in some cases. Integration of these tools and clinical judgment should allow optimal decision making with regard to post-transplantation therapeutic interventions.

  14. Post Treatment With an FGF Chimeric Growth Factor Enhances Epithelial Cell Proliferation to Improve Recovery From Radiation-Induced Intestinal Damage

    SciTech Connect

    Nakayama, Fumiaki; Hagiwara, Akiko; Umeda, Sachiko; Asada, Masahiro; Goto, Megumi; Oki, Junko; Suzuki, Masashi; Imamura, Toru; Akashi, Makoto

    2010-11-01

    Purpose: A fibroblast growth factor (FGF) 1-FGF2 chimera (FGFC) was created previously and showed greater structural stability than FGF1. This chimera was capable of stimulating epithelial cell proliferation much more strongly than FGF1 or FGF2 even without heparin. Therefore FGFC was expected to have greater biologic activity in vivo. This study evaluated and compared the protective activity of FGFC and FGF1 against radiation-induced intestinal injuries. Methods and Materials: We administered FGFC and FGF1 intraperitoneally to BALB/c mice 24 h before or after total-body irradiation (TBI). The numbers of surviving crypts were determined 3.5 days after TBI with gamma rays at doses ranging from 8 to 12 Gy. Results: The effect of FGFC was equal to or slightly superior to FGF1 with heparin. However, FGFC was significantly more effective in promoting crypt survival than FGF1 (p < 0.01) when 10 {mu}g of each FGF was administered without heparin before irradiation. In addition, FGFC was significantly more effective at promoting crypt survival (p < 0.05) than FGF1 even when administered without heparin at 24 h after TBI at 10, 11, or 12 Gy. We found that FGFC post treatment significantly promoted 5-bromo-2'-deoxyuridine incorporation into crypts and increased crypt depth, resulting in more epithelial differentiation. However, the number of apoptotic cells in FGFC-treated mice decreased to almost the same level as that in FGF1-treated mice. Conclusions: These findings suggest that FGFC strongly enhanced radioprotection with the induction of epithelial proliferation without exogenous heparin after irradiation and is useful in clinical applications for both the prevention and post treatment of radiation injuries.

  15. Post-analysis of learned rules

    SciTech Connect

    Liu, Bing; Hsu, W.

    1996-12-31

    Rule induction research implicitly assumes that after producing the rules from a dataset, these rules will be used directly by an expert system or a human user. In real-life applications, the situation may not be as simple as that, particularly, when the user of the rules is a human being. The human user almost always has some previous concepts or knowledge about the domain represented by the dataset. Naturally, he/she wishes to know how the new rules compare with his/her existing knowledge. In dynamic domains where the rules may change over time, it is important to know what the changes are. These aspects of research have largely been ignored in the past. With the increasing use of machine learning techniques in practical applications such as data mining, this issue of post analysis of rules warrants greater emphasis and attention. In this paper, we propose a technique to deal with this problem. A system has been implemented to perform the post analysis of classification rules generated by systems such as C4.5. The proposed technique is general and highly interactive. It will be particularly useful in data mining and data analysis.

  16. POST-PROCESSING ANALYSIS FOR THC SEEPAGE

    SciTech Connect

    Y. SUN

    2004-09-29

    This report describes the selection of water compositions for the total system performance assessment (TSPA) model of results from the thermal-hydrological-chemical (THC) seepage model documented in ''Drift-Scale THC Seepage Model'' (BSC 2004 [DIRS 169856]). The selection has been conducted in accordance with ''Technical Work Plan for: Near-Field Environment and Transport: Coupled Processes (Mountain-Scale TH/THC/THM, Drift-Scale THC Seepage, and Post-Processing Analysis for THC Seepage) Report Integration'' (BSC 2004 [DIRS 171334]). This technical work plan (TWP) was prepared in accordance with AP-2.27Q, ''Planning for Science Activities''. Section 1.2.3 of the TWP describes planning information pertaining to the technical scope, content, and management of this report. The post-processing analysis for THC seepage (THC-PPA) documented in this report provides a methodology for evaluating the near-field compositions of water and gas around a typical waste emplacement drift as these relate to the chemistry of seepage, if any, into the drift. The THC-PPA inherits the conceptual basis of the THC seepage model, but is an independently developed process. The relationship between the post-processing analysis and other closely related models, together with their main functions in providing seepage chemistry information for the Total System Performance Assessment for the License Application (TSPA-LA), are illustrated in Figure 1-1. The THC-PPA provides a data selection concept and direct input to the physical and chemical environment (P&CE) report that supports the TSPA model. The purpose of the THC-PPA is further discussed in Section 1.2. The data selection methodology of the post-processing analysis (Section 6.2.1) was initially applied to results of the THC seepage model as presented in ''Drift-Scale THC Seepage Model'' (BSC 2004 [DIRS 169856]). Other outputs from the THC seepage model (DTN: LB0302DSCPTHCS.002 [DIRS 161976]) used in the P&CE (BSC 2004 [DIRS 169860

  17. Analysis of monoclonal antibody chimeric BR96-doxorubicin immunoconjugate by sodium dodecyl sulfate-capillary gel electrophoresis with ultraviolet and laser-induced fluorescence detection.

    PubMed

    Liu, J; Abid, S; Lee, M S

    1995-08-10

    Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CAGE), the capillary version of SDS-polyacrylamide-based slab gel electrophoresis, has been utilized for the separation and analysis of monoclonal antibody chimeric BR96 and the corresponding immunoconjugate prepared between BR96 and the anticancer drug doxorubicin (BR96-DOX). SDS-CAGE was performed in a coated capillary column filled with a polymer solution-based gel network matrix. Two detection formats, a uv absorbance detector and an argon-ion laser-based fluorescence detector, were incorporated into this system, providing complementary information for the determination of conjugated species. Both monoclonal antibody and immunoconjugates were studied in their native, denatured, and denatured and reduced states, respectively. Six peaks were identified following separation of the denatured BR96-DOX. These peaks were confirmed to correspond to all the possible conjugated species as expected. Analysis of the resulting "fingerprint" maps indicated that the light, heavy, and light-heavy chain conjugates are the predominant species. SDS-CAGE offers an alternative approach to the conventional slab gel electrophoresis and other chromatographic techniques, providing rapid, efficient, sensitive, and accurate information for the analysis of antibody and bioconjugates.

  18. Development of a single probe for documentation of chimerism following bone marrow transplantation.

    PubMed Central

    Yam, P; Petz, L D; Ali, S; Stock, A D; Wallace, R B

    1987-01-01

    discriminate approximately 98% of sibling donor/recipient pairs. This would be accomplished using only one restriction-endonuclease digestion and only one gel electrophoresis. Since other genetic markers, e.g., red blood cell antigens, immunoglobulin allotypes, and chromosome analysis, are not uniformly informative and, in some cases, cannot be used in the early posttransplantation period, the use of synthetic oligonucleotide probes for analysis of DNA RFLP is emerging as the method of choice for studies of post-BMT chimerism. This method will allow for the development of new knowledge that has not been possible with previous methods. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:2890295

  19. Early recipient chimerism testing in the T- and NK-cell lineages for risk assessment of graft rejection in pediatric patients undergoing allogeneic stem cell transplantation.

    PubMed

    Breuer, S; Preuner, S; Fritsch, G; Daxberger, H; Koenig, M; Poetschger, U; Lawitschka, A; Peters, C; Mann, G; Lion, T; Matthes-Martin, S

    2012-03-01

    Timely diagnosis of impending graft rejection is crucial for effective therapeutic intervention after allogeneic hematopoietic stem cell transplantation (SCT). We have investigated the predictive potential of early leukocyte subset-specific chimerism for graft loss in children undergoing SCT. In total, 192 pediatric patients transplanted for the treatment of malignant and non-malignant diseases after reduced-intensity or myeloablative conditioning were investigated. Surveillance of lineage-specific chimerism was initiated upon first appearance of leukocyte counts amenable to cell sorting. Graft rejection occurred in 23 patients between 24 and 492 days post-transplant (median 63 days). The first chimerism analysis of T and NK cells performed at a median of 20 days after SCT identified three different risk groups that were independent from the conditioning regimen: recipient chimerism (RC) levels in T cells below 50% indicated a very low risk of rejection (1.4%), whereas high levels of RC (>90%) both in T and NK cells heralded graft loss in the majority of patients (90%) despite therapeutic interventions. RC >50% in T cells and ≤90% in NK cells defined an intermediate-risk group in which timely immunotherapy frequently prevented rejection. Early assessment of T- and NK-cell chimerism can therefore be instrumental in the risk assessment and therapeutic management of imminent graft rejection.

  20. Production and functional analysis of rat CD59 and chimeric CD59-Crry as active soluble proteins in Pichia pastoris.

    PubMed

    Quigg, R J; He, C; Hack, B K; Alexander, J J; Morgan, B P

    2000-01-01

    Crry (CR1-related gene/protein) is a rodent complement regulator that inhibits C3 convertases. CD59 is a conserved protein inhibitor active towards C8 and C9. We have previously produced rat Crry as a recombinant soluble (rs) protein in Pichia pastoris. In this study we produced functionally active rat rsCD59 and a chimeric rsCD59-Crry protein in P. pastoris. The GPI anchor addition site of rat CD59 (Asn-79) was replaced either by a stop codon to produce rsCD59, or with the sequence of the first five short consensus repeats of Crry to produce rsCD59-Crry. Proteins were generated by fermentation and purified by affinity chromatography on an anti-CD59 column. In a standard classical pathway haemolysis assay, all three rs proteins had inhibitory activity, with 50% inhibition at 0.5 microM (rsCrry and rsCD59-Crry) and 4.4 microM (rsCD59). In an assay examining inhibition of C5b-9, in which C5b-7 was first formed, followed by purified C8 and C9, rsCD59 and rsCD59-Crry were active with 50% inhibition at 0.8 microM (rsCD59-Crry) and 1.3 microM (rsCD59). The degree of inhibition was independent of whether the C8 and C9 were of rat or human origin. Therefore, we have produced rsCD59 and rsCD59-Crry in P. pastoris. The rsCD59 retains its inhibitory activity towards C5b-9, while rsCD59-Crry appears to have the combined activities of Crry and CD59. In a haemolytic assay, the inclusion of CD59 to Crry is of no additional benefit to Crry, which may illustrate the overall importance of the C3 convertase step. Yet, inclusion of Crry to CD59 increases the potency of CD59 towards C5b-9.

  1. Myeloid and lymphoid chimerism after T-cell-depleted bone marrow transplantation: evaluation of conditioning regimens using the polymerase chain reaction to amplify human minisatellite regions of genomic DNA.

    PubMed

    Mackinnon, S; Barnett, L; Bourhis, J H; Black, P; Heller, G; O'Reilly, R J

    1992-12-15

    Determining both myeloid and lymphoid chimerism after T-cell-depleted allogeneic bone marrow transplantation (BMT) could be helpful in the understanding of the biology of engraftment and could provide a rational method of assessing the ability of different conditioning regimens to promote engraftment. We prospectively investigated the role of different pretransplant conditioning regimens in 29 leukemic patients post-BMT by assessing myeloid and T-cell chimerism using a rapid and sensitive polymerase chain reaction (PCR) method. Minisatellites are hypervariable regions of DNA consisting of tandem repeats of a core nucleotide sequence, and allelic polymorphism results from differences in the number of the repeats. We used this variation to distinguish between donor and recipient cells post-BMT. Seventeen patients (9 sibling and 8 unrelated donors) received conditioning with hyperfractionated total body irradiation (TBI), thiotepa, and cyclophosphamide (Cy). Of the other 12 patients (all sibling donors), 11 received TBI plus Cy plus another agent: VP16, carboplatinum, or AZQ. One patient received TBI plus thiotepa plus VP16. All but one of the patients studied received marrow from HLA-identical donors. PCR analysis confirmed donor lymphoid engraftment within 8 days of transplant in six of six patients studied. All granulocyte DNA was of donor origin within the first 4 weeks of transplant, regardless of the conditioning regimen. The day +28 T cells were exclusively of donor origin in 14 of 17 patients who received TBI plus thiotepa plus Cy, but were mixed chimeric in 10 of 12 patients who received other conditioning regimens (P < .001). Early graft rejection was seen in one unrelated transplant recipient conditioned with TBI plus thiotepa plus Cy. Late graft failure was observed in 3 of 12 patients with mixed T-cell chimerism and in none of 16 patients with full donor chimerism at day +28. However, 5 of 16 patients who had complete T-cell chimerism at day +28 developed

  2. Efficiency of CD19 chimeric antigen receptor-modified T cells for treatment of B cell malignancies in phase I clinical trials: a meta-analysis

    PubMed Central

    Shi, Ni; Zhang, Zhen; Luo, Zhenzhen; Yue, Dongli; Zhang, Zimeng; Wang, Liping; Han, Weidong; Xu, Zhongwei; Chen, Hu; Zhang, Yi

    2015-01-01

    Chimeric antigen receptor (CAR) modified T cells targeted CD19 showed promising clinical outcomes in treatment of B cell malignances such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL) and other indolent lymphomas. However, the clinical benefit varies tremendously among different trials. This meta-analysis investigated the efficacy (response rates and survival time) of CD19-CAR T cells in refractory B cell malignances in Phase I clinical trials. We searched publications between 1991 and 2014 from PubMed and Web of Science. Pooled response rates were calculated using random-effects models. Heterogeneity was investigated by subgroup analysis and meta-regression. Fourteen clinical trials including 119 patients were eligible for response rate evaluation, 62 patients in 12 clinical trials were eligible for progression-free survival analysis. The overall pooled response rate of CD19-CAR T cells was 73% (95% confidence interval [CI]: 46-94%). Significant heterogeneity across estimates of response rates was observed (p < 0.001, I2=88.3%). ALL patients have higher response rate (93%, 95% CI: 65-100%) than CLL (62%, 95% CI: 27-93%) and lymphoma patients (36%, 95% CI: 1-83%). Meta-regression analysis identified lymphodepletion and no IL-2 administrated T cells as two key factors associated with better clinical response. Lymphodepletion and higher infused CAR T cell number were associated with better prognosis. In conclusion, this meta-analysis showed a high clinical response rate of CD19-CAR T cell-based immunotherapy in treatment of refractory B cell malignancies. Lymphodepletion and increasing number of infused CD19-CAR T cells have positive correlations with the clinical efficiency, on the contrary, IL-2 administration to T cells is not recommended. PMID:26376680

  3. Frequent hepatocyte chimerism in long-term human liver allografts independent of graft outcome.

    PubMed

    Aini, Wulamujiang; Miyagawa-Hayashino, Aya; Ozeki, Munetaka; Tsuruyama, Tatsuaki; Tamaki, Keiji; Uemoto, Shinji; Haga, Hironori

    2013-03-01

    Microchimerism after liver transplantation is considered to promote graft tolerance or tissue repair, but its significance is controversial. By using multiplex polymerase chain reaction (PCR) of short tandem repeat (STR) loci after laser capture microdissection of hepatocyte nuclei, we compared the proportions of recipient-derived hepatocytes in long-term stable liver allografts and late dysfunctional allografts caused by chronic rejection or idiopathic post-transplantation hepatitis. Through fluorescence in situ hybridization (FISH), we also analyzed the presence of recipient-derived Y-positive hepatocytes in the biopsies of livers transplanted from female donors to male recipients. The study population comprised 24 pediatric liver transplant recipients who survived with the initial graft, whose 10-year protocol biopsy records were available, and who had normal liver function (stable graft, SG; n=13) or a late dysfunctional graft (LDG; n=11) with similar follow-up periods (mean 10.8years in the SG group and 11.2years in the LDG group). STR analysis revealed that hepatocyte chimerism occurred in 7 of 13 (54%) SGs and 5 of 11 (45%) LDGs (p=0.68). The proportion of hepatocyte chimerism was low, with a mean of 3% seen in 2 of 3 female-to-male transplanted livers (one each of SG and LDG). In conclusion, hepatocyte chimerism was a constant event. The extent of engraftment of recipient-derived hepatocytes does not seem to correlate with the degree of hepatic injury in long-term liver allografts.

  4. Bidirectional Brush Seals: Post-Test Analysis

    NASA Technical Reports Server (NTRS)

    Hendricks, Robert C.; Wilson, Jack; Wu, Tom Y.; Flower, Ralph; Mullen, Robert L.

    1997-01-01

    A post-test analysis of a set of inside-diameter/outside-diameter (ID/OD) bidirectional brush seals used in three-port wave rotor tests was undertaken to determine brush bristle and configuration wear, pullout, and rotor coating wear. The results suggest that sharp changes in the pressure profiles were not well reflected in bristle tip configuration patterns or wear. Also, positive-to-negative changes in axial pressure gradients appeared to have little effect on the backing plates. Although the brushes had similar porosities, they had very different unpacked arrays. This difference could explain the departure of experimental data from computational fluid dynamics flow predictions for well-packed arrays at higher pressure drops. The rotor wear led to "car-track" scars (upper and lower wear bands) with a whipped surface between the bands. Those bands may have resulted from bristle stiffening at the fence and gap plates during alternate portions of the rotor cycle. Within the bristle response range the wear surface reflected the pressure distribution effect on bristle motion. No sacrificial metallurgical data were taken. The bristles did wear, with correspondingly more wear on the ID brush configurations than on the OD configurations; the complexity in constructing the ID brush was a factor.

  5. T-and B-lymphocyte chimerism in the marmoset.

    PubMed Central

    Niblack, G D; Kateley, J R; Gengozian, N

    1977-01-01

    Marmosets are natural blood chimeras, this condition resulting from the high frequency of fraternal twinning and the consistent development of placental vasular anastomoses between the two embryos. Identification of chimerism by sex-chromosome analysis of cultured blood lymphocytes provided a means of determining the proportion of chimerism among T and B lymphocytes. Peripheral blood lymphocytes were enriched for T or B cells by filtration through a nylon column (yields greater than 95 per cent T-cells) or inactivation of T lymphocytes by treatment with a goat anti-marmoset thymocyte antiserum in the presence of complement (yeilds greater than 95 per cent B cells). Mitogenic stimulation of these separated, enriched cell populations yielded metaphase plates which could be scored for percentage male and female cells. Tests on five different blood chimeras showed the T- and B-lymphocyte chimerism to be the same. Stimulation of blood lymphocytes with cells from another species of marmoset in a mixed lymphocyte culture test revealed the chimeric T-cell response (i.e., host and co-twin cells) to be similar to that obtained with a mitogenic lectin. The demonstration of equivalent T- and B-cell chimerism in these animals suggests derivation of these cells from a common stem cell pool and the response of both T-cell populations to an antigenic stimulus in proportions similar to their percentage chimerism suggests complete immunologic tolerance exists in this species for co-twin histocompatibility antigens. PMID:139360

  6. Manufacture of diploid/tetraploid chimeric mice.

    PubMed Central

    Lu, T Y; Markert, C L

    1980-01-01

    Tetraploid mouse embryos were produced by cytochalasin B treatment. These embryos usually die before completion of embryonic development and are abnormal morphologically and physiologically. The tetraploid embryos can be rescued to develop to maturity by aggregating them with normal diploid embryos to produce diploid/tetraploid chimeric mice. The diploid/tetraploid chimeric embryos are frequently abnormal: the larger the proportion of tetraploid cells, the greater the abnormality. By karyotype analysis and by the use of appropriate pigment cell markers, we have demonstrated that two of our surviving chimeras are in fact diploid/tetraploid chimeras. One surviving chimera is retarded in growth and displays neurological abnormalities. The coat color chimerism suggests that this chimera is about 50% tetraploid. Another chimera with about 10% tetraploid pigment cells in the coat is only slightly retarded in growth and is a fertile male. Tetraploid cells are distributed in many, if not all, tissues of embryos but evidently are physiologically inadequate to support completely normal development and function in the absence of substantial numbers of normal diploid cells. Images PMID:6934528

  7. Lentiviral Gag Assembly Analyzed through the Functional Characterization of Chimeric Simian Immunodeficiency Viruses Expressing Different Domains of the Feline Immunodeficiency Virus Capsid Protein

    PubMed Central

    Esteva, María J.; Affranchino, José L.; González, Silvia A.

    2014-01-01

    To gain insight into the functional relationship between the capsid (CA) domains of the Gag polyproteins of simian and feline immunodeficiency viruses (SIV and FIV, respectively), we constructed chimeric SIVs in which the CA-coding region was partially or totally replaced by the equivalent region of the FIV CA. The phenotypic characterization of the chimeras allowed us to group them into three categories: the chimeric viruses that, while being assembly-competent, exhibit a virion-associated unstable FIV CA; a second group represented only by the chimeric SIV carrying the N-terminal domain (NTD) of the FIV CA which proved to be assembly-defective; and a third group constituted by the chimeric viruses that produce virions exhibiting a mature and stable FIV CA protein, and which incorporate the envelope glycoprotein and contain wild-type levels of viral genome RNA and reverse transcriptase. Further analysis of the latter group of chimeric SIVs demonstrated that they are non-infectious due to a post-entry impairment, such as uncoating of the viral core, reverse transcription or nuclear import of the preintegration complex. Furthermore, we show here that the carboxyl-terminus domain (CTD) of the FIV CA has an intrinsic ability to dimerize in vitro and form high-molecular-weight oligomers, which, together with our finding that the FIV CA-CTD is sufficient to confer assembly competence to the resulting chimeric SIV Gag polyprotein, provides evidence that the CA-CTD exhibits more functional plasticity than the CA-NTD. Taken together, our results provide relevant information on the biological relationship between the CA proteins of primate and nonprimate lentiviruses. PMID:25462889

  8. Expression and purification of toxic anti-breast cancer p28-NRC chimeric protein

    PubMed Central

    Soleimani, Meysam; Mirmohammad-Sadeghi, Hamid; Sadeghi-Aliabadi, Hojjat; Jahanian-Najafabadi, Ali

    2016-01-01

    Background: Chimeric proteins consisting of a targeting moiety and a cytotoxic moiety are now under intense research focus for targeted therapy of cancer. Here, we report cloning, expression, and purification of such a targeted chimeric protein made up of p28 peptide as both targeting and anticancer moiety fused to NRC peptide as a cytotoxic moiety. However, since the antimicrobial activity of the NRC peptide would intervene expression of the chimeric protein in Escherichia coli, we evaluated the effects of two fusion tags, that is, thioredoxin (Trx) and 6x-His tags, and various expression conditions, on the expression of p28-NRC chimeric protein. Materials and Methods: In order to express the chimeric protein with only 6x-His tag, pET28 expression plasmid was used. Cloning in pET32 expression plasmid was performed to add both Trx and 6x-His tags to the chimeric protein. Expression of the chimeric protein with both plasmids was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis following optimization of expression conditions and host strains. Results: Expression of the chimeric protein in pET28a was performed. However, expression yield of the chimeric protein was low. Optimization of culture conditions and host strains led to reasonable expression yield of the toxic chimeric protein in pET32a vector. In cases of both plasmids, approximately 10 kDa deviation of the apparent molecular weight from the theoretical one was seen in SDS-PAGE of purified chimeric proteins. Conclusions: The study leads to proper expression and purification yield of p28-NRC chimeric protein with Trx tag following optimizing culture conditions and host strains. PMID:27169101

  9. Chimeric enzymes with improved cellulase activities

    SciTech Connect

    Xu, Qi; Baker, John O; Himmel, Michael E

    2015-03-31

    Nucleic acid molecules encoding chimeric cellulase polypeptides that exhibit improved cellulase activities are disclosed herein. The chimeric cellulase polypeptides encoded by these nucleic acids and methods to produce the cellulases are also described, along with methods of using chimeric cellulases for the conversion of cellulose to sugars such as glucose.

  10. Assessment of chimerism in epithelial cancers in transplanted patients.

    PubMed

    Leboeuf, Christophe; Ratajczak, Philippe; Vérine, Jérôme; Elbouchtaoui, Morad; Plassa, François; Legrès, Luc; Ferreira, Irmine; Sandid, Wissam; Varna, Mariana; Bousquet, Guilhem; Verneuil, Laurence; Janin, Anne

    2014-01-01

    Cancer is now the most severe complication in the long term in transplant recipients. As most solid-organ or hematopoietic stem-cell transplantations are allogeneic, chimerism studies can be performed on cancers occurring in recipients. We summarize here the different methods used to study chimerism in cancers developing in allogeneic-transplant recipients, analyze their respective advantages and report the main results obtained from these studies. Chimerism analyses of cancers in transplant recipients require methods suited to tissue samples. In the case of gender-mismatched transplantation, the XY chromosomes can be explored using fluorescent in situ hybridization on whole-tissue sections or Y-sequence-specific PCR after the laser microdissection of tumor cells. For cancers occurring after gender-matched transplantation, laser microdissection of tumor cells enables studies of microsatellite markers and high-resolution melting analysis of mitochondrial DNA on genes with marked polymorphism, provided these are different in the donor and the recipient. The results of different studies address the cancers that develop in both recipients and in transplants. The presence of chimeric cells in these two types of cancer implies an exchange of progenitor/stem-cells between transplant and recipient, and the plasticity of these progenitor/stem-cells contributes to epithelial cancers. The presence of chimeric cells in concomitant cancers and preneoplastic lesions implies that the oncogenesis of these cancers progresses through a multistep process.

  11. Pre/Post Data Analysis - Simple or Is It?

    NASA Technical Reports Server (NTRS)

    Feiveson, Al; Fiedler, James; Ploutz-Snyder, Robert

    2011-01-01

    This slide presentation reviews some of the problems of data analysis in analyzing pre and post data. Using as an example, ankle extensor strength (AES) experiments, to measure bone density loss during bed rest, the presentation discusses several questions: (1) How should we describe change? (2) Common analysis methods for comparing post to pre results. (3) What do we mean by "% change"? and (4) What are we testing when we compare % changes?

  12. Patterns of hematopoietic chimerism following bone marrow transplantation for childhood acute lymphoblastic leukemia from volunteer unrelated donors.

    PubMed

    Molloy, K; Goulden, N; Lawler, M; Cornish, J; Oakhill, A; Pamphilon, D; Potter, M; Steward, C; Langlands, K; Humphries, P; McCann, S R

    1996-04-01

    Hematopoietic chimerism was analyzed in serial bone marrow samples taken from 28 children following T-cell depleted unrelated donor bone marrow transplants (UD BMT) for acute lymphoblastic leukemia (ALL). Chimeric status was determined by polymerase chain reaction (PCR) of simple tandem repeat (STR) sequences (maximal sensitivity, 0.1%). At least two serial samples were examined in 23 patients. Of these, two had evidence of complete donor engraftment at all times and eight showed stable low level mixed chimerism (MC) (<1% recipient hematopoiesis). All 10 of these patients remain in remission with a minimum follow-up of 24 months. By contrast, 13 patients demonstrated a progressive return of recipient hematopoiesis. Five of these relapsed (4 to 9 months post BMT), one died of cytomegalovirus pneumonitis and seven remain in remission with a minimum follow-up of 24 months. Five children were excluded from serial analysis as two serial samples were not collected before either relapse (3) or graft rejection (2). We conclude that as with sibling transplants, ex vivo T depleted UD BMT in children with ALL is associated with a high incidence of MC. Stable donor engraftment and low level MC always correlated with continued remission. However, detection of a progressive return of recipient cells did not universally correlate with relapse, but highlighted those patients at greatest risk. Serial chimerism analysis by PCR of STRs provides a rapid and simple screening technique for the detection of relapse and the identification of patients with progressive MC who might benefit from detailed molecular analysis for minimal residual disease following matched volunteer UD BMT for childhood ALL.

  13. Quantitative chimerism: an independent acute leukemia prognosis indicator following allogeneic hematopoietic SCT.

    PubMed

    Qin, X-Y; Li, G-X; Qin, Y-Z; Wang, Y; Wang, F-R; Liu, D-H; Xu, L-P; Chen, H; Han, W; Wang, J-Z; Zhang, X-H; Li, J-L; Li, L-D; Liu, K-Y; Huang, X-J

    2014-10-01

    This study evaluates the prognostic significance of quantitative chimerism to monitor minimal residual disease and predict relapse in acute leukemia (AL) patients following allogeneic hematopoietic SCT (HSCT). The quantitative chimerism levels of 129 AL patients were measured using RQ-PCR based on 29 sequence polymorphisms. Receiver-operating characteristic curve indicated that the optimal cutoff point to predict an inevitable relapse was 1.0%, which results in 100.0% sensitivity and 79.6% specificity.The relapse rate of patients with chimerism >1.0% at 2 years was 55.0%, whereas that for patients with chimerism <1.0% was 0%(P=0.000). Quantitative chimerism >1.00% indicated a higher probability of relapse. Cox multivariate analysis indicated that quantitative chimerism >1.00% was associated with lower disease-free survival (hazard ratio (HR)=10.825; 95% confidence interval (CI) =4.704-24.912, P=0.000) and lower OS (HR=8.681; 95% CI=3.728-20.212, P=0.000). Patients (24/47 with quantitative chimerism >1.00%) who received preemptive modified DLI immunotherapy had significantly lower relapse rate (37.5%) than those (n=9) who did not (100%; P=0.001). Thus, quantitative chimerism is an independent prognostic factor that predicts clinical outcomes after HSCT and provides a guide for suitable interventions.

  14. Double umbilical cord blood transplantation: relevance of persistent mixed-unit chimerism.

    PubMed

    Hashem, Hasan; Lazarus, Hillard M

    2015-04-01

    Double umbilical cord blood transplantation (UCBT) was developed as a strategy to circumvent the cell dose limitation of single UCBT with a concomitant potential benefit of lowering the rate of leukemia relapse. Sustained hematopoiesis after double UCBT usually is derived from a single donor unit, as only a few patients have been reported to display stable mixed-unit chimerism for varying periods of time. Explanations for the 1 unit dominance, predictors for identifying unit superiority, and persistence of long-term mixed-unit chimerism remain elusive. Review of published literature revealed only 11 of 280 patients (4%) with mixed-unit chimerism for at least 1 year after transplantation, with 3 patients receiving reduced-intensity conditioning regimens. Mixed-unit chimerism was more likely if both units were closely HLA matched to each other. Outcome data for patients with stable mixed-unit chimerism, for the most part, were scarcely reported. Analysis of the small sample size revealed a potential advantage of stable mixed-unit chimerism on enhancing the graft-versus-leukemia effect; however, definitive conclusions cannot be made on the effect of mixed-unit chimerism on the rates of graft-versus-host disease. Therefore, gathering outcome data prospectively in larger clinical series will help answer the question of whether stable mixed-unit chimerism is either beneficial and, therefore, should be strived for, detrimental and, thus, needs to be eliminated, or if it is of no clinical consequence.

  15. T-Shaped Frame Critical and Post-Critical Analysis

    NASA Astrophysics Data System (ADS)

    Doicheva, Albena

    2016-03-01

    The paper shows solution of a T-shaped frame, strength- ened with two linear springs, regarding critical and post-critical analysis. The solution is exact using the Euler elastic approach and the frame of reference, originated in the point of column axis inflexion. The derived Numerical results show the effect of the springs strengthening for the crit- ical and the post-critical system behaviour. The influence of the geometry change is analyzed, as well.

  16. Generation of Chimeric Rhesus Monkeys

    PubMed Central

    Tachibana, Masahito; Sparman, Michelle; Ramsey, Cathy; Ma, Hong; Lee, Hyo-Sang; Penedo, Maria Cecilia T.; Mitalipov, Shoukhrat

    2011-01-01

    Summary Totipotent cells in early embryos are progenitors of all stem cells and are capable of developing into a whole organism, including extraembryonic tissues such as placenta. Pluripotent cells in the inner cell mass (ICM) are the descendants of totipotent cells and can differentiate into any cell type of a body except extraembryonic tissues. The ability to contribute to chimeric animals upon reintroduction into host embryos is the key feature of murine totipotent and pluripotent cells. Here, we demonstrate that rhesus monkey embryonic stem cells (ESCs) and isolated ICMs fail to incorporate into host embryos and develop into chimeras. However, chimeric offspring were produced following aggregation of totipotent cells of the 4-cell embryos. These results provide insights into the species-specific nature of primate embryos and suggest that a chimera assay using pluripotent cells may not be feasible. PMID:22225614

  17. Tolerance of Lung Allografts Achieved in Nonhuman Primates via Mixed Hematopoietic Chimerism.

    PubMed

    Tonsho, M; Lee, S; Aoyama, A; Boskovic, S; Nadazdin, O; Capetta, K; Smith, R-N; Colvin, R B; Sachs, D H; Cosimi, A B; Kawai, T; Madsen, J C; Benichou, G; Allan, J S

    2015-08-01

    While the induction of transient mixed chimerism has tolerized MHC-mismatched renal grafts in nonhuman primates and patients, this approach has not been successful for more immunogenic organs. Here, we describe a modified delayed-tolerance-induction protocol resulting in three out of four monkeys achieving long-term lung allograft survival without ongoing immunosuppression. Two of the tolerant monkeys displayed stable mixed lymphoid chimerism, and the other showed transient chimerism. Serial biopsies and post-mortem specimens from the tolerant monkeys revealed no signs of chronic rejection. The tolerant recipients also exhibited T cell unresponsiveness and a lack of alloantibody. This is the first report of durable mixed chimerism and successful tolerance induction of MHC-mismatched lungs in primates.

  18. Chimerism and tolerance in transplantation.

    PubMed

    Starzl, Thomas E

    2004-10-01

    Studies in experimental models (1953-1956) demonstrated that acquired donor-specific allotolerance in immunologically immature or irradiated animals is strongly associated with donor leukocyte chimerism. Bone marrow transplantation in immune-deficient or cytoablated human recipients was a logical extension (1968). In contrast, clinical (1959) and then experimental organ transplantation was systematically accomplished in the apparent absence of leukocyte chimerism. Consequently, it was assumed for many years that success with organ and bone marrow transplantation involved fundamentally different mechanisms. With the discovery in 1992 of small numbers of donor leukocytes in the tissues or blood of long-surviving organ recipients (microchimerism), we concluded that organ engraftment was a form of leukocyte chimerism-dependent partial tolerance. In this initially controversial paradigm, alloengraftment after both kinds of transplantation is the product of a double immune reaction in which responses, each to the other, of coexisting donor and recipient immune systems results in variable reciprocal clonal exhaustion, followed by peripheral clonal deletion. It was proposed with Rolf Zinkernagel that the individual alloresponses are the equivalent of the MHC-restricted T cell recognition of, and host response to, intracellular parasites and that the mechanisms of immune responsiveness, or nonresponsiveness, are governed by the migration and localization of the respective antigens. Elucidation of the mechanisms of nonresponsiveness (clonal exhaustion-deletion and immune ignorance) and their regulation removed much of the historical mystique of transplantation. The insight was then applied to improve the timing and dosage of immunosuppression of current human transplant recipients.

  19. Donor Chimerism Early after Reduced-intensity Conditioning Hematopoietic Stem Cell Transplantation Predicts Relapse and Survival

    PubMed Central

    Koreth, John; Kim, Haesook T.; Nikiforow, Sarah; Milford, Edgar L.; Armand, Philippe; Cutler, Corey; Glotzbecker, Brett; Ho, Vincent T.; Antin, Joseph H.; Soiffer, Robert J.; Ritz, Jerome; Alyea, Edwin P.

    2015-01-01

    The impact of early donor cell chimerism on outcomes of T-replete reduced-intensity conditioning (RIC) hematopoietic stem cell transplantation (HSCT) is ill-defined. We evaluated day 30 (D30) and 100 (D100) total donor cell chimerism after RIC HSCT undertaken between 2002 and 2010 at our institution, excluding patients who died or relapsed before D30. When available, donor T-cell chimerism was also assessed. The primary outcome was overall survival (OS). Secondary outcomes included progression-free survival (PFS), relapse and non-relapse mortality (NRM). 688 patients with hematologic malignancies (48% myeloid; 52% lymphoid) and a median age of 57 years (range, 18-74) undergoing RIC HSCT with T-replete donor grafts (97% peripheral blood; 92% HLA-matched) and median follow-up of 58.2 months (range, 12.6-120.7) were evaluated. In multivariable analysis total donor cell and T-cell chimerism at D30 and D100 each predicted RIC HSCT outcomes, with D100 total donor cell chimerism most predictive. D100 total donor cell chimerism <90% was associated with increased relapse (HR 2.54, 95% CI 1.83-3.51, p<0.0001), impaired PFS (HR 2.01, 95% CI 1.53-2.65, p<0.0001) and worse OS (1.50, 95% CI 1.11-2.04, p=0.009), but not NRM (HR 0.76; 95% CI 0.44-2.27, p=0.33). There was no additional utility of incorporating sustained D30-D100 total donor cell chimerism, or T-cell chimerism. Low donor chimerism early after RIC HSCT is an independent risk factor for relapse and impaired survival. Donor chimerism assessment early after RIC HSCT can prognosticate for long-term outcomes and help identify high-risk patient cohorts that may benefit from additional therapeutic interventions. PMID:24907627

  20. The concept of the eudicot shoot apical meristem as it applies to four Spiraea (Rosaceae), one Mentha (Lamiaceae) and one Euonymus (Celastraceae) cultivars based on chimeric analysis

    PubMed Central

    Korn, Robert W.

    2013-01-01

    Background and Aims Eversporting eudicots were sought to see if they behave like gymnosperms. Behaviour of eversporting gymnosperm chimeras indicates a single apical cell is present in SAM and it would be of interest to see if eudicot chimeras have the same behaviour. Methods Four eversporting spireas, the pineapple mint and the Silver King euonymus were inspected for the fate of the yellow (mutant)–green (wild type) chimeras. Key Results As with gymnosperms, unstable eudicot chimeras in the four spireas, the pineapple mint and the Silver King euonymus became stable yellow about 80 % or more of the time and 20 % or less became stable green. Conclusions The statistically significant preponderance of chimeric fates becoming all yellow suggests that a single apical cell resides in the yellow tunica. As with gymnosperms, descendent cells of the yellow replacement corpus cell eventually take over the corpus. Here is the first chimeric set of data to support the hypothesis of a one-celled meristem in eudicots rather than the traditional view of a muticellular meristem. PMID:23482330

  1. Digital PCR to assess hematopoietic chimerism after allogeneic stem cell transplantation.

    PubMed

    Stahl, Tanja; Böhme, Manja U; Kröger, Nicolaus; Fehse, Boris

    2015-06-01

    Analysis of hematopoietic chimerism after allogeneic stem cell transplantation represents a crucial method to evaluate donor-cell engraftment. Whereas sensitivity of classical approaches for chimerism monitoring is limited to ≥1%, quantitative polymerase chain reaction (qPCR)-based techniques readily detect one patient cell in >1,000 donor cells, thus facilitating application of chimerism assessment as a surrogate for minimal residual disease. However, due to methodologic specificities, qPCR combines its high sensitivity with limited resolution power in the state of mixed chimerism (e.g., >10% patient cells). Our aim was to overcome this limitation by employing a further development of qPCR, namely digital PCR (dPCR), for chimerism analysis. For proof-of-principle, we established more than 10 dPCR assays detecting Indel polymorphisms or Y-chromosome sequences and tested them on artificial cell mixtures and patient samples. Employing artificial cell mixtures, we found that dPCR allows exact quantification of chimerism over several orders of magnitude. Digital PCR results proved to be highly reproducible (deviation <5%), particularly in the "difficult" range of mixed chimerism. Excellent performance of the new assays was confirmed by analysis of multiple retrospective blood samples from patients after allogeneic stem cell transplantation, in comparison with established qPCR (14 patients) and short-tandem repeat PCR (4 patients) techniques. Finally, dPCR is easy to perform, needs only small amounts of DNA for chimerism assessment (65 ng corresponds to a sensitivity of approximately 0.03%), and does not require the use of standard curves and replicate analysis. In conclusion, dPCR represents a very promising method for routine chimerism monitoring.

  2. Transient mixed chimerism for allograft tolerance.

    PubMed

    Oura, Tetsu; Hotta, Kiyohiko; Cosimi, A B; Kawai, Tatsuo

    2015-04-01

    Mixed chimerism discovered in Freemartin cattle by Ray Owen 70 years ago paved the way for research on immune tolerance. Since his discovery, significant progress has been made in the effort to induce allograft tolerance via mixed chimerism in various murine models. However, induction of persistent mixed chimerism has proved to be extremely difficult in major histocompatibility complex mismatched humans. Chimerism induced in humans tends to either disappear or convert to full donor chimerism, depending on the intensity of the conditioning regimen. Nevertheless, our studies in both NHPs and humans have clearly demonstrated that renal allograft tolerance can be induced by transient mixed chimerism. Our studies have shown that solid organ allograft tolerance via transient mixed chimerism 1) requires induction of multilineage hematologic chimerism, 2) depends on peripheral regulatory mechanisms, rather than thymic deletion, for long-term maintenance, 3) is organ specific (kidney and lung but not heart allograft tolerance are feasible). A major advantage of tolerance induction via transient mixed chimerism is exclusion of the risk of graft-versus-host disease. Our ongoing studies are directed toward improving the consistency of tolerance induction, reducing the morbidity of the conditioning regimen, substituting clinically available agents, such as Belatacept for the now unavailable anti-CD2 monoclonal antibody, and extending the protocol to recipients of deceased donor allografts.

  3. Probabilistic structural analysis of space propulsion system LOX post

    NASA Technical Reports Server (NTRS)

    Newell, J. F.; Rajagopal, K. R.; Ho, H. W.; Cunniff, J. M.

    1990-01-01

    The probabilistic structural analysis program NESSUS (Numerical Evaluation of Stochastic Structures Under Stress; Cruse et al., 1988) is applied to characterize the dynamic loading and response of the Space Shuttle main engine (SSME) LOX post. The design and operation of the SSME are reviewed; the LOX post structure is described; and particular attention is given to the generation of composite load spectra, the finite-element model of the LOX post, and the steps in the NESSUS structural analysis. The results are presented in extensive tables and graphs, and it is shown that NESSUS correctly predicts the structural effects of changes in the temperature loading. The probabilistic approach also facilitates (1) damage assessments for a given failure model (based on gas temperature, heat-shield gap, and material properties) and (2) correlation of the gas temperature with operational parameters such as engine thrust.

  4. A technical application of quantitative next generation sequencing for chimerism evaluation

    PubMed Central

    Aloisio, Michelangelo; Licastro, Danilo; Caenazzo, Luciana; Torboli, Valentina; D'eustacchio, Angela; Severini, Giovanni Maria; Athanasakis, Emmanouil

    2016-01-01

    At present, the most common genetic diagnostic method for chimerism evaluation following hematopoietic stem cell transplantation is microsatellite analysis by capillary electrophoresis. The main objective was to establish, through repeated analysis over time, if a complete chimerism was present, or if the mixed chimerism was stable, increasing or decreasing over time. Considering the recent introduction of next generation sequencing (NGS) in clinical diagnostics, a detailed study evaluating an NGS protocol was conducted, coupled with a custom bioinformatics pipeline, for chimerism quantification. Based on the technology of Ion AmpliSeq, a 44-amplicon custom chimerism panel was designed, and a custom bioinformatics pipeline dedicated to the genotyping and quantification of NGS data was coded. The custom chimerism panel allowed identification of an average of 16 informative recipient alleles. The limit of detection of the protocol was fixed at 1% due to the NGS background (<1%). The protocol followed the standard Ion AmpliSeq library preparation and Ion Torrent Personal Genome Machine guidelines. Overall, the present study added to the scientific literature, identifying novel technical details for a possible future application of NGS for chimerism quantification. PMID:27499173

  5. A technical application of quantitative next generation sequencing for chimerism evaluation.

    PubMed

    Aloisio, Michelangelo; Licastro, Danilo; Caenazzo, Luciana; Torboli, Valentina; D'Eustacchio, Angela; Severini, Giovanni Maria; Athanasakis, Emmanouil

    2016-10-01

    At present, the most common genetic diagnostic method for chimerism evaluation following hematopoietic stem cell transplantation is microsatellite analysis by capillary electrophoresis. The main objective was to establish, through repeated analysis over time, if a complete chimerism was present, or if the mixed chimerism was stable, increasing or decreasing over time. Considering the recent introduction of next generation sequencing (NGS) in clinical diagnostics, a detailed study evaluating an NGS protocol was conducted, coupled with a custom bioinformatics pipeline, for chimerism quantification. Based on the technology of Ion AmpliSeq, a 44‑amplicon custom chimerism panel was designed, and a custom bioinformatics pipeline dedicated to the genotyping and quantification of NGS data was coded. The custom chimerism panel allowed identification of an average of 16 informative recipient alleles. The limit of detection of the protocol was fixed at 1% due to the NGS background (<1%). The protocol followed the standard Ion AmpliSeq library preparation and Ion Torrent Personal Genome Machine guidelines. Overall, the present study added to the scientific literature, identifying novel technical details for a possible future application of NGS for chimerism quantification.

  6. A technical application of quantitative next generation sequencing for chimerism evaluation.

    PubMed

    Aloisio, Michelangelo; Licastro, Danilo; Caenazzo, Luciana; Torboli, Valentina; D'Eustacchio, Angela; Severini, Giovanni Maria; Athanasakis, Emmanouil

    2016-10-01

    At present, the most common genetic diagnostic method for chimerism evaluation following hematopoietic stem cell transplantation is microsatellite analysis by capillary electrophoresis. The main objective was to establish, through repeated analysis over time, if a complete chimerism was present, or if the mixed chimerism was stable, increasing or decreasing over time. Considering the recent introduction of next generation sequencing (NGS) in clinical diagnostics, a detailed study evaluating an NGS protocol was conducted, coupled with a custom bioinformatics pipeline, for chimerism quantification. Based on the technology of Ion AmpliSeq, a 44‑amplicon custom chimerism panel was designed, and a custom bioinformatics pipeline dedicated to the genotyping and quantification of NGS data was coded. The custom chimerism panel allowed identification of an average of 16 informative recipient alleles. The limit of detection of the protocol was fixed at 1% due to the NGS background (<1%). The protocol followed the standard Ion AmpliSeq library preparation and Ion Torrent Personal Genome Machine guidelines. Overall, the present study added to the scientific literature, identifying novel technical details for a possible future application of NGS for chimerism quantification. PMID:27499173

  7. Analysis of the Relevance of Posts in Asynchronous Discussions

    ERIC Educational Resources Information Center

    Azevedo, Breno T.; Reategui, Eliseo; Behar, Patrícia A.

    2014-01-01

    This paper presents ForumMiner, a tool for the automatic analysis of students' posts in asynchronous discussions. ForumMiner uses a text mining system to extract graphs from texts that are given to students as a basis for their discussion. These graphs contain the most relevant terms found in the texts, as well as the relationships between them.…

  8. Evidence for Kidney Rejection after Combined Bone Marrow and Renal Transplantation Despite Ongoing Whole-blood Chimerism in Rhesus Macaques

    PubMed Central

    Ramakrishnan, Swetha K; Page, Andrew; Farris, Alton B.; Singh, Karnail; Leopardi, Frank; Hamby, Kelly; Sen, Sharon; Polnett, Aneesah; Deane, Taylor; Song, Mingqing; Stempora, Linda; Strobert, Elizabeth; Kirk, Allan D.; Larsen, Christian P.; Kean, Leslie S.

    2012-01-01

    Although there is evidence linking hematopoietic chimerism-induction and solid organ transplant tolerance, the mechanistic requirements for chimerism-induced tolerance are not clearly elucidated. To address this, we used an MHC-defined primate model to determine the impact of impermanent, T cell-poor, mixed-chimerism on renal allograft survival. We compared two cohorts: one receiving a bone marrow + renal transplant (“BMT/renal”) and one receiving only a renal transplant. Both cohorts received maintenance immunosuppression with CD28/CD40-directed costimulation blockade and sirolimus. As previously demonstrated, this transplant strategy consistently induced compartmentalized donor chimerism, (significant whole-blood chimerism, lacking T cell chimerism). This chimerism was not sufficient to prolong renal allograft acceptance: the BMT/renal mean survival time (MST, 76 days) was not significantly different than the renal transplant alone MST (85 days, p= 0. 46), with histopathology documenting T-cell mediated rejection. Flow cytometric analysis revealed significant enrichment for CD28-/CD95+ CD4+ and CD8+ Tem cells in the rejected kidney, suggesting a link between CD28-negative Tem and costimulation blockade-resistant rejection. These results suggest that in some settings, transient T cell-poor chimerism is not sufficient to induce tolerance to a concurrently placed renal allograft and that the presence of this chimerism per se is not an independent biomarker to identify tolerance. PMID:22642491

  9. Advanced Post-Irradiation Examination Capabilities Alternatives Analysis Report

    SciTech Connect

    Jeff Bryan; Bill Landman; Porter Hill

    2012-12-01

    An alternatives analysis was performed for the Advanced Post-Irradiation Capabilities (APIEC) project in accordance with the U.S. Department of Energy (DOE) Order DOE O 413.3B, “Program and Project Management for the Acquisition of Capital Assets”. The Alternatives Analysis considered six major alternatives: ? No Action ? Modify Existing DOE Facilities – capabilities distributed among multiple locations ? Modify Existing DOE Facilities – capabilities consolidated at a few locations ? Construct New Facility ? Commercial Partnership ? International Partnerships Based on the alternatives analysis documented herein, it is recommended to DOE that the advanced post-irradiation examination capabilities be provided by a new facility constructed at the Materials and Fuels Complex at the Idaho National Laboratory.

  10. VDJtools: Unifying Post-analysis of T Cell Receptor Repertoires.

    PubMed

    Shugay, Mikhail; Bagaev, Dmitriy V; Turchaninova, Maria A; Bolotin, Dmitriy A; Britanova, Olga V; Putintseva, Ekaterina V; Pogorelyy, Mikhail V; Nazarov, Vadim I; Zvyagin, Ivan V; Kirgizova, Vitalina I; Kirgizov, Kirill I; Skorobogatova, Elena V; Chudakov, Dmitriy M

    2015-11-01

    Despite the growing number of immune repertoire sequencing studies, the field still lacks software for analysis and comprehension of this high-dimensional data. Here we report VDJtools, a complementary software suite that solves a wide range of T cell receptor (TCR) repertoires post-analysis tasks, provides a detailed tabular output and publication-ready graphics, and is built on top of a flexible API. Using TCR datasets for a large cohort of unrelated healthy donors, twins, and multiple sclerosis patients we demonstrate that VDJtools greatly facilitates the analysis and leads to sound biological conclusions. VDJtools software and documentation are available at https://github.com/mikessh/vdjtools. PMID:26606115

  11. VDJtools: Unifying Post-analysis of T Cell Receptor Repertoires

    PubMed Central

    Bolotin, Dmitriy A.; Britanova, Olga V.; Putintseva, Ekaterina V.; Pogorelyy, Mikhail V.; Nazarov, Vadim I.; Zvyagin, Ivan V.; Kirgizova, Vitalina I.; Kirgizov, Kirill I.; Skorobogatova, Elena V.; Chudakov, Dmitriy M.

    2015-01-01

    Despite the growing number of immune repertoire sequencing studies, the field still lacks software for analysis and comprehension of this high-dimensional data. Here we report VDJtools, a complementary software suite that solves a wide range of T cell receptor (TCR) repertoires post-analysis tasks, provides a detailed tabular output and publication-ready graphics, and is built on top of a flexible API. Using TCR datasets for a large cohort of unrelated healthy donors, twins, and multiple sclerosis patients we demonstrate that VDJtools greatly facilitates the analysis and leads to sound biological conclusions. VDJtools software and documentation are available at https://github.com/mikessh/vdjtools. PMID:26606115

  12. Increasing mixed chimerism and the risk of graft loss in children undergoing allogeneic hematopoietic stem cell transplantation for non-malignant disorders.

    PubMed

    Ozyurek, E; Cowan, M J; Koerper, M A; Baxter-Lowe, L-A; Dvorak, C C; Horn, B N

    2008-07-01

    We performed quantitative PCR-based serial chimerism testing of whole blood (WB) and CD3+ cells and retrospectively correlated the results of chimerism tests and the risk of graft loss in children undergoing transplant for non-malignant disorders. Twenty-four children were included in this study. All patients initially engrafted; subsequently, 12% lost the graft, 21% achieved complete donor chimerism and 67% had mixed chimerism (MC). Patients underwent delayed taper of cyclosporine (CsA) if they had MC. Overall survival was 87+/-7% (s.d.) at 5-years post transplant, and it was not affected by chimerism status. Both WB and CD3+ chimerism showed significant fluctuations with a peak in autologous cell signal occurring at a median of 7 months for WB and 2 months for CD3+ cells. Initial post transplant chimerism percentage in either WB or CD3+ lineage was not related to graft loss. Increasing MC to >30% host cells was seen in 33% of patients, and it was related to increased risk of graft loss, as previously published. However, 63% of children with increasing MC did not lose their graft. Additional studies of post transplant chimerism are required to improve our ability to accurately identify children at risk of graft loss following transplant for non-malignant disorders.

  13. Monitoring of chimerism following allogeneic haematopoietic stem cell transplantation (HSCT): technical recommendations for the use of short tandem repeat (STR) based techniques, on behalf of the United Kingdom National External Quality Assessment Service for Leucocyte Immunophenotyping Chimerism Working Group.

    PubMed

    Clark, Jordan R; Scott, Stuart D; Jack, Andrea L; Lee, Helena; Mason, Joanne; Carter, Geoffrey I; Pearce, Laurence; Jackson, Tony; Clouston, Hazel; Sproul, Anne; Keen, Leigh; Molloy, Karen; Folarin, Najeem'deen; Whitby, Liam; Snowden, John A; Reilly, John T; Barnett, David

    2015-01-01

    Analysis of short tandem repeats (STR) is the predominant method for post-transplant monitoring of donor engraftment. It can enable early detection of disease relapse, level of engraftment and provide useful information on the graft-versus-host disease (GVHD)/graft-versus-tumour (GVT) effect, facilitating therapeutic intervention. Harmonization and standardization of techniques and result interpretation is essential to reduce the impact of laboratory variability on both clinical management and the results of multi-centre clinical trials. However, the United Kingdom National External Quality Assessment Service for Leucocyte Immunophenotyping (UK NEQAS LI) has highlighted significant issues inherent in STR testing that impact upon inter- and intra- laboratory variation. We present here consensus best practice guidelines and recommendations for STR chimerism testing, data interpretation and reporting that have been drawn up and agreed by a consortium of 11 UK and Eire clinical laboratories. This document uses data obtained from the UK NEQAS LI Post-Stem Cell Transplant (SCT) Chimerism Monitoring Programme.

  14. Post-Optimality Analysis In Aerospace Vehicle Design

    NASA Technical Reports Server (NTRS)

    Braun, Robert D.; Kroo, Ilan M.; Gage, Peter J.

    1993-01-01

    This analysis pertains to the applicability of optimal sensitivity information to aerospace vehicle design. An optimal sensitivity (or post-optimality) analysis refers to computations performed once the initial optimization problem is solved. These computations may be used to characterize the design space about the present solution and infer changes in this solution as a result of constraint or parameter variations, without reoptimizing the entire system. The present analysis demonstrates that post-optimality information generated through first-order computations can be used to accurately predict the effect of constraint and parameter perturbations on the optimal solution. This assessment is based on the solution of an aircraft design problem in which the post-optimality estimates are shown to be within a few percent of the true solution over the practical range of constraint and parameter variations. Through solution of a reusable, single-stage-to-orbit, launch vehicle design problem, this optimal sensitivity information is also shown to improve the efficiency of the design process, For a hierarchically decomposed problem, this computational efficiency is realized by estimating the main-problem objective gradient through optimal sep&ivity calculations, By reducing the need for finite differentiation of a re-optimized subproblem, a significant decrease in the number of objective function evaluations required to reach the optimal solution is obtained.

  15. Engineering bacterial microcompartment shells: chimeric shell proteins and chimeric carboxysome shells.

    PubMed

    Cai, Fei; Sutter, Markus; Bernstein, Susan L; Kinney, James N; Kerfeld, Cheryl A

    2015-04-17

    Bacterial microcompartments (BMCs) are self-assembling organelles composed entirely of protein. Depending on the enzymes they encapsulate, BMCs function in either inorganic carbon fixation (carboxysomes) or organic carbon utilization (metabolosomes). The hallmark feature of all BMCs is a selectively permeable shell formed by multiple paralogous proteins, each proposed to confer specific flux characteristics. Gene clusters encoding diverse BMCs are distributed broadly across bacterial phyla, providing a rich variety of building blocks with a predicted range of permeability properties. In theory, shell permeability can be engineered by modifying residues flanking the pores (symmetry axes) of hexameric shell proteins or by combining shell proteins from different types of BMCs into chimeric shells. We undertook both approaches to altering shell properties using the carboxysome as a model system. There are two types of carboxysomes, α and β. In both, the predominant shell protein(s) contain a single copy of the BMC domain (pfam00936), but they are significantly different in primary structure. Indeed, phylogenetic analysis shows that the two types of carboxysome shell proteins are more similar to their counterparts in metabolosomes than to each other. We solved high resolution crystal structures of the major shell proteins, CsoS1 and CcmK2, and the presumed minor shell protein CcmK4, representing both types of cyanobacterial carboxysomes and then tested the interchangeability. The in vivo study presented here confirms that both engineering pores to mimic those of other shell proteins and the construction of chimeric shells is feasible.

  16. A development of chimeric VEGFR2 TK inhibitor based on two ligand conformers from PDB: 1Y6A complex--medicinal chemistry consequences of a TKs analysis.

    PubMed

    Lintnerová, Lucia; García-Caballero, Melissa; Gregáň, Fridrich; Melicherčík, Milan; Quesada, Ana R; Dobiaš, Juraj; Lác, Ján; Sališová, Marta; Boháč, Andrej

    2014-01-24

    VEGFR2 is an important mediator of angiogenesis and influences fate of some cancer stem cells. Here we analysed all 34 structures of VEGFR2 TK available from PDB database. From them a complex PDB: 1Y6A has an exceptional AAZ ligand bound to TK in form of two conformers (U- and S-shaped). This observation inspired us to develop three chimeric bispyridyl VEGFR2 inhibitors by combining structural features of both AAZ conformers and/or their relative ligand AAX (PDB: 1Y6B). Our most interesting inhibitor 22SYM has an enzymatic VEGFR2 TK activity (IC50: 15.1 nM) comparable or better to the active compounds from clinical drugs Nexavar and Sutent. 22SYM inhibits growth, migration and tube formation of endothelial cells (EC) and selectively induces EC apoptosis. 22SYM also inhibits in vivo angiogenesis in Zebrafish embryo assay. Additionally to the above results, we proved here that tyrosine kinases in an inactive form possessing Type I inhibitors can adopt both a closed or an opened conformation of kinase A-loop independently on their DFG-out arrangement. We proposed here that an activity of certain Type I inhibitors (e.g. 22SYM-like) in complex with DFG-out TK can be negatively influenced by collisions with a dynamically moving TK A-loop.

  17. In silico analysis of chimeric espA, eae and tir fragments of Escherichia coli O157:H7 for oral immunogenic applications

    PubMed Central

    2009-01-01

    Background In silico techniques are highly suited for both the discovery of new and development of existing vaccines. Enterohemorrhagic Escherichia coli O157:H7 (EHEC) exhibits a pattern of localized adherence to host cells, with the formation of microcolonies, and induces a specific histopathological lesion (attaching/effacing). The genes encoding the products responsible for this phenotype are clustered on a 35-kb pathogenicity island. Among these proteins, Intimin, Tir, and EspA, which are expressed by attaching-effacing genes, are responsible for the attachment to epithelial cell that leads to lesions. Results We designed synthetic genes encoding the carboxy-terminal fragment of Intimin, the middle region of Tir and the carboxy-terminal part of EspA. These multi genes were synthesized with codon optimization for a plant host and were fused together by the application of four repeats of five hydrophobic amino acids as linkers. The structure of the synthetic construct gene, its mRNA and deduced protein and their stabilities were analyzed by bioinformatic software. Furthermore, the immunogenicity of this multimeric recombinant protein consisting of three different domains was predicted. Conclusion a structural model for a chimeric gene from LEE antigenic determinants of EHEC is presented. It may define accessibility, solubility and immunogenecity. PMID:19995413

  18. FARO base case post-test analysis by COMETA code

    SciTech Connect

    Annunziato, A.; Addabbo, C.

    1995-09-01

    The paper analyzes the COMETA (Core Melt Thermal-Hydraulic Analysis) post test calculations of FARO Test L-11, the so-called Base Case Test. The FARO Facility, located at JRC Ispra, is used to simulate the consequences of Severe Accidents in Nuclear Power Plants under a variety of conditions. The COMETA Code has a 6 equations two phase flow field and a 3 phases corium field: the jet, the droplets and the fused-debris bed. The analysis shown that the code is able to pick-up all the major phenomena occurring during the fuel-coolant interaction pre-mixing phase.

  19. Long-term outcome and lineage-specific chimerism in 194 patients with Wiskott-Aldrich syndrome treated by hematopoietic cell transplantation in the period 1980-2009: an international collaborative study

    PubMed Central

    Moratto, Daniele; Giliani, Silvia; Bonfim, Carmem; Mazzolari, Evelina; Fischer, Alain; Ochs, Hans D.; Cant, Andrew J.; Thrasher, Adrian J.; Cowan, Morton J.; Albert, Michael H.; Small, Trudy; Pai, Sung-Yun; Haddad, Elie; Lisa, Antonella; Hambleton, Sophie; Slatter, Mary; Cavazzana-Calvo, Marina; Mahlaoui, Nizar; Picard, Capucine; Torgerson, Troy R.; Burroughs, Lauri; Koliski, Adriana; Neto, Jose Zanis; Porta, Fulvio; Qasim, Waseem; Veys, Paul; Kavanau, Kristina; Hönig, Manfred; Schulz, Ansgar; Friedrich, Wilhelm

    2011-01-01

    In this retrospective collaborative study, we have analyzed long-term outcome and donor cell engraftment in 194 patients with Wiskott-Aldrich syndrome (WAS) who have been treated by hematopoietic cell transplantation (HCT) in the period 1980- 2009. Overall survival was 84.0% and was even higher (89.1% 5-year survival) for those who received HCT since the year 2000, reflecting recent improvement of outcome after transplantation from mismatched family donors and for patients who received HCT from an unrelated donor at older than 5 years. Patients who went to transplantation in better clinical conditions had a lower rate of post-HCT complications. Retrospective analysis of lineage-specific donor cell engraftment showed that stable full donor chimerism was attained by 72.3% of the patients who survived for at least 1 year after HCT. Mixed chimerism was associated with an increased risk of incomplete reconstitution of lymphocyte count and post-HCT autoimmunity, and myeloid donor cell chimerism < 50% was associated with persistent thrombocytopenia. These observations indicate continuous improvement of outcome after HCT for WAS and may have important implications for the development of novel protocols aiming to obtain full correction of the disease and reduce post-HCT complications. PMID:21659547

  20. Long-term outcome and lineage-specific chimerism in 194 patients with Wiskott-Aldrich syndrome treated by hematopoietic cell transplantation in the period 1980-2009: an international collaborative study.

    PubMed

    Moratto, Daniele; Giliani, Silvia; Bonfim, Carmem; Mazzolari, Evelina; Fischer, Alain; Ochs, Hans D; Cant, Andrew J; Thrasher, Adrian J; Cowan, Morton J; Albert, Michael H; Small, Trudy; Pai, Sung-Yun; Haddad, Elie; Lisa, Antonella; Hambleton, Sophie; Slatter, Mary; Cavazzana-Calvo, Marina; Mahlaoui, Nizar; Picard, Capucine; Torgerson, Troy R; Burroughs, Lauri; Koliski, Adriana; Neto, Jose Zanis; Porta, Fulvio; Qasim, Waseem; Veys, Paul; Kavanau, Kristina; Hönig, Manfred; Schulz, Ansgar; Friedrich, Wilhelm; Notarangelo, Luigi D

    2011-08-11

    In this retrospective collaborative study, we have analyzed long-term outcome and donor cell engraftment in 194 patients with Wiskott-Aldrich syndrome (WAS) who have been treated by hematopoietic cell transplantation (HCT) in the period 1980- 2009. Overall survival was 84.0% and was even higher (89.1% 5-year survival) for those who received HCT since the year 2000, reflecting recent improvement of outcome after transplantation from mismatched family donors and for patients who received HCT from an unrelated donor at older than 5 years. Patients who went to transplantation in better clinical conditions had a lower rate of post-HCT complications. Retrospective analysis of lineage-specific donor cell engraftment showed that stable full donor chimerism was attained by 72.3% of the patients who survived for at least 1 year after HCT. Mixed chimerism was associated with an increased risk of incomplete reconstitution of lymphocyte count and post-HCT autoimmunity, and myeloid donor cell chimerism < 50% was associated with persistent thrombocytopenia. These observations indicate continuous improvement of outcome after HCT for WAS and may have important implications for the development of novel protocols aiming to obtain full correction of the disease and reduce post-HCT complications. PMID:21659547

  1. Real-time qPCR for chimerism assessment in allogeneic hematopoietic stem cell transplants from unrelated adult and double umbilical cord blood.

    PubMed

    Frankfurt, Olga; Zitzner, Jennifer R; Tambur, Anat R

    2015-03-01

    Allogeneic hematopoietic stem cell transplantation is the standard therapy for patients with various malignant hematologic disorders. A successful treatment results in complete engraftment of donor cells in the absence of the patient's own hematopoietic system. Chimerism analysis, aimed at determining the coexistence of genetically different cell populations, is considered a useful method to measure treatment success. A new, qPCR based, commercially available chimerism assay was recently introduced. Here we report our results of comparing STR with qPCR-based chimerism analysis, and assessment of sensitivity and reproducibility of the qPCR chimerism assay. A specific emphasis is put on analyzing chimerism in recipients of double cord blood transplantation. We conclude that the qPCR chimerism assay for engraftment monitoring is a reliable and sensitive assay. Advantages and limitations of the assay in its current format are summarized.

  2. Mapping of low-frequency chimeric yeast artificial chromosome libraries from human chromosomes 16 and 21 by fluorescence in situ hybridization and quantitative image analysis

    SciTech Connect

    Marrone, B.L.; Campbell, E.W.; Anzick, S.L.; Shera, K.; Campbell, M.; Yoshida, T.M.; McCormick, M.K.; Deaven, L. )

    1994-05-01

    Yeast artificial chromosome (YAC) clones from low-frequency chimeric libraries of human chromosomes 16 and 21 were mapped onto human diploid fibroblast metaphase chromosomes using fluorescence in situ hybridization (FISH) and digital imaging microscopy. YACs mapped onto chromosome 21 were selected to provide subregional location and ordering of known and unknown markers on the long arm of chromosome 21, particularly in the Down syndrome region (q22). YACs mapped onto chromosome 16 were selected to overlap regions spanning chromosome 16 cosmid maps. YAC clones were indirectly labeled with fluorescein, and the total DNA of the chromosome was counterstained with propidium iodide. A single image containing both the FISH signal and the whole chromosome was acquired for each chromosome of interest containing the fluorescent probe signal in a metaphase spread. From the digitized image, the fluorescence intensity profile through the long axis of the chromosome gave the total chromosome length and the probe position. The map position of the probe was expressed as the fractional length (FL) of the total chromosome relative to the end of the short arm (Flpter). From each clone hybridized, 20-40 chromosome images were analyzed. Thirty-eight YACs were mapped onto chromosome 16, and their FLs were distributed along the short and long arms. On chromosome 21, 47 YACs were mapped, including 12 containing known markers. To confirm the order of a dense population of YACs within the Down syndrome region, a two-color mapping strategy was used in which an anonymous YAC was located relative to one or two known markers on the metaphase chromosome. The chromosome FL maps have a 1- to 2-Mb resolution, and the FL measurement of each probe has a typical standard error of 0.5-1 Mb. 14 refs., 3 figs., 3 tabs.

  3. Characterization and Functional Analysis of scFv-based Chimeric Antigen Receptors to Redirect T Cells to IL13Rα2-positive Glioma.

    PubMed

    Krenciute, Giedre; Krebs, Simone; Torres, David; Wu, Meng-Fen; Liu, Hao; Dotti, Gianpietro; Li, Xiao-Nan; Lesniak, Maciej S; Balyasnikova, Irina V; Gottschalk, Stephen

    2016-02-01

    Immunotherapy with T cells expressing chimeric antigen receptors (CARs) is an attractive approach to improve outcomes for patients with glioblastoma (GBM). IL13Rα2 is expressed at a high frequency in GBM but not in normal brain, making it a promising CAR T-cell therapy target. IL13Rα2-specific CARs generated up to date contain mutated forms of IL13 as an antigen-binding domain. While these CARs target IL13Rα2, they also recognize IL13Rα1, which is broadly expressed. To overcome this limitation, we constructed a panel of IL13Rα2-specific CARs that contain the IL13Rα2-specific single-chain variable fragment (scFv) 47 as an antigen binding domain, short or long spacer regions, a transmembrane domain, and endodomains derived from costimulatory molecules and CD3.ζ (IL13Rα2-CARs). IL13Rα2-CAR T cells recognized IL13Rα2-positive target cells in coculture and cytotoxicity assays with no cross-reactivity to IL13Rα1. However, only IL13Rα2-CAR T cells with a short spacer region produced IL2 in an antigen-dependent fashion. In vivo, T cells expressing IL13Rα2-CARs with short spacer regions and CD28.ζ, 41BB.ζ, and CD28.OX40.ζ endodomains had potent anti-glioma activity conferring a significant survival advantage in comparison to mice that received control T cells. Thus, IL13Rα2-CAR T cells hold the promise to improve current IL13Rα2-targeted immunotherapy approaches for GBM and other IL13Rα2-positive malignancies. PMID:26514825

  4. Regression analysis exploring teacher impact on student FCI post scores

    NASA Astrophysics Data System (ADS)

    Mahadeo, Jonathan V.; Manthey, Seth R.; Brewe, Eric

    2013-01-01

    High School Modeling Workshops are designed to improve high school physics teachers' understanding of physics and how to teach using the Modeling method. The basic assumption is that the teacher plays a critical role in their students' physics education. This study investigated teacher impacts on students' Force Concept Inventory scores, (FCI), with the hopes of identifying quantitative differences between teachers. This study examined student FCI scores from 18 teachers with at least a year of teaching high school physics. This data was then evaluated using a General Linear Model (GLM), which allowed for a regression equation to be fitted to the data. This regression equation was used to predict student post FCI scores, based on: teacher ID, student pre FCI score, gender, and representation. The results show 12 out of 18 teachers significantly impact their student post FCI scores. The GLM further revealed that of the 12 teachers only five have a positive impact on student post FCI scores. Given these differences among teachers it is our intention to extend our analysis to investigate pedagogical differences between them.

  5. Early establishment of hematopoietic chimerism following allogeneic peripheral blood stem cell transplantation in comparison with allogeneic bone marrow transplantation.

    PubMed

    Nakao, S; Zeng, W; Yamazaki, H; Wang, H; Takami, A; Sugimori, N; Miura, Y; Shiobara, S; Matsuda, T; Shinagawa, Y; Harada, M

    1999-04-01

    To characterize the process of the establishment of complete chimerism after allogeneic peripheral blood stem cell transplantation (allo-PBSCT), we determined the origin of leukocytes in peripheral blood (PB) obtained from 23 patients in the very early period after allo-PBSCT using amplification of mini- or microsatellite regions of genomic DNA. Donor-specific alleles were amplified from the PB obtained at day 8 post-transplant for 19 allo-PBSCT patients. Among the 19 patients, 12 showed only donor-specific alleles (complete chimerism) while 7 did both donor and host-specific alleles (mixed chimerism). Although donor specific alleles were amplified in 10 of 12 patients who received allogeneic bone marrow transplantation (allo-BMT) similarly to allo-PBSCT, all of these ten showed mixed chimerism. When the chimeric state was examined in PB samples obtained serially at 2-3-day intervals post-transplant, host-specific alleles in allo-PBSCT patients were not detectable in the PB much earlier than those in allo-BMT patients. These findings indicate that the appearance of donor-derived cells associated with the disappearance of host-derived cells in the circulation occurs earlier after allo-PBSCT as compared with allo-BMT, leading to the rapid establishment of complete chimerism.

  6. A POST ANALYSIS OF A PREVENTIVE AND CHRONIC HEALTHCARE TOOL.

    PubMed

    Borrayo, Brooke D; O'Lawrence, Henry

    2016-01-01

    This study uses the data set from Kaiser Permanente to examine the post implementation of a preventive and chronic care that utilizes clinical information system, delivery system design, and clinical decision support to maximize the office visit. The analysis suggests a significant positive relationship between frequency of utilization rates to address preventive and chronic care gaps. There is no implication of a significant positive relationship with the successfully captured rate, which satisfies closing the care gap within 45 days. The use of the preventive care tool will assist members in satisfying the preventive care gap, cervical cancer screening, within 45 days of the encounter. PMID:27483973

  7. Prognostic utility of routine chimerism testing at 2 to 6 months after allogeneic hematopoietic cell transplantation.

    PubMed

    Mossallam, Ghada I; Kamel, Azza M; Storer, Barry; Martin, Paul J

    2009-03-01

    The utility of routine chimerism analysis as a prognostic indicator of subsequent outcomes after allogeneic hematopoietic cell transplantation (HCT) with myeloablative conditioning regimens remains controversial. To address this controversy, routine chimerism test results at 2 to 6 months after HCT with myeloablative conditioning regimens were evaluated for association with subsequent risk of chronic graft-versus-host disease (GVHD), nonrelapse mortality (NRM), relapse, and overall mortality. Only 70 of 1304 patients (5%) had < 95% donor-derived cells in the marrow. Low donor chimerism in the marrow occurred more often in patients with low-risk diseases compared with those with higher-risk diseases and was significantly associated with a reduced risk of chronic GVHD. Among 673 patients evaluated, 164 (24%) had < 85% donor-derived T cells in the blood. Low donor T cell chimerism was more frequent in patients with low-risk diseases compared with those with higher-risk diseases, in those who received conditioning with busulfan compared with those who received conditioning with total body irradiation, and in those with lower-grade acute GVHD. Low donor T cell chimerism in the blood was significantly associated with a reduced risk of chronic GVHD but not with a reduced risk of relapse, NRM, or overall mortality. Routine testing of chimerism in the marrow and blood at 2 to 6 months after HCT with myeloablative conditioning regimens may be helpful in documenting engraftment in clinical trials, but provides only limited prognostic information in clinical practice.

  8. Generation and evaluation of a chimeric classical swine fever virus expressing a visible marker gene.

    PubMed

    Li, Yongfeng; Wang, Xiao; Sun, Yuan; Li, Lian-Feng; Zhang, Lingkai; Li, Su; Luo, Yuzi; Qiu, Hua-Ji

    2016-03-01

    Classical swine fever virus (CSFV) is a noncytopathogenic virus, and the incorporation of an enhanced green fluorescent protein (EGFP) tag into the viral genome provides a means of direct monitoring of viral infection without immunostaining. It is well established that the 3' untranslated region (3'-UTR) of the CSFV plays an important role in viral RNA replication. Although CSFV carrying a reporter gene and chimeric CSFV have been generated and evaluated, a chimeric CSFV with a visible marker has not yet been reported. Here, we generated and evaluated a chimeric virus containing the EGFP tag and the 3'-UTR from vaccine strain HCLV (C-strain) in the genetic background of the highly virulent CSFV Shimen strain. The chimeric marker CSFV was fluorescent and had an approximately 100-fold lower viral titer, lower replication level of viral genome, and weaker fluorescence intensity than the recombinant CSFV with only the EGFP tag or the parental virus. Furthermore, the marker chimera was avirulent and displayed no viremia in inoculated pigs, which were completely protected from lethal CSFV challenge as early as 15 days post-inoculation. The chimeric marker virus was visible in vitro and attenuated in vitro and in vivo, which suggests that CSFV can be engineered to produce attenuated variants with a visible marker to facilitate in vitro studies of CSFV infection and replication and to develop of novel vaccines against CSF. PMID:26614259

  9. Post mitigation impact risk analysis for asteroid deflection demonstration missions

    NASA Astrophysics Data System (ADS)

    Eggl, Siegfried; Hestroffer, Daniel; Thuillot, William; Bancelin, David; Cano, Juan L.; Cichocki, Filippo

    2015-08-01

    Even though mankind believes to have the capabilities to avert potentially disastrous asteroid impacts, only the realization of mitigation demonstration missions can validate this claim. Such a deflection demonstration attempt has to be cost effective, easy to validate, and safe in the sense that harmless asteroids must not be turned into potentially hazardous objects. Uncertainties in an asteroid's orbital and physical parameters as well as those additionally introduced during a mitigation attempt necessitate an in depth analysis of deflection mission designs in order to dispel planetary safety concerns. We present a post mitigation impact risk analysis of a list of potential kinetic impactor based deflection demonstration missions proposed in the framework of the NEOShield project. Our results confirm that mitigation induced uncertainties have a significant influence on the deflection outcome. Those cannot be neglected in post deflection impact risk studies. We show, furthermore, that deflection missions have to be assessed on an individual basis in order to ensure that asteroids are not inadvertently transported closer to the Earth at a later date. Finally, we present viable targets and mission designs for a kinetic impactor test to be launched between the years 2025 and 2032.

  10. Post-optimality analysis in aerospace vehicle design

    NASA Technical Reports Server (NTRS)

    Braun, Robert D.; Kroo, Ilan M.; Gage, Peter J.

    1993-01-01

    This analysis pertains to the applicability of optimal sensitivity information to aerospace vehicle design. The present analysis demonstrates that post-optimality information generated through first-order computations can be used to accurately predict file effect of constraint and parameter perturbations on the optimal solution. This assessment is based on the solution of an aircraft design problem in which the post-optimality estimates are shown to be within a few percent of the true solution over the practical range of constraint and parameter variations. Through solution of a reusable, single-stage-to-orbit, launch vehicle design problem, this optimal sensitivity information is also shown to improve the efficiency of the design process. For a hierarchically decomposed problem, this computational efficiency is realizable by estimating the main-problem objective gradient through optimal sensitivity calculations. By reducing the need for finite differentiation of a re-optimized subproblem, a significant decrease in the number of objective function evaluations required to reach the optimal solution is obtained.

  11. Chimerism and xenotransplantation. New concepts.

    PubMed

    Starzl, T E; Rao, A S; Murase, N; Demetris, A J; Thomson, A; Fung, J J

    1999-02-01

    In both transplant and infectious circumstances, the immune response is governed by migration and localization of the antigen. If the antigenic epitopes of transgenic xenografts are sufficiently altered to avoid evoking the destructive force of innate immunity, the mechanisms of engraftment should be the same as those that permit the chimerism-dependent immunologic confrontation and resolution that is the basis of allograft acceptance. In addition to "humanizing" the epitopes, one of the unanswered questions is whether the species restriction of complement described in 1994 by Valdivia and colleagues also necessitates the introduction of human complement regulatory genes in animal donors. Because the liver is the principal or sole source of most complement components, the complement quickly is transformed to that of the donor after hepatic transplantation. Thus, the need for complementary regulatory transgenes may vary according to the kind of xenograft used. Much evidence shows that physiologically important peptides produced by xenografts (e.g., insulin, clotting factors, and enzymes) are incorporated into the metabolic machinery of the recipient body. To the extent that this is not true, xenotransplantation could result in the production of diseases that are analogous to inborn errors of metabolism. In the climate of pessimism that followed the failures of baboon to human liver xenotransplantation in 1992-1993, it seemed inconceivable that the use of even more discordant donors, such as the pig, could ever be seriously entertained; however, this preceded insight into the xenogeneic and allogeneic barriers that has brought transplantation infectious immunity to common ground. With this new insight and the increasing ease of producing transgenic donors, the goal of clinical xenotransplantation may not be so distant.

  12. Donor chimerism early after reduced-intensity conditioning hematopoietic stem cell transplantation predicts relapse and survival.

    PubMed

    Koreth, John; Kim, Haesook T; Nikiforow, Sarah; Milford, Edgar L; Armand, Philippe; Cutler, Corey; Glotzbecker, Brett; Ho, Vincent T; Antin, Joseph H; Soiffer, Robert J; Ritz, Jerome; Alyea, Edwin P

    2014-10-01

    The impact of early donor cell chimerism on outcomes of T cell-replete reduced-intensity conditioning (RIC) hematopoietic stem cell transplantation (HSCT) is ill defined. We evaluated day 30 (D30) and 100 (D100) total donor cell chimerism after RIC HSCT undertaken between 2002 and 2010 at our institution, excluding patients who died or relapsed before D30. When available, donor T cell chimerism was also assessed. The primary outcome was overall survival (OS). Secondary outcomes included progression-free survival (PFS), relapse, and nonrelapse mortality (NRM). We evaluated 688 patients with hematologic malignancies (48% myeloid and 52% lymphoid) and a median age of 57 years (range, 18 to 74) undergoing RIC HSCT with T cell-replete donor grafts (97% peripheral blood; 92% HLA-matched), with a median follow-up of 58.2 months (range, 12.6 to 120.7). In multivariable analysis, total donor cell and T cell chimerism at D30 and D100 each predicted RIC HSCT outcomes, with D100 total donor cell chimerism most predictive. D100 total donor cell chimerism <90% was associated with increased relapse (hazard ratio [HR], 2.54; 95% confidence interval [CI], 1.83 to 3.51; P < .0001), impaired PFS (HR, 2.01; 95% CI, 1.53 to 2.65; P < .0001), and worse OS (HR, 1.50; 95% CI, 1.11 to 2.04, P = .009), but not with NRM (HR, .76; 95% CI, .44 to 2.27; P = .33). There was no additional utility of incorporating sustained D30 to D100 total donor cell chimerism or T cell chimerism. Low donor chimerism early after RIC HSCT is an independent risk factor for relapse and impaired survival. Donor chimerism assessment early after RIC HSCT can prognosticate for long-term outcomes and help identify high-risk patient cohorts who may benefit from additional therapeutic interventions.

  13. Induction of tolerance through mixed chimerism.

    PubMed

    Sachs, David H; Kawai, Tatsuo; Sykes, Megan

    2014-01-01

    "Mixed chimerism" refers to a state in which the lymphohematopoietic system of the recipient of allogeneic hematopoietic stem cells comprises a mixture of host and donor cells. This state is usually attained through either bone marrow or mobilized peripheral blood stem cell transplantation. Although numerous treatment regimens have led to transplantation tolerance in mice, the induction of mixed chimerism is currently the only treatment modality that has been successfully extended to large animals and to the clinic. Here we describe and compare the use of mixed chimerism to establish transplantation tolerance in mice, pigs, monkeys, and in the clinic. We also attempt to correlate the mechanisms involved in achieving tolerance with the nature of the tolerance that has resulted in each case.

  14. Pre-treatment of allogeneic bone marrow recipients with the CXCR4 antagonist AMD3100 transiently enhances hematopoietic chimerism without promoting donor-specific skin allograft tolerance.

    PubMed

    Li, Zhanzhuo; Xu, Xin; Weiss, Ido D; Jacobson, Orit; Murphy, Philip M

    2015-10-01

    Hematopoietic chimerism established by allogeneic bone marrow transplantation is known to promote donor-specific organ allograft tolerance; however, clinical application is limited by the need for toxic host conditioning and "megadoses" of donor bone marrow cells. A potential solution to this problem has been suggested by the observation that recipient bone marrow mobilization by the CXCR4 antagonist AMD3100 promotes chimerism in congenic bone marrow transplantation experiments in mice. Here we report that a single subcutaneous dose of 10 mg/kg AMD3100 in recipient C57BL/6 mice was able to enhance hematopoietic chimerism when complete MHC-mismatched BALB/c donor bone marrow cells were transplanted 1h after drug dosing. However, levels of chimerism measured 30 days post-transplantation were not sustained when mice were reexamined on day 90 post-transplantation. Moreover, transient chimerism induced by this protocol did not support robust donor-specific skin allograft tolerance. Using the same transient immunosuppression protocol, we confirmed that "megadoses" of donor bone marrow cells could induce durable chimerism associated with donor-specific skin allograft tolerance without AMD3100 pre-treatment. We conclude that in this protocol AMD3100 pretreatment may empty bone marrow niches that become reoccupied by allogeneic donor hematopoietic progenitor cells but not by true long-lived donor hematopoietic stem cells, resulting in short-lived chimerism and failure to support durable donor-specific allograft tolerance.

  15. A protective chimeric antibody to tick-borne encephalitis virus.

    PubMed

    Baykov, Ivan K; Matveev, Andrey L; Stronin, Oleg V; Ryzhikov, Alexander B; Matveev, Leonid E; Kasakin, Marat F; Richter, Vladimir A; Tikunova, Nina V

    2014-06-17

    The efficiency of several mouse monoclonal antibodies (mAbs) specific to the tick-borne encephalitis virus (TBEV) glycoprotein E in post-exposure prophylaxis was assessed, and mAb14D5 was shown to be the most active of all those studied. It was proven that the hybridoma cell line 14D5 produced one immunoglobulin H chain and two L chains. They were used to construct chimeric antibodies ch14D5a and ch14D5b, the affinity constants of which were 2.6 × 10(10)M(-1) and 1.0 × 10(7)M(-1), respectively, according to the SPR-based ProteOn biosensor assay. The neutralization index (IC50) of ch14D5a was 0.04 μg/ml in the focus reduction neutralization test. In in vivo experiments, ch14D5a at a dose of 10 μg/mouse resulted in a 100% survival of the mice infected with 240 LD50 of TBEV. This chimeric antibody is promising for further development of prevention and therapeutic drugs against TBEV. PMID:24837772

  16. Cost analysis of post-polio certification immunization policies.

    PubMed Central

    Sangrujee, Nalinee; Cáceres, Victor M.; Cochi, Stephen L.

    2004-01-01

    OBJECTIVE: An analysis was conducted to estimate the costs of different potential post-polio certification immunization policies currently under consideration, with the objective of providing this information to policy-makers. METHODS: We analyzed three global policy options: continued use of oral poliovirus vaccine (OPV); OPV cessation with optional inactivated poliovirus vaccine (IPV); and OPV cessation with universal IPV. Assumptions were made on future immunization policy decisions taken by low-, middle-, and high-income countries. We estimated the financial costs of each immunization policy, the number of vaccine-associated paralytic poliomyelitis (VAPP) cases, and the global costs of maintaining an outbreak response capacity. The financial costs of each immunization policy were based on estimates of the cost of polio vaccine, its administration, and coverage projections. The costs of maintaining outbreak response capacity include those associated with developing and maintaining a vaccine stockpile in addition to laboratory and epidemiological surveillance. We used the period 2005-20 as the time frame for the analysis. FINDINGS: OPV cessation with optional IPV, at an estimated cost of US$ 20,412 million, was the least costly option. The global cost of outbreak response capacity was estimated to be US$ 1320 million during 2005-20. The policy option continued use of OPV resulted in the highest number of VAPP cases. OPV cessation with universal IPV had the highest financial costs, but it also had the least number of VAPP cases. Sensitivity analyses showed that global costs were sensitive to assumptions on the cost of the vaccine. Analysis also showed that if the price per dose of IPV was reduced to US$ 0.50 for low-income countries, the cost of OPV cessation with universal IPV would be the same as the costs of continued use of OPV. CONCLUSION: Projections on the vaccine price per dose and future coverage rates were major drivers of the global costs of post

  17. Emoting infertility online: A qualitative analysis of men's forum posts.

    PubMed

    Hanna, Esmée; Gough, Brendan

    2016-07-01

    Relatively little research on infertility focuses exclusively or significantly on men's experiences, particularly in relation to emotional aspects. Evidence that does exist around male infertility suggests that it is a distressing experience for men, due to stigma, threats to masculinity and the perceived need to suppress emotions, and that men and women experience infertility differently. Using thematic analysis, this article examines the online emoting of men in relation to infertility via forum posts from a men-only infertility discussion board. It was noted that men 'talked' to each other about the emotional burdens of infertility, personal coping strategies and relationships with others. Three major themes were identified following in-depth analysis: 'the emotional rollercoaster', 'the tyranny of infertility' and 'infertility paranoia'. This article then offers insights into how men experience infertility emotionally, negotiate the emotional challenges involved (especially pertaining to diagnosis, treatment outcomes and their intimate relationships) and how they share (and find value in doing so) with other men the lived experience of infertility. PMID:27246813

  18. SHEFEX II Flight Instrumentation And Preparation Of Post Flight Analysis

    NASA Astrophysics Data System (ADS)

    Thiele, Thomas; Siebe, Frank; Gulhan, Ali

    2011-05-01

    A main disadvantage of modern TPS systems for re- entry vehicles is the expensive manufacturing and maintenance process due to the complex geometry of these blunt nose configurations. To reduce the costs and to improve the aerodynamic performance the German Aerospace Center (DLR) is following a different approach using TPS structures consisting of flat ceramic tiles. To test these new sharp edged TPS structures the SHEFEX I flight experiment was designed and successfully performed by DLR in 2005. To further improve the reliability of the sharp edged TPS design at even higher Mach numbers, a second flight experiment SHEFEX II will be performed in September 2011. In comparison to SHEFEX I the second flight experiment has a fully symmetrical shape and will reach a maximum Mach number of about 11. Furthermore the vehicle has an active steering system using four canards to control the flight attitude during re-entry, e.g. roll angle, angle of attack and sideslip. After a successful flight the evaluation of the flight data will be performed using a combination of numerical and experimental tools. The data will be used for the improvement of the present numerical analysis tools and to get a better understanding of the aerothermal behaviour of sharp TPS structures. This paper presents the flight instrumentation of the SHEFEX II TPS. In addition the concept of the post flight analysis is presented.

  19. Early donor chimerism levels predict relapse and survival after allogeneic stem-cell transplantation with reduced intensity conditioning

    PubMed Central

    Reshef, Ran; Hexner, Elizabeth O.; Loren, Alison W.; Frey, Noelle V.; Stadtmauer, Edward A.; Luger, Selina M.; Mangan, James K.; Gill, Saar I.; Vassilev, Pavel; Lafferty, Kathryn A.; Smith, Jacqueline; Van Deerlin, Vivianna M.; Mick, Rosemarie; Porter, David L.

    2014-01-01

    The success of hematopoietic stem-cell transplantation (HSCT) with reduced-intensity conditioning (RIC) is limited by a high rate of disease relapse. Early risk assessment could potentially improve outcomes by identifying appropriate patients for pre-emptive strategies that may ameliorate this high risk. Using a series of landmark analyses, we investigated the predictive value of early (day-30) donor chimerism measurements on disease relapse, graft-versus-host disease and survival in a cohort of 121 patients who were allografted with a uniform RIC regimen. Chimerism levels were analyzed as continuous variables. In multivariate analysis, day-30 whole blood chimerism levels were significantly associated with relapse (HR=0.90, p<0.001), relapse-free survival (HR=0.89, p<0.001) and overall survival (HR=0.94, p=0.01). Day-30 T-cell chimerism levels were also significantly associated with relapse (HR=0.97, p=0.002), relapse-free survival (HR=0.97, p<0.001) and overall survival (HR=0.99, p=0.05). Multivariate models that included T-cell chimerism provided a better prediction for these outcomes compared to whole blood chimerism. Day-30 chimerism levels were not associated with acute or chronic graft-versus-host disease. We found that high donor chimerism levels were significantly associated with a low lymphocyte count in the recipient prior to transplant, highlighting the impact of pre-transplant lymphopenia on the kinetics of engraftment after RIC HSCT. In summary, low donor chimerism levels are associated with relapse and mortality and can potentially be used as an early predictive and prognostic marker. These findings can be used to design novel approaches to prevent relapse and to improve survival after RIC HSCT. PMID:25016197

  20. Paucity of chimeric gene-transposable element transcripts in the Drosophila melanogaster genome

    PubMed Central

    Lipatov, Mikhail; Lenkov, Kapa; Petrov, Dmitri A; Bergman, Casey M

    2005-01-01

    Background Recent analysis of the human and mouse genomes has shown that a substantial proportion of protein coding genes and cis-regulatory elements contain transposable element (TE) sequences, implicating TE domestication as a mechanism for the origin of genetic novelty. To understand the general role of TE domestication in eukaryotic genome evolution, it is important to assess the acquisition of functional TE sequences by host genomes in a variety of different species, and to understand in greater depth the population dynamics of these mutational events. Results Using an in silico screen for host genes that contain TE sequences, we identified a set of 63 mature "chimeric" transcripts supported by expressed sequence tag (EST) evidence in the Drosophila melanogaster genome. We found a paucity of chimeric TEs relative to expectations derived from non-chimeric TEs, indicating that the majority (~80%) of TEs that generate chimeric transcripts are deleterious and are not observed in the genome sequence. Using a pooled-PCR strategy to assay the presence of gene-TE chimeras in wild strains, we found that over half of the observed chimeric TE insertions are restricted to the sequenced strain, and ~15% are found at high frequencies in North American D. melanogaster populations. Estimated population frequencies of chimeric TEs did not differ significantly from non-chimeric TEs, suggesting that the distribution of fitness effects for the observed subset of chimeric TEs is indistinguishable from the general set of TEs in the genome sequence. Conclusion In contrast to mammalian genomes, we found that fewer than 1% of Drosophila genes produce mRNAs that include bona fide TE sequences. This observation can be explained by the results of our population genomic analysis, which indicates that most potential chimeric TEs in D. melanogaster are deleterious but that a small proportion may contribute to the evolution of novel gene sequences such as nested or intercalated gene

  1. Analysis of heat generation using ultrasonic vibration for post removal.

    PubMed

    Dominici, John T; Clark, Stephen; Scheetz, James; Eleazer, Paul D

    2005-04-01

    This study measured the temperature of the root surface and post during the application of ultrasonic vibration to cemented posts to simulate post removal procedure. Root canal therapy was performed on ten extracted maxillary incisors. A stainless steel Parapost was cemented into each prepared post space. Ultrasonic vibration was applied to the post and temperatures were recorded at the coronal post and the cervical root surface. Data were analyzed with ANOVA using the independent variables of (a) time of ultrasonic application (15, 30, 45 and 60 s) and 2) location (post and root surface). Greater temperature increase was observed at the post (52.6 degrees C, SD 11.1; 82.6 degrees C, SD 20.1; 111.0 degrees C, SD 29.1; 125.3 degrees C, SD 33.2) compared to the root surface (9.5 degrees C, SD 4.6; 17.5 degrees C, SD 4.8; 25.4 degrees C, SD 7.3; 32.2 degrees C, SD 8.1) for each time period, P < 0.001. Ultrasonic application to the post for longer than 15 s generates high temperature on the root surface.

  2. Outcomes of donor lymphocyte infusion for treatment of mixed donor chimerism after a reduced-intensity preparative regimen for pediatric patients with nonmalignant diseases.

    PubMed

    Haines, Hilary L; Bleesing, Jack J; Davies, Stella M; Hornung, Lindsey; Jordan, Michael B; Marsh, Rebecca A; Filipovich, Alexandra H

    2015-02-01

    Mixed donor chimerism is increasingly common in the pediatric hematopoietic stem cell transplantation (HSCT) setting because of the increased use of reduced-intensity preparative regimens for nonmalignant diseases. Donor lymphocyte infusion (DLI) is potentially useful in the treatment of mixed donor chimerism, but little are data available on the use of DLI in this setting. We conducted a retrospective review of 27 pediatric patients who received DLI for mixed donor chimerism between January 2006 and December 2010 after receiving a preparative regimen of alemtuzumab, fludarabine, and melphalan. Twenty-one patients (78%) were alive at a median of 35 months post-transplant. Seven patients (26%) sustained full donor chimerism after DLI only at a median of 35 months post-HSCT. Nine patients (33%) continued with mixed donor chimerism (median, 38% [range, 18% to 70%]) at a median of 37 months after DLI only. Five patients underwent unconditioned stem cell boosts or second conditioned transplants after no improvement in donor chimerism was seen following DLI. Donor source appeared to contribute to outcomes after DLI; patients with mismatched unrelated donors had earlier first decline in chimerism and timing of first DLI, a higher response rate to DLI, and an increased rate of graft-versus-host disease (GVHD). There was no response to DLI in patients with matched sibling donors. Ten patients, all with improvement in chimerism after DLI, developed acute GVHD after DLI, with 3 having grade III GVHD. Three patients developed chronic GVHD after DLI. These data illustrate the potential efficacy of DLI in the treatment of mixed donor chimerism after a reduced-intensity preparative regimen.

  3. Outcomes of donor lymphocyte infusion for treatment of mixed donor chimerism after a reduced-intensity preparative regimen for pediatric patients with nonmalignant diseases.

    PubMed

    Haines, Hilary L; Bleesing, Jack J; Davies, Stella M; Hornung, Lindsey; Jordan, Michael B; Marsh, Rebecca A; Filipovich, Alexandra H

    2015-02-01

    Mixed donor chimerism is increasingly common in the pediatric hematopoietic stem cell transplantation (HSCT) setting because of the increased use of reduced-intensity preparative regimens for nonmalignant diseases. Donor lymphocyte infusion (DLI) is potentially useful in the treatment of mixed donor chimerism, but little are data available on the use of DLI in this setting. We conducted a retrospective review of 27 pediatric patients who received DLI for mixed donor chimerism between January 2006 and December 2010 after receiving a preparative regimen of alemtuzumab, fludarabine, and melphalan. Twenty-one patients (78%) were alive at a median of 35 months post-transplant. Seven patients (26%) sustained full donor chimerism after DLI only at a median of 35 months post-HSCT. Nine patients (33%) continued with mixed donor chimerism (median, 38% [range, 18% to 70%]) at a median of 37 months after DLI only. Five patients underwent unconditioned stem cell boosts or second conditioned transplants after no improvement in donor chimerism was seen following DLI. Donor source appeared to contribute to outcomes after DLI; patients with mismatched unrelated donors had earlier first decline in chimerism and timing of first DLI, a higher response rate to DLI, and an increased rate of graft-versus-host disease (GVHD). There was no response to DLI in patients with matched sibling donors. Ten patients, all with improvement in chimerism after DLI, developed acute GVHD after DLI, with 3 having grade III GVHD. Three patients developed chronic GVHD after DLI. These data illustrate the potential efficacy of DLI in the treatment of mixed donor chimerism after a reduced-intensity preparative regimen. PMID:25464116

  4. Minimal residual disease after allogeneic stem cell transplant: a comparison among multiparametric flow cytometry, Wilms tumor 1 expression and chimerism status (Complete chimerism versus Low Level Mixed Chimerism) in acute leukemia.

    PubMed

    Rossi, Giovanni; Carella, Angelo Michele; Minervini, Maria Marta; Savino, Lucia; Fontana, Andrea; Pellegrini, Fabio; Greco, Michele Mario; Merla, Emanuela; Quarta, Gianni; Loseto, Giacomo; Capalbo, Silvana; Palumbo, Gaetano; Cascavilla, Nicola

    2013-12-01

    Relapse represents the main cause of treatment failure after allogeneic stem cell transplant (allo-SCT). The detection of minimal residual disease (MRD) by multiparametric flow cytometry (MFC), chimerism, cytogenetics and molecular analysis may be critical to prevent relapse. Therefore, we assessed the overall agreement among chimerism (low level mixed chimerism [LL-MC] vs. complete chimerism [CC]), MFC and Wilms tumor 1 (WT1) mRNA to detect MRD and investigated the impact of MRD obtained from the three methods on patient outcome. Sixty-seven fresh bone marrow (BM) samples from 24 patients (17 acute myeloid leukemia [AML], seven acute lymphoblastic leukemia [ALL]) in complete remission (CR) after allo-SCT were investigated at different time points. A moderate agreement was found among the three techniques investigated. A higher concordance between positive results from MFC (75.0% vs. 32.7%, p = 0.010) and WT1 (58.3% vs. 29.1%, p = 0.090) was detected among LL-MC rather than CC samples. Relapse-free survival (RFS) and overall survival (OS) were found to be higher in MRD negative patients than in MRD positive patients analyzed with MFC and WT1. Our results discourage the use of low autologous signals as the only marker of MRD, and suggest the usefulness of MFC and WT1 real-time quantitative polymerase chain reaction (RQ-PCR) in stratifying patients with respect to risk of relapse.

  5. Evaluation of epithelial chimerism after bone marrow mesenchymal stromal cell infusion in intestinal transplant patients.

    PubMed

    Kilinc, S; Gurkan, U A; Guven, S; Koyuncu, G; Tan, S; Karaca, C; Ozdogan, O; Dogan, M; Tugmen, C; Pala, E E; Bayol, U; Baran, M; Kurtulmus, Y; Pirim, I; Kebapci, E; Demirci, U

    2014-01-01

    Intestinal transplantation is the most effective treatment for patients with short bowel syndrome and small bowel insufficiencies. We evaluated epithelial chimerism after infusion of autologous bone marrow mesenchymal stromal cells (BMSCs) in patients undergoing cadaveric donor isolated intestinal transplantation (I-ITx). BMSCs were isolated from patients' bone marrow via iliac puncture and expanded in vitro prior to infusion. Two out of the 3 patients were infused with autologous BMSCs, and small intestine tissue biopsies collected post-operatively were analyzed for epithelial chimerism using XY fluorescent in situ hybridization and short tandem repeat polymerase chain reaction. We observed epithelial chimeric effect in conditions both with and without BMSC infusion. Although our results suggest a higher epithelial chimerism effect with autologous BMSC infusion in I-ITx, the measurements in multiple biopsies at different time points that demonstrate the reproducibility of this finding and its stability or changes in the level over time would be beneficial. These approaches may have potential implications for improved graft survival, lower immunosuppressant doses, superior engraftment of the transplanted tissue, and higher success rates in I-ITx.

  6. Finite element analysis and fracture resistance testing of a new intraradicular post

    PubMed Central

    YAMAMOTO, Eron Toshio Colauto; PAGANI, Clovis; da SILVA, Eduardo Galera; NORITOMI, Pedro Yoshito; UEHARA, André Yugou; KEMMOKU, Daniel Takanori

    2012-01-01

    Objectives The objective of the present study was to evaluate a prefabricated intraradicular threaded pure titanium post, designed and developed at the São José dos Campos School of Dentistry - UNESP, Brazil. This new post was designed to minimize stresses observed with prefabricated post systems and to improve cost-benefits. Material and methods Fracture resistance testing of the post/core/root complex, fracture analysis by microscopy and stress analysis by the finite element method were used for post evaluation. The following four prefabricated metal post systems were analyzed: group 1, experimental post; group 2, modification of the experimental post; group 3, Flexi Post, and group 4, Para Post. For the analysis of fracture resistance, 40 bovine teeth were randomly assigned to the four groups (n=10) and used for the fabrication of test specimens simulating the situation in the mouth. The test specimens were subjected to compressive strength testing until fracture in an EMIC universal testing machine. After fracture of the test specimens, their roots were sectioned and analyzed by microscopy. For the finite element method, specimens of the fracture resistance test were simulated by computer modeling to determine the stress distribution pattern in the post systems studied. Results The fracture test presented the following averages and standard deviation: G1 (45.63±8.77), G2 (49.98±7.08), G3 (43.84±5.52), G4 (47.61±7.23). Stress was homogenously distributed along the body of the intraradicular post in group 1, whereas high stress concentrations in certain regions were observed in the other groups. These stress concentrations in the body of the post induced the same stress concentration in root dentin. Conclusions The experimental post (original and modified versions) presented similar fracture resistance and better results in the stress analysis when compared with the commercial post systems tested (08/2008-PA/CEP). PMID:23032204

  7. Large human YACs constructed in a rad52 strain show a reduced rate of chimerism

    SciTech Connect

    Haldi, M.; Perrot, V.; Saumier, M.

    1994-12-01

    Current YAC libraries are plagued by a high frequency of chimeras - that is, clones containing fragments from multiple genomic regions. Chimeras are thought to arise largely through recombination in the yeast host cell. If so, the use of recombination-deficient yeast strains, such as rad52 mutants, might be expected to alleviate the problem. Here, we report the construction of megabase-sized human YACs in the rad52 strain MHY5201 and the determination of their rate of chimerism by fluorescence in situ hybridization analysis. Examination of 48 YACs showed a rate of chimerism of at most 8%, whereas YACs constructed in the wildtype host AB1380 showed a rate of about 50%. These results show that it is possible to significantly decrease the rate of YAC chimerism through the use of appropriate yeast host strains. 27 refs., 3 figs., 3 tabs.

  8. Measurement of chimerism in cynomolgus monkeys using human-specific short tandem repeat-based assay.

    PubMed

    Akpinar, Edip; Keary, Jodie M; Kurlander, Roger; Hale, Douglas A

    2005-01-27

    Preclinical testing of a mixed chimerism mediated organ transplant tolerance strategy, in a cynomolgus macaque model, would be facilitated by the establishment of a reliable technique for quantitative assessment of chimerism. Among various techniques used for measurement of chimerism in humans, microsatellite DNA profiling has been considered the most versatile one that can discriminate between two individuals. We adopted a commercially available short tandem repeat profiling methodology to cynomolgus monkeys using two human specific alleles, TPOX and CSF1PO. Polymerase chain reaction (PCR) was used to amplify these alleles, and the analysis of the PCR products was performed by capillary electrophoresis. Of 54 cynomolgus macaques investigated, only one pair with the same ABO blood type demonstrated identity at both alleles. This implies that this technique should interfere minimally with the assignment of donor-recipient pairs based upon molecular tissue typing or mixed lymphocyte cultures.

  9. Chimerism in DNA of buccal swabs from recipients after allogeneic hematopoietic stem cell transplantations: implications for forensic DNA testing.

    PubMed

    Berger, Burkhard; Parson, Roswitha; Clausen, Johannes; Berger, Cordula; Nachbaur, David; Parson, Walther

    2013-01-01

    We attempted to quantitatively determine the chimeric state in a total of 162 buccal swabs from 77 adult recipients aged 19-74 (median 50 years) after allogeneic hematopoietic cell transplantation by estimating the chimeric recipient/donor DNA ratios through analysis of 15 autosomal short tandem repeat markers. From each individual between one and nine, buccal swabs were taken at known time intervals after transplantation, ranging from 17 to 3,361 days (median 394 days). In buccal cells, the determined recipient/donor DNA ratios turned out to be highly variable between individuals and also within an individual. Relative donor chimerism levels (%Ch) between 0 and 100 % were detected with maximal frequencies between 10 and 30 %. Blood was always found to show the donor's genotype while hair samples in all cases gave the recipient's genotype. We examine chimerism levels with respect to age, gender, and posttransplantation period and discuss the results in the context of forensic identity testing.

  10. An Analysis of Access Barriers to Post-Secondary Education

    ERIC Educational Resources Information Center

    Vaccaro, Angelo

    2012-01-01

    Post-Secondary Education (PSE) in Ontario and in Canada has expanded on both the demand and supply sides in the last couple of decades. As of 2007, 50% of the population aged 18 to 24 was enrolled in post-secondary institutions. Enrolment in Ontario universities grew from 10,000 in 1960 to approximately 400,000 in 2007 (Clark, Moran, Skolnik, &…

  11. Three-dimensional finite element analysis of the effects of posts on stress distribution in dentin.

    PubMed

    Ho, M H; Lee, S Y; Chen, H H; Lee, M C

    1994-10-01

    A finite element analysis was conducted to study the influence of posts on dentinal stress in pulpless teeth. Three-dimensional models of an intact Chinese maxillary central incisor with and without post restoration were analyzed. When the tooth was subjected to masticatory and traumatic loads, stress distributions in dentin were similar whether or not the post was present. Maximal dentinal stresses were reduced by only 7% to 10% and 10% to 14.5%, respectively, with gold alloy and stainless steel posts. Thus the reinforcement effects from posts appeared limited in pulpless incisors.

  12. Validation of chimerism in pediatric recipients of allogeneic hematopoietic stem cell transplantation (HSCT) a comparison between two methods: real-time PCR (qPCR) vs. variable number tandem repeats PCR (VNTR PCR).

    PubMed

    Kletzel, Morris; Huang, Wei; Olszewski, Marie; Khan, Sana

    2013-01-01

    Post-hematopoietic stem cell transplantation (HSCT) chimerism monitoring is important to assess relapse and therapeutic intervention. The purpose of our study is to compare two methods variable number tandem repeats (VNTR) vs. quantitative real- time polymerase chain reaction (qPCR) in terms of determining chimerism. 127 (peripheral blood n=112, bone marrow n=15) samples were simultaneously tested by VNTR using APO-B, D1S80, D1S111, D17S30, gene loci SRY and ZP3 and qPCR using 34 assays (CA001-CA034) that are designed to a bi-allelic insertion/deletion (indel) polymorphism in the human genome. Samples were separated in three subsets: total WBC, T-cell and Myeloid cells. Extraction of DNA was performed then quantified. We analyzed column statistics, paired t-test and regression analysis for both methods. There was complete correlation between the two methods. The simplicity and rapidity of the test results from the qPCR method is more efficient and accurate to assess chimerism.

  13. Engineering of chimeric class II polyhydroxyalkanoate synthases.

    PubMed

    Niamsiri, Nuttawee; Delamarre, Soazig C; Kim, Young-Rok; Batt, Carl A

    2004-11-01

    PHA synthase is a key enzyme involved in the biosynthesis of polyhydroxyalkanoates (PHAs). Using a combinatorial genetic strategy to create unique chimeric class II PHA synthases, we have obtained a number of novel chimeras which display improved catalytic properties. To engineer the chimeric PHA synthases, we constructed a synthetic phaC gene from Pseudomonas oleovorans (phaC1Po) that was devoid of an internal 540-bp fragment. Randomly amplified PCR products (created with primers based on conserved phaC sequences flanking the deleted internal fragment) were generated using genomic DNA isolated from soil and were substituted for the 540-bp internal region. The chimeric genes were expressed in a PHA-negative strain of Ralstonia eutropha, PHB(-)4 (DSM 541). Out of 1,478 recombinant clones screened for PHA production, we obtained five different chimeric phaC1Po genes that produced more PHA than the native phaC1Po. Chimeras S1-71, S4-8, S5-58, S3-69, and S3-44 exhibited 1.3-, 1.4-, 2.0-, 2.1-, and 3.0-fold-increased levels of in vivo activity, respectively. All of the mutants mediated the synthesis of PHAs with a slightly increased molar fraction of 3-hydroxyoctanoate; however, the weight-average molecular weights (Mw) of the PHAs in all cases remained almost the same. Based upon DNA sequence analyses, the various phaC fragments appear to have originated from Pseudomonas fluorescens and Pseudomonas aureofaciens. The amino acid sequence analyses showed that the chimeric proteins had 17 to 20 amino acid differences from the wild-type phaC1Po, and these differences were clustered in the same positions in the five chimeric clones. A threading model of PhaC1Po, developed based on homology of the enzyme to the Burkholderia glumae lipase, suggested that the amino acid substitutions found in the active chimeras were located mostly on the protein model surface. Thus, our combinatorial genetic engineering strategy proved to be broadly useful for improving the catalytic

  14. Recognition of chimeric small-subunit ribosomal DNAs composed of genes from uncultivated microorganisms

    NASA Technical Reports Server (NTRS)

    Kopczynski, E. D.; Bateson, M. M.; Ward, D. M.

    1994-01-01

    When PCR was used to recover small-subunit (SSU) rRNA genes from a hot spring cyanobacterial mat community, chimeric SSU rRNA sequences which exhibited little or no secondary structural abnormality were recovered. They were revealed as chimeras of SSU rRNA genes of uncultivated species through separate phylogenetic analysis of short sequence domains.

  15. Designing a recombinant chimeric construct contain MUC1 and HER2 extracellular domain for prediagnostic breast cancer.

    PubMed

    Gheybi, Elaheh; Amani, Jafar; Salmanian, Ali Hatef; Mashayekhi, Farhad; Khodi, Samaneh

    2014-11-01

    Breast cancer is the most common cancer among women in the world. One of the approaches for diagnosis of breast cancer is detection of its tumor-associated markers. Mucin 1 (MUC1), a tumor-associated antigen, is a transmembrane glycoprotein expressed by normal epithelial cells and overexpressed by carcinomas of epithelial origin. Also, human epidermal growth factor receptor-2 (HER2/erbB-2) belongs to the one of four members of tyrosin kinase type 1 family in which overexpression of HER2 is associated with malignancy in breast cancer. This study was aimed to bioinformatics analysis and designing a recombinant chimeric protein containing MUC1 and HER2 antigens to express in prokaryotic host (Escherichia coli) as breast cancer diagnosis tools. The immunogenic sequences of MUC1 and HER2 were extracted and fused together by a linker. The chimeric construct was analyzed by bioinformatics softwares. The optimization and purification, evaluation of the expression of chimeric protein was performed using Western blotting, ELISA, and immunohistochemistry. The results showed that the chimeric construct was stable and immunogenic domains were exposed. The pET-28a vector containing chimeric gene had high level of protein expression. The recombinant chimeric protein was confirmed using Western blotting, and it was investigated using ELISA and IHC. Then, the MUC1 and HER2 combined peptides can be used as coating antigens in ELISA for detection of antibodies against MUC1 or HER2 in human serum.

  16. Induction of mixed chimerism depletes pre-existing and de novo-developed autoreactive B cells in autoimmune NOD mice.

    PubMed

    Racine, Jeremy J; Wang, Miao; Zhang, Mingfeng; Zeng, Defu

    2014-06-01

    Destruction of pancreatic islet β-cells in type 1 diabetes (T1D) is mainly mediated by autoimmune T and B lymphocytes. We reported that induction of major histocompatibility complex (MHC)-mismatched mixed chimerism reversed autoimmunity and reestablished thymic negative selection of autoreactive T cells in NOD mice, but it is still unclear how mixed chimerism tolerizes autoreactive B cells. The current studies were designed to reveal the mechanisms on how mixed chimerism tolerizes autoreactive B cells in T1D. Accordingly, mixed chimerism was induced in NOD mice through radiation-free nonmyeloablative anti-CD3/CD8 conditioning and infusion of donor CD4(+) T cell-depleted spleen and whole bone marrow (BM) cells or through myeloablative total body irradiation conditioning and reconstitution with T cell-depleted BM cells from donor and host. Kinetic analysis of percentage and yield of preplasma and plasma B cells, newly developed B-cell subsets, and their apoptosis was performed 30-60 days after transplantation. Induction of MHC-mismatched mixed chimerism results in depleting host-type pre-existing preplasma and plasma B cells as well as augmenting apoptosis of immature transitional T1 B cells, including insulin-specific B cells in a donor B cell-dependent manner. Therefore, induction of MHC-mismatched mixed chimerism depletes pre-existing and de novo-developed autoreactive B cells.

  17. Donor Chimerism of B Cells and Nature Killer Cells Provides Useful Information to Predict Hematologic Relapse following Allogeneic Hematopoietic Stem Cell Transplantation.

    PubMed

    Jiang, Ying; Wan, Liping; Qin, Youwen; Wang, Xiaorui; Yan, Shike; Xie, Kuangcheng; Wang, Chun

    2015-01-01

    In this study we investigated the correlation between donor chimerism status and disease relapse following allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism of Fluorescence-activated cell sorter (FACS) sorted CD3+T lymphocytes of 153 cases, CD56+CD16+NK lymphocytes of 153 cases and CD19+B lymphocytes of 31 cases with acute B lymphoblastic leukemia (B-ALL) was analyzed post-transplant utilizing polymerase chain reaction amplification of short tandem repeats (PCR-STR). A total of 33 patients (33/153, 21.6%) had recurrent disease. The positive predictive values of declining donor chimerism for hematologic and isolated extramedullary relapse were 58.8% and 10% (P=0.018, Chi-Square). The positive predictive values of declining donor chimerism in BMB, BMT, BMNK and PBB for hematologic relapse were 11.6%, 0%, 0% and 0% under close monitoring in patients with B-ALL. Only the donor chimerism in BMB significantly decreased in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.00, Independent-samples T test) in patients with B-ALL. The median drop of donor chimerism in PBT, BMT, PBNK and BMNK were 0%, 0%, 5.9% and 2.8% one or two weeks prior to hematologic relapse in patients with non-B-ALL. The donor chimerism in PBNK significantly decreased prior to hematologic relapse in the group with hematologic relapse as compared with the group without hematologic relapse (P=0.022, Independent-samples T test).These data suggest donor chimerism of BMB can be used to predict the occurrence of hematologic relapse in patients with B-ALL. Donor chimerism decrease in PBNK was associated with a somewhat increased risk of hematologic relapse in patients with non-B-ALL. Therefore, our results reveal a more effective path to individually predict for hematologic relapse by dynamic monitoring different cell lineages in different disease.

  18. Analysis of processing and polyadenylation signals of the hepatitis B virus surface antigen gene by using simian virus 40-hepatitis B virus chimeric plasmids

    SciTech Connect

    Simonsen, C.C.; Levinson, A.D.

    1983-12-01

    The authors examined the transcription of the hepatitis B virus surface antigen (HBsAg) gene in COs cells transfected with simian virus 40-based recombinant plasmids. When positioned behind the simian virus 40 late promoter, three transcripts were identified which hybridized to the HBsAg gene: a 2,000-nucleotide transcript colinear with a gene, a 1,100-nucleotide transcript representing a spliced molecule in which a major portion of the sequences encoding HBsAg were deleted, and an 800-nucleotide transcript derived primarily from sequences 3' to the HBsAg gene. The splice acceptor site utilized by the 1,100-nucleotide transcript is located immediately upstream of an open reading frame of unknown function contained within the 3' nontranslated region of the HBsAg gene. The HBsAg-specific mRNA species terminate 12 to 19 base pairs 3' of the sequence UAUAAA, similar to the concensus hexanucleotide which is thought to promote polyadenylation (AAUAAA). They constructed a series of plasmids with progressive deletions from the region surrounding where these transcripts terminate. Analysis of mRNA produced by cells transfected with these plasmids indicated that the signal hexanucleotide is in itself unable to promote the efficient processing of mRNA in the absence of downstream hepatitis B virus sequences. Processing proceeds properly, however, from plasmids containing an additional 30 nucleotides 3' of this signal.

  19. Xenogenic oogenesis of chicken (Gallus domesticus) female primordial germ cells in germline chimeric quail (Coturnix japonica) ovary.

    PubMed

    Liu, C H; Chang, I K; Sasse, J; Dumatol, C J; Basker, J V; Wernery, U

    2007-10-01

    In present study, chicken primordial germ cells (PGCs) were transferred into quail embryos to investigate the development of these germ cells in quail ovary. Briefly, 2 microl of chicken embryonic blood (stage 14) or about 100 purified circulating PGCs were transferred into quail embryo. Contribution of chicken PGCs were detected in gonads of chimeric quail embryos (stage 28) by immunocytochemical staining of cell surface antigen SSEA-1, and by in situ hybridization (ISH) with female chicken specific DNA probe. As a result, 52.0+/-43.2 (n=18) and 42.7+/-27.3 (n=17) chicken PGCs were found in the gonads of chimeric quail embryo that was injected with chicken embryonic blood (stage 14) and about 100 purified circulating PGCs, respectively. Furthermore, the ovaries of 81.8% (9/11) 12 days post incubation (dpi) chimeric quail embryos were observed with a mean of 457.6+/-237.1 female chicken PGCs-derived oogonia scattered in ovarian cortex area. In 9 out of 12 newly hatched and one week old chimeric quail chicks, on average of 2883.0+/-1924.1 primary oocytes and 3 follicles derived from chicken PGCs were found, respectively. The present results suggest that chicken female PGCs are able to migrate, colonize, proliferate and differentiate into oogonia, primary oocytes in chimeric quail ovary.

  20. An MHC-defined primate model reveals significant rejection of bone marrow after mixed chimerism induction despite full MHC matching.

    PubMed

    Larsen, C P; Page, A; Linzie, K H; Russell, M; Deane, T; Stempora, L; Strobert, E; Penedo, M C T; Ward, T; Wiseman, R; O'Connor, D; Miller, W; Sen, S; Singh, K; Kean, L S

    2010-11-01

    In murine models, mixed hematopoietic chimerism induction leads to robust immune tolerance. However, translation to primates and to patients has been difficult. In this study, we used a novel MHC-defined rhesus macaque model to examine the impact of MHC matching on the stability of costimulation blockade-/sirolimus-mediated chimerism, and to probe possible mechanisms of bone marrow rejection after nonmyeloablative transplant. Using busulfan-based pretransplant preparation and maintenance immunosuppression with sirolimus, as well as CD28 and CD154 blockade, all recipients demonstrated donor engraftment after transplant. However, the mixed chimerism that resulted was compartmentalized, with recipients demonstrating significantly higher whole blood chimerism compared to T cell chimerism. Thus, the vast majority of T cells presenting posttransplant were recipient-rather than donor-derived. Surprisingly, even in MHC-matched transplants, rejection of donor hematopoiesis predominated after immunosuppression withdrawal. Weaning of immunosuppression was associated with a surge of antigen-experienced T cells, and transplant rejection was associated with the acquisition of donor-directed T cell alloreactivity. These results suggest that a reservoir of alloreactive cells was present despite prior costimulation blockade and sirolimus, and that the post-immunosuppression lymphocytic rebound may have lead to a phenotypic shift in these recipient T cells towards an activated, antigen-experienced phenotype, and ultimately, to transplant rejection. PMID:20849552

  1. Chimeric Measles Viruses with a Foreign Envelope

    PubMed Central

    Spielhofer, Pius; Bächi, Thomas; Fehr, Thomas; Christiansen, Gudrun; Cattaneo, Roberto; Kaelin, Karin; Billeter, Martin A.; Naim, Hussein Y.

    1998-01-01

    Measles virus (MV) and vesicular stomatitis virus (VSV) are both members of the Mononegavirales but are only distantly related. We generated two genetically stable chimeric viruses. In MGV, the reading frames of the MV envelope glycoproteins H and F were substituted by a single reading frame encoding the VSV G glycoprotein; MG/FV is similar but encodes a G/F hybrid in which the VSV G cytoplasmic tail was replaced by that of MV F. In contrast to MG/FV, MGV virions do not contain the MV matrix (M) protein. This demonstrates that virus assembly is possible in the absence of M; conversely, the cytoplasmic domain of F allows incorporation of M and enhances assembly. The formation of chimeric viruses was substantially delayed and the titers obtained were reduced about 50-fold in comparison to standard MV. In the novel chimeras, transcription and replication are mediated by the MV ribonucleoproteins but the envelope glycoproteins dictate the host range. Mice immunized with the chimeric viruses were protected against lethal doses of wild-type VSV. These findings suggest that it is feasible to construct MV variants bearing a variety of different envelopes for use as vaccines or for gene therapeutic purposes. PMID:9499071

  2. Repository post-sealing risk analysis using MACRO. [MACRO

    SciTech Connect

    Kaufman, A.M.; Edwards, L.L.; O'Connell, W.J.

    1980-02-14

    MACRO, a code to propagate probability distributions through a set of linked models, is currently under development at Lawrence Livermore Laboratory. An early version of this code, MACRO1, has been used to assess post-sealing dose to man for simple repository and site models based on actual site data.

  3. Understanding Students' Online Interaction: Analysis of Discussion Board Postings

    ERIC Educational Resources Information Center

    Song, Liyan; McNary, Scot W.

    2011-01-01

    The purpose of this paper was to report on the findings of a study examining students' online interaction patterns. The context of the study was a graduate online class delivered via Blackboard[R]. The primary data for the study came from students' discussion board postings, online learning journals, and course grades. Various data analysis…

  4. Engineering Post-Graduate Programmes: A Quality and Productivity Analysis

    ERIC Educational Resources Information Center

    Soares de Mello, Joao Carlos C. B.; Gomes, Eliane Goncalves; Meza, Lidia Angulo; Soares de Mello, Maria Helena C.; Soares de Mello, Adelino Jose R.

    2006-01-01

    Post-graduate courses are periodically evaluated in accordance with not always very clear criteria by CAPES (Co-ordinating Committee for Teaching Staff Improvement). These criteria try to measure, among other things, academic productivity (including quantity and quality of scientific production), number of classes given by teachers, time to…

  5. reSpect: software for identification of high and low abundance ion species in chimeric tandem mass spectra.

    PubMed

    Shteynberg, David; Mendoza, Luis; Hoopmann, Michael R; Sun, Zhi; Schmidt, Frank; Deutsch, Eric W; Moritz, Robert L

    2015-11-01

    Most shotgun proteomics data analysis workflows are based on the assumption that each fragment ion spectrum is explained by a single species of peptide ion isolated by the mass spectrometer; however, in reality mass spectrometers often isolate more than one peptide ion within the window of isolation that contribute to additional peptide fragment peaks in many spectra. We present a new tool called reSpect, implemented in the Trans-Proteomic Pipeline (TPP), which enables an iterative workflow whereby fragment ion peaks explained by a peptide ion identified in one round of sequence searching or spectral library search are attenuated based on the confidence of the identification, and then the altered spectrum is subjected to further rounds of searching. The reSpect tool is not implemented as a search engine, but rather as a post-search engine processing step where only fragment ion intensities are altered. This enables the application of any search engine combination in the iterations that follow. Thus, reSpect is compatible with all other protein sequence database search engines as well as peptide spectral library search engines that are supported by the TPP. We show that while some datasets are highly amenable to chimeric spectrum identification and lead to additional peptide identification boosts of over 30% with as many as four different peptide ions identified per spectrum, datasets with narrow precursor ion selection only benefit from such processing at the level of a few percent. We demonstrate a technique that facilitates the determination of the degree to which a dataset would benefit from chimeric spectrum analysis. The reSpect tool is free and open source, provided within the TPP and available at the TPP website. Graphical Abstract ᅟ.

  6. reSpect: software for identification of high and low abundance ion species in chimeric tandem mass spectra.

    PubMed

    Shteynberg, David; Mendoza, Luis; Hoopmann, Michael R; Sun, Zhi; Schmidt, Frank; Deutsch, Eric W; Moritz, Robert L

    2015-11-01

    Most shotgun proteomics data analysis workflows are based on the assumption that each fragment ion spectrum is explained by a single species of peptide ion isolated by the mass spectrometer; however, in reality mass spectrometers often isolate more than one peptide ion within the window of isolation that contribute to additional peptide fragment peaks in many spectra. We present a new tool called reSpect, implemented in the Trans-Proteomic Pipeline (TPP), which enables an iterative workflow whereby fragment ion peaks explained by a peptide ion identified in one round of sequence searching or spectral library search are attenuated based on the confidence of the identification, and then the altered spectrum is subjected to further rounds of searching. The reSpect tool is not implemented as a search engine, but rather as a post-search engine processing step where only fragment ion intensities are altered. This enables the application of any search engine combination in the iterations that follow. Thus, reSpect is compatible with all other protein sequence database search engines as well as peptide spectral library search engines that are supported by the TPP. We show that while some datasets are highly amenable to chimeric spectrum identification and lead to additional peptide identification boosts of over 30% with as many as four different peptide ions identified per spectrum, datasets with narrow precursor ion selection only benefit from such processing at the level of a few percent. We demonstrate a technique that facilitates the determination of the degree to which a dataset would benefit from chimeric spectrum analysis. The reSpect tool is free and open source, provided within the TPP and available at the TPP website. Graphical Abstract ᅟ. PMID:26419769

  7. reSpect: Software for Identification of High and Low Abundance Ion Species in Chimeric Tandem Mass Spectra

    NASA Astrophysics Data System (ADS)

    Shteynberg, David; Mendoza, Luis; Hoopmann, Michael R.; Sun, Zhi; Schmidt, Frank; Deutsch, Eric W.; Moritz, Robert L.

    2015-11-01

    Most shotgun proteomics data analysis workflows are based on the assumption that each fragment ion spectrum is explained by a single species of peptide ion isolated by the mass spectrometer; however, in reality mass spectrometers often isolate more than one peptide ion within the window of isolation that contribute to additional peptide fragment peaks in many spectra. We present a new tool called reSpect, implemented in the Trans-Proteomic Pipeline (TPP), which enables an iterative workflow whereby fragment ion peaks explained by a peptide ion identified in one round of sequence searching or spectral library search are attenuated based on the confidence of the identification, and then the altered spectrum is subjected to further rounds of searching. The reSpect tool is not implemented as a search engine, but rather as a post-search engine processing step where only fragment ion intensities are altered. This enables the application of any search engine combination in the iterations that follow. Thus, reSpect is compatible with all other protein sequence database search engines as well as peptide spectral library search engines that are supported by the TPP. We show that while some datasets are highly amenable to chimeric spectrum identification and lead to additional peptide identification boosts of over 30% with as many as four different peptide ions identified per spectrum, datasets with narrow precursor ion selection only benefit from such processing at the level of a few percent. We demonstrate a technique that facilitates the determination of the degree to which a dataset would benefit from chimeric spectrum analysis. The reSpect tool is free and open source, provided within the TPP and available at the TPP website.

  8. reSpect: Software for Identification of High and Low Abundance Ion Species in Chimeric Tandem Mass Spectra

    PubMed Central

    Shteynberg, David; Mendoza, Luis; Hoopmann, Michael R.; Sun, Zhi; Schmidt, Frank; Deutsch, Eric W.; Moritz, Robert L.

    2016-01-01

    Most shotgun proteomics data analysis workflows are based on the assumption that each fragment ion spectrum is explained by a single species of peptide ion isolated by the mass spectrometer; however, in reality mass spectrometers often isolate more than one peptide ion within the window of isolation that contributes to additional peptide fragment peaks in many spectra. We present a new tool called reSpect, implemented in the Trans-Proteomic Pipeline (TPP), that enables an iterative workflow whereby fragment ion peaks explained by a peptide ion identified in one round of sequence searching or spectral library search are attenuated based on the confidence of the identification, and then the altered spectrum is subjected to further rounds of searching. The reSpect tool is not implemented as a search engine, but rather as a post search engine processing step where only fragment ion intensities are altered. This enables the application of any search engine combination in the following iterations. Thus, reSpect is compatible with all other protein sequence database search engines as well as peptide spectral library search engines that are supported by the TPP. We show that while some datasets are highly amenable to chimeric spectrum identification and lead to additional peptide identification boosts of over 30% with as many as four different peptide ions identified per spectrum, datasets with narrow precursor ion selection only benefit from such processing at the level of a few percent. We demonstrate a technique that facilitates the determination of the degree to which a dataset would benefit from chimeric spectrum analysis. The reSpect tool is free and open source, provided within the TPP and available at the TPP website. PMID:26419769

  9. Design and analysis of post-marketing research.

    PubMed

    Zhou, Xiao-Hua Andrew; Yang, Wei

    2013-07-01

    A post-marketing study is an integral part of research that helps to ensure a favorable risk-benefit profile for approved drugs used in the market. Because most of post-marketing studies use observational designs, which are liable to confounding, estimation of the causal effect of a drug versus a comparative one is very challenging. This article focuses on methodological issues of importance in designing and analyzing studies to evaluate the safety of marketed drugs, especially marketed traditional Chinese medicine (TCM) products. Advantages and limitations of the current designs and analytic methods for postmarketing studies are discussed, and recommendations are given for improving the validity of postmarketing studies in TCM products.

  10. Stress distribution of endodontically treated teeth with titanium alloy post and carbon fiber post with different alveolar bone height: A three-dimensional finite element analysis

    PubMed Central

    Singh, S. Vijay; Bhat, Manohar; Gupta, Saurabh; Sharma, Deepak; Satija, Harsha; Sharma, Sumeet

    2015-01-01

    Objective: A three-dimensional (3D) finite element analysis (FEA) on the stress distribution of endodontically treated teeth with titanium alloy post and carbon fiber post with different alveolar bone height. Materials and Methods: The 3D model was fabricated using software to represent an endodontically treated mandibular second premolar with post and restored with a full ceramic crown restoration, which was then analyzed using FEA using FEA ANSYS Workbench V13.0 (ANSYS Inc., Canonsburg, Pennsylvania, U.S.A) software. Results: The FEA showed the maximum stresses of 137.43 Mpa in dentin with alveolar bone height of 4 mm when the titanium post was used, 138.48 Mpa when carbon fiber post was used as compared to 105.91 Mpa in the model with alveolar bone height of 2 mm from the cement enamel junction (CEJ) when the titanium post was used and 107.37 Mpa when the carbon fiber post was used. Conclusions: Stress was observed more in alveolar bone height level of 4 mm from CEJ than 2 mm from CEJ. Stresses in the dentin were almost similar when the carbon fiber post was compared to titanium post. However, stresses in the post and the cement were much higher when titanium post was used as compared to carbon fiber post. PMID:26430375

  11. Donor-specific tolerance induction in organ transplantation via mixed splenocytes chimerism.

    PubMed

    Yamazaki, S; Kanamoto, A; Takayama, T

    2013-08-01

    We have shown previously that donor-derived splenocytes can replace recipients' bone marrow and induce donor-specific tolerance (DST). We have also shown the usefulness of the chimeric state for the induction of DST. Further analysis of mixed splenocytes chimera, especially the role of each T cells in mixed splenocytes chimera, is indispensable issue for its clinical use. A chimeric state has been shown to achieve long-term survival in major histocompatibility complex (MHC)-mismatched grafts. The donor-derived splenocytes can replace recipients' bone marrow and induce DST. The long-term survival of allogeneic skin grafts was achieved without immunosuppressants. In this study we show the role of each T cell type in a splenocyte mixed chimera. This review provides a short summary of our original work, adding some supplemental interpretations. Mixed chimerism is thus considered an attractive approach for the induction of DST without the use of immunosuppressants. In this paper, we summarize some of the findings on mixed splenocyte chimeras and review mixed chimerism in recent organ transplantation.

  12. A case of leucocyte chimerism (78,XX/78,XY) in a dog with a disorder of sexual development.

    PubMed

    Szczerbal, I; Nowacka-Woszuk, J; Nizanski, W; Salamon, S; Ochota, M; Dzimira, S; Atamaniuk, W; Switonski, M

    2014-06-01

    A 1-year-old Shih Tzu dog was presented for examination because of abnormal external genitalia. A residual penis with a prepuce was located in a position typical of a male. The dog had no palpable testicles or scrotum. The ultrasound examination revealed the presence of the prostate, but the gonads remained undetectable. Cytogenetic analysis performed on chromosome preparations obtained from lymphocyte culture showed two cell lines - 78,XX and 78,XY. Molecular analysis of 14 polymorphic microsatellite markers allowed us to distinguish leucocyte chimerism from whole body chimerism. The presence of 3 or 4 alleles was confirmed in DNA isolated from blood, while in DNA isolated from hair follicles only 1 or 2 alleles were detected. The case was classified as leucocyte 78,XX/78,XY chimerism. Our study showed that XX/XY leucocyte chimerism might be associated with disorder of sexual development in dogs. Furthermore, it is emphasized that the use of cytogenetic study, in combination with analysis of polymorphic markers in DNA isolated from different somatic cells, facilitates distinguishing between leucocyte and whole body chimerism.

  13. Regional atmospheric composition modeling with CHIMERE

    NASA Astrophysics Data System (ADS)

    Menut, L.; Bessagnet, B.; Khvorostyanov, D.; Beekmann, M.; Colette, A.; Coll, I.; Curci, G.; Foret, G.; Hodzic, A.; Mailler, S.; Meleux, F.; Monge, J.-L.; Pison, I.; Turquety, S.; Valari, M.; Vautard, R.; Vivanco, M. G.

    2013-01-01

    Tropospheric trace gas and aerosol pollutants have adverse effects on health, environment and climate. In order to quantify and mitigate such effects, a wide range of processes leading to the formation and transport of pollutants must be considered, understood and represented in numerical models. Regional scale pollution episodes result from the combination of several factors: high emissions (from anthropogenic or natural sources), stagnant meteorological conditions, velocity and efficiency of the chemistry and the deposition. All these processes are highly variable in time and space, and their relative importance to the pollutants budgets can be quantified within a chemistry-transport models (CTM). The offline CTM CHIMERE model uses meteorological model fields and emissions fluxes and calculates deterministically their behavior in the troposphere. The calculated three-dimensional fields of chemical concentrations can be compared to measurements to analyze past periods or used to make air quality forecasts and CHIMERE has enabled a fine understanding of pollutants transport during numerous measurements campaigns. It is a part of the PREVAIR french national forecast platform, delivering pollutant concentrations up to three days in advance. The model also allows scenario studies and long term simulations for pollution trends. The modelling of photochemical air pollution has reached a good level of maturity, and the latest projects involving CHIMERE now aim at increasing our understanding of pollution impact on health at the urban scale or at the other end of the spectrum for long term air quality and climate change interlinkage studies, quantifying the emissions and transport of pollen, but also, at a larger scale, analyzing the transport of pollutants plumes emitted by volcanic eruptions and forest fires.

  14. Post Buckling Progressive Failure Analysis of Composite Laminated Stiffened Panels

    NASA Astrophysics Data System (ADS)

    Anyfantis, Konstantinos N.; Tsouvalis, Nicholas G.

    2012-06-01

    The present work deals with the numerical prediction of the post buckling progressive and final failure response of stiffened composite panels based on structural nonlinear finite element methods. For this purpose, a progressive failure model (PFM) is developed and applied to predict the behaviour of an experimentally tested blade-stiffened panel found in the literature. Failure initiation and propagation is calculated, owing to the accumulation of the intralaminar failure modes induced in fibre reinforced composite materials. Hashin failure criteria have been employed in order to address the fiber and matrix failure modes in compression and tension. On the other hand, the Tsai-Wu failure criterion has been utilized for addressing shear failure. Failure detection is followed with the introduction of corresponding material degradation rules depending on the individual failure mechanisms. Failure initiation and failure propagation as well as the post buckling ultimate attained load have been numerically evaluated. Final failure behaviour of the simulated stiffened panel is due to sudden global failure, as concluded from comparisons between numerical and experimental results being in good agreement.

  15. A Post-Monte-Carlo Sensitivity Analysis Code

    2000-04-04

    SATOOL (Sensitivity Analysis TOOL) is a code for sensitivity analysis, following an uncertainity analysis with Monte Carlo simulations. Sensitivity analysis identifies those input variables, whose variance contributes dominatly to the variance in the output. This analysis can be used to reduce the variance in the output variables by redefining the "sensitive" variables with greater precision, i.e. with lower variance. The code identifies a group of sensitive variables, ranks them in the order of importance andmore » also quantifies the relative importance among the sensitive variables.« less

  16. Ethical acceptability of research on human-animal chimeric embryos: summary of opinions by the Japanese Expert Panel on Bioethics.

    PubMed

    Mizuno, Hiroshi; Akutsu, Hidenori; Kato, Kazuto

    2015-01-01

    Human-animal chimeric embryos are embryos obtained by introducing human cells into a non-human animal embryo. It is envisaged that the application of human-animal chimeric embryos may make possible many useful research projects including producing three-dimensional human organs in animals and verification of the pluripotency of human ES cells or iPS cells in vivo. The use of human-animal chimeric embryos, however, raises several ethical and moral concerns. The most fundamental one is that human-animal chimeric embryos possess the potential to develop into organisms containing human-derived tissue, which may lead to infringing upon the identity of the human species, and thus impairing human dignity. The Japanese Expert Panel on Bioethics in the Cabinet Office carefully considered the scientific significance and ethical acceptability of the issue and released its "Opinions regarding the handling of research using human-animal chimeric embryos". The Panel proposed a framework of case-by-case review, and suggested that the following points must be carefully reviewed from the perspective of ethical acceptability: (a) Types of animal embryos and types of animals receiving embryo transfers, particularly in dealing with non-human primates; (b) Types of human cells and organs intended for production, particularly in dealing with human nerve or germ cells; and (c) Extent of the period required for post-transfer studies. The scientific knowledge that can be gained from transfer into an animal uterus and from the production of an individual must be clarified to avoid unnecessary generation of chimeric animals. The time is ripe for the scientific community and governments to start discussing the ethical issues for establishing a global consensus. PMID:26694481

  17. Ethical acceptability of research on human-animal chimeric embryos: summary of opinions by the Japanese Expert Panel on Bioethics.

    PubMed

    Mizuno, Hiroshi; Akutsu, Hidenori; Kato, Kazuto

    2015-01-01

    Human-animal chimeric embryos are embryos obtained by introducing human cells into a non-human animal embryo. It is envisaged that the application of human-animal chimeric embryos may make possible many useful research projects including producing three-dimensional human organs in animals and verification of the pluripotency of human ES cells or iPS cells in vivo. The use of human-animal chimeric embryos, however, raises several ethical and moral concerns. The most fundamental one is that human-animal chimeric embryos possess the potential to develop into organisms containing human-derived tissue, which may lead to infringing upon the identity of the human species, and thus impairing human dignity. The Japanese Expert Panel on Bioethics in the Cabinet Office carefully considered the scientific significance and ethical acceptability of the issue and released its "Opinions regarding the handling of research using human-animal chimeric embryos". The Panel proposed a framework of case-by-case review, and suggested that the following points must be carefully reviewed from the perspective of ethical acceptability: (a) Types of animal embryos and types of animals receiving embryo transfers, particularly in dealing with non-human primates; (b) Types of human cells and organs intended for production, particularly in dealing with human nerve or germ cells; and (c) Extent of the period required for post-transfer studies. The scientific knowledge that can be gained from transfer into an animal uterus and from the production of an individual must be clarified to avoid unnecessary generation of chimeric animals. The time is ripe for the scientific community and governments to start discussing the ethical issues for establishing a global consensus.

  18. Chimerism in monochorionic dizygotic twins: case study and review.

    PubMed

    Chen, Kristen; Chmait, Ramen H; Vanderbilt, Douglas; Wu, Samuel; Randolph, Linda

    2013-07-01

    Chimerism occurs when an organism contains cells derived from more than one distinct zygote. We focus on monochorionic dizygotic twin blood chimerism, and particularly twin-twin transfusion syndrome in such pregnancies. For years, researchers have understood chimerism to be a common phenomenon in cattle. Although, this review will not delve deeply into animal chimerism, an understanding of chimerism in the animal world can provide clues regarding health implications for human chimeras. This report serves two purposes: an update and assessment of the twins we reported previously in 2010 [Assaf et al., 2010] and a review on dizygotic monochorionic chimeric twins. First, our updated assessment of the twins shows no identifiable regression of Müllerian sex derivatives in the female, and normal neurodevelopment was documented in both. Our research has suggested several key points; one that blood chimerism persists from fetal life to at least age two years. Second, chimerism in humans is not as rare as previously thought, although it has been studied only recently. Third, assisted reproductive technologies appear to increase the risk of monochorionic dizygotic twin pregnancies.

  19. Hybridization accompanying FRET event in labeled natural nucleoside-unnatural nucleoside containing chimeric DNA duplexes.

    PubMed

    Bag, Subhendu Sekhar; Das, Suman K; Pradhan, Manoj Kumar; Jana, Subhashis

    2016-09-01

    Förster resonance energy transfer (FRET) is a highly efficient strategy in illuminating the structures, structural changes and dynamics of DNA, proteins and other biomolecules and thus is being widely utilized in studying such phenomena, in designing molecular/biomolecular probes for monitoring the hybridization event of two single stranded DNA to form duplex, in gene detection and in many other sensory applications in chemistry, biology and material sciences. Moreover, FRET can give information about the positional status of chromophores within the associated biomolecules with much more accuracy than other methods can yield. Toward this end, we want to report here the ability of fluorescent unnatural nucleoside, triazolylphenanthrene ((TPhen)BDo) to show FRET interaction upon hybridization with fluorescently labeled natural nucleosides, (Per)U or (OxoPy)U or (Per)U, forming two stable chimeric DNA duplexes. The pairing selectivity and the thermal duplex stability of the chimeric duplexes are higher than any of the duplexes with natural nucleoside formed. The hybridization results in a Förster resonance energy transfer (FRET) from donor triazolylphenanthrene of (TPhen)BDo to acceptor oxopyrene of (OxoPy)U and/or to perylene chromophore of (Per)U, respectively, in two chimeric DNA duplexes. Therefore, we have established the FRET process in two chimeric DNA duplexes wherein a fluorescently labeled natural nucleoside ((OxoPy)U or (Per)U) paired against an unnatural nucleoside ((TPhen)BDo) without sacrificing the duplex stability and B-DNA conformation. The hybridization accompanying FRET event in these classes of interacting fluorophores is new. Moreover, there is no report of such designed system of chimeric DNA duplex. Our observed phenomenon and the design can potentially be exploited in designing more of such efficient FRET pairs for useful application in the detection and analysis of biomolecular interactions and in material science application. PMID:27498231

  20. Hybridization accompanying FRET event in labeled natural nucleoside-unnatural nucleoside containing chimeric DNA duplexes.

    PubMed

    Bag, Subhendu Sekhar; Das, Suman K; Pradhan, Manoj Kumar; Jana, Subhashis

    2016-09-01

    Förster resonance energy transfer (FRET) is a highly efficient strategy in illuminating the structures, structural changes and dynamics of DNA, proteins and other biomolecules and thus is being widely utilized in studying such phenomena, in designing molecular/biomolecular probes for monitoring the hybridization event of two single stranded DNA to form duplex, in gene detection and in many other sensory applications in chemistry, biology and material sciences. Moreover, FRET can give information about the positional status of chromophores within the associated biomolecules with much more accuracy than other methods can yield. Toward this end, we want to report here the ability of fluorescent unnatural nucleoside, triazolylphenanthrene ((TPhen)BDo) to show FRET interaction upon hybridization with fluorescently labeled natural nucleosides, (Per)U or (OxoPy)U or (Per)U, forming two stable chimeric DNA duplexes. The pairing selectivity and the thermal duplex stability of the chimeric duplexes are higher than any of the duplexes with natural nucleoside formed. The hybridization results in a Förster resonance energy transfer (FRET) from donor triazolylphenanthrene of (TPhen)BDo to acceptor oxopyrene of (OxoPy)U and/or to perylene chromophore of (Per)U, respectively, in two chimeric DNA duplexes. Therefore, we have established the FRET process in two chimeric DNA duplexes wherein a fluorescently labeled natural nucleoside ((OxoPy)U or (Per)U) paired against an unnatural nucleoside ((TPhen)BDo) without sacrificing the duplex stability and B-DNA conformation. The hybridization accompanying FRET event in these classes of interacting fluorophores is new. Moreover, there is no report of such designed system of chimeric DNA duplex. Our observed phenomenon and the design can potentially be exploited in designing more of such efficient FRET pairs for useful application in the detection and analysis of biomolecular interactions and in material science application.

  1. Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy.

    PubMed

    Destouni, Aspasia; Zamani Esteki, Masoud; Catteeuw, Maaike; Tšuiko, Olga; Dimitriadou, Eftychia; Smits, Katrien; Kurg, Ants; Salumets, Andres; Van Soom, Ann; Voet, Thierry; Vermeesch, Joris R

    2016-05-01

    Dramatic genome dynamics, such as chromosome instability, contribute to the remarkable genomic heterogeneity among the blastomeres comprising a single embryo during human preimplantation development. This heterogeneity, when compatible with life, manifests as constitutional mosaicism, chimerism, and mixoploidy in live-born individuals. Chimerism and mixoploidy are defined by the presence of cell lineages with different parental genomes or different ploidy states in a single individual, respectively. Our knowledge of their mechanistic origin results from indirect observations, often when the cell lineages have been subject to rigorous selective pressure during development. Here, we applied haplarithmisis to infer the haplotypes and the copy number of parental genomes in 116 single blastomeres comprising entire preimplantation bovine embryos (n = 23) following in vitro fertilization. We not only demonstrate that chromosome instability is conserved between bovine and human cleavage embryos, but we also discovered that zygotes can spontaneously segregate entire parental genomes into different cell lineages during the first post-zygotic cleavage division. Parental genome segregation was not exclusively triggered by abnormal fertilizations leading to triploid zygotes, but also normally fertilized zygotes can spontaneously segregate entire parental genomes into different cell lineages during cleavage of the zygote. We coin the term "heterogoneic division" to indicate the events leading to noncanonical zygotic cytokinesis, segregating the parental genomes into distinct cell lineages. Persistence of those cell lines during development is a likely cause of chimerism and mixoploidy in mammals. PMID:27197242

  2. Analysis of post-harvest fungicides by micellar electrokinetic chromatography.

    PubMed

    Rodríguez, R; Picó, Y; Font, G; Mañes, J

    2001-07-27

    A method based on solid-phase extraction (SPE) and micellar electrokinetic chromatography (MEKC) was developed for the simultaneous determination of carbendazim, imazalil, methylthiophanate, O-phenylphenol, prochloraz, procimidone, thiabendazole and triadimefon residues in grape, lettuce, orange and tomato. Selectivity and resolution were studied changing the pH and the concentration of the buffer, the type and concentration of surfactant and the methanol content in the mobile phase. A buffer consisting of 4 mM borate with 75 mM sodium cholate (pH 9.2) gave the best results. The recoveries of the fungicides in spiked fruit and vegetable samples ranged from 30 to 105%, and the limits of detection were between 0.1 and 1 mg kg(-1). The reproducibility and repeatability of the combination of SPE pretreatment and MEKC were good for all the compounds, except for imazalil and O-phenylphenol in oranges, due to some matrix compounds interfering with the separation. The method was applied to post harvest treated samples, and the fungicides were sometimes detected at concentration levels lower than maximum residue limits (MRLs). PMID:11521888

  3. Chimeric conundra: are nucleomorphs and chromists monophyletic or polyphyletic?

    PubMed Central

    Cavalier-Smith, T; Allsopp, M T; Chao, E E

    1994-01-01

    All algae with chloroplasts located not freely in the cytosol, but inside two extra membranes, probably arose chimerically by the permanent fusion of two different eukaryote cells: a protozoan host and a eukaryotic algal symbiont. Two such groups, cryptomonads (phylum Cryptista) and Chlorarachniophyta, still retain a DNA-containing relic of the nucleus of the algal endosymbiont, known as the nucleomorph, as well as the host nucleus. These two phyla were traditionally assumed to have obtained their chloroplasts separately by two independent symbioses. We have sequenced the nuclear and the nucleomorph 18S rRNA genes of the nonphotosynthetic cryptomonad Chilomonas paramecium. Our phylogenetic analysis suggests that cryptomonad and chlorarachniophyte nucleomorphs may be related to each other and raises the possibility that both phyla may have diverged from a common ancestral chimeric cell that originated by a single endosymbiosis involving an algal endosymbiont related to the ancestor of red algae. But, because of the instability of the molecular trees when different taxa are added, there is insufficient evidence to overturn the traditional view that Chlorarachnion nucleomorphs evolved separately from a relative of green algae. The four phyla that contain chromophyte algae (those with chlorophyll c--i.e., Cryptista, Heterokonta, Haptophyta, Dinozoa) are distantly related to each other and to Chlorarachniophyta on our trees. However, all of the photosynthetic taxa within each of these four phyla radiate from each other very substantially after the radiation of the four phyla themselves. This favors the view that the common ancestor of these four phyla was not photosynthetic and that chloroplasts were implanted separately into each much more recently. This probable polyphyly of the chromophyte algae, if confirmed, would make it desirable to treat Cryptista, Heterokonta, and Haptophyta as separate kingdoms, rather than to group them together in the single kingdom

  4. An MHC-defined primate model reveals significant rejection of bone marrow after mixed-chimerism induction despite full MHC matching

    PubMed Central

    Larsen, Christian P.; Page, Andrew; Linzie, Kelly Hamby; Russell, Maria; Deane, Taylor; Stempora, Linda; Strobert, Elizabeth; Penedo, Maria Cecilia T.; Ward, Thea; Wiseman, Roger; O'Connor, David; Miller, Weston; Sen, Sharon; Singh, Karnail; Kean, Leslie S.

    2010-01-01

    In murine models, mixed hematopoietic chimerism-induction leads to robust immune tolerance. However, translation to primates and to patients has been difficult. In this study, we used a novel MHC-defined rhesus macaque model to examine the impact of MHC matching on the stability of costimulation blockade/sirolimus-mediated chimerism, and to probe possible mechanisms of bone marrow rejection after non-myeloablative transplant. Using busulfan-based pre-transplant preparation and maintenance immunosuppression with sirolimus, as well as CD28- and CD154-blockade, all recipients demonstrated donor engraftment after transplant. However, the mixed-chimerism that resulted was compartmentalized, with recipients demonstrating significantly higher whole blood chimerism compared to T cell chimerism Thus, the vast majority of T cells present post-transplant were recipient- rather than donor-derived. Surprisingly, even in MHC-matched transplants, rejection of donor hematopoiesis predominated after immunosuppression withdrawal. Weaning of immunosuppression was associated with a surge of antigen-experienced T cells, and transplant rejection was associated with the acquisition of donor-directed T cell alloreactivity. These results suggest that a reservoir of alloreactive cells was present despite prior costimulation blockade and sirolimus, and that the post-immunosuppression lymphocytic rebound may have lead to a phenotypic shift in these recipient T cells towards an activated, antigen experienced phenotype, and ultimately, to transplant rejection. PMID:20849552

  5. Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.

    PubMed

    Aghazadeh, Rustam; Zamani, Mohammadreza; Motallebi, Mostafa; Moradyar, Mehdi; Moghadassi Jahromi, Zahra

    2016-09-01

    Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant. PMID:27430511

  6. Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.

    PubMed

    Aghazadeh, Rustam; Zamani, Mohammadreza; Motallebi, Mostafa; Moradyar, Mehdi; Moghadassi Jahromi, Zahra

    2016-09-01

    Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant.

  7. Utility of saliva and hair follicles in donor selection for hematopoietic stem cell transplantation and chimerism monitoring.

    PubMed

    Kaur, Gurvinder; Kumar, Neeraj; Nandakumar, Ramya; Rapthap, Chowphi C; Sharma, Gaurav; Neolia, Shekhar; Kumra, Heena; Mahalwar, Prateek; Garg, Abhinav; Kumar, Sunil; Kaur, Jasmeet; Hakim, Mrinali; Kumar, Lalit; Mehra, Narinder K

    2012-01-01

    Selection of an HLA identical donor is a critical pre-requisite for successful hematopoietic stem cell transplantation (HSCT). Most transplant centers utilize blood as the most common source of DNA for HLA testing. However, obtaining blood through phlebotomy is often challenging in patients with conditions like severe leucopenia or hemophilia, pediatric and elderly patients. We have used a simple in-house protocol and shown that HLA genotypes obtained on DNA extracted from saliva or hair are concordant with blood and hence can be used for selection of donors for HSCT or organ transplantation. Similarly, for post-HSCT chimerism monitoring, non-availability of pre-transplant DNA samples poses a major limitation of reference STR fingerprints. This study shows that DNA obtained post-HSCT from hair follicles can be used to generate pre-transplant patient specific fingerprints while the STR profiles obtained in saliva samples cannot as these display a mixed state of chimerism.

  8. Shear Force at the Cell-Matrix Interface: Enhanced Analysis for Microfabricated Post Array Detectors

    PubMed Central

    Lemmon, Christopher A.; Sniadecki, Nathan J.; Ruiz, Sami Alom; Tan, John L.; Romer, Lewis H.; Chen, Christopher S.

    2006-01-01

    The interplay of mechanical forces between the extracellular environment and the cytoskeleton drives development, repair, and senescence in many tissues. Quantitative definition of these forces is a vital step in understanding cellular mechanosensing. Microfabricated post array detectors (mPADs) provide direct measurements of cell-generated forces during cell adhesion to extracellular matrix. A new approach to mPAD post labeling, volumetric imaging, and an analysis of post bending mechanics determined that cells apply shear forces and not point moments at the matrix interface. In addition, these forces could be accurately resolved from post deflections by using images of post tops and bases. Image analysis tools were then developed to increase the precision and throughput of post centroid location. These studies resulted in an improved method of force measurement with broad applicability and concise execution using a fully automated force analysis system. The new method measures cell-generated forces with less than 5%error and less than 90 seconds of computational time. Using this approach, we demonstrated direct and distinct relationships between cellular traction force and spread cell surface area for fibroblasts, endothelial cells, epithelial cells and smooth muscle cells. PMID:16708468

  9. Finite element analysis of weakened roots restored with composite resin and posts.

    PubMed

    Coelho, Carla Santina de Miranda; Biffi, João Carlos Gabrielli; Silva, Gisele Rodrigues da; Abrahão, Anthony; Campos, Roberto Elias; Soares, Carlos José

    2009-11-01

    Finite element analysis (FEA) was used to investigate the influence of different post systems on the stress distribution of weakened teeth under oblique-load application. A maxillary central incisor root obtained from a sound tooth was weakened by partial removal of dentin inside the root canal. Seven two-dimensional numerical models, one from the sound tooth and six from the weakened root restored with composite resin and post systems were created as follows - ST: sound tooth; CPC: cast CuAl post and core; SSP: stainless steel post + composite core; GP: fiberglass + composite core; CP: carbon fiber + composite core; ZP: zirconium dioxide post + composite core; TP: titanium post + composite core. The numerical models were considered to be restored with a leucite-reinforced all-ceramic crown and received a 45 masculine occlusal load (10 N) on the lingual surface.All the materials and structures were considered linear elastic, homogeneous, and isotropic, with the exception of fiberglass and carbon fiber posts which assumed orthotropic behavior. The numerical models were plotted and meshed with isoparametric elements, and the results were analyzed using von Mises and Sy stress criteria. When compared with the sound tooth, FEA revealed differences in stress distribution when post systems were used. Among the restored teeth, the use of CPC, SSP, ZP, and TP resulted in higher stress concentration in the post itself when compared to GP and CP. Therefore, results from the FEA images suggested that the use of non-metallic post systems could result in improved mechanical behavior for the weakened restored teeth. PMID:20019417

  10. Chimerism and use of mesenchymal stem cells in umbilical cord blood transplantation.

    PubMed

    Berglund, Sofia; Uhlin, Michael; Mattsson, Jonas

    2013-01-01

    We performed a retrospective single-center analysis of 50 umbilical cord blood transplantations (UCBTs), focusing on chimerism development. Complete donor chimerism (DC) was associated with acute graft-vs.-host disease (aGVHD) grades II-IV for the CD3 (+) cell lineage (p = 0.01) and, in multivariate analysis, with total body irradiation (TBI) for all lineages (p < 0.01). Overall survival (OS) was negatively associated with patient age, (p < 0.001); aGVHD grades III-IV, (p < 0.001); and treatment with mesenchymal stem cells (MSCs) (p = 0.027). In conclusion, though multiple factors may have contributed, the association of TBI and DC might be worthy of consideration in the treatment of patients with malignant disease in the UCBT setting. The negative influence of MSCs on OS may be a reason for more careful usage of this treatment modality in combination with UCBT.

  11. Chimerism of allogeneic mesenchymal cells in bone marrow, liver, and spleen after mesenchymal stem cells infusion.

    PubMed

    Meleshko, Alexander; Prakharenia, Irina; Kletski, Semen; Isaikina, Yanina

    2013-12-01

    Although an infusion of culture-expanded MSCs is applied in clinic to improve results of HSCs transplantation and for a treatment of musculoskeletal disorders, homing, and engraftment potential of culture-expanded MSC in humans is still obscure. We report two female patients who received allogeneic BM transplantation as a treatment of hematological diseases and a transplantation of MSCs from third-party male donors. Both patients died within one yr of infectious complications. Specimens of paraffin-embedded blocks of tissues from transplanted patients were taken. The aim of the study was to estimate possible homing and engraftment of allogeneic BM-derived MSCs in some tissues/organs of recipient. Sensitive real-time quantitative PCR analysis was applied with SRY gene as a target. MSC chimerism was found in BM, liver, and spleen of both patients. We conclude that sensitive RQ-PCR analysis is acceptable for low-level chimerism evaluation even in paraffin-embedded tissue specimens.

  12. Chimerism of buccal membrane cells in a monochorionic dizygotic twin.

    PubMed

    Fumoto, Seiko; Hosoi, Kenichiro; Ohnishi, Hiroaki; Hoshina, Hiroaki; Yan, Kunimasa; Saji, Hiroh; Oka, Akira

    2014-04-01

    No monochorionic dizygotic twins (MCDZTs) with cellular chimerism involving cells other than blood cells have been reported in the literature to date. Here we report a probable first case of MCDZTs with buccal cell chimerism. A 32-year-old woman conceived twins by in vitro fertilization by using 2 cryopreserved blastocysts that were transferred into her uterus. An ultrasound scan at 8 weeks' gestation showed signs indicative of monochorionic twins. A healthy boy and a healthy girl were born, showing no sexual ambiguity. Cytogenetic analyses and microsatellite studies demonstrated chimerism in blood cells of both twins. Notably, repeated fluorescence in situ hybridization and microsatellite studies revealed chimerism in buccal cells obtained from 1 of the twins. Although the mechanism through which buccal cell chimerism was generated remains to be elucidated, ectopic differentiation of chimeric hematopoietic cells that migrated to the buccal membrane or the cellular transfer between the 2 embryos at the early stage of development might be responsible for the phenomenon. This hypothesis raises an interesting issue regarding embryonic development and cellular differentiation into organs during fetal development. Given the possibility of cryptic chimerism in various organs including gonadal tissues in MCDZTs, close observation will be required to determine whether complications develop in the course of the patients' growth.

  13. Complex chimerism: pregnancy after solid organ transplantation.

    PubMed

    Ma, Kimberly K; Petroff, Margaret G; Coscia, Lisa A; Armenti, Vincent T; Adams Waldorf, Kristina M

    2013-01-01

    Thousands of women with organ transplantation have undergone successful pregnancies, however little is known about how the profound immunologic changes associated with pregnancy might influence tolerance or rejection of the allograft. Pregnant women with a solid organ transplant are complex chimeras with multiple foreign cell populations from the donor organ, fetus, and mother of the pregnant woman. We consider the impact of complex chimerism and pregnancy-associated immunologic changes on tolerance of the allograft both during pregnancy and the postpartum period. Mechanisms of allograft tolerance are likely dynamic during pregnancy and affected by the influx of fetal microchimeric cells, HLA relationships (between the fetus, pregnant woman and/or donor), peripheral T cell tolerance to fetal cells, and fetal minor histocompatibility antigens. Further research is necessary to understand the complex immunology during pregnancy and the postpartum period of women with a solid organ transplant.

  14. Practical post-calibration uncertainty analysis: Yucca Mountain, Nevada, USA

    NASA Astrophysics Data System (ADS)

    James, S. C.; Doherty, J.; Eddebbarh, A.

    2009-12-01

    The values of parameters in a groundwater flow model govern the precision of predictions of future system behavior. Predictive precision, thus, typically depends on an ability to infer values of system properties from historical measurements through calibration. When such data are scarce, or when their information content with respect to parameters that are most relevant to predictions of interest is weak, predictive uncertainty may be high, even if the model is “calibrated.” Recent advances help recognize this condition, quantitatively evaluate predictive uncertainty, and suggest a path toward improved predictive accuracy by identifying sources of predictive uncertainty and by determining what observations will most effectively reduce this uncertainty. We demonstrate linear and nonlinear predictive error/uncertainty analyses as applied to a groundwater flow model of Yucca Mountain, Nevada, the US’s proposed site for disposal of high-level radioactive waste. Both of these types uncertainty analysis are readily implemented as an adjunct to model calibration with medium to high parameterization density. Linear analysis yields contributions made by each parameter to a prediction’s uncertainty and the worth of different observations, both existing and yet-to-be-gathered, toward reducing this uncertainty. Nonlinear analysis provides more accurate characterization of the uncertainty of model predictions while yielding their (approximate) probability distribution functions. This paper applies the above methods to a prediction of specific discharge and confirms the uncertainty bounds on specific discharge supplied in the Yucca Mountain Project License Application. Furthermore, Monte Carlo simulations confirm that hydrogeologic units thought to be flow barriers have probability distributions skewed toward lower permeabilities.

  15. Early peripheral blood and T-cell chimerism dynamics after umbilical cord blood transplantation supported with haploidentical cells.

    PubMed

    Kwon, M; Martínez-Laperche, C; Balsalobre, P; Serrano, D; Anguita, J; Gayoso, J; Díez-Martín, J L; Buño, I

    2014-02-01

    Single-unit umbilical cord blood (CB) SCT is limited by low total nucleated cell (TNC) dose. Co-infusion of CD34+ cells from a third party HLA-mismatched donor, known as dual or haplo-cord transplant, reduces the period of post-transplant neutropenia and related complications. The aim of this study was to analyze the value of early post-transplant peripheral blood (PB) and T cell chimerism after 28 dual transplants regarding CB engraftment. Cumulative incidence of myeloid engraftment at 30 days was 93% with a median time to engraftment of 14 days (10-29). Patients who developed CB graft failure (n=5) showed very low percentages of CB cells on days +14, +21 and +28 with decreasing dynamics. On the other hand, percentages of CB cells in patients who achieved CB engraftment increased over time. Interestingly, such patients showed two distinct chimerism dynamics in PB, but all of them showed a predominance of CB T cells early after SCT with increasing dynamics over time. Early post-transplant chimerism dynamics in PB and T cells predicts CB graft failure enabling rapid therapeutic measures to be applied. On the other hand, early increasing percentages of CB T cells correlates with ultimate CB engraftment.

  16. Expansion of recipient-derived antiviral T cells may influence donor chimerism after allogeneic stem cell transplantation.

    PubMed

    Borchers, S; Weissinger, E M; Pabst, B; Ganzenmueller, T; Dammann, E; Luther, S; Diedrich, H; Ganser, A; Stadler, M

    2013-12-01

    Donor chimerism (DC) analysis is an important marker in the hematopoietic stem cell transplant follow-up. Here, we present evidence for a possible relationship of infectious complications and declines in DC. We analyzed the DC in patients experiencing cytomegalovirus (CMV) reactivation. In addition, in some patients chimerism analyses of T-cell subsets were performed. CMV-specific cytotoxic T-lymphocytes (CMV-CTL) were monitored using human leukocyte antigen-restricted multimer staining. Interestingly, CMV reactivation was accompanied by changes in DC in 11 of 67 patients transplanted. For example, DC declined in a cord blood recipient, in both total leukocytes and CD4 and CD8 T-cell subsets upon CMV reactivation. The latter was controlled after only 5 days through expanding CMV-CTL of 96% recipient origin, according to chimerism analysis of CMV-CTL (enriched beyond 50%). In another patient, transplanted after reduced-intensity conditioning from a DQB1 mismatched, CMV seronegative donor, incipient CMV reactivation was completely aborted by CMV-CTL of recipient origin. However, at the same time, mixed chimerism dropped from 51% to 0% donor type, resulting in late graft rejection. Our data indicate that chimerism analyses in subset populations lead to a better understanding of declining total leukocyte chimerism. Furthermore, recipient-derived CMV-CTL may be able to control CMV reactivation after reduced-intensity conditioning. We speculate that autologous CMV-CTL may be instrumental to overcome recurrent CMV reactivations, especially in patients transplanted from CMV-seronegative donors. In addition, the expansion of recipient-derived CMV-CTL may contribute to both, graft failure or to conversion to full DC.

  17. Prediction of hospital failure: a post-PPS analysis.

    PubMed

    Gardiner, L R; Oswald, S L; Jahera, J S

    1996-01-01

    This study investigates the ability of discriminant analysis to provide accurate predictions of hospital failure. Using data from the period following the introduction of the Prospective Payment System, we developed discriminant functions for each of two hospital ownership categories: not-for-profit and proprietary. The resulting discriminant models contain six and seven variables, respectively. For each ownership category, the variables represent four major aspects of financial health (liquidity, leverage, profitability, and efficiency) plus county marketshare and length of stay. The proportion of closed hospitals misclassified as open one year before closure does not exceed 0.05 for either ownership type. Our results show that discriminant functions based on a small set of financial and nonfinancial variables provide the capability to predict hospital failure reliably for both not-for-profit and proprietary hospitals.

  18. Diversification of catalytic function in a synthetic family of chimeric cytochrome p450s.

    PubMed

    Landwehr, Marco; Carbone, Martina; Otey, Christopher R; Li, Yougen; Arnold, Frances H

    2007-03-01

    We report initial characterization of a synthetic family of more than 3000 cytochrome P450s made by SCHEMA recombination of 3 bacterial CYP102s. A total of 16 heme domains and their holoenzyme fusions with each of the 3 parental reductase domains were tested for activity on 11 different substrates. The results show that the chimeric enzymes have acquired significant functional diversity, including the ability to accept substrates not accepted by the parent enzymes. K-means clustering analysis of the activity data allowed the enzymes to be classified into five distinct groups based on substrate specificity. The substrates can also be grouped such that one can be a "surrogate" for others in the group. Fusion of a functional chimeric heme domain with a parental reductase domain always reconstituted a functional holoenzyme, indicating that key interdomain interactions are conserved upon reductase swapping. PMID:17379142

  19. Calcium-stimulated autophosphorylation site of plant chimeric calcium/calmodulin-dependent protein kinase

    NASA Technical Reports Server (NTRS)

    Sathyanarayanan, P. V.; Siems, W. F.; Jones, J. P.; Poovaiah, B. W.

    2001-01-01

    The existence of two molecular switches regulating plant chimeric Ca(2+)/calmodulin-dependent protein kinase (CCaMK), namely the C-terminal visinin-like domain acting as Ca(2+)-sensitive molecular switch and calmodulin binding domain acting as Ca(2+)-stimulated autophosphorylation-sensitive molecular switch, has been described (Sathyanarayanan, P. V., Cremo, C. R., and Poovaiah, B. W. (2000) J. Biol. Chem. 275, 30417-30422). Here we report the identification of Ca(2+)-stimulated autophosphorylation site of CCaMK by matrix-assisted laser desorption ionization time of flight-mass spectrometry. Thr(267) was confirmed as the Ca(2+)-stimulated autophosphorylation site by post-source decay experiments and by site-directed mutagenesis. The purified T267A mutant form of CCaMK did not show Ca(2+)-stimulated autophosphorylation, autophosphorylation-dependent variable calmodulin affinity, or Ca(2+)/calmodulin stimulation of kinase activity. Sequence comparison of CCaMK from monocotyledonous plant (lily) and dicotyledonous plant (tobacco) suggests that the autophosphorylation site is conserved. This is the first identification of a phosphorylation site specifically responding to activation by second messenger system (Ca(2+) messenger system) in plants. Homology modeling of the kinase and calmodulin binding domain of CCaMK with the crystal structure of calcium/calmodulin-dependent protein kinase 1 suggests that the Ca(2+)-stimulated autophosphorylation site is located on the surface of the kinase and far from the catalytic site. Analysis of Ca(2+)-stimulated autophosphorylation with increasing concentration of CCaMK indicates the possibility that the Ca(2+)-stimulated phosphorylation occurs by an intermolecular mechanism.

  20. Does inversion abolish the left chimeric face processing advantage?

    PubMed

    Butler, Stephen H; Harvey, Monika

    2005-12-19

    Experiments using chimeric stimuli have shown that the right hemisphere is more influential in processing facial information. Here, again, we found clear evidence that study participants used the information from the left side of the face to inform their gender decisions when chimeric male/female, female/male stimuli were presented. Most interestingly though, this effect was not only present for upright faces but also for inverted (flipped) faces (although the effect was significantly reduced). We propose that the chimeric bias effects found here argue against the idea that inversion destroys the right hemisphere superiority for faces. If this was indeed the case, flipping the chimeric faces should have resulted in a loss of the left face bias. This was not the case. PMID:16317340

  1. Stable mixed chimerism and tolerance to human organ transplants.

    PubMed

    Strober, Samuel

    2015-04-01

    Tolerance to combined kidney and hematopoietic cell transplant has been achieved in humans after establishment of mixed chimerism allowing for the withdrawal of immunosuppressive drugs. The seminal contributions of Ray Owen provided the scientific basis for the human protocol.

  2. Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation.

    PubMed

    Stahl, Tanja; Rothe, Caroline; Böhme, Manja U; Kohl, Aloisa; Kröger, Nicolaus; Fehse, Boris

    2016-01-01

    Accurate and sensitive determination of hematopoietic chimerism is a crucial diagnostic measure after allogeneic stem cell transplantation to monitor engraftment and potentially residual disease. Short tandem repeat (STR) amplification, the current "gold standard" for chimerism assessment facilitates reliable accuracy, but is hampered by its limited sensitivity (≥1%). Digital PCR (dPCR) has been shown to combine exact quantification and high reproducibility over a very wide measurement range with excellent sensitivity (routinely ≤0.1%) and thus represents a promising alternative to STR analysis. We here aimed at developing a whole panel of digital-PCR based assays for routine diagnostic. To this end, we tested suitability of 52 deletion/insertion polymorphisms (DIPs) for duplex analysis in combination with either a reference gene or a Y-chromosome specific PCR. Twenty-nine DIPs with high power of discrimination and good performance were identified, optimized and technically validated. We tested the newly established assays on retrospective patient samples that were in parallel also measured by STR amplification and found excellent correlation. Finally, a screening plate for initial genotyping with DIP-specific duplex dPCR assays was designed for convenient assay selection. In conclusion, we have established a comprehensive dPCR system for precise and high-sensitivity measurement of hematopoietic chimerism, which should be highly useful for clinical routine diagnostics. PMID:27618030

  3. Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation

    PubMed Central

    Stahl, Tanja; Rothe, Caroline; Böhme, Manja U.; Kohl, Aloisa; Kröger, Nicolaus; Fehse, Boris

    2016-01-01

    Accurate and sensitive determination of hematopoietic chimerism is a crucial diagnostic measure after allogeneic stem cell transplantation to monitor engraftment and potentially residual disease. Short tandem repeat (STR) amplification, the current “gold standard” for chimerism assessment facilitates reliable accuracy, but is hampered by its limited sensitivity (≥1%). Digital PCR (dPCR) has been shown to combine exact quantification and high reproducibility over a very wide measurement range with excellent sensitivity (routinely ≤0.1%) and thus represents a promising alternative to STR analysis. We here aimed at developing a whole panel of digital-PCR based assays for routine diagnostic. To this end, we tested suitability of 52 deletion/insertion polymorphisms (DIPs) for duplex analysis in combination with either a reference gene or a Y-chromosome specific PCR. Twenty-nine DIPs with high power of discrimination and good performance were identified, optimized and technically validated. We tested the newly established assays on retrospective patient samples that were in parallel also measured by STR amplification and found excellent correlation. Finally, a screening plate for initial genotyping with DIP-specific duplex dPCR assays was designed for convenient assay selection. In conclusion, we have established a comprehensive dPCR system for precise and high-sensitivity measurement of hematopoietic chimerism, which should be highly useful for clinical routine diagnostics. PMID:27618030

  4. Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation.

    PubMed

    Stahl, Tanja; Rothe, Caroline; Böhme, Manja U; Kohl, Aloisa; Kröger, Nicolaus; Fehse, Boris

    2016-01-01

    Accurate and sensitive determination of hematopoietic chimerism is a crucial diagnostic measure after allogeneic stem cell transplantation to monitor engraftment and potentially residual disease. Short tandem repeat (STR) amplification, the current "gold standard" for chimerism assessment facilitates reliable accuracy, but is hampered by its limited sensitivity (≥1%). Digital PCR (dPCR) has been shown to combine exact quantification and high reproducibility over a very wide measurement range with excellent sensitivity (routinely ≤0.1%) and thus represents a promising alternative to STR analysis. We here aimed at developing a whole panel of digital-PCR based assays for routine diagnostic. To this end, we tested suitability of 52 deletion/insertion polymorphisms (DIPs) for duplex analysis in combination with either a reference gene or a Y-chromosome specific PCR. Twenty-nine DIPs with high power of discrimination and good performance were identified, optimized and technically validated. We tested the newly established assays on retrospective patient samples that were in parallel also measured by STR amplification and found excellent correlation. Finally, a screening plate for initial genotyping with DIP-specific duplex dPCR assays was designed for convenient assay selection. In conclusion, we have established a comprehensive dPCR system for precise and high-sensitivity measurement of hematopoietic chimerism, which should be highly useful for clinical routine diagnostics.

  5. Plasma Fractionation Enriches Post-Myocardial Infarction Samples Prior to Proteomics Analysis

    PubMed Central

    de Castro Brás, Lisandra E.; DeLeon, Kristine Y.; Ma, Yonggang; Dai, Qiuxia; Hakala, Kevin; Weintraub, Susan T.; Lindsey, Merry L.

    2012-01-01

    Following myocardial infarction (MI), matrix metalloproteinase-9 (MMP-9) levels increase, and MMP-9 deletion improves post-MI remodeling of the left ventricle (LV). We provide here a technical report on plasma-analysis from wild type (WT) and MMP-9 null mice using fractionation and mass-spectrometry-based proteomics. MI was induced by coronary artery ligation in male WT and MMP-9 null mice (4–8 months old; n = 3/genotype). Plasma was collected on days 0 (pre-) and 1 post-MI. Plasma proteins were fractionated and proteins in the lowest (fraction 1) and highest (fraction 12) molecular weight fractions were separated by 1-D SDS-PAGE, digested in-gel with trypsin and analyzed by HPLC-ESI-MS/MS on an Orbitrap Velos. We tried five different fractionation protocols, before reaching an optimized protocol that allowed us to identify over 100 proteins. Serum amyloid A substantially increased post-MI in both genotypes, while alpha-2 macroglobulin increased only in the null samples. In fraction 12, extracellular matrix proteins were observed only post-MI. Interestingly, fibronectin-1, a substrate of MMP-9, was identified at both day 0 and day 1 post-MI in the MMP-9 null mice but was only identified post-MI in the WT mice. In conclusion, plasma fractionation offers an improved depletion-free method to evaluate plasma changes following MI. PMID:22778955

  6. Chimerism status after unrelated donor bone marrow transplantation with fludarabine-melphalan conditioning is affected by the melphalan dose and is predictive of relapse.

    PubMed

    Imahashi, Nobuhiko; Ohashi, Haruhiko; Terakura, Seitaro; Miyao, Kotaro; Sakemura, Reona; Kato, Tomonori; Sawa, Masashi; Yokohata, Emi; Kurahashi, Shingo; Ozawa, Yukiyasu; Nishida, Tetsuya; Kiyoi, Hitoshi; Watamoto, Koichi; Kohno, Akio; Kasai, Masanobu; Kato, Chiaki; Iida, Hiroatsu; Naoe, Tomoki; Miyamura, Koichi; Murata, Makoto

    2015-07-01

    Little is known regarding the chimerism status after reduced-intensity conditioning transplantation when bone marrow is used as a stem cell source. We prospectively analyzed lineage-specific chimerism and retrospectively evaluated clinical outcomes in 80 adult patients who underwent unrelated donor bone marrow transplantation (URBMT) with fludarabine plus melphalan (FM) as the conditioning regimen. Mixed donor chimerism (MDC) was seen in 43 and 10 % of patients at days 14 and 28, respectively. Melphalan at ≤130 mg/m(2) was associated with an increased incidence of MDC at day 28 (P = 0.03). Patients with MDC at day 14 showed a marginally increased risk of primary graft failure and a marginally decreased risk of graft-versus-host disease. In multivariate analysis, MDC at day 14 was associated with higher overall mortality (hazard ratio (HR) = 2.1; 95 % confidence interval (CI), 1.1-4.2; P = 0.04) and relapse rate (HR = 3.0; 95 % CI, 1.2-7.5; P = 0.02), but not with non-relapse mortality (HR = 1.8; 95 % CI, 0.70-4.6; P = 0.23). Thus, the FM regimen yields prompt complete donor chimerism after URBMT, but the melphalan dose significantly impacts the kinetics of chimerism. Chimerism status evaluation at day 14 may be instrumental in predicting relapse after URBMT with the FM regimen. PMID:25680895

  7. Chimerism status after unrelated donor bone marrow transplantation with fludarabine-melphalan conditioning is affected by the melphalan dose and is predictive of relapse.

    PubMed

    Imahashi, Nobuhiko; Ohashi, Haruhiko; Terakura, Seitaro; Miyao, Kotaro; Sakemura, Reona; Kato, Tomonori; Sawa, Masashi; Yokohata, Emi; Kurahashi, Shingo; Ozawa, Yukiyasu; Nishida, Tetsuya; Kiyoi, Hitoshi; Watamoto, Koichi; Kohno, Akio; Kasai, Masanobu; Kato, Chiaki; Iida, Hiroatsu; Naoe, Tomoki; Miyamura, Koichi; Murata, Makoto

    2015-07-01

    Little is known regarding the chimerism status after reduced-intensity conditioning transplantation when bone marrow is used as a stem cell source. We prospectively analyzed lineage-specific chimerism and retrospectively evaluated clinical outcomes in 80 adult patients who underwent unrelated donor bone marrow transplantation (URBMT) with fludarabine plus melphalan (FM) as the conditioning regimen. Mixed donor chimerism (MDC) was seen in 43 and 10 % of patients at days 14 and 28, respectively. Melphalan at ≤130 mg/m(2) was associated with an increased incidence of MDC at day 28 (P = 0.03). Patients with MDC at day 14 showed a marginally increased risk of primary graft failure and a marginally decreased risk of graft-versus-host disease. In multivariate analysis, MDC at day 14 was associated with higher overall mortality (hazard ratio (HR) = 2.1; 95 % confidence interval (CI), 1.1-4.2; P = 0.04) and relapse rate (HR = 3.0; 95 % CI, 1.2-7.5; P = 0.02), but not with non-relapse mortality (HR = 1.8; 95 % CI, 0.70-4.6; P = 0.23). Thus, the FM regimen yields prompt complete donor chimerism after URBMT, but the melphalan dose significantly impacts the kinetics of chimerism. Chimerism status evaluation at day 14 may be instrumental in predicting relapse after URBMT with the FM regimen.

  8. Assessing the technical efficiency of health posts in rural Guatemala: a data envelopment analysis

    PubMed Central

    Hernández, Alison R.; Sebastián, Miguel San

    2014-01-01

    Introduction Strengthening health service delivery to the rural poor is an important means of redressing inequities. Meso-level managers can help enhance efficiency in the utilization of existing resources through the application of practical tools to analyze routinely collected data reflecting inputs and outputs. This study aimed to assess the efficiency and change in productivity of health posts over two years in a rural department of Guatemala. Methods Data envelopment analysis was used to measure health posts’ technical efficiency and productivity change for 2008 and 2009. Input/output data were collected from the regional health office of Alta Verapaz for 34 health posts from the 19 districts comprising the health region. Results Technical efficiency varied widely across health posts, with mean scores of 0.78 (SD=0.24) and 0.75 (SD=0.21) in 2008 and 2009, respectively. Overall, productivity increased by 4%, though 47% of health posts experienced a decline in productivity. Results were combined on a bivariate plot to identify health posts at the high and low extremes of efficiency, which should be followed up to determine how and why their production processes are operating differently. Conclusions Assessing efficiency using the data that are available at the meso-level can serve as a first step in strengthening performance. Further work is required to support managers in the routine application of efficiency analysis and putting the results to use in guiding efforts to improve service delivery and increase utilization. PMID:24461356

  9. The Impact of Guided Notes on Post-Secondary Student Achievement: A Meta-Analysis

    ERIC Educational Resources Information Center

    Larwin, Karen H.; Larwin, David A.

    2013-01-01

    The common practice of using of guided notes in the post-secondary classroom is not fully appreciated or understood. In an effort to add to the existing research about this phenomenon, the current investigation expands on previously published research and one previously published meta-analysis that examined the impact of guided notes on…

  10. The School-to-Work Transition of Canadian Post-Secondary Graduates: A Dynamic Analysis

    ERIC Educational Resources Information Center

    Finnie, Ross

    2004-01-01

    This paper reports the results of an empirical analysis of the school-to-work transition of Canadian post-secondary graduates based on three waves of the National Graduates Surveys, representing those who successfully completed their programmes at Canadian colleges and universities in 1982, 1986, and 1990. Information was gathered during…

  11. An Analysis of Foster Care Placement History and Post-Secondary Graduation Rates

    ERIC Educational Resources Information Center

    Day, Angelique; Dworsky, Amy; Feng, Wenning

    2013-01-01

    Prior research has document significant disparities in post-secondary educational attainment between young adults who had been in foster care and their peers in the general population. This study uses survival analysis to compare the four-year college graduation rate of students who had been in foster care to the graduation rate of first…

  12. Campus Sustainability Governance in Canada: A Content Analysis of Post-Secondary Institutions' Sustainability Policies

    ERIC Educational Resources Information Center

    Vaughter, Philip; McKenzie, Marcia; Lidstone, Lauri; Wright, Tarah

    2016-01-01

    Purpose: This paper aims to provide an overview of a content analysis of sustainability policies from Canadian post-secondary education institutions. The paper reports findings on the orientations to sustainability evident in the policies; references to other policies within the documents; and other key themes on how sustainability is engaged in…

  13. TOPP: A post-processor for TOPAZ, the one dimensional pipe flow analysis code

    SciTech Connect

    Martin, R.W.

    1987-07-01

    TOPP is a Lawrence Livermore National Laboratory (LLNL) post-processor for producing graphical results from the one dimensional pipe flow analysis code, TOPAZ. TOPAZ was written by W. S. Winters of Sandia National Laboratory, Livermore (SNLL) and is available on the CRAY computers at LLNL. The SNLL version of TOPAZ produces a very limited set of variables that can be used as input to a post-processor. The version at LLNL has been modified to output every time-dependent variable to an absolute binary file at the user specified minor edit frequency. TOPP reads this absolute binary file and produces a variety of graphical results. 2 refs.

  14. TRINITY II: A post-processing program for two-dimensional finite element analysis data

    SciTech Connect

    Glick, J.H.; Gartling, D.K.

    1988-05-01

    TRINITY II is a program for post-processing data from two-dimensional finite element analyses. The program provides graphical display of mesh and solution data as well as data manipulation, file editing and selective printing of data. TRINITY II accepts data from any analysis code that employs the EXODUS file format; post-processing can be done interactively or in batch mode using any graphics device supported by the Sandia Virtual Device Interface. The capabilities and use of the program are described. 4 refs.

  15. Analysis of dentinal stress distribution of maxillary central incisors subjected to various post-and-core applications.

    PubMed

    Toksavul, Suna; Zor, Mehmet; Toman, Muhitten; Güngör, Mehmet Ali; Nergiz, Ibrahim; Artunç, Celal

    2006-01-01

    This study evaluated the stress distribution on an endodontically treated maxillary central incisor restored with different post-and-core systems by using a three-dimensional finite element analysis model. Seven three-dimensional finite element models were created. Each model contained cortical bone, cancelous bone, periodontal ligament, 3 mm apical root canal filling, post-and-core and all-ceramic crowns. Two different pre-fabricated zirconia ceramic post systems, a glass fiber-reinforced post system and a titanium post system were modeled. As a control, an all-ceramic crown on an endodontically treated maxillary central incisor without a post-and-core was modeled. Each model received a 45 degrees oblique occlusal load at a constant intensity of 100 N. In each model, the ratio of Von Mises stress distribution was compared. The greatest stresses were observed in the coronal third of the roots on facial surfaces. The ratio of Von Mises stress distribution in dentin for the zirconia ceramic post (CosmoPost) and ceramic core (Cosmo Ingot), zirconia ceramic post (CosmoPost) and composite core (Tetric Ceram), glass fiber-reinforced post (FRC Postec) and composite core (Tetric Ceram), titanium post (Er post) and composite core (Tetric Ceram), zirconia ceramic post (Cerapost) and ceramic core (Cosmo Ingot), zirconia ceramic post (Cerapost) and composite core (Tetric Ceram) and the control group were 0.886, 0.889, 0.988, 0.924, 0.889, 0.893 and 1, respectively. The stress concentrations in dentin created by two different zirconia ceramic post systems were nearly the same. The zirconia ceramic post systems created slightly less stress concentration in dentin than the glass fiber-reinforced and titanium posts.

  16. Overexpression of a Chimeric Gene, OsDST-SRDX, Improved Salt Tolerance of Perennial Ryegrass

    PubMed Central

    Cen, Huifang; Ye, Wenxing; Liu, Yanrong; Li, Dayong; Wang, Kexin; Zhang, Wanjun

    2016-01-01

    The Drought and Salt Tolerance gene (DST) encodes a C2H2 zinc finger transcription factor, which negatively regulates salt tolerance in rice (Oryza sativa). Phylogenetic analysis of six homologues of DST genes in different plant species revealed that DST genes were conserved evolutionarily. Here, the rice DST gene was linked to an SRDX domain for gene expression repression based on the Chimeric REpressor gene-Silencing Technology (CRES-T) to make a chimeric gene (OsDST-SRDX) construct and introduced into perennial ryegrass by Agrobacterium-mediated transformation. Integration and expression of the OsDST-SRDX in transgenic plants were tested by PCR and RT-PCR, respectively. Transgenic lines overexpressing the OsDST-SRDX fusion gene showed obvious phenotypic differences and clear resistance to salt-shock and to continuous salt stresses compared to non-transgenic plants. Physiological analyses including relative leaf water content, electrolyte leakage, proline content, malondialdehyde (MDA) content, H2O2 content and sodium and potassium accumulation indicated that the OsDST-SRDX fusion gene enhanced salt tolerance in transgenic perennial ryegrass by altering a wide range of physiological responses. To our best knowledge this study is the first report of utilizing Chimeric Repressor gene-Silencing Technology (CRES-T) in turfgrass and forage species for salt-tolerance improvement. PMID:27251327

  17. Overexpression of a Chimeric Gene, OsDST-SRDX, Improved Salt Tolerance of Perennial Ryegrass.

    PubMed

    Cen, Huifang; Ye, Wenxing; Liu, Yanrong; Li, Dayong; Wang, Kexin; Zhang, Wanjun

    2016-01-01

    The Drought and Salt Tolerance gene (DST) encodes a C2H2 zinc finger transcription factor, which negatively regulates salt tolerance in rice (Oryza sativa). Phylogenetic analysis of six homologues of DST genes in different plant species revealed that DST genes were conserved evolutionarily. Here, the rice DST gene was linked to an SRDX domain for gene expression repression based on the Chimeric REpressor gene-Silencing Technology (CRES-T) to make a chimeric gene (OsDST-SRDX) construct and introduced into perennial ryegrass by Agrobacterium-mediated transformation. Integration and expression of the OsDST-SRDX in transgenic plants were tested by PCR and RT-PCR, respectively. Transgenic lines overexpressing the OsDST-SRDX fusion gene showed obvious phenotypic differences and clear resistance to salt-shock and to continuous salt stresses compared to non-transgenic plants. Physiological analyses including relative leaf water content, electrolyte leakage, proline content, malondialdehyde (MDA) content, H2O2 content and sodium and potassium accumulation indicated that the OsDST-SRDX fusion gene enhanced salt tolerance in transgenic perennial ryegrass by altering a wide range of physiological responses. To our best knowledge this study is the first report of utilizing Chimeric Repressor gene-Silencing Technology (CRES-T) in turfgrass and forage species for salt-tolerance improvement. PMID:27251327

  18. A chimeric human-mouse model of Sjögren's syndrome.

    PubMed

    Young, Nicholas A; Wu, Lai-Chu; Bruss, Michael; Kaffenberger, Benjamin H; Hampton, Jeffrey; Bolon, Brad; Jarjour, Wael N

    2015-01-01

    Despite recent advances in the understanding of Sjögren's Syndrome (SjS), the pathogenic mechanisms remain elusive and an ideal model for early drug discovery is not yet available. To establish a humanized mouse model of SjS, peripheral blood mononuclear cells (PBMCs) from healthy volunteers or patients with SjS were transferred into immunodeficient NOD-scid IL-2rγ(null) mouse recipients to produce chimeric mice. While no difference was observed in the distribution of cells, chimeric mice transferred with PBMCs from SjS patients produced enhanced cytokine levels, most significantly IFN-γ and IL-10. Histological examination revealed enhanced inflammatory responses in the lacrimal and salivary glands of SjS chimeras, as measured by digital image analysis and blinded histopathological scoring. Infiltrates were primarily CD4+, with minimal detection of CD8+ T-cells and B-cells. These results demonstrate a novel chimeric mouse model of human SjS that provides a unique in vivo environment to test experimental therapeutics and investigate T-cell disease pathology.

  19. Neutrophils and lymphoid chimerism after adult living-related liver transplantation from a homozygous donor.

    PubMed

    Hajeer, A H; Issa, S; Alaskar, A; Abdullah, K; Awad, M; Tbakhi, A; Alabdulkareem, A

    2005-12-01

    Chimerism and graft-versus-host disease (GVHD) pose significant risks to liver transplant patients. The risk of chimerism and GVHD is higher among cases of living-related liver transplant (LRLT). Donors homozygous at all HLA loci carry a higher risk for GVHD. Herein we present a case of LRLT. The recipient suffered from end-stage liver disease and received a right lobe graft from his son. After 8 months posttransplant, the patient developed profound bone marrow depression. The patient was negative for CMV, Brucella, HHV6, HHV8, HBV, HCV, and parvovirus. No skin or GI signs of GVHD were noted. The patient and donor were HLA typed by SSP. The donor was homozygous for all HLA loci while the patient shared the class II homozygosity and was class I heterozygous. Chimerism studies were prompted after noting that the neutrophil compartment of the patient was homozygous for all HLA loci. This initiated further studies of the PMN and lymphocytes by microsatellite analysis. A total 15 microsatellites were analyzed. The results suggest that the majority (75%) of the PMNs and 45% of the lymphocytes were of donor origin. The patient was treated with G-CSF; his WBC counts returned to normal. At 2.5 years posttransplant the patient had not developed GVHD, despite the large number of donor lymphocytes circulating in his bloodstream. The only complaint he had was severe arthritis, which was treated with steroids. It must be investigated whether this was the result of GVHD.

  20. Long duration exposure facility post-flight thermal analysis, part 1

    NASA Technical Reports Server (NTRS)

    Berrios, William M.; Sampair, Thomas R.

    1992-01-01

    Results of the post-flight thermal analysis of the Long Duration Exposure Facility (LDEF) mission are presented. The LDEF mission thermal analysis was verified by comparing the thermal model results to flight data from the LDEF Thermal Measurements System (THERM). Post-flight calculated temperature uncertainties have been reduced to under +/- 18 F from the pre-flight uncertainties of +/- 40 F. The THERM consisted of eight temperature sensors, a shared tape recorder, a standard LDEF flight battery, and an electronics control box. The temperatures were measured at selected locations on the LDEF structure interior during the first 390 days of flight and recorded for post-flight analysis. After the LDEF retrieval from Space on 12 Jan. 1990, the tape recorder was recovered from the spacecraft and the data reduced for comparison to the LDEF predicted temperatures. The LDEF mission temperatures were calculated prior to the LDEF deployment on 7 Apr. 1980, and updated after the LDEF retrieval with the following actual flight parameter data: including thermal fluxes, spacecraft attitudes, thermal coatings degradation, and contamination effects. All updated data used for the calculation of post-flight temperatures is also presented in this document.

  1. Long duration exposure facility post-flight thermal analysis, part 2

    NASA Technical Reports Server (NTRS)

    Berrios, William M.; Sampair, Thomas R.

    1992-01-01

    Results of the post-flight thermal analysis for the Long Duration Exposure Facility (LDEF) mission are presented. The LDEF mission thermal analysis was verified by comparing the thermal model results to flight data from the LDEF Thermal Measurements System (THERM). Post-flight calculated temperature uncertainties have been reduced to under +/- 18 F from the pre-flight uncertainties of +/- 40 F. The THERM consisted of eight temperature sensors, a shared tape recorder, a standard LDEF flight battery, and an electronics control box. The temperatures were measured at selected locations on the LDEF structure interior during the first 390 days of flight and recorded for post-flight analysis. After the LDEF retrieval from Space on 12 Jan. 1990, the tape recorder was recovered from the spacecraft and the data reduced for comparison to the LDEF predicted temperatures. The LDEF mission temperatures were calculated prior to the LDEF deployment on 7 Apr. 1980, and updated after the LDEF retrieval with the following actual flight parameter data: thermal fluxes, spacecraft attitudes, thermal coatings degradation, and contamination effects. All updated data used for calculation of post-flight temperatures is also presented in this document.

  2. Parallel evolution of chimeric fusion genes.

    PubMed

    Jones, Corbin D; Begun, David J

    2005-08-01

    To understand how novel functions arise, we must identify common patterns and mechanisms shaping the evolution of new genes. Here, we take advantage of data from three Drosophila genes, jingwei, Adh-Finnegan, and Adh-Twain, to find evolutionary patterns and mechanisms governing the evolution of new genes. All three of these genes are independently derived from Adh, which enabled us to use the extensive literature on Adh in Drosophila to guide our analyses. We discovered a fundamental similarity in the temporal, spatial, and types of amino acid changes that occurred. All three genes underwent rapid adaptive amino acid evolution shortly after they were formed, followed by later quiescence and functional constraint. These genes also show striking parallels in which amino acids change in the Adh region. We showed that these early changes tend to occur at amino acid residues that seldom, if ever, evolve in Drosophila Adh. Changes at these slowly evolving sites are usually associated with loss of function or hypomorphic mutations in Drosophila melanogaster. Our data indicate that shifting away from ancestral functions may be a critical step early in the evolution of chimeric fusion genes. We suggest that the patterns we observed are both general and predictive.

  3. Chimeric antigen receptor therapy for cancer.

    PubMed

    Barrett, David M; Singh, Nathan; Porter, David L; Grupp, Stephan A; June, Carl H

    2014-01-01

    Improved outcomes for patients with cancer hinge on the development of new targeted therapies with acceptable short-term and long-term toxicity. Progress in basic, preclinical, and clinical arenas spanning cellular immunology, synthetic biology, and cell-processing technologies has paved the way for clinical applications of chimeric antigen receptor-based therapies. This new form of targeted immunotherapy merges the exquisite targeting specificity of monoclonal antibodies with the potent cytotoxicity and long-term persistence provided by cytotoxic T cells. Although this field is still in its infancy, clinical trials have already shown clinically significant antitumor activity in neuroblastoma, chronic lymphocytic leukemia, and B cell lymphoma, and trials targeting a variety of other adult and pediatric malignancies are under way. Ongoing work is focused on identifying optimal tumor targets and on elucidating and manipulating both cell- and host-associated factors to support expansion and persistence of the genetically engineered cells in vivo. The potential to target essentially any tumor-associated cell-surface antigen for which a monoclonal antibody can be made opens up an entirely new arena for targeted therapy of cancer.

  4. Analysis of Post-Deployment Cognitive Performance and Symptom Recovery in U.S. Marines

    PubMed Central

    Haran, F. J.; Alphonso, Aimee L.; Creason, Alia; Campbell, Justin S.; Johnson, Dagny; Young, Emily; Tsao, Jack W.

    2013-01-01

    Background Computerized neurocognitive testing (NCAT) has been proposed to be useful as a screening tool for post-deployment cognitive deficits in the setting of mild traumatic brain injury (mTBI). We assessed the clinical utility of post-injury/post-deployment Automated Neurocognitive Assessment Metric (ANAM) testing, using a longitudinal design to compare baseline ANAM tests with two post-deployment ANAM tests in a group of Marines who experienced combat during deployment. Methods and Findings Post-deployment cognitive performance and symptom recovery were compared in a subsample of 1324 U.S. Marines with high rates of combat exposure during deployment. Of the sample, 169 Marines had available baseline and twice repeated post-deployment ANAM results. A retrospective analysis of the ANAM data, which consisted of a self-report questionnaire about deployment-related blast exposure, recent history of mTBI, current clinical symptoms, and cognitive performance. Self-reported concussion sustained anytime during deployment was associated with a decrease in cognitive performance measured between 2–8 weeks post-deployment. At the second post-deployment test conducted on average eight months later, performance on the second simple reaction time test, in particular, remained impaired and was the most consistent and sensitive indicator of the cognitive decrements. Additionally, post-concussive symptoms were shown to persist in injured Marines with a self-reported history of concussion for an additional five months after most cognitive deficits resolved. Results of this study showed a measurable deployment effect on cognitive performance, although this effect appears to resolve without lasting clinical sequelae in those without history of deployment-related concussion. Conclusions These results highlight the need for a detailed clinical examination for service members with history of concussion and persistent clinical symptoms. Reliance solely upon computerized

  5. Motivational Interviewing Post-Stroke: An Analysis of Stroke Survivors' Concerns and Adjustment.

    PubMed

    Auton, Malcolm F; Patel, Kulsum; Carter, Bernie; Hackett, Maree; Thornton, Tim; Lightbody, Catherine E; Leathley, Michael J; Watkins, Caroline L

    2016-01-01

    Our earlier research demonstrated that participation in four sessions of motivational interviewing (MI) early post-stroke has a positive impact on stroke survivors' mood. However, the theoretical underpinnings of MI in supporting adjustment (rather than its traditional use in supporting behavior change) require clarification. This article describes a content analysis of MI transcripts for 10 participants in our previous study, to identify the focus of discussions (patient "concerns") and potential effective components of our MI approach. Patients' post-stroke concerns were shown in 16 categories, including frustration, family impact, and getting well. There was a pattern of change discourse across sessions: "Sustain talk" (reasons for not changing) reduced from Session 1 onward, "change talk" (intent to change) increased then reduced, and "change expressed" (changes achieved) increased from Sessions 1 to 4. MI facilitates healthy adjustment post-stroke in some patients, in turn affecting mood, but clarification of how this effect is achieved requires further exploration. PMID:25904673

  6. Failure mode analysis of a post-tension anchored dam using linear finite element analysis

    NASA Astrophysics Data System (ADS)

    Corn, Aimee

    There are currently over 84,000 dams in the United States, and the average age of those dams is 52 years. Concrete gravity dams are the second most common dam type, with more than 3,000 in the United States. Current engineering technology and technical understanding of hydrologic and seismic events has resulted in significant increases to the required design loads for most dams; therefore, many older dams do not have adequate safety for extreme loading events. Concrete gravity dams designed and constructed in the early 20th century did not consider uplift pressures beneath the dam, which reduces the effective weight of the structure. One method that has been used to enhance the stability of older concrete gravity dams includes the post-tension anchor (PTA) system. Post-tensioning infers modifying cured concrete and using self-equilibrating elements to increase the weight of the section, which provides added stability. There is a lack of historical evidence regarding the potential failure mechanisms for PTA concrete gravity dams. Of particular interest, is how these systems behave during large seismic events. The objective of this thesis is to develop a method by which the potential failure modes during a seismic event for a PTA dam can be evaluated using the linear elastic finite element method of analysis. The most likely potential failure modes (PFM) for PTA designs are due to tensile failure and shear failure. A numerical model of a hypothetical project was developed to simulate PTAs in the dam. The model was subjected to acceleration time-history motions that simulated the seismic loads. The results were used to evaluate the likelihood of tendon failure due to both tension and shear. The results from the analysis indicated that the PTA load increased during the seismic event; however, the peak load in the tendons was less than the gross ultimate tensile strength (GUTS) and would not be expected to result in tensile failure at the assumed project. The analysis

  7. Prophylactic clipping and post-polypectomy bleeding: a meta-analysis and systematic review

    PubMed Central

    Boumitri, Christine; Mir, Fazia A.; Ashraf, Imran; Matteson-Kome, Michelle L.; Nguyen, Douglas L.; Puli, Srinivas R.; Bechtold, Matthew L.

    2016-01-01

    Background Bleeding after polypectomy is a common issue associated with colonoscopy. To help prevent post-polypectomy bleeding, many endoscopists place clips at the site. However, this practice remains controversial. Therefore, we performed a meta-analysis of the efficacy of clip placement in the prevention of post-polypectomy bleeding. Methods Multiple databases, including Embase, Scopus, MEDLINE/PubMed, CINAHL, Cochrane databases, and recent abstracts from major American meetings were searched in April 2016. Using the DerSimonian and Laird (random effects) model with odds ratio (OR), a meta-analysis was performed of post-polypectomy bleeding with prophylactic clip versus no prophylactic clip. Results Five hundred and thirty potential articles and abstracts were discovered. Thirty-five articles were reviewed, with 12 studies satisfying the inclusion criteria. No statistically significant difference in prophylactic clipping versus no prophylactic clipping for post-polypectomy bleeding in all polyps was found when all studies (OR 1.49; 95% CI: 0.56–4.00; P=0.42), only peer-reviewed studies where abstracts were excluded (OR 0.84; 95% CI: 0.42–1.69; P=0.63), and only randomized controlled trials (OR 1.24; 95% CI: 0.69–2.24; P=0.47) were analyzed. Conclusions The use of prophylactic clipping for all polypectomies does not seem to prevent post-polypectomy bleeding and should not be a routine practice. However, for large polyps (>2 cm), prophylactic clipping may or may not be beneficial in preventing post-polypectomy bleeding. Further studies are required to fully evaluate this subgroup.

  8. Retrospective analysis of post-mortem findings in 1,444 aged donkeys.

    PubMed

    Morrow, L D; Smith, K C; Piercy, R J; du Toit, N; Burden, F A; Olmos, G; Gregory, N G; Verheyen, K L P

    2011-01-01

    The aim of this study was to describe and report the prevalence of conditions found at necropsy examination of UK donkeys. Records from 1,444 donkeys over a 7-year period were included in the analysis. Sixty-one categories of post-mortem finding were identified from 9,744 observations. The four most prevalent conditions noted were dental disorder (78.7%), vascular disease other than aneurysm (60.9%), arthritis (55.4%) and foot disorder (44.8%). Gastric ulceration was found in 42% of the donkeys and gastrointestinal impaction in 18.6%. The most frequent combination of two post-mortem findings in the same animal was arthritis and dental disorder. The most common disorders were associated with age, body weight and/or body condition post mortem and, for some disorders, gender. For many of the post-mortem findings, crude associations were found between the presence of one finding and the odds of also having certain other post-mortem findings. This study is the first to summarize all conditions noted at necropsy examination for a large group of donkeys. The findings increase knowledge of diseases and conditions of this species and may be useful when investigating the relevance of various pathological conditions in the live animal.

  9. A Juggling Act: A Feminist Post-Structuralist Analysis of Girls' and Boys' Talk in the Secondary Classroom.

    ERIC Educational Resources Information Center

    Baxter, Judith

    2002-01-01

    Argues that post-structuralist discourse analysis has a strong contribution to make to feminist and educational research, examining how the use of feminist post-structuralist discourse analysis (FPDA) can be tricky. Highlights an ethnographic study of secondary school girls' and boys' speech in class. Suggests that FPDA can produce powerful…

  10. Steroid metabolism in chimeric mice with humanized liver.

    PubMed

    Lootens, Leen; Van Eenoo, Peter; Meuleman, Philip; Pozo, Oscar J; Van Renterghem, Pieter; Leroux-Roels, Geert; Delbeke, Frans T

    2009-11-01

    Anabolic androgenic steroids are considered to be doping agents and are prohibited in sports. Their metabolism needs to be elucidated to allow for urinary detection by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS). Steroid metabolism was assessed using uPA(+/+) SCID mice with humanized livers (chimeric mice). This study presents the results of 19-norandrost-4-ene-3,17-dione (19-norAD) administration to these in vivo mice. As in humans, 19-norandrosterone and 19-noretiocholanolone are the major detectable metabolites of 19-norAD in the urine of chimeric mice.A summary is given of the metabolic pathways found in chimeric mice after administration of three model steroid compounds (methandienone, androst-4-ene-3,17-dione and 19-norandrost-4-ene-3,17-dione). From these studies we can conclude that all major metabolic pathways for anabolic steroids in humans are present in the chimeric mouse. It is hoped that, in future, this promising chimeric mouse model might assist the discovery of new and possible longer detectable metabolites of (designer) steroids. PMID:20355169

  11. Effect of reverse chimerism on rejection in clinical transplantation.

    PubMed

    Bolado, Pedro; Landin, Luis

    2013-11-01

    Chimerism may enable allografts to survive when immunosuppressive therapy is administered at low levels or is even absent. Reverse chimerism (RC) is focused on intragraft chimerism that repopulates the allograft with cells of recipient origin. We aimed to identify and analyze current clinical evidence on RC and the presence of endothelial RC and tissue-specific RC. A total of 33 clinical reports on cardiac, kidney, liver, and lung transplants published between 1972 and 2012 that focused on RC were included in a systematic review. Liver allografts presented with the highest percentage of endothelial RC and lung allografts by far the lowest. Tissue-specific RC was present in most of the recipients, but at very low levels. There were also cardiac and kidney allografts with chimerism, but the functionality of the cells of recipient origin was questionable. We were unable to determine whether RC was a trigger for or a result of acute rejection. Further clinical research should focus on outcomes to evaluate the clinical relevance of this form of chimerism in transplantation.

  12. Mixed chimerism and split tolerance: mechanisms and clinical correlations.

    PubMed

    Al-Adra, David P; Anderson, Colin C

    2011-01-01

    Establishing hematopoietic mixed chimerism can lead to donor-specific tolerance to transplanted organs and may eliminate the need for long-term immunosuppressive therapy, while also preventing chronic rejection. In this review, we discuss central and peripheral mechanisms of chimerism induced tolerance. However, even in the long-lasting presence of a donor organ or donor hematopoietic cells, some allogeneic tissues from the same donor can be rejected; a phenomenon known as split tolerance. With the current goal of creating mixed chimeras using clinically feasible amounts of donor bone marrow and with minimal conditioning, split tolerance may become more prevalent and its mechanisms need to be explored. Some predisposing factors that may increase the likelihood of split tolerance are immunogenicity of the graft, certain donor-recipient combinations, prior sensitization, location and type of graft and minimal conditioning chimerism induction protocols. Additionally, split tolerance may occur due to a differential susceptibility of various types of tissues to rejection. The mechanisms involved in a tissue's differential susceptibility to rejection include the presence of polymorphic tissue-specific antigens and variable sensitivity to indirect pathway effector mechanisms. Finally, we review the clinical attempts at allograft tolerance through the induction of chimerism; studies that are revealing the complex relationship between chimerism and tolerance. This relationship often displays split tolerance, and further research into its mechanisms is warranted.

  13. Porcine induced pluripotent stem cells produce chimeric offspring.

    PubMed

    West, Franklin D; Terlouw, Steve L; Kwon, Dae Jin; Mumaw, Jennifer L; Dhara, Sujoy K; Hasneen, Kowser; Dobrinsky, John R; Stice, Steven L

    2010-08-01

    Ethical and moral issues rule out the use of human induced pluripotent stem cells (iPSCs) in chimera studies that would determine the full extent of their reprogrammed state, instead relying on less rigorous assays such as teratoma formation and differentiated cell types. To date, only mouse iPSC lines are known to be truly pluripotent. However, initial mouse iPSC lines failed to form chimeric offspring, but did generate teratomas and differentiated embryoid bodies, and thus these specific iPSC lines were not completely reprogrammed or truly pluripotent. Therefore, there is a need to address whether the reprogramming factors and process used eventually to generate chimeric mice are universal and sufficient to generate reprogrammed iPSC that contribute to chimeric offspring in additional species. Here we show that porcine mesenchymal stem cells transduced with 6 human reprogramming factors (POU5F1, SOX2, NANOG, KLF4, LIN28, and C-MYC) injected into preimplantation-stage embryos contributed to multiple tissue types spanning all 3 germ layers in 8 of 10 fetuses. The chimerism rate was high, 85.3% or 29 of 34 live offspring were chimeras based on skin and tail biopsies harvested from 2- to 5-day-old pigs. The creation of pluripotent porcine iPSCs capable of generating chimeric offspring introduces numerous opportunities to study the facets significantly affecting cell therapies, genetic engineering, and other aspects of stem cell and developmental biology.

  14. Teeth restored using fiber-reinforced posts: in vitro fracture tests and finite element analysis.

    PubMed

    Schmitter, M; Rammelsberg, P; Lenz, J; Scheuber, S; Schweizerhof, K; Rues, S

    2010-09-01

    In dentistry the restoration of decayed teeth is challenging and makes great demands on both the dentist and the materials. Hence, fiber-reinforced posts have been introduced. The effects of different variables on the ultimate load on teeth restored using fiber-reinforced posts is controversial, maybe because the results are mostly based on non-standardized in vitro tests and, therefore, give inhomogeneous results. This study combines the advantages of in vitro tests and finite element analysis (FEA) to clarify the effects of ferrule height, post length and cementation technique used for restoration. Sixty-four single rooted premolars were decoronated (ferrule height 1 or 2 mm), endodontically treated and restored using fiber posts (length 2 or 7 mm), composite fillings and metal crowns (resin bonded or cemented). After thermocycling and chewing simulation the samples were loaded until fracture, recording first damage events. Using UNIANOVA to analyze recorded fracture loads, ferrule height and cementation technique were found to be significant, i.e. increased ferrule height and resin bonding of the crown resulted in higher fracture loads. Post length had no significant effect. All conventionally cemented crowns with a 1-mm ferrule height failed during artificial ageing, in contrast to resin-bonded crowns (75% survival rate). FEA confirmed these results and provided information about stress and force distribution within the restoration. Based on the findings of in vitro tests and computations we concluded that crowns, especially those with a small ferrule height, should be resin bonded. Finally, centrally positioned fiber-reinforced posts did not contribute to load transfer as long as the bond between the tooth and composite core was intact. PMID:20227533

  15. Construction of bacterial ghosts for transfer and expression of a chimeric hepatitis C virus gene in macrophages.

    PubMed

    Miri, M R; Behzad-Behbahani, A; Fardaei, M; Farhadi, A; Talebkhan, Y; Mohammadi, M; Tayebinia, M; Farokhinejad, F; Alavi, P; Fanian, M; Zare, F; Saberzade, J; Nikouyan, N; Okhovat, M A; Ranjbaran, R; Rafiei Dehbidi, G; Naderi, S

    2015-12-01

    The bacterial ghost (BG) production is a field of biotechnology for applications in vaccine and drug delivery. We assessed the capacity of BG for delivery of a recombinant gene encoded for both cell mediated and antibody dependent epitopes of hepatitis C virus (HCV) into murine macrophages. Escherichia coli (E. coli) cells were transformed with the lysis plasmid (pHH43). To produce chimeric gene, NS3 (non-structural protein 3) and core regions of HCV genome were fused together by splicing by overlap extension (SOEing) PCR and were cloned into plasmid pEGFP-C1. Bacterial ghosts were loaded with recombinant pEGFP-C1 and then were transferred to murine macrophages (RAW 264.7). To investigate plasmid transfection and chimeric mRNA transcription, fluorescent microscopy and RT-PCR were used. In vitro studies indicated that bacterial ghosts loaded with pEGFP-C1 plasmid were efficiently taken up by murine macrophages and indicated a high transfection rate (62%), as shown by fluorescent microscopy. RT-PCR from extracted intracellular mRNAs for chimeric Core-NS3 gene showed a specific 607 bp fragment of the gene. The sequence analysis of purified PCR products demonstrated the expected unique mRNA sequence. We constructed a chimeric HCV gene containing both cell mediated and antibody dependent epitopes with a significant expression in murine macrophages delivered by bacterial ghost.

  16. Flow cytometry with anti HLA-antibodies: a simple but highly sensitive method for monitoring chimerism and minimal residual disease after HLA-mismatched stem cell transplantation.

    PubMed

    Schumm, M; Feuchtinger, T; Pfeiffer, M; Hoelle, W; Bethge, W; Ebinger, M; Kuci, S; Handgretinger, R; Lang, P

    2007-06-01

    Transplantation of HLA-mismatched stem cells may allow determination of chimerism status of single cells by differential expression of HLA molecules. Monoclonal antibodies against HLA antigens can be used to determine the HLA type of sub-populations by standard flow cytometry. Blood samples from 23 patients transplanted from HLA-mismatched family donors were monitored using HLA-specific antibodies. Suitable antibodies could be found for all donor recipient pairs by using differences in HLA Bw4 and Bw6 groups or other serological antigens. Pretransplant controls of donor and recipient were used to correct for variable fluorescence intensities of the antibodies and sub-populations. Owing to the high sensitivity, cell populations with a minimum frequency of 0.1% were detectable. Flow-cytometric analysis was confirmed by chimerism analysis of immunomagnetically isolated T cells by standard PCR technique. In addition to chimerism evaluation, HLA antibodies improved the detection of leukemic cells after transplantation with aberrant phenotype. In conclusion, flow cytometry using antibodies against HLA antigens is an interesting tool for determination of chimerism and minimal residual disease after HLA-mismatched transplantation. Information about the chimerism status is given on a single-cell level and allows fast and convenient analysis of sub-populations.

  17. TC-2 post Helios experiment data review. [postflight systems analysis of spacecraft performance

    NASA Technical Reports Server (NTRS)

    1975-01-01

    Data are presented from a systems postflight analysis of the Centaur Launch Vehicle and Helios. Also given is a comparison of data from preflight analyses. Topics examined are: (1) propellant behavior; (2) helium usage; (3) propellant tank pressurization; (4) propellant tank thermodynamics; (5) component heating; thermal control; and thermal protection system; (6) main engine system; (7) H2O2 consumption; (8) boost pump post-meco performance; and (9) an overview of other systems.

  18. Ethical issues in the documentary data analysis of Internet posts and archives.

    PubMed

    Sixsmith, J; Murray, C D

    2001-05-01

    The documentary analysis of e-mail posts and archives for qualitative research has been outlined elsewhere. Although there is an increase in the number of studies being conducted on listserv and newsgroup material in health research, this has not always been accompanied by a careful, in-depth consideration of the concomitant ethical issues. Therefore, this article outlines the ethical considerations surrounding this form of research, including issues of accessing voices, consent, privacy, anonymity, interpretation, and ownership and authorship of research material.

  19. Molecular analysis of post-harvest withering in grape by AFLP transcriptional profiling

    PubMed Central

    Zamboni, Anita; Minoia, Leone; Ferrarini, Alberto; Tornielli, Giovanni Battista; Zago, Elisa; Delledonne, Massimo; Pezzotti, Mario

    2008-01-01

    Post-harvest withering of grape berries is used in the production of dessert and fortified wines to alter must quality characteristics and increase the concentration of simple sugars. The molecular processes that occur during withering are poorly understood, so a detailed transcriptomic analysis of post-harvest grape berries was carried out by AFLP-transcriptional profiling analysis. This will help to elucidate the molecular mechanisms of berry withering and will provide an opportunity to select markers that can be used to follow the drying process and evaluate different drying techniques. AFLP-TP identified 699 withering-specific genes, 167 and 86 of which were unique to off-plant and on-plant withering, respectively. Although similar molecular events were revealed in both withering processes, it was apparent that off-plant withering induced a stronger dehydration stress response resulting in the high level expression of genes involved in stress protection mechanisms, such as dehydrin and osmolite accumulation. Genes involved in hexose metabolism and transport, cell wall composition, and secondary metabolism (particularly the phenolic and terpene compound pathways) were similarly regulated in both processes. This work provides the first comprehensive analysis of the molecular events underpinning post-harvest withering and could help to define markers for different withering processes. PMID:19010774

  20. Micromechanics Analysis Code Post-Processing (MACPOST) User Guide. 1.0

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Comiskey, Michele D.; Bednarcyk, Brett A.

    1999-01-01

    As advanced composite materials have gained wider usage. the need for analytical models and computer codes to predict the thermomechanical deformation response of these materials has increased significantly. Recently, a micromechanics technique called the generalized method of cells (GMC) has been developed, which has the capability to fulfill this -oal. Tc provide a framework for GMC, the Micromechanics Analysis Code with Generalized Method of Cells (MAC/GMC) has been developed. As MAC/GMC has been updated, significant improvements have been made to the post-processing capabilities of the code. Through the MACPOST program, which operates directly within the MSC/PATRAN graphical pre- and post-processing package, a direct link between the analysis capabilities of MAC/GMC and the post-processing capabilities of MSC/PATRAN has been established. MACPOST has simplified the production, printing. and exportation of results for unit cells analyzed by MAC/GMC. MACPOST allows different micro-level quantities to be plotted quickly and easily in contour plots. In addition, meaningful data for X-Y plots can be examined. MACPOST thus serves as an important analysis and visualization tool for the macro- and micro-level data generated by MAC/GMC. This report serves as the user's manual for the MACPOST program.

  1. Facilitating cells: Translation of hematopoietic chimerism to achieve clinical tolerance.

    PubMed

    Ildstad, Suzanne T; Leventhal, Joseph; Wen, Yujie; Yolcu, Esma

    2015-04-01

    For over 50 y the association between hematopoietic chimerism and tolerance has been recognized. This originated with the brilliant observation by Dr. Ray Owen that freemartin cattle twins that shared a common placental blood supply were red blood cell chimeras, which led to the discovery that hematopoietic chimerism resulted in actively acquired tolerance. This was first confirmed in neonatal mice by Medawar et al. and subsequently in adult rodents. Fifty years later this concept has been successfully translated to solid organ transplant recipients in the clinic. The field is new, but cell-based therapies are being used with increasing frequency to induce tolerance and immunomodulation. The future is bright. This review focuses on chimerism and tolerance: past, present and prospects for the future.

  2. Symptotic detection of chimerism: Y does it matter?

    PubMed

    Geck, Peter

    2013-01-01

    Microchimerism (MC), transplacental acquisition of allogeneic cells from the mother (maternofetal MC) or from the fetus (fetomaternal MC) has been in the focus of research recently. Amplicons using Y-chromosome specific SRY and DYS14 sequences have been used as markers to trace cells from a male fetus in the mother. The sensitivity of these markers in formaldehyde fixed paraffin embedded samples, however, is less than optimal. To study chimerism in breast cancer we took advantage of the evolutionary history of the Y chromosome and designed amplicons on gene repeats to generate additive PCR signals. The increased sensitivity detected high incidence of male chimerism in normal breast tissues. We also showed correlation with protection from cancer with unique quantitative biology. Accumulating data from biology and medicine indicate that natural chimerism is astonishingly frequent and may affect human conditions. We hypothesize that it has significant evolutionary ramifications as well.

  3. Chimerism in the immunohematology laboratory in the molecular biology era.

    PubMed

    Bluth, Martin H; Reid, Marion E; Manny, Noga

    2007-04-01

    Dual or multiple cell populations, induced by chimeras, have been the subject of many studies. This long-standing fascination with chimeras has revealed a good deal of knowledge about human inheritance. Although historically most chimeras were caused by natural events, certain current medical intervention therapies are increasing the number of situations that can lead to a mixed cell population, that is, the chimeric condition, in humans. Medical therapies such as transfusion, stem cell transplantation, kidney transplantation, and artificial insemination induce temporary and sometimes permanent chimeras. Such natural or therapeutically induced presentations of chimerism can present challenging issues to the clinical immunohematology laboratory with regard to interpretation of results and subsequent patient management. The purpose of this review was to highlight some of these chimeric states and hypothesize how testing DNA from various tissues can cause apparent discrepancies between phenotype and genotype results.

  4. Chimeric antigen receptors: driving immunology towards synthetic biology.

    PubMed

    Sadelain, Michel

    2016-08-01

    The advent of second generation chimeric antigen receptors and the CD19 paradigm have ushered a new therapeutic modality in oncology. In contrast to earlier forms of adoptive cell therapy, which were based on the isolation and expansion of naturally occurring T cells, CAR therapy is based on the design and manufacture of engineered T cells with optimized properties. A new armamentarium, comprising not only CARs but also chimeric costimulatory receptors, chimeric cytokine receptors, inhibitory receptors and synthetic Notch receptors, expressed in naïve, central memory or stem cell-like memory T cells, is being developed for clinical use in a wide range of cancers. Immunological principles are thus finding a new purpose thanks to advances in genetic engineering, synthetic biology and cell manufacturing sciences. PMID:27372731

  5. Chimeric antigen receptors: driving immunology towards synthetic biology.

    PubMed

    Sadelain, Michel

    2016-08-01

    The advent of second generation chimeric antigen receptors and the CD19 paradigm have ushered a new therapeutic modality in oncology. In contrast to earlier forms of adoptive cell therapy, which were based on the isolation and expansion of naturally occurring T cells, CAR therapy is based on the design and manufacture of engineered T cells with optimized properties. A new armamentarium, comprising not only CARs but also chimeric costimulatory receptors, chimeric cytokine receptors, inhibitory receptors and synthetic Notch receptors, expressed in naïve, central memory or stem cell-like memory T cells, is being developed for clinical use in a wide range of cancers. Immunological principles are thus finding a new purpose thanks to advances in genetic engineering, synthetic biology and cell manufacturing sciences.

  6. Chimerism studies in HLA-identical nonmyeloablative hematopoietic stem cell transplantation point to the donor CD8(+) T-cell count on day + 14 as a predictor of acute graft-versus-host disease.

    PubMed

    Petersen, Søren L; Madsen, Hans O; Ryder, Lars P; Svejgaard, A; Masmas, Tania N; Dickmeiss, Ebbe; Heilmann, Carsten; Vindeløv, Lars L

    2004-05-01

    Chimerism analysis of hematopoietic cells has emerged as an essential tool in nonmyeloablative hematopoietic stem cell transplantation. We have investigated the development of donor chimerism in granulocytes and CD4(+) and CD8(+) T cells in blood and bone marrow of 24 patients with hematologic malignancies who received HLA-identical sibling peripheral blood stem cell grafts after conditioning with fludarabine and 2 Gy of total body irradiation. The T-cell chimerism of blood and bone marrow was tightly correlated. Complete donor chimerism was reached earlier in the granulocytes than in the T cells. Mixed T-cell chimerism was common at the time of onset of acute graft-versus-host disease (aGVHD), and both CD4(+) and CD8(+) donor T-cell chimerism increased with the occurrence of aGVHD grades II to IV (P =.0002 and P =.019, respectively). The rate of disappearance of recipient CD8(+) T cells was faster in patients with aGVHD grades II to IV than in patients without clinically significant aGVHD (P =.016). This observation indicates a role of graft-versus-lymphohematopoietic tissue reactions in creating complete donor T-cell chimerism. A donor CD8(+) T-cell count above the median on day +14 increased the risk of subsequent development of aGVHD grades II to IV (P =.003).

  7. Measurement uncertainty analysis of low-dose-rate prostate seed brachytherapy: post-implant dosimetry.

    PubMed

    Gregory, Kent J; Pattison, John E; Bibbo, Giovanni

    2015-03-01

    The minimal dose covering 90 % of the prostate volume--D 90--is arguably the most important dosimetric parameter in low-dose-rate prostate seed brachytherapy. In this study an analysis of the measurement uncertainties in D 90 from low-dose-rate prostate seed brachytherapy was conducted for two common treatment procedures with two different post-implant dosimetry methods. The analysis was undertaken in order to determine the magnitude of D 90 uncertainty, how the magnitude of the uncertainty varied when D 90 was calculated using different dosimetry methods, and which factors were the major contributors to the uncertainty. The analysis considered the prostate as being homogeneous and tissue equivalent and made use of published data, as well as original data collected specifically for this analysis, and was performed according to the Guide to the expression of uncertainty in measurement (GUM). It was found that when prostate imaging and seed implantation were conducted in two separate sessions using only CT images for post-implant analysis, the expanded uncertainty in D 90 values were about 25 % at the 95 % confidence interval. When prostate imaging and seed implantation were conducted during a single session using CT and ultrasound images for post-implant analysis, the expanded uncertainty in D 90 values were about 33 %. Methods for reducing these uncertainty levels are discussed. It was found that variations in contouring the target tissue made the largest contribution to D 90 uncertainty, while the uncertainty in seed source strength made only a small contribution. It is important that clinicians appreciate the overall magnitude of D 90 uncertainty and understand the factors that affect it so that clinical decisions are soundly based, and resources are appropriately allocated.

  8. Uncertainty characterization and quantification in air pollution models. Application to the CHIMERE model

    NASA Astrophysics Data System (ADS)

    Debry, Edouard; Mallet, Vivien; Garaud, Damien; Malherbe, Laure; Bessagnet, Bertrand; Rouïl, Laurence

    2010-05-01

    probability density function (PDF) is associated with an input parameter, according to its assumed uncertainty. Then the combined PDFs are propagated into the model, by means of several simulations with randomly perturbed input parameters. One may then obtain an approximation of the PDF of modeled concentrations, provided the Monte Carlo process has reasonably converged. The uncertainty analysis with CHIMERE has been led with a Monte Carlo method on the French domain and on two periods : 13 days during January 2009, with a focus on particles, and 28 days during August 2009, with a focus on ozone. The results show that for the summer period and 500 simulations, the time and space averaged standard deviation for ozone is 16 µg/m3, to be compared with an averaged concentration of 89 µg/m3. It is noteworthy that the space averaged standard deviation for ozone is relatively constant over time (the standard deviation of the timeseries itself is 1.6 µg/m3). The space variation of the ozone standard deviation seems to indicate that emissions have a significant impact, followed by western boundary conditions. Monte Carlo simulations are then post-processed by both ensemble [4] and Bayesian [5] methods in order to assess the quality of the uncertainty estimation. (1) Rao, K.S. Uncertainty Analysis in Atmospheric Dispersion Modeling, Pure and Applied Geophysics, 2005, 162, 1893-1917. (2) Beekmann, M. and Derognat, C. Monte Carlo uncertainty analysis of a regional-scale transport chemistry model constrained by measurements from the Atmospheric Pollution Over the Paris Area (ESQUIF) campaign, Journal of Geophysical Research, 2003, 108, 8559-8576. (3) Hanna, S.R. and Lu, Z. and Frey, H.C. and Wheeler, N. and Vukovich, J. and Arunachalam, S. and Fernau, M. and Hansen, D.A. Uncertainties in predicted ozone concentrations due to input uncertainties for the UAM-V photochemical grid model applied to the July 1995 OTAG domain, Atmospheric Environment, 2001, 35, 891-903. (4) Mallet, V., and B

  9. Chimeric gag-V3 virus-like particles of human immunodeficiency virus induce virus-neutralizing antibodies.

    PubMed Central

    Luo, L; Li, Y; Cannon, P M; Kim, S; Kang, C Y

    1992-01-01

    A 41-kDa unprocessed human immunodeficiency virus 2 (HIV-2) gag precursor protein that has a deletion of a portion of the viral protease assembles as virus-like particles by budding through the cytoplasmic membrane of recombinant baculovirus-infected insect cells. We have constructed six different combinations of chimeric genes by coupling the truncated HIV-2 gag gene to the neutralizing domain (V3) or the neutralizing and the CD4 binding domains (V3+CD4BD) of gp120 env gene sequences from HIV-1 or HIV-2. The env gene sequences were inserted either into the middle of the gag gene or at the 3' terminus of the gag gene. Virus-like particles were formed by chimeric gene products only when the env gene sequences were linked to the 3' terminus of the gag gene. Insertion of env gene sequence in the middle of the gag gene resulted in high-level chimeric gene expression but without the formation of virus-like particles. Three different chimeric genes [gag gene with HIV-1 V3 (1V3), gag gene with HIV-2 V3 (2V3), and gag gene with HIV-2 V3+CD4BD (2V3+CD4BD)] formed virus-like particles that were secreted into the cell culture medium. In contrast, the HIV-1 V3+CD4BD/HIV-2 gag construct did not form virus-like particles. The chimeric gag-env particles had spherical morphology and the size was slightly larger than that of the gag particles, but the chimeric particles were similar to the mature HIV particles. Western blot analysis showed that the gag-env chimeric proteins were recognized by antibodies in HIV-positive human serum and rabbit anti-gp120 serum. Rabbit anti-gag 1V3 and anti-gag 2V3 sera reacted with authentic gp120 of HIV-1 and HIV-2, respectively, and neutralized homologous HIV infectivity. Our results show that precursor gag protein has potential as a carrier for the presentation of foreign epitopes in good immunological context. The gag protein is highly immunogenic and has the ability to carry large foreign inserts; as such, it offers an attractive approach for

  10. Association between hepatitis B virus and MHC class I polypeptide-related chain A in human hepatocytes derived from human-mouse chimeric mouse liver.

    PubMed

    Sasaki, Reina; Kanda, Tatsuo; Wu, Shuang; Nakamoto, Shingo; Haga, Yuki; Jiang, Xia; Nakamura, Masato; Shirasawa, Hiroshi; Yokosuka, Osamu

    2015-09-01

    Due to the lack of efficient hepatitis B virus (HBV) infection systems, progress in understanding the role of innate immunity in HBV infection has remained challenging. Here we used human hepatocytes from a humanized severe combined immunodeficiency albumin promoter/enhancer driven-urokinase-type plasminogen activator mouse model for HBV infection. HBV DNA levels in culture medium from these human hepatocytes were 4.8-5.7 log IU/mL between day 16 and day 66 post-infection by HBV genotype C inoculum. HBV surface antigen (HBsAg) was also detected by chemiluminescent immunoassay from day 7 to day 66 post-infection. Western blot analysis revealed that major histocompatibility complex class I-related chain A (MICA), which plays a role in the innate immune system, was induced in HBV-infected human hepatocytes 27 days after infection compared with the uninfected control. MICA was reduced at day 62 and undetectable at day 90. Of interest, MICA expression by human hepatocytes increased after HBV infection and decreased before HBsAg loss. Human hepatocytes derived from chimeric mice with hepatocyte-humanized liver could support HBV genome replication. Further studies of the association between HBV replication and MICA induction should be conducted.

  11. Post-Mortem Tissue Biopsies Obtained at Minimally Invasive Autopsy: An RNA-Quality Analysis

    PubMed Central

    Kap, Marcel; Weustink, Annick C.; Riegman, Peter H. J.; Oosterhuis, J. Wolter

    2014-01-01

    Introduction Bereaved relatives often refuse to give consent for post-mortem investigation of deceased cancer patients, mainly because of the mutilation due to conventional autopsy (CA). Minimally invasive autopsy (MIA) may be a more acceptable alternative and, if implemented in clinical practice, creates an opportunity to more often obtain post-mortem tissue samples of (recurred) primary tumors and metastases for molecular research. As a measure for tissue quality for molecular studies, we hereby present a feasibility study, comparing the RNA quality of MIA and CA samples, and fresh frozen samples as reference. Materials and methods Tissue samples of heart, liver and kidney were prospectively collected from 24 MIAs followed by CA, and compared to corresponding archival fresh frozen tissue. After RNA isolation and RT-qPCR, RNA integrity numbers (RIN) and GAPDH expression (six amplicon sizes ranging from 71 to 530 base pairs) were measured. RIN values and GAPDH Cq values were analyzed and compared between all sample groups and post-mortem intervals (PMI). Results RIN values in MIA samples were significantly higher than those in CA samples. GAPDH was expressed significantly higher in MIA samples than in CA samples and 530 bp PCR products could be measured in all cases. GAPDH expression was significantly lower in samples with PMI >15 hours. As expected, the samples of the fresh frozen reference standard performed best in all analyses. Conclusion MIA samples showed better RNA quality than CA samples, probably due to shorter PMI. Both had lower RNA quality and expression levels than fresh frozen tissue, however, remaining GAPDH RNA was still sufficiently intact. Therefore, other highly expressed genes are most likely also detectable. Gene array analysis should be performed to gain insight into the quality of entire post-mortem genomes. Reducing PMI will further improve the feasibility of demanding molecular research on post-mortem tissues, this is most likely more

  12. Post-traumatic growth following acquired brain injury: a systematic review and meta-analysis

    PubMed Central

    Grace, Jenny J.; Kinsella, Elaine L.; Muldoon, Orla T.; Fortune, Dónal G.

    2015-01-01

    The idea that acquired brain injury (ABI) caused by stroke, hemorrhage, infection or traumatic insult to the brain can result in post-traumatic growth (PTG) for individuals is increasingly attracting psychological attention. However, PTG also attracts controversy as a result of ambiguous empirical findings. The extent that demographic variables, injury factors, subjective beliefs, and psychological health are associated with PTG following ABI is not clear. Consequently, this systematic review and meta-analysis explores the correlates of variables within these four broad areas and PTG. From a total of 744 published studies addressing PTG in people with ABI, eight studies met inclusion criteria for detailed examination. Meta-analysis of these studies indicated that growth was related to employment, longer education, subjective beliefs about change post-injury, relationship status, older age, longer time since injury, and lower levels of depression. Results from homogeneity analyses indicated significant inter-study heterogeneity across variables. There is general support for the idea that people with ABI can experience growth, and that various demographics, injury-related variables, subjective beliefs and psychological health are related to growth. The contribution of social integration and the forming of new identities post-ABI to the experience of PTG is explored. These meta-analytic findings are however constrained by methodological limitations prevalent in the literature. Clinical and research implications are discussed with specific reference to community and collective factors that enable PTG. PMID:26321983

  13. Post-Buckling and Ultimate Strength Analysis of Stiffened Composite Panel Base on Progressive Damage

    NASA Astrophysics Data System (ADS)

    Zhang, Guofan; Sun, Xiasheng; Sun, Zhonglei

    Stiffened composite panel is the typical thin wall structure applied in aerospace industry, and its main failure mode is buckling subjected to compressive loading. In this paper, the development of an analysis approach using Finite Element Method on post-buckling behavior of stiffened composite structures under compression was presented. Then, the numerical results of stiffened panel are obtained by FE simulations. A thorough comparison were accomplished by comparing the load carrying capacity and key position strains of the specimen with test. The comparison indicates that the FEM results which adopted developed methodology could meet the demand of engineering application in predicting the post-buckling behavior of intact stiffened structures in aircraft design stage.

  14. Sex-discordant monochorionic twins with blood and tissue chimerism.

    PubMed

    Rodriguez-Buritica, David; Rojnueangnit, Kitiwan; Messiaen, Ludwine M; Mikhail, Fady M; Robin, Nathaniel H

    2015-04-01

    We report on a pair of normally conceived monochorionic/dizygotic (MC/DZ) sex discordant twins. The comparison of blood and skin genotypes revealed that the chimerism was also present in the skin. We conjecture about the developmental origins of this case.

  15. Therapeutic use of chimeric bacteriophage (phage) lysins in staphylococcal endophthalmitis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purpose: Phage endolysins are peptidoglycan hydrolases that are produced at the end of the phage lytic cycle to digest the host bacterial cell wall, facilitating the release of mature phage progeny. The aim of this study is to determine the antimicrobial activity of chimeric phage lysins against cli...

  16. Computational and statistical methods for high-throughput analysis of post-translational modifications of proteins.

    PubMed

    Schwämmle, Veit; Verano-Braga, Thiago; Roepstorff, Peter

    2015-11-01

    The investigation of post-translational modifications (PTMs) represents one of the main research focuses for the study of protein function and cell signaling. Mass spectrometry instrumentation with increasing sensitivity improved protocols for PTM enrichment and recently established pipelines for high-throughput experiments allow large-scale identification and quantification of several PTM types. This review addresses the concurrently emerging challenges for the computational analysis of the resulting data and presents PTM-centered approaches for spectra identification, statistical analysis, multivariate analysis and data interpretation. We furthermore discuss the potential of future developments that will help to gain deep insight into the PTM-ome and its biological role in cells. This article is part of a Special Issue entitled: Computational Proteomics.

  17. Signature Based Detection of User Events for Post-mortem Forensic Analysis

    NASA Astrophysics Data System (ADS)

    James, Joshua Isaac; Gladyshev, Pavel; Zhu, Yuandong

    This paper introduces a novel approach to user event reconstruction by showing the practicality of generating and implementing signature-based analysis methods to reconstruct high-level user actions from a collection of low-level traces found during a post-mortem forensic analysis of a system. Traditional forensic analysis and the inferences an investigator normally makes when given digital evidence, are examined. It is then demonstrated that this natural process of inferring high-level events from low-level traces may be encoded using signature-matching techniques. Simple signatures using the defined method are created and applied for three popular Windows-based programs as a proof of concept.

  18. Continuous Versus Group Sequential Analysis for Post-Market Drug and Vaccine Safety Surveillance

    PubMed Central

    Silva, I. R.; Kulldorff, M.

    2016-01-01

    Summary The use of sequential statistical analysis for post-market drug safety surveillance is quickly emerging. Both continuous and group sequential analysis have been used, but consensus is lacking as to when to use which approach. We compare the statistical performance of continuous and group sequential analysis in terms of type I error probability; statistical power; expected time to signal when the null hypothesis is rejected; and the sample size required to end surveillance without rejecting the null. We present a mathematical proposition to show that for any group sequential design there always exists a continuous sequential design that is uniformly better. As a consequence, it is shown that more frequent testing is always better. Additionally, for a Poisson based probability model and a flat rejection boundary in terms of the log likelihood ratio, we compare the performance of various continuous and group sequential designs. Using exact calculations, we found that, for the parameter settings used, there is always a continuous design with shorter expected time to signal than the best group design. The two key conclusions from this article are (i) that any post-market safety surveillance system should attempt to obtain data as frequently as possible, and (ii) that sequential testing should always be performed when new data arrives without deliberately waiting for additional data. PMID:26011024

  19. Continuous versus group sequential analysis for post-market drug and vaccine safety surveillance.

    PubMed

    Silva, I R; Kulldorff, M

    2015-09-01

    The use of sequential statistical analysis for post-market drug safety surveillance is quickly emerging. Both continuous and group sequential analysis have been used, but consensus is lacking as to when to use which approach. We compare the statistical performance of continuous and group sequential analysis in terms of type I error probability; statistical power; expected time to signal when the null hypothesis is rejected; and the sample size required to end surveillance without rejecting the null. We present a mathematical proposition to show that for any group sequential design there always exists a continuous sequential design that is uniformly better. As a consequence, it is shown that more frequent testing is always better. Additionally, for a Poisson based probability model and a flat rejection boundary in terms of the log likelihood ratio, we compare the performance of various continuous and group sequential designs. Using exact calculations, we found that, for the parameter settings used, there is always a continuous design with shorter expected time to signal than the best group design. The two key conclusions from this article are (i) that any post-market safety surveillance system should attempt to obtain data as frequently as possible, and (ii) that sequential testing should always be performed when new data arrives without deliberately waiting for additional data.

  20. Analysis of nonstandard and home-made explosives and post-blast residues in forensic practice

    NASA Astrophysics Data System (ADS)

    Kotrlý, Marek; Turková, Ivana

    2014-05-01

    Nonstandard and home-made explosives may constitute a considerable threat and as well as a potential material for terrorist activities. Mobile analytical devices, particularly Raman, or also FTIR spectrometers are used for the initial detection. Various sorts of phlegmatizers (moderants) to decrease sensitivity of explosives were tested, some kinds of low viscosity lubricants yielded very good results. If the character of the substance allows it, phlegmatized samples are taken in the amount of approx.0.3g for a laboratory analysis. Various separation methods and methods of concentrations of samples from post-blast scenes were tested. A wide range of methods is used for the laboratory analysis. XRD techniques capable of a direct phase identification of the crystalline substance, namely in mixtures, have highly proved themselves in practice for inorganic and organic phases. SEM-EDS/WDS methods are standardly employed for the inorganic phase. In analysing post-blast residues, there are very important techniques allowing analysis at the level of separate particles, not the overall composition in a mixed sample.

  1. Development of transgenic watermelon resistant to Cucumber mosaic virus and Watermelon mosaic virus by using a single chimeric transgene construct.

    PubMed

    Lin, Ching-Yi; Ku, Hsin-Mei; Chiang, Yi-Hua; Ho, Hsiu-Yin; Yu, Tsong-Ann; Jan, Fuh-Jyh

    2012-10-01

    Watermelon, an important fruit crop worldwide, is prone to attack by several viruses that often results in destructive yield loss. To develop a transgenic watermelon resistant to multiple virus infection, a single chimeric transgene comprising a silencer DNA from the partial N gene of Watermelon silver mottle virus (WSMoV) fused to the partial coat protein (CP) gene sequences of Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV) and Watermelon mosaic virus (WMV) was constructed and transformed into watermelon (cv. Feeling) via Agrobacterium-mediated transformation. Single or multiple transgene copies randomly inserted into various locations in the genome were confirmed by Southern blot analysis. Transgenic watermelon R(0) plants were individually challenged with CMV, CGMMV or WMV, or with a mixture of these three viruses for resistance evaluation. Two lines were identified to exhibit resistance to CMV, CGMMV, WMV individually, and a mixed inoculation of the three viruses. The R(1) progeny of the two resistant R(0) lines showed resistance to CMV and WMV, but not to CGMMV. Low level accumulation of transgene transcripts in resistant plants and small interfering (si) RNAs specific to CMV and WMV were readily detected in the resistant R(1) plants by northern blot analysis, indicating that the resistance was established via RNA-mediated post-transcriptional gene silencing (PTGS). Loss of the CGMMV CP-transgene fragment in R1 progeny might be the reason for the failure to resistant CGMMV infection, as shown by the absence of a hybridization signal and no detectable siRNA specific to CGMMV in Southern and northern blot analyses. In summary, this study demonstrated that fusion of different viral CP gene fragments in transgenic watermelon contributed to multiple virus resistance via PTGS. The construct and resistant watermelon lines developed in this study could be used in a watermelon breeding program for resistance to multiple viruses.

  2. Lesson Learned from AGILE and LARES ASI Projects About MATED Data Collection and Post Analysis

    NASA Astrophysics Data System (ADS)

    Carpentiero, Rita; Mrchetti, Ernesto; Natalucci, Silvia; Portelli, Claudio

    2012-07-01

    ASI has managed and collected data on project development of two scientific all-Italian missions: AGILE and LARES. Collection of the Model And Test Effectiveness Database (MATED) data, concerning Project, AIV (Assembly Integration and Verification) and NCR (Non Conformance Report) aspects has been performed by the Italian Space Agency (ASI), using available technical documentation of both AGILE e LARES projects. In this paper some consideration on the needs of 'real time' data collection is made, together with proposal of front end improvement of this tool. In addition a preliminary analysis of MATED effectiveness related to the above ASI projects will be presented in a bottom-up and post verification approach.

  3. Application of the Post-Boiling-Transition Standard to Licensing Analysis

    SciTech Connect

    Mizokami, Shinya; Kitamura, Hideya; Kudo, Yoshiro; Komura, Seiichi; Nagata, Yoshifumi; Morooka, Shinichi

    2005-10-15

    To ensure fuel integrity, light water reactor cores are designed to avoid the onset of boiling transition (BT) inside the fuel assembly that leads to a deterioration of the heat transfer characteristics and subsequent excessive rise of the fuel-cladding temperature in the anticipated operational occurrences (AOOs). However, some boiling water reactors' AOO events result in immediate scram or suppression of the reactor power due to an increase in the reactor coolant void fraction. Recent studies show that a short duration of dryout inside the fuel assembly only leads to a small rise in the fuel-cladding temperature and thus does not pose a threat to fuel integrity. Many tests on BT and an improved comprehension of its mechanism have led to the development of a methodology to appropriately assess the fuel-cladding temperature after BT has been reached. The Standards Committee of the Atomic Energy Society of Japan has therefore proposed a cladding temperature criterion after BT. Applying the post-BT standard enables the value of the operating limit minimum critical power ratio (OLMCPR) to be decreased by allowing for a short duration of dryout. We calculated the fuel-cladding temperature and dryout duration in the load rejection condition without a bypass event. The calculated results show that both the fuel-cladding temperature and dryout duration meet the post-BT standard in the case of a small OLMCPR, which is determined by the loss of feedwater heating. This enables a more efficient reactor core to be designed by applying the post-BT standard to licensing analysis. The possibility of applying a post-BT standard is demonstrated from the results of this work.

  4. Chimeric aptamers in cancer cell-targeted drug delivery

    PubMed Central

    Kanwar, Jagat R; Roy, Kislay; Kanwar, Rupinder K

    2011-01-01

    Aptamers are single-stranded structured oligonucleotides (DNA or RNA) that can bind to a wide range of targets ("apatopes") with high affinity and specificity. These nucleic acid ligands, generated from pools of random-sequence by an in vitro selection process referred to as systematic evolution of ligands by exponential enrichment (SELEX), have now been identified as excellent tools for chemical biology, therapeutic delivery, diagnosis, research, and monitoring therapy in real-time imaging. Today, aptamers represent an interesting class of modern Pharmaceuticals which with their low immunogenic potential mimic extend many of the properties of monoclonal antibodies in diagnostics, research, and therapeutics. More recently, chimeric aptamer approach employing many different possible types of chimerization strategies has generated more stable and efficient chimeric aptamers with aptamer-aptamer, aptamer-nonaptamer biomacromolecules (siRNAs, proteins) and aptamer-nanoparticle chimeras. These chimeric aptamers when conjugated with various biomacromolecules like locked nucleic acid (LNA) to potentiate their stability, biodistribution, and targeting efficiency, have facilitated the accurate targeting in preclinical trials. We developed LNA-aptamer (anti-nucleolin and EpCAM) complexes which were loaded in iron-saturated bovine lactofeerin (Fe-blf)-coated dopamine modified surface of superparamagnetic iron oxide (Fe3O4) nanoparticles (SPIONs). This complex was used to deliver the specific aptamers in tumor cells in a co-culture model of normal and cancer cells. This review focuses on the chimeric aptamers, currently in development that are likely to find future practical applications in concert with other therapeutic molecules and modalities. PMID:21955150

  5. Toward minimal conditioning protocols for allogeneic chimerism in tolerance resistant recipients.

    PubMed

    Al-Adra, David P; Anderson, Colin C

    2013-01-01

    Mixed chimerism is a promising approach toward generating donor-specific immunological tolerance. However, chimerism induction can be toxic; therefore, there is an effort to develop non-myeloablative, minimal intensity protocols that can generate chimerism without the toxic side effects. Recently, with the goal of creating a minimalistic chimerism induction protocol in the tolerance resistant non-obese diabetic (NOD) mouse model, we identified pre-existing T cells as cells that resist fully allogeneic chimerism. With monoclonals targeting NOD T cells, we showed that long-term chimerism and tolerance toward donor islets could be established. However, this promising new protocol relied on the administration of a single dose of anti-CD40 ligand, which is not clinically applicable. In refining protocols to move even closer to clinical utility, we report here initial success at generating fully allogeneic mixed chimerism in NOD mice by adding cyclophosphamide to the conditioning regimen in place of anti-CD40 ligand antibodies.

  6. Three-dimensional finite element analysis of stress distribution in composite resin cores with fiber posts of varying diameters.

    PubMed

    Okamoto, Kazuhiko; Ino, Teruno; Iwase, Naoki; Shimizu, Eitaroh; Suzuki, Megumi; Satoh, Goh; Ohkawa, Shuji; Fujisawa, Masanori

    2008-01-01

    Using three-dimensional finite element analysis (3D-FEA), stress distributions in the remaining radicular tooth structure were investigated under the condition of varying diameters of fiber post for fiber post-reinforced composite resin cores (fiber post and core) in maxillary central incisors. Four 3D-FEA models were constructed: (1) fiber post (ø1.2, ø1.4, and ø1.6 mm) and composite resin core; and (2) gold-cast post and core. Maximum stresses in the tooth structure for fiber post and core were higher than that for gold-cast post and core. In the former models, stresses in the tooth structure as well as in the composite resin were slightly reduced with increase in fiber post diameter. These results thus suggested that to reduce stress in the remaining radicular tooth with a large coronal defect, it is recommended to accompany a composite resin core with a fiber post of a large diameter. PMID:18309611

  7. Ex post power economic analysis of record of decision operational restrictions at Glen Canyon Dam.

    SciTech Connect

    Veselka, T. D.; Poch, L. A.; Palmer, C. S.; Loftin, S.; Osiek, B; Decision and Information Sciences; Western Area Power Administration

    2010-07-31

    On October 9, 1996, Bruce Babbitt, then-Secretary of the U.S. Department of the Interior signed the Record of Decision (ROD) on operating criteria for the Glen Canyon Dam (GCD). Criteria selected were based on the Modified Low Fluctuating Flow (MLFF) Alternative as described in the Operation of Glen Canyon Dam, Colorado River Storage Project, Arizona, Final Environmental Impact Statement (EIS) (Reclamation 1995). These restrictions reduced the operating flexibility of the hydroelectric power plant and therefore its economic value. The EIS provided impact information to support the ROD, including an analysis of operating criteria alternatives on power system economics. This ex post study reevaluates ROD power economic impacts and compares these results to the economic analysis performed prior (ex ante) to the ROD for the MLFF Alternative. On the basis of the methodology used in the ex ante analysis, anticipated annual economic impacts of the ROD were estimated to range from approximately $15.1 million to $44.2 million in terms of 1991 dollars ($1991). This ex post analysis incorporates historical events that took place between 1997 and 2005, including the evolution of power markets in the Western Electricity Coordinating Council as reflected in market prices for capacity and energy. Prompted by ROD operational restrictions, this analysis also incorporates a decision made by the Western Area Power Administration to modify commitments that it made to its customers. Simulated operations of GCD were based on the premise that hourly production patterns would maximize the economic value of the hydropower resource. On the basis of this assumption, it was estimated that economic impacts were on average $26.3 million in $1991, or $39 million in $2009.

  8. Modeling birch pollen emission and transport with the chemistry-transport model CHIMERE

    NASA Astrophysics Data System (ADS)

    Potier, Aurelie; Khvorostyanov, Dmitry; Menut, Laurent; Sofiev, Mikhail; Viovy, Nicolas; Vautard, Robert; Thibaudon, Michel; Tao, Phikune

    2013-04-01

    Among pollen species, birch pollen is recognized to have one of the highest allergenic effects. Its emission as well as its transport with air masses depend on several meteorological parameters. If the conditions are favourable (typically sunny and windy days), the pollen can travel at distances of hundred kilometers in only one day. For analysis and source-oriented forecast, the chemistry-transport models are promising tools to simulate emissions and concentrations over large domains such as Europe. In addition to pollution gaseous and particulate species, the birch pollen related processes were recently added in the chemistry-transport model CHIMERE. This first includes an emission module based on a double-threshold temperature sum concept which describes the onset of the flowering season as well as its propagation using a birch pollen source emission. The parameterization is defined following Sofiev et al. (2012). Second, the processes such as transport, turbulent vertical mixing, dry deposition, wash out and resuspension were updated in CHIMERE to account for the specificities of the pollen grains. In this study, we present a simulation of pollen emissions and transport over Europe with an horizontal resolution of 15km. The CHIMERE model is driven by the WRF meteorological fields and the simulation covers the complete spring of 2008. The modeled pollen concentrations are compared to the R.N.S.A. french national aerobiological survey network measurements. The strength and weaknesses of the modeled results are discussed in terms of emissions data available, meteorology and all specific processes added in the model.

  9. Design and Construction of Chimeric VP8-S2 Antigen for Bovine Rotavirus and Bovine Coronavirus

    PubMed Central

    Nasiri, Khadijeh; Nassiri, Mohammadreza; Tahmoorespur, Mojtaba; Haghparast, Alireza; Zibaee, Saeed

    2016-01-01

    Purpose: Bovine Rotavirus and Bovine Coronavirus are the most important causes of diarrhea in newborn calves and in some other species such as pigs and sheep. Rotavirus VP8 subunit is the major determinant of the viral infectivity and neutralization. Spike glycoprotein of coronavirus is responsible for induction of neutralizing antibody response. Methods: In the present study, several prediction programs were used to predict B and T-cells epitopes, secondary and tertiary structures, antigenicity ability and enzymatic degradation sites. Finally, a chimeric antigen was designed using computational techniques. The chimeric VP8-S2 antigen was constructed. It was cloned and sub-cloned into pGH and pET32a(+) expression vector. The recombinant pET32a(+)-VP8-S2 vector was transferred into E.oli BL21CodonPlus (DE3) as expression host. The recombinant VP8-S2 protein was purified by Ni-NTA chromatography column. Results: The results of colony PCR, enzyme digestion and sequencing showed that the VP8-S2 chimeric antigen has been successfully cloned and sub-cloned into pGH and pET32a(+).The results showed that E.coli was able to express VP8-S2 protein appropriately. This protein was expressed by induction of IPTG at concentration of 1mM and it was confirmed by Ni–NTA column, dot-blotting analysis and SDS-PAGE electrophoresis. Conclusion: The results of this study showed that E.coli can be used as an appropriate host to produce the recombinant VP8-S2 protein. This recombinant protein may be suitable to investigate to produce immunoglobulin, recombinant vaccine and diagnostic kit in future studies after it passes biological activity tests in vivo in animal model and or other suitable procedure. PMID:27123423

  10. Analysis of post-blood meal flight distances in mosquitoes utilizing zoo animal blood meals

    PubMed Central

    Greenberg, Jacob A.; DiMenna, Mark A.; Hanelt, Ben

    2012-01-01

    We assessed the post-blood meal flight distance of four mosquito species in a unique environment using blood meal analysis. Mosquitoes were trapped at the Rio Grande Zoo in Albuquerque, NM, and the blood source of blood-engorged mosquitoes was identified. The distance from the enclosure of the animal serving as a blood source to the trap site was then determined. We found that mosquitoes captured at the zoo flew no more than 170 m with an average distance of 106.7 m after taking a blood meal. This is the first study in which the flight distance of wild mosquitoes has been assessed using blood meal analysis and the first in which zoo animals have served as the exclusive source of blood meals. PMID:22548540

  11. Post-game analysis: An initial experiment for heuristic-based resource management in concurrent systems

    NASA Technical Reports Server (NTRS)

    Yan, Jerry C.

    1987-01-01

    In concurrent systems, a major responsibility of the resource management system is to decide how the application program is to be mapped onto the multi-processor. Instead of using abstract program and machine models, a generate-and-test framework known as 'post-game analysis' that is based on data gathered during program execution is proposed. Each iteration consists of (1) (a simulation of) an execution of the program; (2) analysis of the data gathered; and (3) the proposal of a new mapping that would have a smaller execution time. These heuristics are applied to predict execution time changes in response to small perturbations applied to the current mapping. An initial experiment was carried out using simple strategies on 'pipeline-like' applications. The results obtained from four simple strategies demonstrated that for this kind of application, even simple strategies can produce acceptable speed-up with a small number of iterations.

  12. Dynamic analysis of a pre-and-post ice impacted blade

    NASA Technical Reports Server (NTRS)

    Abumeri, G. H.; Reddy, E. S.; Murthy, P. L. N.; Chamis, C. C.

    1992-01-01

    The dynamic characteristics of an engine blade are evaluated under pre-and-post ice impact conditions using the NASA in-house computer code BLASIM. The ice impacts the leading edge of the blade causing severe local damage. The local structural response of the blade due to the ice impact is predicted via a transient response analysis by modeling only a local patch around the impact region. After ice impact, the global geometry of the blade is updated using deformations of the local patch and a free vibration analysis is performed. The effects of ice impact location, size and ice velocity on the blade mode shapes and natural frequencies are investigated. The results indicate that basic nature of the mode shapes remains unchanged after impact and that the maximum variation in natural frequencies occurs for the twisting mode of the blade.

  13. Dynamic analysis of a pre-and-post ice impacted blade

    NASA Technical Reports Server (NTRS)

    Abumeri, G. H.; Reddy, E. S.; Murthy, P. L. N.; Chamis, C. C.

    1992-01-01

    The dynamic characteristics of an engine blade are evaluated under pre-and-post ice impact conditions using the NASA in-house computer code BLASIM. The ice impacts the leading edge of the blade causing severe local damage. The local structural response of the blade due to the ice impact is predicted via a transient response analysis by modeling only a local patch around the impact region. After ice impact, the global geometry of the blade is updated using deformations of the local patch and a free vibration analysis is performed. The effects of ice impact location, ice size and ice velocity on the blade mode shapes and natural frequencies are investigated. The results indicate that basic nature of the mode shapes remains unchanged after impact and that the maximum variation in natural frequencies occurs for the twisting mode of the blade.

  14. Post-buckling and Large Amplitude Free Vibration Analysis of Composite Beams: Simple Intuitive Formulation

    NASA Astrophysics Data System (ADS)

    Gunda, Jagadish Babu; Venkateswara Rao, Gundabathula

    2016-04-01

    Post-buckling and large amplitude free vibration analysis of composite beams with axially immovable ends is investigated in the present study using a simple intuitive formulation. Geometric nonlinearity of Von-Karman type is considered in the analysis which accounts for mid-plane stretching action of the beam. Intuitive formulation uses only two parameters: the critical bifurcation point and the axial stretching force developed due to membrane stretching action of the beam. Hinged-hinged, clamped-clamped and clamped-hinged boundary conditions are considered. Numerical accuracy of the proposed analytical closed-form solutions obtained from the intuitive formulation are compared to available finite element solutions for symmetric and asymmetric layup schemes of laminated composite beam which indicates the confidence gained on the present formulation.

  15. Correlative scanning-transmission electron microscopy reveals that a chimeric flavivirus is released as individual particles in secretory vesicles.

    PubMed

    Burlaud-Gaillard, Julien; Sellin, Caroline; Georgeault, Sonia; Uzbekov, Rustem; Lebos, Claude; Guillaume, Jean-Marc; Roingeard, Philippe

    2014-01-01

    The intracellular morphogenesis of flaviviruses has been well described, but flavivirus release from the host cell remains poorly documented. We took advantage of the optimized production of an attenuated chimeric yellow fever/dengue virus for vaccine purposes to study this phenomenon by microscopic approaches. Scanning electron microscopy (SEM) showed the release of numerous viral particles at the cell surface through a short-lived process. For transmission electron microscopy (TEM) studies of the intracellular ultrastructure of the small number of cells releasing viral particles at a given time, we developed a new correlative microscopy method: CSEMTEM (for correlative scanning electron microscopy - transmission electron microscopy). CSEMTEM analysis suggested that chimeric flavivirus particles were released as individual particles, in small exocytosis vesicles, via a regulated secretory pathway. Our morphological findings provide new insight into interactions between flaviviruses and cells and demonstrate that CSEMTEM is a useful new method, complementary to SEM observations of biological events by intracellular TEM investigations.

  16. Characterization of oligosaccharide structures on a chimeric respiratory syncytial virus protein expressed in insect cell line Sf9

    SciTech Connect

    Wathen, M.W.; Aeed, P.A.; Elhammer, A.P. )

    1991-03-19

    The oligosaccharide structures added to a chimeric protein (FG) composed of the extracellular domains of respiratory syncytial virus F and G proteins, expressed in the insect cell line Sf9, were investigated. Cells were labeled in vivo with ({sup 3}H)glucosamine and infected wit a recombinant baculovirus containing the FG gene. The secreted chimeric protein was isolated by immunoprecipitation and subjected to oligosaccharide analysis. The FG protein contains two types of O-linked oligosaccharides: GalNAc and Gal{beta}1-3GalNAc constituting 17 and 66% of the total number of structures respectively. Only one type of N-linked oligosaccharide, constituting the remaining 17% of the structures on FG, was detected: a trimannosyl core structure with a fucose residue linked {alpha}1-6 to the asparagine-linked N-acetylglucosamine.

  17. Mixed T Lymphocyte Chimerism after Allogeneic Hematopoietic Transplantation Is Predictive for Relapse of Acute Myeloid Leukemia and Myelodysplastic Syndromes.

    PubMed

    Lee, Hans C; Saliba, Rima M; Rondon, Gabriela; Chen, Julianne; Charafeddine, Yasmeen; Medeiros, L Jeffrey; Alatrash, Gheath; Andersson, Borje S; Popat, Uday; Kebriaei, Partow; Ciurea, Stefan; Oran, Betul; Shpall, Elizabeth; Champlin, Richard

    2015-11-01

    Chimerism testing after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) represents a promising tool for predicting disease relapse, although its precise role in this setting remains unclear. We investigated the predictive value of T lymphocyte chimerism analysis at 90 to 120 days after allo-HSCT in 378 patients with AML/MDS who underwent busulfan/fludarabine-based myeloablative preparative regimens. Of 265 (70%) patients with available T lymphocyte chimerism data, 43% of patients in first or second complete remission (CR1/CR2) at the time of transplantation had complete (100%) donor T lymphocytes at day +90 to +120 compared with 60% of patients in the non-CR1/CR2 cohort (P = .005). In CR1/CR2 patients, donor T lymphocyte chimerism ≤ 85% at day +90 to +120 was associated with a higher frequency of 3-year disease progression (29%; 95% confidence interval [CI], 18% to 46% versus 15%; 95% CI, 9% to 23%; hazard ratio [HR], 2.1; P = .04). However, in the more advanced, non-CR1/CR2 cohort, mixed T lymphocyte chimerism was not associated with relapse (37%; 95% CI, 20% to 66% versus 34%; 95% CI, 25% to 47%; HR, 1.3; P = .60). These findings demonstrate that early T lymphocyte chimerism testing at day +90 to +120 is a useful approach for predicting AML/MDS disease recurrence in patients in CR1/CR2 at the time of transplantation. PMID:26183077

  18. Mixed T Lymphocyte Chimerism after Allogeneic Hematopoietic Transplantation Is Predictive for Relapse of Acute Myeloid Leukemia and Myelodysplastic Syndromes.

    PubMed

    Lee, Hans C; Saliba, Rima M; Rondon, Gabriela; Chen, Julianne; Charafeddine, Yasmeen; Medeiros, L Jeffrey; Alatrash, Gheath; Andersson, Borje S; Popat, Uday; Kebriaei, Partow; Ciurea, Stefan; Oran, Betul; Shpall, Elizabeth; Champlin, Richard

    2015-11-01

    Chimerism testing after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) represents a promising tool for predicting disease relapse, although its precise role in this setting remains unclear. We investigated the predictive value of T lymphocyte chimerism analysis at 90 to 120 days after allo-HSCT in 378 patients with AML/MDS who underwent busulfan/fludarabine-based myeloablative preparative regimens. Of 265 (70%) patients with available T lymphocyte chimerism data, 43% of patients in first or second complete remission (CR1/CR2) at the time of transplantation had complete (100%) donor T lymphocytes at day +90 to +120 compared with 60% of patients in the non-CR1/CR2 cohort (P = .005). In CR1/CR2 patients, donor T lymphocyte chimerism ≤ 85% at day +90 to +120 was associated with a higher frequency of 3-year disease progression (29%; 95% confidence interval [CI], 18% to 46% versus 15%; 95% CI, 9% to 23%; hazard ratio [HR], 2.1; P = .04). However, in the more advanced, non-CR1/CR2 cohort, mixed T lymphocyte chimerism was not associated with relapse (37%; 95% CI, 20% to 66% versus 34%; 95% CI, 25% to 47%; HR, 1.3; P = .60). These findings demonstrate that early T lymphocyte chimerism testing at day +90 to +120 is a useful approach for predicting AML/MDS disease recurrence in patients in CR1/CR2 at the time of transplantation.

  19. Comparing Social Network Analysis of Posts with Counting of Posts as a Measurement of Learners' Participation in Facebook Discussions

    ERIC Educational Resources Information Center

    Lee, Hye Yeon; Lee, Hyeon Woo

    2016-01-01

    With the currently growing interest in social network services, many college courses use social network services as platforms for discussions, and a number of studies have been conducted on the use of social network analysis to measure students' participation in online discussions. This study aims to demonstrate the difference between counting…

  20. Sharing feelings online: studying emotional well-being via automated text analysis of Facebook posts.

    PubMed

    Settanni, Michele; Marengo, Davide

    2015-01-01

    Digital traces of activity on social network sites represent a vast source of ecological data with potential connections with individual behavioral and psychological characteristics. The present study investigates the relationship between user-generated textual content shared on Facebook and emotional well-being. Self-report measures of depression, anxiety, and stress were collected from 201 adult Facebook users from North Italy. Emotion-related textual indicators, including emoticon use, were extracted form users' Facebook posts via automated text analysis. Correlation analyses revealed that individuals with higher levels of depression, anxiety expressed negative emotions on Facebook more frequently. In addition, use of emoticons expressing positive emotions correlated negatively with stress level. When comparing age groups, younger users reported higher frequency of both emotion-related words and emoticon use in their posts. Also, the relationship between online emotional expression and self-report emotional well-being was generally stronger in the younger group. Overall, findings support the feasibility and validity of studying individual emotional well-being by means of examination of Facebook profiles. Implications for online screening purposes and future research directions are discussed.

  1. The Orion Exploration Flight Test Post Flight Solid Particle Flight Environment Inspection and Analysis

    NASA Technical Reports Server (NTRS)

    Miller, Joshua E.

    2016-01-01

    Orbital debris in the millimeter size range can pose a hazard to current and planned spacecraft due to the high relative impact speeds in Earth orbit. Fortunately, orbital debris has a relatively short life at lower altitudes due to atmospheric effects; however, at higher altitudes orbital debris can survive much longer and has resulted in a band of high flux around 700 to 1,500 km above the surface of the Earth. While large orbital debris objects are tracked via ground based observation, little information can be gathered about small particles except by returned surfaces, which until the Orion Exploration Flight Test number one (EFT-1), has only been possible for lower altitudes (400 to 500 km). The EFT-1 crew module backshell, which used a porous, ceramic tile system with surface coatings, has been inspected post-flight for potential micrometeoroid and orbital debris (MMOD) damage. This paper describes the pre- and post-flight activities of inspection, identification and analysis of six candidate MMOD impact craters from the EFT-1 mission.

  2. Forest Fires and Post - Fire Regeneration in Algeria Analysis with Satellite Data

    NASA Astrophysics Data System (ADS)

    Zegrar, Ahmed

    2016-07-01

    The Algerian forests are characterized by a particularly flammable material and fuel. The wind, the relief and the slope facilitates the propagation of fire. The use of remote sensing data multi-­dates, combined with other types of data of various kinds on the environment and forest burned, opens up interesting perspectives for the management of post-­fire regeneration. In this study the use of multi-­temporal remote sensing image Alsat-­1 and Landsat combined with other types of data concerning both background and burned down forest appears to be promising in evaluating and spatial and temporal effects of post fire regeneration. A spatial analysis taking into consideration the characteristics of the burned down site in the North West of Algeria, allowed to better account new factors to explain the regeneration and its temporal and spatial variation. We intended to show the potential use of remote sensing data from satellite ALSAT-­1, of spatial resolution of 32 m. . This approach allows showing the contribution of the data of Algerian satellite ALSAT in the detection and the well attended some forest fires in Algeria.

  3. Post failure localization instabilities in chemically active creeping faults: Steady-state bifurcation and transient analysis

    NASA Astrophysics Data System (ADS)

    Alevizos, S.; Poulet, T.; Veveakis, M.

    2014-12-01

    In this study we emphasize in the post failure evolution of a creeping fault, and provide temporal and spatial modes of evolution. In particular we study the behavior of a fluid-saturated fault under shear, based on the assumption that the fabric presents rate- and temperature dependent response to shear loading. A creeping fault of this type can, under certain conditions, produce excess heat due to shear heating, reaching temperatures which are high enough for triggering endothermic chemical reactions. We focus on the decomposition reactions and incorporate excess pore pressure generation and variations of the porosity due to the chemical effects (a process called chemical pressurization). After deriving the corresponding system of equations in the region of the ultra-cataclastic core, we study the influence of the model parameters, namely the frictional, hydraulic and chemical properties of the material, along with the boundary conditions of the problem, on the behavior of the fault and through a non-linear bifurcation analysis we provide regimes of stable-frictional sliding and pressurization. Furthermore, the system is integrated in time to extract its temporal behavior, providing regimes of stable creep, non-periodic and periodic seismic slip events due to chemical pressurization, depending on their frictional properties. It is shown that this chemically induced seismic slip is an ultra-localized event in the post failure regime. It takes place in an extremely narrow band, 2 orders of magnitude narrower than the initial one, verifying the field observations.

  4. Gravitational waves from compact binaries inspiralling along post-Newtonian accurate eccentric orbits: Data analysis implications

    SciTech Connect

    Tessmer, Manuel; Gopakumar, Achamveedu

    2008-10-15

    Compact binaries inspiralling along eccentric orbits are plausible gravitational-wave (GW) sources for the ground-based laser interferometers. We explore the losses in the event rates incurred when searching for GWs from compact binaries inspiralling along post-Newtonian accurate eccentric orbits with certain obvious nonoptimal search templates. For the present analysis, GW signals having 2.5 post-Newtonian (PN) accurate orbital evolution are modeled following the phasing formalism, presented by T. Damour, A. Gopakumar, and B. R. Iyer [Phys. Rev. D 70, 064028 (2004)]. We demonstrate that the search templates that model in a gauge-invariant manner GWs from compact binaries inspiralling under quadrupolar radiation reaction along 2PN accurate circular orbits are very efficient in capturing our somewhat realistic GW signals. However, three types of search templates based on the adiabatic, complete adiabatic, and gauge-dependent complete nonadiabatic approximants, detailed in P. Ajith, B. R. Iyer, C. A. K. Robinson, and B. S. Sathyaprakash, Phys. Rev. D 71, 044029 (2005), relevant for the circular inspiral under the quadrupolar radiation reaction were found to be inefficient in capturing the above-mentioned eccentric signal. We conclude that further investigations will be required to probe the ability of various types of PN accurate circular templates, employed to analyze the LIGO/VIRGO data, to capture GWs from compact binaries having tiny orbital eccentricities.

  5. Gravitational waves from compact binaries inspiralling along post-Newtonian accurate eccentric orbits: Data analysis implications

    NASA Astrophysics Data System (ADS)

    Tessmer, Manuel; Gopakumar, Achamveedu

    2008-10-01

    Compact binaries inspiralling along eccentric orbits are plausible gravitational-wave (GW) sources for the ground-based laser interferometers. We explore the losses in the event rates incurred when searching for GWs from compact binaries inspiralling along post-Newtonian accurate eccentric orbits with certain obvious nonoptimal search templates. For the present analysis, GW signals having 2.5 post-Newtonian (PN) accurate orbital evolution are modeled following the phasing formalism, presented by T. Damour, A. Gopakumar, and B. R. Iyer [Phys. Rev. D 70, 064028 (2004)PRVDAQ0556-282110.1103/PhysRevD.70.064028]. We demonstrate that the search templates that model in a gauge-invariant manner GWs from compact binaries inspiralling under quadrupolar radiation reaction along 2PN accurate circular orbits are very efficient in capturing our somewhat realistic GW signals. However, three types of search templates based on the adiabatic, complete adiabatic, and gauge-dependent complete nonadiabatic approximants, detailed in P. Ajith, B. R. Iyer, C. A. K. Robinson, and B. S. Sathyaprakash, Phys. Rev. D 71, 044029 (2005)PRVDAQ0556-282110.1103/PhysRevD.71.044029, relevant for the circular inspiral under the quadrupolar radiation reaction were found to be inefficient in capturing the above-mentioned eccentric signal. We conclude that further investigations will be required to probe the ability of various types of PN accurate circular templates, employed to analyze the LIGO/VIRGO data, to capture GWs from compact binaries having tiny orbital eccentricities.

  6. Sharing feelings online: studying emotional well-being via automated text analysis of Facebook posts

    PubMed Central

    Settanni, Michele; Marengo, Davide

    2015-01-01

    Digital traces of activity on social network sites represent a vast source of ecological data with potential connections with individual behavioral and psychological characteristics. The present study investigates the relationship between user-generated textual content shared on Facebook and emotional well-being. Self-report measures of depression, anxiety, and stress were collected from 201 adult Facebook users from North Italy. Emotion-related textual indicators, including emoticon use, were extracted form users’ Facebook posts via automated text analysis. Correlation analyses revealed that individuals with higher levels of depression, anxiety expressed negative emotions on Facebook more frequently. In addition, use of emoticons expressing positive emotions correlated negatively with stress level. When comparing age groups, younger users reported higher frequency of both emotion-related words and emoticon use in their posts. Also, the relationship between online emotional expression and self-report emotional well-being was generally stronger in the younger group. Overall, findings support the feasibility and validity of studying individual emotional well-being by means of examination of Facebook profiles. Implications for online screening purposes and future research directions are discussed. PMID:26257692

  7. Sharing feelings online: studying emotional well-being via automated text analysis of Facebook posts.

    PubMed

    Settanni, Michele; Marengo, Davide

    2015-01-01

    Digital traces of activity on social network sites represent a vast source of ecological data with potential connections with individual behavioral and psychological characteristics. The present study investigates the relationship between user-generated textual content shared on Facebook and emotional well-being. Self-report measures of depression, anxiety, and stress were collected from 201 adult Facebook users from North Italy. Emotion-related textual indicators, including emoticon use, were extracted form users' Facebook posts via automated text analysis. Correlation analyses revealed that individuals with higher levels of depression, anxiety expressed negative emotions on Facebook more frequently. In addition, use of emoticons expressing positive emotions correlated negatively with stress level. When comparing age groups, younger users reported higher frequency of both emotion-related words and emoticon use in their posts. Also, the relationship between online emotional expression and self-report emotional well-being was generally stronger in the younger group. Overall, findings support the feasibility and validity of studying individual emotional well-being by means of examination of Facebook profiles. Implications for online screening purposes and future research directions are discussed. PMID:26257692

  8. Physical rehabilitation in post-conflict settings: analysis of public policy and stakeholder networks.

    PubMed

    Blanchet, Karl; Girois, Susan; Urseau, Isabelle; Smerdon, Christine; Drouet, Yann; Jama, Ali

    2014-01-01

    Physical rehabilitation plays a determinant role in post-conflict contexts to restore disabled citizens' mobility and independence. While the main objectives of any physical rehabilitation programme are to ensure that the services provided are accessible and of good quality to meet existing needs, it is intended that the services need to be supported over the long term by public health and social welfare authorities. This article presents the results of a study conducted in three post-conflict countries on the relationships between the level of commitment of national governments to rehabilitation services and the influence of social networks on national policy related to physical rehabilitation. From a policy and resource standpoint, the environment in Nepal is the most favourable for creating leverage at the national level to influence the commitment of ministries in the rehabilitation sector, compared with Cambodia and Somaliland. Stakeholder network analysis in Nepal, furthermore, reveals a dominant civil society and private sector supporting rehabilitation services, including intense involvement of local organisations and user groups. Implications for Rehabilitation Physical rehabilitation is not on the top of the agenda of governments in fragile states. The commitment and involvement of national authorities in the rehabilitation sector is positively influenced by civil society and international organisations. The denser the social network of the rehabilitation sector is, the more influence the actors can exert influence over national authorities. PMID:23672208

  9. Chimeric Plantibody Passively Protects Mice against Aerosolized Ricin Challenge

    PubMed Central

    Sully, Erin K.; Whaley, Kevin J.; Bohorova, Natasha; Bohorov, Ognian; Goodman, Charles; Kim, Do H.; Pauly, Michael H.; Velasco, Jesus; Hiatt, Ernie; Morton, Josh; Swope, Kelsi; Roy, Chad J.; Zeitlin, Larry

    2014-01-01

    Recent incidents in the United States and abroad have heightened concerns about the use of ricin toxin as a bioterrorism agent. In this study, we produced, using a robust plant-based platform, four chimeric toxin-neutralizing monoclonal antibodies that were then evaluated for the ability to passively protect mice from a lethal-dose ricin challenge. The most effective antibody, c-PB10, was further evaluated in mice as a therapeutic following ricin exposure by injection and inhalation. PMID:24574537

  10. Chimeric plantibody passively protects mice against aerosolized ricin challenge.

    PubMed

    Sully, Erin K; Whaley, Kevin J; Bohorova, Natasha; Bohorov, Ognian; Goodman, Charles; Kim, Do H; Pauly, Michael H; Velasco, Jesus; Hiatt, Ernie; Morton, Josh; Swope, Kelsi; Roy, Chad J; Zeitlin, Larry; Mantis, Nicholas J

    2014-05-01

    Recent incidents in the United States and abroad have heightened concerns about the use of ricin toxin as a bioterrorism agent. In this study, we produced, using a robust plant-based platform, four chimeric toxin-neutralizing monoclonal antibodies that were then evaluated for the ability to passively protect mice from a lethal-dose ricin challenge. The most effective antibody, c-PB10, was further evaluated in mice as a therapeutic following ricin exposure by injection and inhalation. PMID:24574537

  11. Immunogenicity of candidate chimeric DNA vaccine against tuberculosis and leishmaniasis.

    PubMed

    Dey, Ayan; Kumar, Umesh; Sharma, Pawan; Singh, Sarman

    2009-08-13

    Mycobacterium tuberculosis and Leishmania donovani are important intracellular pathogens, especially in Indian context. In India and other South East Asian countries, both these infections are highly endemic and in about 20% cases co-infection of these pathogens is reported. For both these pathogens cell mediated immunity plays most important role. The available treatment of these infections is either prolonged or cumbersome or it is ineffective in controlling the outbreaks and spread. Therefore, potentiation of a common host defense mechanism can be used to prevent both the infections simultaneously. In this study we have developed a novel chimeric DNA vaccine candidate comprising the esat-6 gene of M. tuberculosis and kinesin motor domain gene of L. donovani. After developing this novel chimera, its immunogenicity was studied in mouse model. The immune response was compared with individual constructs of esat-6 and kinesin motor domain. The results showed that immunization with chimeric DNA vaccine construct resulted in stronger IFN-gamma and IL-2 response against kinesin (3012+/-102 and 367.5+/-8.92pg/ml) and ESAT-6 (1334+/-46.5 and 245.1+/-7.72pg/ml) in comparison to the individual vaccine constructs. The reciprocal immune response (IFN-gamma and IL-2) against individual construct was lower (kinesin motor domain: 1788+/-36.48 and 341.8+/-9.801pg/ml and ESAT-6: 867.0+/-47.23 and 170.8+/-4.578pg/ml, respectively). The results also suggest that using the chimeric construct both proteins yielded a reciprocal adjuvant affect over each other as the IFN-gamma production against chimera vaccination is statistically significant (p<0.0001) than individual construct vaccination. From this pilot study we could envisage that the chimeric DNA vaccine construct may offer an attractive strategy in controlling co-infection of leishmaniasis and tuberculosis and have important implication in future vaccine design. PMID:19559111

  12. Chimeric Protein Complexes in Hybrid Species Generate Novel Phenotypes

    PubMed Central

    Piatkowska, Elzbieta M.; Naseeb, Samina; Knight, David; Delneri, Daniela

    2013-01-01

    Hybridization between species is an important mechanism for the origin of novel lineages and adaptation to new environments. Increased allelic variation and modification of the transcriptional network are the two recognized forces currently deemed to be responsible for the phenotypic properties seen in hybrids. However, since the majority of the biological functions in a cell are carried out by protein complexes, inter-specific protein assemblies therefore represent another important source of natural variation upon which evolutionary forces can act. Here we studied the composition of six protein complexes in two different Saccharomyces “sensu stricto” hybrids, to understand whether chimeric interactions can be freely formed in the cell in spite of species-specific co-evolutionary forces, and whether the different types of complexes cause a change in hybrid fitness. The protein assemblies were isolated from the hybrids via affinity chromatography and identified via mass spectrometry. We found evidence of spontaneous chimericity for four of the six protein assemblies tested and we showed that different types of complexes can cause a variety of phenotypes in selected environments. In the case of TRP2/TRP3 complex, the effect of such chimeric formation resulted in the fitness advantage of the hybrid in an environment lacking tryptophan, while only one type of parental combination of the MBF complex allowed the hybrid to grow under respiratory conditions. These phenotypes were dependent on both genetic and environmental backgrounds. This study provides empirical evidence that chimeric protein complexes can freely assemble in cells and reveals a new mechanism to generate phenotypic novelty and plasticity in hybrids to complement the genomic innovation resulting from gene duplication. The ability to exchange orthologous members has also important implications for the adaptation and subsequent genome evolution of the hybrids in terms of pattern of gene loss. PMID

  13. Immunogenicity of candidate chimeric DNA vaccine against tuberculosis and leishmaniasis.

    PubMed

    Dey, Ayan; Kumar, Umesh; Sharma, Pawan; Singh, Sarman

    2009-08-13

    Mycobacterium tuberculosis and Leishmania donovani are important intracellular pathogens, especially in Indian context. In India and other South East Asian countries, both these infections are highly endemic and in about 20% cases co-infection of these pathogens is reported. For both these pathogens cell mediated immunity plays most important role. The available treatment of these infections is either prolonged or cumbersome or it is ineffective in controlling the outbreaks and spread. Therefore, potentiation of a common host defense mechanism can be used to prevent both the infections simultaneously. In this study we have developed a novel chimeric DNA vaccine candidate comprising the esat-6 gene of M. tuberculosis and kinesin motor domain gene of L. donovani. After developing this novel chimera, its immunogenicity was studied in mouse model. The immune response was compared with individual constructs of esat-6 and kinesin motor domain. The results showed that immunization with chimeric DNA vaccine construct resulted in stronger IFN-gamma and IL-2 response against kinesin (3012+/-102 and 367.5+/-8.92pg/ml) and ESAT-6 (1334+/-46.5 and 245.1+/-7.72pg/ml) in comparison to the individual vaccine constructs. The reciprocal immune response (IFN-gamma and IL-2) against individual construct was lower (kinesin motor domain: 1788+/-36.48 and 341.8+/-9.801pg/ml and ESAT-6: 867.0+/-47.23 and 170.8+/-4.578pg/ml, respectively). The results also suggest that using the chimeric construct both proteins yielded a reciprocal adjuvant affect over each other as the IFN-gamma production against chimera vaccination is statistically significant (p<0.0001) than individual construct vaccination. From this pilot study we could envisage that the chimeric DNA vaccine construct may offer an attractive strategy in controlling co-infection of leishmaniasis and tuberculosis and have important implication in future vaccine design.

  14. Post-16 Physics and Chemistry Uptake: Combining Large-Scale Secondary Analysis with In-Depth Qualitative Methods

    ERIC Educational Resources Information Center

    Hampden-Thompson, Gillian; Lubben, Fred; Bennett, Judith

    2011-01-01

    Quantitative secondary analysis of large-scale data can be combined with in-depth qualitative methods. In this paper, we discuss the role of this combined methods approach in examining the uptake of physics and chemistry in post compulsory schooling for students in England. The secondary data analysis of the National Pupil Database (NPD) served…

  15. Human natural chimerism: an acquired character or a vestige of evolution?

    PubMed

    Rinkevich, B

    2001-06-01

    Analysis on five common classes of human natural chimeras (cytomictical, whole body, fetal-maternal, germ cell, and tumor chimeras) reveals that (1) they initiate only during pregnancy, (2) the most common class are chimeras which contain maternal cells, and (3) the primary mechanisms that are involved in their formation and establishment are still elusive. These classes of natural chimerism, are involved only with maladaptive phenomena such as malignancy and autoimmune diseases and without any documented benefit. A recent review has challenged the accepted dogma that the evolution of immunity is pathogen-directed and asserted that preserving individuality from littering the soma and the germline by conspecific alien cells might have been the original function of the innate immunity. Following this tenet, I propose here that human natural chimerism is a by-product of the new role evolved from primitive components of immunity to "educate" the developing embryo with the armamentarium of effector mechanisms, dedicated to purge the individual from pervasive somatic and germline variants, and is not a vestige of evolution.

  16. Chimeric piggyBac transposases for genomic targeting in human cells.

    PubMed

    Owens, Jesse B; Urschitz, Johann; Stoytchev, Ilko; Dang, Nong C; Stoytcheva, Zoia; Belcaid, Mahdi; Maragathavally, Kommineni J; Coates, Craig J; Segal, David J; Moisyadi, Stefan

    2012-08-01

    Integrating vectors such as viruses and transposons insert transgenes semi-randomly and can potentially disrupt or deregulate genes. For these techniques to be of therapeutic value, a method for controlling the precise location of insertion is required. The piggyBac (PB) transposase is an efficient gene transfer vector active in a variety of cell types and proven to be amenable to modification. Here we present the design and validation of chimeric PB proteins fused to the Gal4 DNA binding domain with the ability to target transgenes to pre-determined sites. Upstream activating sequence (UAS) Gal4 recognition sites harbored on recipient plasmids were preferentially targeted by the chimeric Gal4-PB transposase in human cells. To analyze the ability of these PB fusion proteins to target chromosomal locations, UAS sites were randomly integrated throughout the genome using the Sleeping Beauty transposon. Both N- and C-terminal Gal4-PB fusion proteins but not native PB were capable of targeting transposition nearby these introduced sites. A genome-wide integration analysis revealed the ability of our fusion constructs to bias 24% of integrations near endogenous Gal4 recognition sequences. This work provides a powerful approach to enhance the properties of the PB system for applications such as genetic engineering and gene therapy. PMID:22492708

  17. Design and production of a chimeric resilin-, elastin-, and collagen-like engineered polypeptide.

    PubMed

    Bracalello, Angelo; Santopietro, Valentina; Vassalli, Massimo; Marletta, Giovanni; Del Gaudio, Rosanna; Bochicchio, Brigida; Pepe, Antonietta

    2011-08-01

    Protein-inspired biomaterials have gained great interest as an alternative to synthetic polymers, in particular, for their potential use as biomedical devices. The potential inspiring models are mainly proteins able to confer mechanical properties to tissues and organs, such as elasticity (elastin, resilin, spider silk) and strength (collagen, silk). The proper combination of repetitive sequences, each of them derived from different proteins, represents a useful tool for obtaining biomaterials with tailored mechanical properties and biological functions. In this report we describe the design, the production, and the preliminary characterization of a chimeric polypeptide, based on sequences derived from the highly resilient proteins resilin and elastin and from collagen-like sequences. The results show that the obtained chimeric recombinant material exhibits promising self-assembling properties. Young's modulus of the fibers was determined by AFM image analysis and lies in the range of 0.1-3 MPa in agreement with the expectations for elastin-like and resilin-like materials. PMID:21707089

  18. Engineered fumarate sensing Escherichia coli based on novel chimeric two-component system.

    PubMed

    Ganesh, Irisappan; Ravikumar, Sambandam; Lee, Seung Hwan; Park, Si Jae; Hong, Soon Ho

    2013-12-01

    DcuS/DcuR two component system (TCS) was firstly employed for the expression of the gfp gene under the dcuB gene promoter in aerobic condition to develop high throughput screening system able to screen microorganisms producing high amount of fumarate. However, the DcuS/DcuR TCS could not produce a signal strong enough to mediate the expression of the gfp gene responding fumarate concentration. Thus, DcuS/DucR TCS was engineered by recruiting the EnvZ/OmpR system, the most-studied TCS in E. coli. A chimeric DcuS/EnvZ (DcuSZ) TCS was constructed by fusing the sensor histidine kinase of DcuS with the cytoplasmic catalytic domain of EnvZ, in which the expression of the gfp gene or the ompC gene was mediated by the ompC gene promoter through the cognate response regulator, OmpR. The output signals produced by the chimeric DcuSZ TCS were enough to detect fumarate concentration quantatively, in which the expressions of the gfp gene and the ompC gene were proportional to the fumarate concentration in the medium. Moreover, principal component analysis of C4-dicarboxylates showed that DcuSZ chimera was highly specific to fumarate but could also respond to other C4-dicarboxylates, which strongly suggests that TCS-based high throughput screening system able to screen microorganisms producing target chemicals can be developed.

  19. Chimeric piggyBac transposases for genomic targeting in human cells

    PubMed Central

    Owens, Jesse B.; Urschitz, Johann; Stoytchev, Ilko; Dang, Nong C.; Stoytcheva, Zoia; Belcaid, Mahdi; Maragathavally, Kommineni J.; Coates, Craig J.; Segal, David J.; Moisyadi, Stefan

    2012-01-01

    Integrating vectors such as viruses and transposons insert transgenes semi-randomly and can potentially disrupt or deregulate genes. For these techniques to be of therapeutic value, a method for controlling the precise location of insertion is required. The piggyBac (PB) transposase is an efficient gene transfer vector active in a variety of cell types and proven to be amenable to modification. Here we present the design and validation of chimeric PB proteins fused to the Gal4 DNA binding domain with the ability to target transgenes to pre-determined sites. Upstream activating sequence (UAS) Gal4 recognition sites harbored on recipient plasmids were preferentially targeted by the chimeric Gal4–PB transposase in human cells. To analyze the ability of these PB fusion proteins to target chromosomal locations, UAS sites were randomly integrated throughout the genome using the Sleeping Beauty transposon. Both N- and C-terminal Gal4-PB fusion proteins but not native PB were capable of targeting transposition nearby these introduced sites. A genome-wide integration analysis revealed the ability of our fusion constructs to bias 24% of integrations near endogenous Gal4 recognition sequences. This work provides a powerful approach to enhance the properties of the PB system for applications such as genetic engineering and gene therapy. PMID:22492708

  20. Report of a chimeric origin of transposable elements in a bovine-coding gene.

    PubMed

    Almeida, L M; Amaral, M E J; Silva, I T; Silva, W A; Riggs, P K; Carareto, C M

    2008-02-01

    Despite the wide distribution of transposable elements (TEs) in mammalian genomes, part of their evolutionary significance remains to be discovered. Today there is a substantial amount of evidence showing that TEs are involved in the generation of new exons in different species. In the present study, we searched 22,805 genes and reported the occurrence of TE-cassettes in coding sequences of 542 cow genes using the RepeatMasker program. Despite the significant number (542) of genes with TE insertions in exons only 14 (2.6%) of them were translated into protein, which we characterized as chimeric genes. From these chimeric genes, only the FAST kinase domains 3 (FASTKD3) gene, present on chromosome BTA 20, is a functional gene and showed evidence of the exaptation event. The genome sequence analysis showed that the last exon coding sequence of bovine FASTKD3 is approximately 85% similar to the ART2A retrotransposon sequence. In addition, comparison among FASTKD3 proteins shows that the last exon is very divergent from those of Homo sapiens, Pan troglodytes and Canis familiares. We suggest that the gene structure of bovine FASTKD3 gene could have originated by several ectopic recombinations between TE copies. Additionally, the absence of TE sequences in all other species analyzed suggests that the TE insertion is clade-specific, mainly in the ruminant lineage.

  1. An In silico Chimeric Vaccine Targeting Breast Cancer Containing Inherent Adjuvant

    PubMed Central

    Imani Fooladi, Abbas Ali; Mahmoodzadeh Hosseini, Hamideh; Amani, Jafar

    2015-01-01

    Background: Today, Lack of efficient therapeutic strategy for breast cancer (the most common cause of death in women) is one of the momentous problematic topics for all health care committees. Designing new specific vaccine, based on antigens located on the surface of cancer cells can be useful. Over expression of ROR1, lacked of HER2/neu, and hormone receptors on cell surface in the breast cancer, introduce this protein as an appropriate candidate for designing cancer vaccine. Objectives: We hypothesized the extracellular domain of receptor tyrosine kinase like orphan receptor 1 (ROR-1) along with a super antigen such as staphylococcal enterotoxin B could be a potent vaccine for drug resistant breast cancer. Materials and Methods: Here, we assessed the findings of bioinformatics analysis to identify the antitumor immune properties of this chimeric construct. In addition, the stability, physic-chemical properties and allergic potency of designed fusion protein were investigated by valid bioinformatics software. Results: Our result suggested that chimeric model is capable to be a stimulant of both T-cell and B- cell mediated immune responses with an acceptable accessibility and solubility but without any allergenicity. Conclusions: The ROR-1 with an enterotoxin B could be a potent vaccine for breast cancer. PMID:26413246

  2. Error analysis in post linac to driver linac transport beam line of RAON

    NASA Astrophysics Data System (ADS)

    Kim, Chanmi; Kim, Eun-San

    2016-07-01

    We investigated the effects of magnet errors in the beam transport line connecting the post linac to the driver linac (P2DT) in the Rare Isotope Accelerator in Korea (RAON). The P2DT beam line is bent by 180-degree to send the radioactive Isotope Separation On-line (ISOL) beams accelerated in Linac-3 to Linac-2. This beam line transports beams with multi-charge state 132Sn45,46,47. The P2DT beam line includes 42 quadrupole, 4 dipole and 10 sextupole magnets. We evaluate the effects of errors on the trajectory of the beam by using the TRACK code, which includes the translational and the rotational errors of the quadrupole, dipole and sextupole magnets in the beam line. The purpose of this error analysis is to reduce the rate of beam loss in the P2DT beam line. The distorted beam trajectories can be corrected by using six correctors and seven monitors.

  3. A pilot DTI analysis in patients with recent onset post-traumatic stress disorder

    NASA Astrophysics Data System (ADS)

    Liu, Yang; Li, Liang; Li, Baojuan; Zhang, Xi; Lu, Hongbing

    2016-03-01

    To explore the alteration in white matter between survivors with recent onset post-traumatic stress disorder (PTSD) and without PTSD, who survived from the same coal mine flood disaster, the diffusion tensor imaging (DTI) sequences were analyzed using DTI studio and statistical parametric mapping (SPM) packages in this paper. From DTI sequence, the fractional anisotropy (FA) value describes the degree of anisotropy of a diffusion process, while the apparent diffusion coefficient (ADC) value reflects the magnitude of water diffusion. The DTI analyses between PTSD and non-PTSD indicate lower FA values in the right caudate nucleus, right middle temporal gyrus, right fusiform gyrus, and right superior temporal gyrus, and higher ADC values in the right superior temporal gyrus and right corpus callosum of the subjects with PTSD. These results are partly in line with our previous volume and cortical thickness analyses, indicating the importance of multi-modality analysis for PTSD.

  4. A guide to genome-wide association analysis and post-analytic interrogation.

    PubMed

    Reed, Eric; Nunez, Sara; Kulp, David; Qian, Jing; Reilly, Muredach P; Foulkes, Andrea S

    2015-12-10

    This tutorial is a learning resource that outlines the basic process and provides specific software tools for implementing a complete genome-wide association analysis. Approaches to post-analytic visualization and interrogation of potentially novel findings are also presented. Applications are illustrated using the free and open-source R statistical computing and graphics software environment, Bioconductor software for bioinformatics and the UCSC Genome Browser. Complete genome-wide association data on 1401 individuals across 861,473 typed single nucleotide polymorphisms from the PennCATH study of coronary artery disease are used for illustration. All data and code, as well as additional instructional resources, are publicly available through the Open Resources in Statistical Genomics project: http://www.stat-gen.org.

  5. Construction of chimeric vaccinia viruses by molecular cloning and packaging.

    PubMed Central

    Scheiflinger, F; Dorner, F; Falkner, F G

    1992-01-01

    Foreign DNA was inserted into unique restriction endonuclease cleavage sites (Sma I or Not I) of the 200,000-base-pair vaccinia virus genome by direct molecular cloning. The modified vaccinia virus DNA was packaged in fowlpox virus-infected avian cells, and chimeric vaccinia virus was isolated from mammalian cells not supporting the growth of the fowlpox helper virus. In contrast to the classical "in vivo" recombination technique, chimeric viruses with inserts in both possible orientations and families of chimeras with multiple inserts were obtained. The different genomic configurations of chimeric viruses provide a broader basis for screening of optimal viruses. In addition to packaging in avian cells, a second packaging procedure for vaccinia DNA, based on the abortive infection of mammalian cells with the fowlpox helper virus, was developed. This procedure permits simultaneous packaging and host-range selection for the packaged virus. The cloning/packaging procedure allows the direct insertion of foreign DNA without the need for plasmids having flanking regions homologous to viral nonessential regions and is independent of inefficient in vivo recombination events. By direct cloning and packaging, about 5-10% of the total vaccinia virus yield consisted of chimeras. The procedure is, therefore, a useful tool in molecular virology. Images PMID:1438247

  6. Chimeric creatures in Greek mythology and reflections in science.

    PubMed

    Bazopoulou-Kyrkanidou, E

    2001-04-15

    "The Chimaera" in Homer's Iliad, "was of divine stock, not of men, in the forepart a lion, in the hinder a serpent, and in the midst a goat, ellipsis Bellerophon slew her, trusting in the signs of the gods." In Hesiod's Theogony it is emphasized that "Chimaera ellipsis had three heads, one of a grim-eyed lion, another of a goat, and another of a snakeellipsis". In addition to this interspecies animal chimera, human/animal chimeras are referred to in Greek mythology, preeminent among them the Centaurs and the Minotaur. The Centaurs, as horse/men, first appear in Geometric and early Archaic art, but in the literature not until early in the fifth century B.C. The bullheaded-man Minotaur, who is not certainly attested in the literary evidence until circa 500 B.C., first appears in art about 650 B.C. Attempts, in the fourth century B.C. and thereafter, to rationalize their mythical appearance were in vain; their chimeric nature retained its fascinating and archetypal form over the centuries. Early in the 1980s, experimental sheep/goat chimeras were produced removing the reproductive barrier between these two animal species. Late in the 1990s, legal, political, ethical, and moral fights loomed over a patent bid on human/animal chimeras. Chimeric technology is recently developed; however, the concept of chimerism has existed in literary and artistic form in ancient mythology. This is yet another example where art and literature precede scientific research and development.

  7. Systemic chimerism in human female recipients of male livers.

    PubMed

    Starzl, T E; Demetris, A J; Trucco, M; Ramos, H; Zeevi, A; Rudert, W A; Kocova, M; Ricordi, C; Ildstad, S; Murase, N

    1992-10-10

    We have previously reported data from clinical and laboratory animal observations which suggest that organ tolerance after transplantation depends on a state of balanced lymphodendritic cell chimerism between the host and donor graft. We have sought further evidence to support this hypothesis by investigating HLA-mismatched liver allograft recipients. 9 of 9 female recipients of livers from male donors had chimerism in their allografts and extrahepatic tissues, according to in-situ hybridisation and molecular techniques 10 to 19 years posttransplantation. In 8 women with good graft function, evidence of the Y chromosome was found in the blood (6/8), skin (8/8), and lymph nodes (7/8). A ninth patient whose transplant failed after 12 years from recurrent chronic viral hepatitis had chimerism in her lymph nodes, skin, jejunum, and aorta at the time of retransplantation. Although cell migration is thought to take place after all types of transplantation, the large population of migratory cells in, and the extent of their seeding from, hepatic grafts may explain the privileged tolerogenicity of the liver compared with other organs.

  8. Enhancement by dimethyl myleran of donor type chimerism in murine recipients of bone marrow allografts

    SciTech Connect

    Lapidot, T.; Terenzi, A.; Singer, T.S.; Salomon, O.; Reisner, Y. )

    1989-05-15

    A major problem in using murine models for studies of bone marrow allograft rejection in leukemia patients is the narrow margin in which graft rejection can be analyzed. In mice irradiated with greater than 9 Gy total body irradiation (TBI) rejection is minimal, whereas after administration of 8 Gy TBI, which spares a significant number of clonable T cells, a substantial frequency of host stem cells can also be detected. In current murine models, unlike in humans, bone marrow allograft rejection is generally associated with full autologous hematopoietic reconstitution. In the present study, we investigated the effect of the myeloablative drug dimethyl myleran (DMM) on chimerism status following transplantation of T cell-depleted allogenic bone marrow (using C57BL/6 donors and C3H/HeJ recipients, conditioned with 8 Gy TBI). Donor type chimerism 1 to 2 months post-transplant of 1 to 3 x 10(6) bone marrow cells was markedly enhanced by using DMM one day after TBI and prior to transplantation. Conditioning with cyclophosphamide instead of DMM, in combination with 8 Gy TBI, did not enhance engraftment of donor type cells. Artificial reconstitution of T cells, after conditioning with TBI plus DMM, by adding mature thymocytes, or presensitization with irradiated donor type spleen cells 1 week before TBI and DMM, led to strong graft rejection and consequently to severe anemia. The anti-donor responses in these models were proportional to the number of added T cells and to the number of cells used for presensitization, and they could be neutralized by increasing the bone marrow inoculum.

  9. Rotavirus VP7 epitope chimeric proteins elicit cross-immunoreactivity in guinea pigs.

    PubMed

    Zhao, Bingxin; Pan, Xiaoxia; Teng, Yumei; Xia, Wenyue; Wang, Jing; Wen, Yuling; Chen, Yuanding

    2015-10-01

    VP7 of group A rotavirus (RVA) contains major neutralizing epitopes. Using the antigenic protein VP6 as the vector, chimeric proteins carrying foreign epitopes have been shown to possess good immunoreactivity and immunogenicity. In the present study, using modified VP6 as the vector, three chimeric proteins carrying epitopes derived from VP7 of RVA were constructed. The results showed that the chimeric proteins reacted with anti-VP6 and with SA11 and Wa virus strains. Antibodies from guinea pigs inoculated with the chimeric proteins recognized VP6 and VP7 of RVA and protected mammalian cells from SA11 and Wa infection in vitro. The neutralizing activities of the antibodies against the chimeric proteins were significantly higher than those against the vector protein VP6F. Thus, development of chimeric vaccines carrying VP7 epitopes using VP6 as a vector could be a promising alternative to enhance immunization against RVAs.

  10. Effect of IL-2-Bax, a novel interleukin-2-receptor-targeted chimeric protein, on bleomycin lung injury1

    PubMed Central

    Segel, Michael J; Aqeilan, Rami; Zilka, Keren; Lorberboum-Galski, Haya; Wallach-Dayan, Shulamit B; Conner, Michael W; Christensen, Thomas G; Breuer, Raphael

    2005-01-01

    The role of lymphocytes in the pathogenesis of lung fibrosis is not clear, but the weight of the evidence supports a pro-fibrotic effect for lymphocytes. The high-affinity interleukin-2 receptor (haIL-2R) is expressed on activated, but not quiescent, T lymphocytes. This selective expression of haIL-2R provides the basis for therapeutic strategies that target IL-2R-expressing cells. We hypothesized that elimination of activated lymphocytes by IL-2R-targeted chimeric proteins might ameliorate lung fibrosis. We investigated the effects of IL-2-Bax, a novel apoptosis-inducing IL-2R-targeted chimeric protein, on bleomycin-induced lung injury in mice. Treatment groups included (i) a single intratracheal instillation of bleomycin and twice-daily intraperitoneal injections of IL-2-Bax; (ii) intratracheal bleomycin and intraperitoneal IL-2-PE664Glu, an older-generation chimeric protein; (iii) intratracheal bleomycin/intraperitoneal PBS; (iv) intratracheal saline/intraperitoneal PBS. Lung injury was evaluated 14 days after intratracheal instillation by cell count in bronchoalveolar lavage (BAL) fluid, semi-quantitative and quantitative histomorphological measurements and by biochemical analysis of lung hydroxyproline. Bleomycin induced a BAL lymphocytosis that was significantly attenuated by IL-2-Bax and IL-2-PE664Glu. However, morphometric parameters and lung hydroxyproline were unaffected by the chimeric proteins. These results show that IL-2-Bax reduces the lymphocytic infiltration of the lungs in response to bleomycin, but this effect is not accompanied by a decrease in lung fibrosis. PMID:16191100

  11. ProteomeScout: a repository and analysis resource for post-translational modifications and proteins

    PubMed Central

    Matlock, Matthew K.; Holehouse, Alex S.; Naegle, Kristen M.

    2015-01-01

    ProteomeScout (https://proteomescout.wustl.edu) is a resource for the study of proteins and their post-translational modifications (PTMs) consisting of a database of PTMs, a repository for experimental data, an analysis suite for PTM experiments, and a tool for visualizing the relationships between complex protein annotations. The PTM database is a compendium of public PTM data, coupled with user-uploaded experimental data. ProteomeScout provides analysis tools for experimental datasets, including summary views and subset selection, which can identify relationships within subsets of data by testing for statistically significant enrichment of protein annotations. Protein annotations are incorporated in the ProteomeScout database from external resources and include terms such as Gene Ontology annotations, domains, secondary structure and non-synonymous polymorphisms. These annotations are available in the database download, in the analysis tools and in the protein viewer. The protein viewer allows for the simultaneous visualization of annotations in an interactive web graphic, which can be exported in Scalable Vector Graphics (SVG) format. Finally, quantitative data measurements associated with public experiments are also easily viewable within protein records, allowing researchers to see how PTMs change across different contexts. ProteomeScout should prove useful for protein researchers and should benefit the proteomics community by providing a stable repository for PTM experiments. PMID:25414335

  12. Cochlear neuropathy in human presbycusis: confocal analysis of hidden hearing loss in post-mortem tissue

    PubMed Central

    Viana, Lucas M.; O’Malley, Jennifer T.; Burgess, Barbara J.; Jones, Dianne D.; Oliveira, Carlos A.C.P.; Santos, Felipe; Merchant, Saumil N.; Liberman, Leslie D.; Liberman, M. Charles

    2015-01-01

    Recent animal work has suggested that cochlear synapses are more vulnerable than hair cells in both noise-induced and age-related hearing loss. This synaptopathy is invisible in conventional histopathological analysis, because cochlear nerve cell bodies in the spiral ganglion survive for years, and synaptic analysis requires special immunostaining or serial-section electron microscopy. Here, we show that the same quadruple-immunostaining protocols that allow synaptic counts, hair cell counts, neuronal counts and differentiation of afferent and efferent fibers in mouse can be applied to human temporal bones, when harvested within 9 hrs post-mortem and prepared as dissected whole mounts of the sensory epithelium and osseous spiral lamina. Quantitative analysis of five “normal” ears, aged 54 to 89 yrs, without any history of otologic disease, suggests that cochlear synaptopathy and the degeneration of cochlear nerve peripheral axons, despite a near-normal hair cell population, may be an important component of human presbycusis. Although primary cochlear nerve degeneration is not expected to affect audiometric thresholds, it may be key to problems with hearing in noise that are characteristic of declining hearing abilities in the aging ear. PMID:26002688

  13. Construction and use of chimeric SPR/phi 3T DNA methyltransferases in the definition of sequence recognizing enzyme regions.

    PubMed Central

    Balganesh, T S; Reiners, L; Lauster, R; Noyer-Weidner, M; Wilke, K; Trautner, T A

    1987-01-01

    Multispecific DNA methyltransferases (Mtases) of temperate Bacillus subtilis phages SPR and phi 3T methylate the internal cytosine of the sequence GGCC. They differ in their capacity to methylate additional sequences. These are CCGG and CC(A/T)GG in SPR and GCNGC in phi 3T. Introducing unique restriction sites at equivalent locations within the two genes facilitated the construction of chimeric genes. These expressed Mtase activity at a level comparable to that of the parental genes. The methylation specificity of chimeric enzymes was correlated with the location of chimeric fusions. This analysis, which also included the use of mutant genes, showed that domains involved in the recognition of target sequences unique to each enzyme [CCGG, CC(A/T)GG or GCNGC] are represented by the central non-conserved parts of the proteins, whilst recognition of the sequence (GGCC), which is a target for both enzymes, is determined by an adjacent conserved region. Images Fig. 4. Fig. 5. PMID:2828032

  14. Non-myeloablative conditioning is sufficient to induce mixed chimerism and subsequent acceptance of donor specific cardiac and skin grafts.

    PubMed

    Liu, Chi; Zhu, Ping; Saito, Taro; Isaka, Yoshitaka; Nagahara, Yukitoshi; Zhuang, Jian; Li, Xiao-Kang

    2013-07-01

    Organ transplant recipients have elevated cancer and viral infection risks due to immunosuppression and long-term results of organ transplantation remain unsatisfactory, mainly because of chronic rejection. The purpose of the current study is to establish a nonmyeloablative perioperative regimen, able to induce mixed chimerism and tolerance of allografts. To establish a nonmyeloablative perioperative regimen, we used Busulfan, an important component of many bone marrow transplantation preparative regimens for a variety of non-neoplastic diseases as an alternative to total body irradiation (TBI), and FTY720, a unique immunosuppression agent, inhibition lymphocyte homing. We found that creating a lymphohematopoietic chimera in which donor and recipient hematopoiesis coexist resulted in prolongation of the donor specific heart and skin allografts. Consistent with graft survival, pathological analysis indicated that the allografts from tolerant recipients were free of myocardial injury and had only a few interstitial infiltrates, and obliterative vasculopathy was not observed. Furthermore, we found that Treg cells were increased in the long-term graft acceptance recipients. Our data revealed that the therapeutic potential for using hematopoietic chimerism in non-myeloablated recipients hope the advances in rodent models described above in the development of minimal, nontoxic host conditioning regimens for mixed chimerism induction and subsequent acceptance of donor specific grafts. PMID:23428909

  15. Chimeric Filoviruses for Identification and Characterization of Monoclonal Antibodies

    PubMed Central

    Ilinykh, Philipp A.; Shen, Xiaoli; Flyak, Andrew I.; Kuzmina, Natalia; Ksiazek, Thomas G.; Crowe, James E.

    2016-01-01

    ABSTRACT Recent experiments suggest that some glycoprotein (GP)-specific monoclonal antibodies (MAbs) can protect experimental animals against the filovirus Ebola virus (EBOV). There is a need for isolation of MAbs capable of neutralizing multiple filoviruses. Antibody neutralization assays for filoviruses frequently use surrogate systems such as the rhabdovirus vesicular stomatitis Indiana virus (VSV), lentiviruses or gammaretroviruses with their envelope proteins replaced with EBOV GP or pseudotyped with EBOV GP. It is optimal for both screening and in-depth characterization of newly identified neutralizing MAbs to generate recombinant filoviruses that express a reporter fluorescent protein in order to more easily monitor and quantify the infection. Our study showed that unlike neutralization-sensitive chimeric VSV, authentic filoviruses are highly resistant to neutralization by MAbs. We used reverse genetics techniques to replace EBOV GP with its counterpart from the heterologous filoviruses Bundibugyo virus (BDBV), Sudan virus, and even Marburg virus and Lloviu virus, which belong to the heterologous genera in the filovirus family. This work resulted in generation of multiple chimeric filoviruses, demonstrating the ability of filoviruses to tolerate swapping of the envelope protein. The sensitivity of chimeric filoviruses to neutralizing MAbs was similar to that of authentic biologically derived filoviruses with the same GP. Moreover, disabling the expression of the secreted GP (sGP) resulted in an increased susceptibility of an engineered virus to the BDBV52 MAb isolated from a BDBV survivor, suggesting a role for sGP in evasion of antibody neutralization in the context of a human filovirus infection. IMPORTANCE The study demonstrated that chimeric rhabdoviruses in which G protein is replaced with filovirus GP, widely used as surrogate targets for characterization of filovirus neutralizing antibodies, do not accurately predict the ability of antibodies to

  16. Lassa-Vesicular Stomatitis Chimeric Virus Safely Destroys Brain Tumors

    PubMed Central

    Wollmann, Guido; Drokhlyansky, Eugene; Davis, John N.; Cepko, Connie

    2015-01-01

    ABSTRACT High-grade tumors in the brain are among the deadliest of cancers. Here, we took a promising oncolytic virus, vesicular stomatitis virus (VSV), and tested the hypothesis that the neurotoxicity associated with the virus could be eliminated without blocking its oncolytic potential in the brain by replacing the neurotropic VSV glycoprotein with the glycoprotein from one of five different viruses, including Ebola virus, Marburg virus, lymphocytic choriomeningitis virus (LCMV), rabies virus, and Lassa virus. Based on in vitro infections of normal and tumor cells, we selected two viruses to test in vivo. Wild-type VSV was lethal when injected directly into the brain. In contrast, a novel chimeric virus (VSV-LASV-GPC) containing genes from both the Lassa virus glycoprotein precursor (GPC) and VSV showed no adverse actions within or outside the brain and targeted and completely destroyed brain cancer, including high-grade glioblastoma and melanoma, even in metastatic cancer models. When mice had two brain tumors, intratumoral VSV-LASV-GPC injection in one tumor (glioma or melanoma) led to complete tumor destruction; importantly, the virus moved contralaterally within the brain to selectively infect the second noninjected tumor. A chimeric virus combining VSV genes with the gene coding for the Ebola virus glycoprotein was safe in the brain and also selectively targeted brain tumors but was substantially less effective in destroying brain tumors and prolonging survival of tumor-bearing mice. A tropism for multiple cancer types combined with an exquisite tumor specificity opens a new door to widespread application of VSV-LASV-GPC as a safe and efficacious oncolytic chimeric virus within the brain. IMPORTANCE Many viruses have been tested for their ability to target and kill cancer cells. Vesicular stomatitis virus (VSV) has shown substantial promise, but a key problem is that if it enters the brain, it can generate adverse neurologic consequences, including death. We

  17. Nitrogen Species in the Post-Pinatubo Stratosphere: Model Analysis Utilizing UARS Measurements. Appendix F

    NASA Technical Reports Server (NTRS)

    Danilin, Michael Y.; Rodriguez, Jose M.; Hu, Wenjie; Ko, Malcolm K. W.; Weisenstein, Debra K.; Kumer, John B.; Mergenthaler, John L.; Russell, James M., III; Koike, Makoto; Yue, Glenn K.

    1999-01-01

    We present an analysis of the impact of heterogeneous chemistry on the partitioning of nitrogen species measured by the Upper Atmosphere Research Satellite (UARS) instruments. The UARS measurements utilized include N2O, HNO3, and ClONO2 from the cryogenic limb array etalon spectrometer (CLAES), version 7 (v.7), and temperature, methane, ozone, H2O, HCl, NO and NO2 from the halogen occultation experiment (HALOE), version 18. The analysis is carried out for the UARS data obtained between January 1992 and September 1994 in the 100- to 1-mbar (approx. 17-47 km) altitude range and over 10 deg latitude bins from 70 deg S to 70 deg N. The spatiotemporal evolution of aerosol surface area density (SAD) is adopted from analysis of the Stratospheric Aerosol and Gas Experiment (SAGE) II data. A diurnal steady state photochemical box model, constrained by the temperature, ozone, H2O, CH4, aerosol SAD, and columns of O2 and O3 above the point of interest, has been used as the main tool to analyze these data. Total inorganic nitrogen (NOY) is obtained by three different methods: (1) as a sum of the UARS-measured NO, NO2, HNO3, and ClONO2; (2) from the N2O-NOY correlation; and (3) from the CH4-NOY correlation. To validate our current understanding of stratospheric heterogeneous chemistry for post-Pinatubo conditions, the model-calculated monthly averaged NO(x)/NO(y) ratios and the NO, NO2, and HNO3 profiles are compared with the UARS-derived data. In general, the UARS-constrained box model captures the main features of nitrogen species partitioning in the post-Pinatubo years, such as recovery of NO(x) after the eruption, their seasonal variability and vertical profiles. However, the model underestimates the NO2 content, particularly in the 30- to 7-mbar (approx. 23-32 km) range. Comparisons of the calculated temporal behavior of the partial columns of NO2 and HNO3 and ground-based measurements at 45 deg S and 45 deg N are also presented. Our analysis indicates that ground

  18. Nitrogen Species in the Post-Pinatubo Stratosphere: Model Analysis Utilizing UARS Measurements

    NASA Technical Reports Server (NTRS)

    Danilin, Michael Y.; Rodriguez, Jose M.; Hu, Wen-Jie; Ko, Malcolm K. W.; Weisenstein, Debra K.; Kumer, John B.; Mergenthaler, John L.; Russel, James M., III; Koike, Makoto; Yue, Glenn K.

    1999-01-01

    We present an analysis of the impact of heterogeneous chemistry on the partitioning of nitrogen species measured by the Upper Atmosphere Research Satellite (UARS) instruments. The UARS measurements utilized include N2O, HNO3, and ClONO2 from the cryogenic limb array etalon spectrometer (CLAES), version 7 (v.7), and temperature, methane, ozone, H2O, HCl, NO and NO2 from the halogen occultation experiment (HALOE), version 18. The analysis is carried out for the UARS data obtained between January 1992 and September 1994 in the 100-to 1-mbar (approx. 17-47 km) altitude range and over 10 degrees latitude bins from 70 S to 70 N. The spatiotemporal evolution of aerosol surface area density (SAD) is adopted from analysis of the Stratospheric Aerosol and Gas Experiment (SAGE) II data. A diurnal steady state photochemical box model, constrained by the temperature, ozone, H2O, CH4, aerosol SAD, and columns of O2 and O3 above the point of interest, has been used as the main tool to analyze these data. Total inorganic nitrogen (NOy) is obtained by three different methods: (1) as a sum of the UARS-measured NO, NO2, HNO3, and ClONO2; (2) from the N2O-NOy correlation, and (3) from the CH4-NOy correlation. To validate our current understanding of stratospheric heterogeneous chemistry for post-Pinatubo conditions, the model-calculated monthly averaged NOx/NOy ratios and the NO, NO2, and HNO3 profiles are compared with the UARS-derived data. In general, the UARS-constrained box model captures the main features of nitrogen species partitioning in the post-Pinatubo years, such as recovery of NOx after the eruption, their seasonal variability and vertical profiles. However, the model underestimates the NO2 content, particularly in the 30- to 7-mbar (approx.23-32 km) range. Comparisons of the calculated temporal behavior of the partial columns of NO2 and HNO3 and ground-based measurements at 45 S and 45 N are also presented. Our analysis indicates that ground-based and HALOE v.18

  19. Post-mastectomy Radiation Therapy for T3N0: A SEER Analysis

    PubMed Central

    Johnson, Matthew E.; Handorf, Elizabeth A.; Martin, Jeffrey M.; Hayes, Shelly B.

    2015-01-01

    Background There is conflicting evidence regarding the benefit of post-mastectomy radiation therapy (PMRT) for pathologic stage T3N0M0 breast cancers. We analyzed data from the Surveillance, Epidemiology, and End Results (SEER) database to investigate the benefit of PMRT in these patients. Methods We queried the SEER database for T3N0M0 breast cancer patients diagnosed from 2000–2010 who underwent modified radical mastectomy. We excluded males, patients with unknown radiation timing/type, other primary tumors, or survival <6 months. 2525 patients were included in this analysis. We performed univariate and multivariate statistical analysis using Chi-squared tests, log rank test, and Cox proportional hazards regression. Primary endpoints were overall survival (OS) and breast cancer-specific survival (CSS). Results Of the 2525 patients identified, 1063 received PMRT. The median follow-up was 56 months (range: 6–131). On univariate analysis, PMRT improved OS (76.5% vs. 61.8%, p<0.01) and CSS (85.0% vs. 82.4%, p<0.01) at 8 years. The use of PMRT remained significant on multivariate analysis: PMRT improved OS (HR 0.63, p<0.001) and CSS (HR 0.77, p=0.045). Low tumor grade (p<0.01) and marital status "married" (p=0.01) also predicted for improved CSS on multivariate analysis. Conclusion(s) PMRT was associated with significant improvements in both CSS and OS in patients with T3N0M0 breast cancers treated with modified radical mastectomy from 2000 to 2010. PMRT should be strongly considered in T3N0M0 patients. PMID:24985911

  20. Immigration control in the vertebrate body with special reference to chimerism.

    PubMed

    Davies, Anthony J S

    2012-01-01

    The phenomenon of chimerism is reviewed against an understanding of adaptive immunity in vertebrates. It is shown that chimerism can be regarded as a ubiquitous condition and this suggests that monophylesis has played little part in evolution. It is suggested that the adaptive immune response has a special role in facilitating the development of chimerism and that the consensus view of adaptive immunity as a rejection mechanism should be revised.

  1. Regional analysis of distribution of pre and post 2015 Nepal Earthquake landslides

    NASA Astrophysics Data System (ADS)

    Valagussa, Andrea; Frattini, Paolo; Crosta, Giovanni; Valbuzzi, Elena

    2016-04-01

    A magnitude 7.8 earthquake struck Nepal on April 25, 2015. Three landslide inventories have been prepared in four districts: Dhading (1885 km2), Sindhupalchok (2488 km2), Rasuwa (1522 km2) and Nuwakot (1194 km2), that are located north of Kathmandu. These inventories extend 14 to 138 km SE from the epicenter of the main shock (April 25, 2015), 4.5 to 143 km NW from the epicenter of the main aftershock (May 12, 2015), and 34 to 136 km from the Main Frontal Thrust. The first inventory is a coseismic and post-seismic landslide inventory based on multi-temporal images (Google Earth, Google Crisis maps, Bing maps), and helicopter-based video. The second one is a pre-event shallow landslide inventory. In these two inventories the most abundant landslide types are: debris flows, shallow translational slides, and rockfalls. The third is a deep seated landslide inventory, in which the most represented landslide types are rock avalanches, slumps, rockslides and deep-seated gravitational slope deformations (DSGSD). All the landslides have been mapped as individual polygons. For the analysis we focus our attention on four districts: First we studied how the landslide frequency density changes as a function of topographic parameters (i.e. slope gradient, slope aspect, and elevation). The analyses have been based on the ASTER Global Digital Elevation Model (ASTER GDEM). For coseismic and post-seismic landslides we observed that the mean slope gradient at which the landslide occurs is higher with respect to the two other inventories (50° and 30/40° respectively). The slope aspect of coseismic and post-seismic landslides is also different, with a larger frequency of landslides towards SW, whereas in pre-event landslides the most common slope aspect is SE. This could be related to the direction of the seismic wave. At least the coseismic and post-seismic landslides occur, in mean, at an elevation lower than the pre-event landslides. We also analyzed the relationship between the

  2. Influence of perceptual and semantic conflicts between the two halves of chimeric stimuli on the expression of visuo-spatial neglect.

    PubMed

    Peru, A; Moro, V; Avesani, R; Aglioti, S

    1997-05-01

    We investigated how perceptual and semantic relationships between the left and right half of chimeric stimuli influence overt and covert visual processing by asking eight right brain damaged (RBD) patients with hemispatial neglect to identify complete, half-, and chimeric drawings. Chimeric stimuli belonged in one of four categories defined according to the perceptual and semantic relatedness between the two compounding hemi-figures. Thus, the hemi-figures could be related both perceptually and semantically, only perceptually, only semantically, or neither perceptually nor semantically. Although patients often appeared to base their report on the right part of the chimerics, the number of errors was minimal when conflicts between the two hemi-figures were maximal. Moreover, perceptual conflicts, which mainly affect the perception of the shape, appeared to influence the performance more than semantic conflicts. Since the analysis of shape incongruency is probably accomplished at early levels of information processing, the result suggests that preattentive analysis is largely spared in the experimental patients and that, in our task, bottom-up factors more than top-down factors modulate the expression of left neglect. PMID:9153020

  3. Chimeric Proteins to Detect DNA Damage and Mismatches

    SciTech Connect

    McCutchen-Maloney, S; Malfatti, M; Robbins, K M

    2002-01-14

    The goal of this project was to develop chimeric proteins composed of a DNA mismatch or damage binding protein and a nuclease, as well as methods to detect DNA mismatches and damage. We accomplished this through protein engineering based on using polymerase chain reactions (PCRs) to create chimeras with novel functions for damage and mismatch detection. This project addressed fundamental questions relating to disease susceptibility and radiation-induced damage in cells. It also supported and enhanced LLNL's competency in the emerging field of proteomics. In nature, DNA is constantly being subjected to damaging agents such as exposure to ultraviolet (UV) radiation and various environmental and dietary carcinogens. If DNA damage is not repaired however, mutations in DNA result that can eventually manifest in cancer and other diseases. In addition to damage-induced DNA mutations, single nucleotide polymorphisms (SNPs), which are variations in the genetic sequence between individuals, may predispose some to disease. As a result of the Human Genome Project, the integrity of a person's DNA can now be monitored. Therefore, methods to detect DNA damage, mutations, and SNPs are useful not only in basic research but also in the health and biotechnology industries. Current methods of detection often use radioactive labeling and rely on expensive instrumentation that is not readily available in many research settings. Our methods to detect DNA damage and mismatches employ simple gel electrophoresis and flow cytometry, thereby alleviating the need for radioactive labeling and expensive equipment. In FY2001, we explored SNP detection by developing methods based on the ability of the chimeric proteins to detect mismatches. Using multiplex assays with flow cytometry and fluorescent beads to which the DNA substrates where attached, we showed that several of the chimeras possess greater affinity for damaged and mismatched DNA than for native DNA. This affinity was demonstrated in

  4. Post-analysis report on Chesapeake Bay data processing. [spectral analysis and recognition computer signature extension

    NASA Technical Reports Server (NTRS)

    Thomson, F.

    1972-01-01

    The additional processing performed on data collected over the Rhode River Test Site and Forestry Site in November 1970 is reported. The techniques and procedures used to obtain the processed results are described. Thermal data collected over three approximately parallel lines of the site were contoured, and the results color coded, for the purpose of delineating important scene constituents and to identify trees attacked by pine bark beetles. Contouring work and histogram preparation are reviewed and the important conclusions from the spectral analysis and recognition computer (SPARC) signature extension work are summarized. The SPARC setup and processing records are presented and recommendations are made for future data collection over the site.

  5. Characterization of a chimeric enzyme comprising feruloyl esterase and family 42 carbohydrate-binding module.

    PubMed

    Koseki, Takuya; Mochizuki, Keiji; Kisara, Hiroe; Miyanaga, Akimasa; Fushinobu, Shinya; Murayama, Tetsuya; Shiono, Yoshihito

    2010-03-01

    We engineered a chimeric enzyme (AwFaeA-CBM42) comprising of type-A feruloyl esterase from Aspergillus awamori (AwFaeA) and family 42 carbohydrate-binding module (AkCBM42) from glycoside hydrolase family 54 alpha-L-arabinofuranosidase of Aspergillus kawachii. The chimeric enzyme was successfully produced in Pichia pastoris and accumulated in the culture broth. The purified chimeric enzyme had an apparent relative molecular mass (M(r)) of 53,000. The chimeric enzyme binds to arabinoxylan; this indicates that the AkCBM42 in AwFaeA-CBM42 binds to arabinofuranose side chain moiety of arabinoxylan. The thermostability of the chimeric enzyme was greater than that of AwFaeA. No significant difference of the specific activity toward methyl ferulate was observed between the AwFaeA and chimeric enzyme, but the release of ferulic acid from insoluble arabinoxylan by the chimeric enzyme was approximately 4-fold higher than that achieved by AwFaeA alone. In addition, the chimeric enzyme and xylanase acted synergistically for the degradation of arabinoxylan. In conclusion, the findings of our study demonstrated that the components of the AwFaeA-CBM42 chimeric enzyme act synergistically to bring about the degradation of complex substrates and that the family 42 carbohydrate-binding module has potential for application in the degradation of polysaccharides.

  6. The need for complementary hydraulic analysis in post-restoration monitoring of river restoration projects

    NASA Astrophysics Data System (ADS)

    Endreny, T. A.; Soulman, M. M.

    2011-03-01

    River restoration design methods are incrementally improved by studying and learning from monitoring data in previous projects. In this paper, we report post-restoration monitoring data for a Natural Channel Design (NCD) restoration project along 1600 m (10 channel wavelengths) of the Batavia Kill in the Catskill Mountains, NY, implemented in 2001 and 2002. The NCD project used a reference-reach to determine channel form, empirical relations between the project site and reference site bankfull dimensions to size channel geometry, and hydraulic and sediment computations to test channel capacity and sediment stability. In addition 12 cross-vanes and 48 j-hook vanes used in NCD for river training were installed to protect against bank erosion and maintain scour pools for fish habitat. Changes in pool depths were monitored with surveys from 2002-2004, and then after the channel-altering April 2005 flood. Aggradation in pools was attributed to cross-vane arms not concentrating flow in the center of the channel, which subsequently caused flow splitting and 4 partial point bar avulsions during the 2005 flood. Hydrodynamic simulation at the 18 m3s-1 bankfull flow suggested avulsions occurred where vanes allowed erosive bank scour to initiate the avulsion cut, and once the flow was split, the diminished in-channel flow caused more aggradation in the pools. In this project post-restoration monitoring had detected aggradation and considered it a problem. The lesson for the larger river restoration community is monitoring protocol should include complementary hydraulic and sediment analysis to comprehend potential consequences and develop preventative maintenance. River restoration and monitoring teams should be trained in robust hydraulic and sediment analytical methods that help them extend project restoration goals.

  7. BOT3P: Bologna Transport Analysis Pre-Post-Processors Version 1.0

    SciTech Connect

    Orsi, Roberto

    2002-11-15

    BOT3P is a set of standard FORTRAN 77 language programs that were developed at the ENEA-Bologna Nuclear Data Centre. BOT3P aims to give the users of the DORT and TORT deterministic transport codes some useful diagnostic tools to prepare and to check their input data files. It also includes some plotting programs that can be employed both as preprocessors and as postprocessors of a transport analysis. The BOT3P plotting programs use the RSCORS Graphics System subroutines, included in the DOORS-3.3 and earlier software package together with the Oak Ridge National Laboratory DORT and TORT transport codes.BOT3P was developed on a DIGITAL UNIX ALPHA 500/333 workstation and successfully used in some complex applications such as the VENUS-1 and VENUS-3 neutron shielding benchmarks. BOT3P was tested on an IBM RS/6000 workstation also and is designed to run on most UNIX platforms.The following programs are included in the package: GGDM, DDM, GGTM, DTM2, DTM3, and RVARSCL. GGDM and GGTM generate the geometrical and material entries for DORT and TORT, respectively. DDM is a DORT graphics pre/post processor. DTM2 and DTM3 are graphics pre/post processors showing cuts and three-dimensional views of the TORT model, respectively. RVARSCL reads DORT/TORT 'VARSCL' sequential format files and selects and writes the data required by the user in new files to be visualized by DDM, DTM2, and DTM3 as postprocessor applications.The package is publicly available from the Organization for Economic Cooperation and Development/Nuclear Energy Agency Data Bank (NEA-1627)

  8. Evaluation of a post-processing approach for multiscale analysis of biphasic mechanics of chondrocytes.

    PubMed

    Sibole, Scott C; Maas, Steve; Halloran, Jason P; Weiss, Jeffrey A; Erdemir, Ahmet

    2013-10-01

    Understanding the mechanical behaviour of chondrocytes as a result of cartilage tissue mechanics has significant implications for both evaluation of mechanobiological function and to elaborate on damage mechanisms. A common procedure for prediction of chondrocyte mechanics (and of cell mechanics in general) relies on a computational post-processing approach where tissue-level deformations drive cell-level models. Potential loss of information in this numerical coupling approach may cause erroneous cellular-scale results, particularly during multiphysics analysis of cartilage. The goal of this study was to evaluate the capacity of first- and second-order data passing to predict chondrocyte mechanics by analysing cartilage deformations obtained for varying complexity of loading scenarios. A tissue-scale model with a sub-region incorporating representation of chondron size and distribution served as control. The post-processing approach first required solution of a homogeneous tissue-level model, results of which were used to drive a separate cell-level model (same characteristics as the sub-region of control model). The first-order data passing appeared to be adequate for simplified loading of the cartilage and for a subset of cell deformation metrics, for example, change in aspect ratio. The second-order data passing scheme was more accurate, particularly when asymmetric permeability of the tissue boundaries was considered. Yet, the method exhibited limitations for predictions of instantaneous metrics related to the fluid phase, for example, mass exchange rate. Nonetheless, employing higher order data exchange schemes may be necessary to understand the biphasic mechanics of cells under lifelike tissue loading states for the whole time history of the simulation.

  9. [Photoelastic stress analysis of root dentin with different composite resin post and core systems and crowns].

    PubMed

    Takei, Hidenori

    2010-03-01

    Much research has been reported about post and core systems with composite resin, but the influence of the different types of prefabricated posts on the distribution of stress in the root has not yet been elucidated. It is necessary to clarify the influence of the relationship between core and crown materials to obtain combined restorations. The aim of this study is to analyze the influence of the combination of various post and core systems and different kinds of crown material on the stress distribution in the root. Six 2-dimensional photoelastic premolar models were designed. Three types of post and core systems (composite resin post and core, composite resin core with the fiber post, and composite resin core with a prefabricated stainless steel post) and two kinds of crown materials (metal and hybrid-type hard composite resin) were fabricated and cemented to each model. In these models, we applied a load of 200 N at an angle of 45 degrees to the tooth axis and analyzed the fringe order using a transmission polariscope. As a result, it has been clarified that the combination of the post and core and the crown plays an important role in preventing stress concentration within root Stress concentration can be prevented using a crown fabricated with a high-elastic modulus for the post and core with a high-elastic modulus, and a crown fabricated with a low-elastic modulus for the post and core with a low-elastic modulus.

  10. Nitrogen Species in the Post-Pinatubo Stratosphere: Model Analysis Utilizing UARS Measurements

    NASA Technical Reports Server (NTRS)

    Danilin, M. Y.; Rodriquez, J. M.; Hu, W.; Ko, M. K. W.; Weisenstein, D. K.; Mergenthaler, J. L.; Russell, J. M., III; Koike, M.; Yue, G. K.

    1998-01-01

    We present an analysis of the impact of heterogeneous chemistry on the partitioning of nitrogen species measured by the Upper Atmosphere Research Satellite (UARS) instruments. The UARS measurements utilized include: N2O, HNO3 and ClONO2 (Cryogen Limb Array Etalon Spectrometer (CLAES), version 7), temperature, methane, ozone, H2O, HCl, NO and NO2 (HALogen Occultation Experiment (HALOE), version 18). The analysis is carried out for the data from January 1992 to September 1994 in the 100-1 mbar (approx.17-47 km) altitude range and over 10 degree latitude bins from 70degS to 70degN. Temporal-spatial evolution of aerosol surface area density (SAD) is adopted according to the Stratospheric Aerosol and Gas Experiment (SAGE) 11 data. A diurnal steady-state photochemical box model, constrained by the temperature, ozone, H2O, CH4, aerosol SAD and columns of O2 and O3 above the point of interest, has been used as the main tool to analyze these data. Total inorganic nitrogen (NO(y)) is obtained by three different methods: (1) as a sum of the UARS measured NO, NO2, HNO3, and ClONO2; (2) from the N2O-NO(y) correlation, and (3) from the CH4-NO(y) correlation. To validate our current understanding of stratospheric heterogeneous chemistry for post-Pinatubo conditions, the model-calculated NO(x)/NO(y) ratios and the NO, NO2, and HNO3 profiles are compared to the UARS-derived data. In general, the UARS-constrained box model captures the main features of nitrogen species partitioning in the post-Pinatubo years. However, the model underestimates the NO2 content, particularly, in the 30-7 mbar (approx. 23-32 km) range. Comparisons of the calculated temporal behavior of the partial columns of NO2 and HNO3 and ground based measurements at 45degS and 45degN are also presented. Our analysis indicates that ground-based and HALOE v. 18 measurements of the NO2 vertical columns are consistent within the range of their uncertainties and are systematically higher (up to 50%) than the model

  11. Nitrogen Species in the Post-Pinatubo Stratosphere: Model Analysis Utilizing UARS Measurements

    NASA Technical Reports Server (NTRS)

    Danilin, M. Y.; Rodriguez, J. M.; Hu, W.; Ko, M. K. W.; Weisenstein, D. K.; Kumer, J. B.; Mergenthaler, J. L.; Russell, J. M., III; Koike, M.; Yue, G. K.

    1998-01-01

    We present an analysis of the impact of heterogeneous chemistry on the partitioning of nitrogen species measured by the Upper Atmosphere Research Satellite (UARS) instruments. The UARS measurements utilized include: N2O, HNO3 and ClONO2 (Cryogen Limb Array Etalon Spectrometer (CLAES), version 7), temperature, methane, ozone, H2O, HCI, NO and NO2 (HALogen Occultation Experiment (HALOE), version 18). The analysis is carried out for the data from January 1992 to September 1994 in the 100-1 mbar (approximately 17-47 km) altitude range and over 10 degree latitude bins from 70 deg S to 70 deg N. Temporal-spatial evolution of aerosol surface area density (SAD) is adopted according to the Stratospheric Aerosol and Gas Experiment (SAGE) II data. A diurnal steady-state photochemical box model, constrained by the temperature, ozone, H2O, CH4, aerosol SAD and columns of O2 and O3 above the point of interest, has been used as the main tool to analyze these data. Total inorganic nitrogen (NOy) is obtained by three different methods: (1) as a sum of the UARS measured NO, NO2, HNO3, and CIONO2; (2) from the N2O-NOy correlation, (3) from the CH4-NOy correlation. To validate our current understanding of stratospheric heterogeneous chemistry for post-Pinatubo conditions, the model-calculated NOx/NOy ratios and the NO, NO2, and HNO3 profiles are compared to the UARS-derived data. In general, the UARS-constrained box model captures the main features of nitrogen species partitioning in the post-Pinatubo years. However, the model underestimates the NO2 content, particularly, in the 30-7 mbar (approximately 23-32 km) range. Comparisons of the calculated temporal behavior of the partial columns of NO2 and HNO3 and ground based measurements at 45 deg S and 45 deg N are also presented. Our analysis indicates that ground-based and HALOE v.18 measurements of the NO2 vertical columns are consistent within the range of their uncertainties and are systematically higher (up to 50%) than the model

  12. Case of 46,XX/47,XY, +21 chimerism in a newborn infant with ambiguous genitalia

    SciTech Connect

    Sawai, Tomoko; Yoshimoto, Masaaki; Kinoshita, Ei-ichi; Baba, Tsuneyoshi; Matsumoto, Tadashi; Tsuji, Yoshiro, Niikawa, Norio; Fukuda, Shinpei; Harada, Naoki

    1994-02-15

    The authors describe the whole-body chimerism in a newborn infant with small phallus, pseudo-vaginal perineal hypospadias, and a bifid scrotum containing gonads. The human testis determining factor gene (SRY) was detected by PCR amplification. GTG-banding chromosome analysis in peripheral blood lymphocytes and cultured fibroblasts derived from right cubital skin showed a 46,XX/47,XY, +21 karyotype. Their ratios in each cell line were 294:5 and 178:7, respectively. QFQ-banding chromosome analysis documented 3 heteromorphic satellites on trisomic chromsomes 21 in the 47,XY,+21 cell line and a homozygous satellite pattern in the 46,XX cell line. Heteromorphic patterns of chromsomes 4, 13, 14, and 22 were also different between the two cell lines. To our knowledge, such disomy/trisomy chimeras have not been described previously. 10 refs., 3 figs.

  13. Gait analysis in a pre- and post-ischemic stroke biomedical pig model.

    PubMed

    Duberstein, Kylee Jo; Platt, Simon R; Holmes, Shannon P; Dove, C Robert; Howerth, Elizabeth W; Kent, Marc; Stice, Steven L; Hill, William D; Hess, David C; West, Franklin D

    2014-02-10

    Severity of neural injury including stroke in human patients, as well as recovery from injury, can be assessed through changes in gait patterns of affected individuals. Similar quantification of motor function deficits has been measured in rodent animal models of such injuries. However, due to differences in fundamental structure of human and rodent brains, there is a need to develop a large animal model to facilitate treatment development for neurological conditions. Porcine brain structure is similar to that of humans, and therefore the pig may make a more clinically relevant animal model. The current study was undertaken to determine key gait characteristics in normal biomedical miniature pigs and dynamic changes that occur post-neural injury in a porcine middle cerebral artery (MCA) occlusion ischemic stroke model. Yucatan miniature pigs were trained to walk through a semi-circular track and were recorded with high speed cameras to detect changes in key gait parameters. Analysis of normal pigs showed overall symmetry in hindlimb swing and stance times, forelimb stance time, along with step length, step velocity, and maximum hoof height on both fore and hindlimbs. A subset of pigs were again recorded at 7, 5 and 3 days prior to MCA occlusion and then at 1, 3, 5, 7, 14 and 30 days following surgery. MRI analysis showed that MCA occlusion resulted in significant infarction. Gait analysis indicated that stroke resulted in notable asymmetries in both temporal and spatial variables. Pigs exhibited lower maximum front hoof height on the paretic side, as well as shorter swing time and longer stance time on the paretic hindlimb. These results support that gait analysis of stroke injury is a highly sensitive detection method for changes in gait parameters in pig.

  14. Chimerism-based pre-emptive immunotherapy with fast withdrawal of immunosuppression and donor lymphocyte infusions after allogeneic stem cell transplantation for pediatric hematologic malignancies.

    PubMed

    Horn, Biljana; Petrovic, Aleksandra; Wahlstrom, Justin; Dvorak, Christopher C; Kong, Denice; Hwang, Jimmy; Expose-Spencer, Jueleah; Gates, Michael; Cowan, Morton J

    2015-04-01

    The presence of increasing host chimerism or persistent mixed chimerism (MC) after hematopoietic stem cell transplantation for leukemia in children is a predictor of relapse. To reduce the risk of relapse, we prospectively studied post-transplantation chimerism-based immunotherapy (IT) using fast withdrawal of immunosuppression (FWI) and donor lymphocyte infusions (DLI) in children with early post-transplantation MC. Forty-three children with hematologic malignancies at 2 institutions were enrolled prospectively in this study from 2009 until 2012 and were followed for a mean of 42 (SD, 10) months. Twelve patients (28%) were assigned to the observation arm based on the presence of graft-versus-host disease (GVHD) or full donor chimerism (FDC), and 5 (12%) sustained early events and could not undergo intervention. Twenty-six (60%) patients with MC were assigned to IT with FWI, which started at a median of 49 days (range, 35 to 85 days) after transplantation. Fourteen patients proceeded to DLI after FWI. Toxicities of treatment included GVHD, which developed in 19% of patients undergoing intervention, with 1 of 26 (4%) dying from GVHD and 1 (4%) still requiring therapy for chronic GVHD 21 months after DLI. Patients with MC undergoing IT had similar 2-year event-free survival (EFS) (73%; 95% confidence interval (CI), 55% to 91%) compared with patients who achieved FDC spontaneously (83%; 95% CI, 62% to 100%); however, because 50% of all relapses in the IT occurred later than 2 years after transplantation, the EFS declined to 55% (95% CI, 34% to 76%) at 42 (SD, 11) months. There were no late relapses in the observation group. EFS in the entire cohort was 58% (95% CI, 42% to 73%) at 42 (SD, 11) months after transplantation. Evidence of disease before transplantation remained a significant predictor of relapse, whereas development of chronic GVHD was protective against relapse. PMID:25644958

  15. Software Analysis of Uncorrelated MS1 Peaks for Discovery of Post-Translational Modifications.

    PubMed

    Pascal, Bruce D; West, Graham M; Scharager-Tapia, Catherina; Flefil, Ricardo; Moroni, Tina; Martinez-Acedo, Pablo; Griffin, Patrick R; Carvalloza, Anthony C

    2015-12-01

    The goal in proteomics to identify all peptides in a complex mixture has been largely addressed using various LC MS/MS approaches, such as data dependent acquisition, SRM/MRM, and data independent acquisition instrumentation. Despite these developments, many peptides remain unsequenced, often due to low abundance, poor fragmentation patterns, or data analysis difficulties. Many of the unidentified peptides exhibit strong evidence in high resolution MS(1) data and are frequently post-translationally modified, playing a significant role in biological processes. Proteomics Workbench (PWB) software was developed to automate the detection and visualization of all possible peptides in MS(1) data, reveal candidate peptides not initially identified, and build inclusion lists for subsequent MS(2) analysis to uncover new identifications. We used this software on existing data on the autophagy regulating kinase Ulk1 as a proof of concept for this method, as we had already manually identified a number of phosphorylation sites Dorsey, F. C. et al (J. Proteome. Res. 8(11), 5253-5263 (2009)). PWB found all previously identified sites of phosphorylation. The software has been made freely available at http://www.proteomicsworkbench.com . Graphical Abstract ᅟ.

  16. Software Analysis of Uncorrelated MS1 Peaks for Discovery of Post-Translational Modifications

    NASA Astrophysics Data System (ADS)

    Pascal, Bruce D.; West, Graham M.; Scharager-Tapia, Catherina; Flefil, Ricardo; Moroni, Tina; Martinez-Acedo, Pablo; Griffin, Patrick R.; Carvalloza, Anthony C.

    2015-12-01

    The goal in proteomics to identify all peptides in a complex mixture has been largely addressed using various LC MS/MS approaches, such as data dependent acquisition, SRM/MRM, and data independent acquisition instrumentation. Despite these developments, many peptides remain unsequenced, often due to low abundance, poor fragmentation patterns, or data analysis difficulties. Many of the unidentified peptides exhibit strong evidence in high resolution MS1 data and are frequently post-translationally modified, playing a significant role in biological processes. Proteomics Workbench (PWB) software was developed to automate the detection and visualization of all possible peptides in MS1 data, reveal candidate peptides not initially identified, and build inclusion lists for subsequent MS2 analysis to uncover new identifications. We used this software on existing data on the autophagy regulating kinase Ulk1 as a proof of concept for this method, as we had already manually identified a number of phosphorylation sites Dorsey, F. C. et al (J. Proteome. Res. 8(11), 5253-5263 (2009)). PWB found all previously identified sites of phosphorylation. The software has been made freely available at http://www.proteomicsworkbench.com .

  17. Nonlinear Analysis and Post-Test Correlation for a Curved PRSEUS Panel

    NASA Technical Reports Server (NTRS)

    Gould, Kevin; Lovejoy, Andrew E.; Jegley, Dawn; Neal, Albert L.; Linton, Kim, A.; Bergan, Andrew C.; Bakuckas, John G., Jr.

    2013-01-01

    The Pultruded Rod Stitched Efficient Unitized Structure (PRSEUS) concept, developed by The Boeing Company, has been extensively studied as part of the National Aeronautics and Space Administration's (NASA s) Environmentally Responsible Aviation (ERA) Program. The PRSEUS concept provides a light-weight alternative to aluminum or traditional composite design concepts and is applicable to traditional-shaped fuselage barrels and wings, as well as advanced configurations such as a hybrid wing body or truss braced wings. Therefore, NASA, the Federal Aviation Administration (FAA) and The Boeing Company partnered in an effort to assess the performance and damage arrestments capabilities of a PRSEUS concept panel using a full-scale curved panel in the FAA Full-Scale Aircraft Structural Test Evaluation and Research (FASTER) facility. Testing was conducted in the FASTER facility by subjecting the panel to axial tension loads applied to the ends of the panel, internal pressure, and combined axial tension and internal pressure loadings. Additionally, reactive hoop loads were applied to the skin and frames of the panel along its edges. The panel successfully supported the required design loads in the pristine condition and with a severed stiffener. The panel also demonstrated that the PRSEUS concept could arrest the progression of damage including crack arrestment and crack turning. This paper presents the nonlinear post-test analysis and correlation with test results for the curved PRSEUS panel. It is shown that nonlinear analysis can accurately calculate the behavior of a PRSEUS panel under tension, pressure and combined loading conditions.

  18. Glycoproteomic analysis of seven major allergenic proteins reveals novel post-translational modifications.

    PubMed

    Halim, Adnan; Carlsson, Michael C; Madsen, Caroline Benedicte; Brand, Stephanie; Møller, Svenning Rune; Olsen, Carl Erik; Vakhrushev, Sergey Y; Brimnes, Jens; Wurtzen, Peter Adler; Ipsen, Henrik; Petersen, Bent L; Wandall, Hans H

    2015-01-01

    Allergenic proteins such as grass pollen and house dust mite (HDM) proteins are known to trigger hypersensitivity reactions of the immune system, leading to what is commonly known as allergy. Key allergenic proteins including sequence variants have been identified but characterization of their post-translational modifications (PTMs) is still limited. Here, we present a detailed PTM(1) characterization of a series of the main and clinically relevant allergens used in allergy tests and vaccines. We employ Orbitrap-based mass spectrometry with complementary fragmentation techniques (HCD/ETD) for site-specific PTM characterization by bottom-up analysis. In addition, top-down mass spectrometry is utilized for targeted analysis of individual proteins, revealing hitherto unknown PTMs of HDM allergens. We demonstrate the presence of lysine-linked polyhexose glycans and asparagine-linked N-acetylhexosamine glycans on HDM allergens. Moreover, we identified more complex glycan structures than previously reported on the major grass pollen group 1 and 5 allergens, implicating important roles for carbohydrates in allergen recognition and response by the immune system. The new findings are important for understanding basic disease-causing mechanisms at the cellular level, which ultimately may pave the way for instigating novel approaches for targeted desensitization strategies and improved allergy vaccines.

  19. Glycoproteomic Analysis of Seven Major Allergenic Proteins Reveals Novel Post-translational Modifications*

    PubMed Central

    Halim, Adnan; Carlsson, Michael C.; Madsen, Caroline Benedicte; Brand, Stephanie; Møller, Svenning Rune; Olsen, Carl Erik; Vakhrushev, Sergey Y.; Brimnes, Jens; Wurtzen, Peter Adler; Ipsen, Henrik; Petersen, Bent L.; Wandall, Hans H.

    2015-01-01

    Allergenic proteins such as grass pollen and house dust mite (HDM) proteins are known to trigger hypersensitivity reactions of the immune system, leading to what is commonly known as allergy. Key allergenic proteins including sequence variants have been identified but characterization of their post-translational modifications (PTMs) is still limited. Here, we present a detailed PTM1 characterization of a series of the main and clinically relevant allergens used in allergy tests and vaccines. We employ Orbitrap-based mass spectrometry with complementary fragmentation techniques (HCD/ETD) for site-specific PTM characterization by bottom-up analysis. In addition, top-down mass spectrometry is utilized for targeted analysis of individual proteins, revealing hitherto unknown PTMs of HDM allergens. We demonstrate the presence of lysine-linked polyhexose glycans and asparagine-linked N-acetylhexosamine glycans on HDM allergens. Moreover, we identified more complex glycan structures than previously reported on the major grass pollen group 1 and 5 allergens, implicating important roles for carbohydrates in allergen recognition and response by the immune system. The new findings are important for understanding basic disease-causing mechanisms at the cellular level, which ultimately may pave the way for instigating novel approaches for targeted desensitization strategies and improved allergy vaccines. PMID:25389185

  20. Effects of 3D PIV post-processing on impulse and force analysis in vortical flows

    NASA Astrophysics Data System (ADS)

    Mendelson, Leah; Techet, Alexandra

    2014-11-01

    Vortical flows measured using 3D PIV techniques are fundamentally filtered versions of physical phenomena, with velocity information lost below the length and time scales of the measurement system. In the context of propulsive vortices, such as those generated during biological locomotion, these factors, combined with experimental noise and error, can lead to inaccuracies in analysis of the vortex momentum and net thrust. As a result, while 3D velocity measurements remove many of the assumptions required to analyze planar PIV data, they should not be considered absolute physical quantities. Our work focuses on post-processing for 3D PIV data sets to enable the extraction of accurate, quantitative 3D force measurements for unsteady vortical propulsion. In this study, we compare utilizing measurement signal processing techniques, orthogonal decomposition, and identification of coherent structures to measure the impulse of a canonical vortex ring generated by a mechanical piston. In particular, we consider the ability of these methods to confront the influences of limited spatial resolution and arbitrary geometries, and make recommendations for a general procedure for propulsion analysis from 3D PIV data, regardless of which PIV technique is used to obtain the velocity fields.

  1. Struggles for recognition: a content analysis of messages posted on the Internet

    PubMed Central

    Andersen, Anders Johan W; Svensson, Tommy

    2012-01-01

    Background The Internet has enlarged the possibilities of human communication and opened new ways of exploring perceptions of mental health. This study is part of a research project aiming to explore, describe, and analyze different discourses of mental health in Norway and Sweden, using material from Internet-based services. Aim To examine messages posed by users of publicly available question-and-answer services and to describe their content. Methods A Web search was used to identify Norwegian and Swedish Websites offering mental health services by email or posted messages. A total of 601 messages from 20 services, 10 Norwegian and 10 Swedish, were analyzed by means of qualitative content analysis and further interpreted in light of the social theory of recognition by Honneth. Results Eight categories emerged from the analysis: family life, couples, others, violence, the ungovernable, self-image, negotiating normality, and life struggles. These categories were then grouped into three themes: (1) relationship to significant others, (2) relationship to self, and (3) relationship to the social community. The themes promoted an understanding of mental health as closely connected to political and social factors. Conclusions The results showed a variety of concerns from various parts of life and empowered the view that mental health should be understood broadly, at a conceptual level. Mental health emerged as a deeply relational concept that emphasized the equal distribution of chances in life. It strengthened the moral grammar of social inclusion and the acceptance of plurality in social life. PMID:22888257

  2. Analysis of the real EADGENE data set: Multivariate approaches and post analysis (Open Access publication)

    PubMed Central

    Sørensen, Peter; Bonnet, Agnès; Buitenhuis, Bart; Closset, Rodrigue; Déjean, Sébastien; Delmas, Céline; Duval, Mylène; Glass, Liz; Hedegaard, Jakob; Hornshøj, Henrik; Hulsegge, Ina; Jaffrézic, Florence; Jensen, Kirsty; Jiang, Li; de Koning, Dirk-Jan; Cao, Kim-Anh Lê; Nie, Haisheng; Petzl, Wolfram; Pool, Marco H; Robert-Granié, Christèle; San Cristobal, Magali; Lund, Mogens Sandø; van Schothorst, Evert M; Schuberth, Hans-Joachim; Seyfert, Hans-Martin; Tosser-Klopp, Gwenola; Waddington, David; Watson, Michael; Yang, Wei; Zerbe, Holm

    2007-01-01

    The aim of this paper was to describe, and when possible compare, the multivariate methods used by the participants in the EADGENE WP1.4 workshop. The first approach was for class discovery and class prediction using evidence from the data at hand. Several teams used hierarchical clustering (HC) or principal component analysis (PCA) to identify groups of differentially expressed genes with a similar expression pattern over time points and infective agent (E. coli or S. aureus). The main result from these analyses was that HC and PCA were able to separate tissue samples taken at 24 h following E. coli infection from the other samples. The second approach identified groups of differentially co-expressed genes, by identifying clusters of genes highly correlated when animals were infected with E. coli but not correlated more than expected by chance when the infective pathogen was S. aureus. The third approach looked at differential expression of predefined gene sets. Gene sets were defined based on information retrieved from biological databases such as Gene Ontology. Based on these annotation sources the teams used either the GlobalTest or the Fisher exact test to identify differentially expressed gene sets. The main result from these analyses was that gene sets involved in immune defence responses were differentially expressed. PMID:18053574

  3. Detection of looseness degree of dental posts using natural frequency analysis.

    PubMed

    Hsieh, Sung-Chih; Lin, Shu-Li; Huang, Yung-Kai; Chang, Wei-Jen; Yuan, Chia-Chun; Lo, Yi-June; Lee, Sheng-Yang; Huang, Haw-Ming

    2013-08-01

    This study examines the relationship between the retention force and natural frequency of dental posts cemented with zinc phosphate cement. Forty-two decoronated maxillary incisors were collected and embedded in resin blocks. The 7-mm post spaces were prepared for all test samples prior to cementing the 13-mm Para-Posts with zinc phosphate cement. The natural frequency values and retention forces of the zinc phosphate cement-cemented Para-Posts in various liquid/powder mixing ratios were measured. Thereafter, the natural frequency values of the zinc phosphate cement-cemented posts were continuously detected during ultrasonic vibration. Moreover, both the natural frequency values and the dislodgement forces in response to ultrasonic vibrations were measured for the pretreated post samples. Results showed that the natural frequency values of the posts significantly decreased from 8.8 ± 0.8 to 5.0 ± 0.5 kHz (P < 0.05) before and after ultrasound treatment, respectively. The natural frequency values and the dislodgement forces of tested posts had a highly linear relationship (R² = 0.99, P < 0.05). These findings suggest that natural frequency is a variable for monitoring the stability status of zinc phosphate cement-cemented dental posts.

  4. 3D-FE analysis of functionally graded structured dental posts.

    PubMed

    Abu Kasim, Noor H; Madfa, Ahmed A; Hamdi, Mohd; Rahbari, Ghahnavyeh R

    2011-01-01

    This study aimed to compare the biomechanical behaviour of functionally graded structured posts (FGSPs) and homogenous-type posts in simulated models of a maxillary central incisor. Two models of FGSPs consisting of a multilayer xTi-yHA composite design, where zirconia and alumina was added as the first layer for models A and B respectively were compared to homogenous zirconia post (model C) and a titanium post (model D). The amount of Ti and HA in the FGSP models was varied in gradations. 3D-FEA was performed on all models and stress distributions were investigated along the dental post. In addition, interface stresses between the posts and their surrounding structures were investigated under vertical, oblique, and horizontal loadings. Strain distribution along the post-dentine interface was also investigated. The results showed that FGSPs models, A and B demonstrated better stress distribution than models C and D, indicating that dental posts with multilayered structure dissipate localized and interfacial stress and strain more efficiently than homogenous-type posts. PMID:22123011

  5. Understanding Impact and Implications of Data Standards on Post Disaster Risk Analysis

    NASA Astrophysics Data System (ADS)

    Stevenson, Robert

    2010-05-01

    Although the physical and humanitarian effects of a natural catastrophe are often bound to the locality of the event the financial impacts can have global effects. This is particularly prominent in the re/insurance community, where through a number of market mechanisms and re/insurance structures financial loss is mitigated amongst many companies across the globe. The level of risk a company wishes to retain, given an event, represents the level of risk decision makers deem acceptable. Catastrophe risk modelling tools aid the estimation of risk retention and transfer mechanisms, and increasingly the level of capital required to withstand a catastrophic event. These tools rely on appropriate representations hazard, exposure, vulnerability and insurance conditions that reflect the reality of risk. In addition, accurate estimation of loss potential in the aftermath of a catastrophic event equally relies on the data available to assess the scale of damages experienced and to provide views on the likely scale of loss. A coherent and focussed data and modelling strategy is required to ensure that the risk assessment made is as accurate as possible. A fundamental factor in determining the accuracy of catastrophe output, is the quality of data entered. It is of vital importance, therefore, to have an understanding of both the data used as well as the standard of this data, which will so powerfully impact upon the decision making process. This is perhaps best illustrated through the study of historical events, such as Hurricane Katrina and Ike. The extent of data variance in post disaster analysis clearly demonstrates issues of data discrepancies, vintage, resolution and uncertainty propagation, and reflects on the standard of the original data utilized for modelling purposes and decision making. Using experience gained from recent events, this paper will explore current data variabilities, and the impacts on effective loss estimation, both in relation to reinsurance

  6. Radar rainfall estimation in the context of post-event analysis of flash-flood events

    NASA Astrophysics Data System (ADS)

    Bouilloud, Ludovic; Delrieu, Guy; Boudevillain, Brice; Kirstetter, Pierre-Emmanuel

    2010-11-01

    SummaryA method to estimate rainfall from radar data for post-event analysis of flash-flood events has been developed within the EC-funded HYDRATE project. It follows a pragmatic approach including careful analysis of the observation conditions for the radar system(s) available for the considered case. Clutter and beam blockage are characterised by dry-weather observations and simulations based on a digital terrain model of the region of interest. The vertical profile of reflectivity (VPR) is either inferred from radar data if volume scanning data are available or simply defined using basic meteorological parameters (idealised VPR). Such information is then used to produce correction factor maps for each elevation angle to correct for range-dependent errors. In a second step, an effective Z-R relationship is optimised to remove the bias over the hit region. Due to limited data availability, the optimisation is carried out with reference to raingauge rain amounts measured at the event time scale. Sensitivity tests performed with two well-documented rain events show that a number of Z = aRb relationships, organised along hyperbolic curves in the (a and b) parameter space, lead to optimum assessment results in terms of the Nash coefficient between the radar and raingauge estimates. A refined analysis of these equifinality patterns shows that the “total additive conditional bias” can be used to discriminate between the Nash coefficient equifinal solutions. We observe that the optimisation results are sensitive to the VPR description and also that the Z-R optimisation procedure can largely compensate for range-dependent errors, although this shifts the optimal coefficients in the parameter space. The time-scale dependency of the equifinality patterns is significant, however near-optimal Z-R relationships can be obtained at all time scales from the event time step optimisation.

  7. Combination of liquid-chromatography tandem mass spectrometry in different scan modes with human and chimeric mouse urine for the study of steroid metabolism.

    PubMed

    Pozo, Oscar J; Lootens, Leen; Van Eenoo, Peter; Deventer, Koen; Meuleman, Philip; Leroux-Roels, Geert; Parr, Maria K; Schänzer, Wilhelm; Delbeke, Frans T

    2009-11-01

    Anabolic steroids are among the most frequently detected compounds in doping analysis. They are extensively metabolized and therefore an in-depth knowledge about steroid metabolism is needed. In this study, a liquid chromatography tandem mass spectometry (LC-MS/MS) method based on a precursor ion scan with a uPA-SCID mouse with humanized liver (a chimeric mouse) was explored for the detection of steroid metabolism. Methandienone was used as a model compound. The application of the precursor ion scan method in positive human samples and chimeric mice samples after methandienone administration allowed the detection of most steroid metabolites without any structural restriction. Three hitherto unreported metabolites were found using this approach. These metabolites were characterized using LC-MS/MS and feasible structures were proposed. The structure of one of them, 6-ene-epimethandienone, was confirmed by the synthesis of the reference compound. A selected reaction monitoring (SRM) method for the specific detection of all these metabolites has been developed. The application of this method to several human and chimeric mouse samples confirmed that more than 80% of the steroid metabolites were found in both samples. Only metabolites that are poorly detectable by LC-MS/MS were not detected in some urine samples. The metabolic nature of the unreported metabolites was also confirmed. A global strategy for the detection of steroid metabolites combining both human and chimeric mouse urine is proposed. PMID:20355172

  8. Reflections on the unique tolerogenicity of bone marrow, the enigma of chimerism and clinical tolerance.

    PubMed

    Monaco, Anthony P

    2013-01-01

    Since the discovery of acquired immunological tolerance, chimerism has always been associated with tolerance. There is, however, a frequent dichotomy between chimerism and tolerance. Many experimental strategies that produce chimerism do not induce tolerance. In addition, some types of chimerism frequently occur after solid organ transplantation, but rarely result in tolerance. In experimental models of transient lymphocyte depletion with antilymphocyte serum, bone marrow cells exhibit a unique ability to induce allograft tolerance that is superior to that of other lymphoid cells. This tolerance can be augmented with standard immunosuppressive agents used in clinical transplantation. There are currently four ongoing clinical trials of tolerance induction to renal allografts that employ various protocols of non-myeloablative conditioning and donor bone marrow infusion. All four trials have been remarkably successful in achieving short- and moderate-term duration tolerance with minimal morbidity and complications. Persistent tolerance (total drug withdrawal) has been achieved in recipients with durable substantial chimerism. Durable tolerance has also been achieved in recipients who have lost chimerism before or after drug withdrawal has been initiated, as well as in recipients in whom only transient (less than three weeks) or no chimerism at all has been achieved. Although chimeric recipients have rejected grafts during drug withdrawal, durable chimerism is thus far the most positive biomarker for likely successful tolerance induction. At present, there is no proof that chimerism causes tolerance per se; the data are also consistent with another etiological mechanism that causes tolerance and thereby permits chimerism to persist. The current experimental protocols for tolerance induction are safe. More transplant programs should consider doing clinical tolerance research.

  9. Chimeras taking shape: Potential functions of proteins encoded by chimeric RNA transcripts

    PubMed Central

    Frenkel-Morgenstern, Milana; Lacroix, Vincent; Ezkurdia, Iakes; Levin, Yishai; Gabashvili, Alexandra; Prilusky, Jaime; del Pozo, Angela; Tress, Michael; Johnson, Rory; Guigo, Roderic; Valencia, Alfonso

    2012-01-01

    Chimeric RNAs comprise exons from two or more different genes and have the potential to encode novel proteins that alter cellular phenotypes. To date, numerous putative chimeric transcripts have been identified among the ESTs isolated from several organisms and using high throughput RNA sequencing. The few corresponding protein products that have been characterized mostly result from chromosomal translocations and are associated with cancer. Here, we systematically establish that some of the putative chimeric transcripts are genuinely expressed in human cells. Using high throughput RNA sequencing, mass spectrometry experimental data, and functional annotation, we studied 7424 putative human chimeric RNAs. We confirmed the expression of 175 chimeric RNAs in 16 human tissues, with an abundance varying from 0.06 to 17 RPKM (Reads Per Kilobase per Million mapped reads). We show that these chimeric RNAs are significantly more tissue-specific than non-chimeric transcripts. Moreover, we present evidence that chimeras tend to incorporate highly expressed genes. Despite the low expression level of most chimeric RNAs, we show that 12 novel chimeras are translated into proteins detectable in multiple shotgun mass spectrometry experiments. Furthermore, we confirm the expression of three novel chimeric proteins using targeted mass spectrometry. Finally, based on our functional annotation of exon organization and preserved domains, we discuss the potential features of chimeric proteins with illustrative examples and suggest that chimeras significantly exploit signal peptides and transmembrane domains, which can alter the cellular localization of cognate proteins. Taken together, these findings establish that some chimeric RNAs are translated into potentially functional proteins in humans. PMID:22588898

  10. An analysis of Social Work Oncology Network Listserv Postings on the Commission of Cancer's distress screening guidelines.

    PubMed

    Burg, Mary Ann; Adorno, Gail; Hidalgo, Jorge

    2012-01-01

    This is a qualitative study of listserv postings by members of the Social Work Oncology Network (SWON) in response to the Commission on Cancer's 2011 guidelines for distress screening of cancer patients. Archived listserv postings for the period of December 2010 to November 2011 were deidentified and a sample was derived by a list of keywords for the analysis. Aims of the study included describing the general categories and themes of the postings devoted to the new distress screening standard and examining the process of facilitation of mutual support and information exchange by oncology social workers in response to the new screening standards. During the 12-month timeframe there were 242 unique listserv postings sampled for the analysis. Oncology social worker (OSW) discussion of the distress screening guidelines remained a constant topic over the 12 months, and major themes that emerged from the data included processes of implementation of distress screening in cancer centers, screening policies and protocols, screening tool choice, and oncology social worker professional identity. The SWON listserv members used the listserv as a mechanism to post their requests for information on screening, to share their experiences in the beginning stages of implementing the guidelines, and to build support for legitimizing oncology social workers as the lead profession in the implementation of the guidelines in member cancer centers.

  11. Identifying Multimedia Production Competencies and Skills of Instructional Design and Technology Professionals: An Analysis of Recent Job Postings

    ERIC Educational Resources Information Center

    Sugar, William; Hoard, Brent; Brown, Abbie; Daniels, Lee

    2012-01-01

    In an effort to document necessary multimedia production competencies of Instructional Design and Technology graduates, a recent analysis of over 7 months' worth of Instructional Design and Technology job advertisements (n = 615) were conducted. Specific job skills from these postings were categorized and analyzed. The data set includes three job…

  12. Analysis of Post-Fire Vegetation Recovery in the Mediterranean Basin using MODIS Derived Vegetation Indices

    NASA Astrophysics Data System (ADS)

    Hawtree, Daniel; San Miguel, Jesus; Sedano, Fernando; Kempeneers, Pieter

    2010-05-01

    The Mediterranean basin region is highly susceptible to wildfire, with approximately 60,000 individual fires and half a million ha of natural vegetation burnt per year. Of particular concern in this region is the impact of repeated wildfires on the ability of natural lands to return to a pre-fire state, and of the possibility of desertification of semi-arid areas. Given these concerns, understanding the temporal patterns of vegetation recovery is important for the management of environmental resources in the region. A valuable tool for evaluating these recovery patterns are vegetation indices derived from remote sensing data. Previous research on post-fire vegetation recovery conducted in this region has found significant variability in recovery times across different study sites. It is unclear what the primary variables are affecting the differences in the rates of recovery, and if any geographic patterns of behavior exist across the Mediterranean basin. This research has primarily been conducted using indices derived from Landsat imagery. However, no extensive analysis of vegetation regeneration for large regions has been published, and assessment of vegetation recovery on the basis of medium-spatial resolution imagery such as that of MODIS has not yet been analyzed. This study examines the temporal pattern of vegetation recovery in a number of fire sites in the Mediterranean basin, using data derived from MODIS 16 -day composite vegetation indices. The intent is to develop a more complete picture of the temporal sequence of vegetation recovery, and to evaluate what additional factors impact variations in the recovery sequence. In addition, this study evaluates the utility of using MODIS derived vegetation indices for regeneration studies, and compares the findings to earlier studies which rely on Landsat data. Wildfires occurring between the years 2000 and 2004 were considered as potential study sites for this research. Using the EFFIS dataset, all wildfires

  13. Novel nanocomposites from spider silk–silica fusion (chimeric) proteins

    PubMed Central

    Wong Po Foo, Cheryl; Patwardhan, Siddharth V.; Belton, David J.; Kitchel, Brandon; Anastasiades, Daphne; Huang, Jia; Naik, Rajesh R.; Perry, Carole C.; Kaplan, David L.

    2006-01-01

    Silica skeletal architectures in diatoms are characterized by remarkable morphological and nanostructural details. Silk proteins from spiders and silkworms form strong and intricate self-assembling fibrous biomaterials in nature. We combined the features of silk with biosilica through the design, synthesis, and characterization of a novel family of chimeric proteins for subsequent use in model materials forming reactions. The domains from the major ampullate spidroin 1 (MaSp1) protein of Nephila clavipes spider dragline silk provide control over structural and morphological details because it can be self-assembled through diverse processing methods including film casting and fiber electrospinning. Biosilica nanostructures in diatoms are formed in aqueous ambient conditions at neutral pH and low temperatures. The R5 peptide derived from the silaffin protein of Cylindrotheca fusiformis induces and regulates silica precipitation in the chimeric protein designs under similar ambient conditions. Whereas mineralization reactions performed in the presence of R5 peptide alone form silica particles with a size distribution of 0.5–10 μm in diameter, reactions performed in the presence of the new fusion proteins generate nanocomposite materials containing silica particles with a narrower size distribution of 0.5–2 μm in diameter. Furthermore, we demonstrate that composite morphology and structure could be regulated by controlling processing conditions to produce films and fibers. These results suggest that the chimeric protein provides new options for processing and control over silica particle sizes, important benefits for biomedical and specialty materials, particularly in light of the all aqueous processing and the nanocomposite features of these new materials. PMID:16769898

  14. Chimeric creatures in Greek mythology and reflections in science.

    PubMed

    Bazopoulou-Kyrkanidou, E

    2001-04-15

    "The Chimaera" in Homer's Iliad, "was of divine stock, not of men, in the forepart a lion, in the hinder a serpent, and in the midst a goat, ellipsis Bellerophon slew her, trusting in the signs of the gods." In Hesiod's Theogony it is emphasized that "Chimaera ellipsis had three heads, one of a grim-eyed lion, another of a goat, and another of a snakeellipsis". In addition to this interspecies animal chimera, human/animal chimeras are referred to in Greek mythology, preeminent among them the Centaurs and the Minotaur. The Centaurs, as horse/men, first appear in Geometric and early Archaic art, but in the literature not until early in the fifth century B.C. The bullheaded-man Minotaur, who is not certainly attested in the literary evidence until circa 500 B.C., first appears in art about 650 B.C. Attempts, in the fourth century B.C. and thereafter, to rationalize their mythical appearance were in vain; their chimeric nature retained its fascinating and archetypal form over the centuries. Early in the 1980s, experimental sheep/goat chimeras were produced removing the reproductive barrier between these two animal species. Late in the 1990s, legal, political, ethical, and moral fights loomed over a patent bid on human/animal chimeras. Chimeric technology is recently developed; however, the concept of chimerism has existed in literary and artistic form in ancient mythology. This is yet another example where art and literature precede scientific research and development. PMID:11337752

  15. Generation of Novel Chimeric Mice with Humanized Livers by Using Hemizygous cDNA-uPA/SCID Mice.

    PubMed

    Tateno, Chise; Kawase, Yosuke; Tobita, Yoshimi; Hamamura, Satoko; Ohshita, Hiroki; Yokomichi, Hiroshi; Sanada, Harumi; Kakuni, Masakazu; Shiota, Akira; Kojima, Yuha; Ishida, Yuji; Shitara, Hiroshi; Wada, Naoko A; Tateishi, Hiromi; Sudoh, Masayuki; Nagatsuka, Shin-Ichiro; Jishage, Kou-Ichi; Kohara, Michinori

    2015-01-01

    producing chimeric mice for use in future long-term studies, including hepatitis virus infection analysis or drug toxicity studies. PMID:26536627

  16. Generation of Novel Chimeric Mice with Humanized Livers by Using Hemizygous cDNA-uPA/SCID Mice.

    PubMed

    Tateno, Chise; Kawase, Yosuke; Tobita, Yoshimi; Hamamura, Satoko; Ohshita, Hiroki; Yokomichi, Hiroshi; Sanada, Harumi; Kakuni, Masakazu; Shiota, Akira; Kojima, Yuha; Ishida, Yuji; Shitara, Hiroshi; Wada, Naoko A; Tateishi, Hiromi; Sudoh, Masayuki; Nagatsuka, Shin-Ichiro; Jishage, Kou-Ichi; Kohara, Michinori

    2015-01-01

    producing chimeric mice for use in future long-term studies, including hepatitis virus infection analysis or drug toxicity studies.

  17. The History of Post-Secondary Finance in Alberta - An Analysis

    ERIC Educational Resources Information Center

    Hauserman, Calvin P.; Stick, Sheldon L.

    2005-01-01

    Post-secondary systems throughout Canada and the United States have struggled with funding issues during most of the last decade of the 20th Century, and the new millennium did not open with great enthusiasm for change. This article examines the impact of post-secondary education funding changes in Alberta, Canada, by tracing the historical…

  18. Asynchronous Electronic Discussion Group: Analysis of Postings and Perception of In-Service Teachers

    ERIC Educational Resources Information Center

    Kim, Tina Lim Swee; Wah, Wong Kiet; Lee, Tan Ai

    2007-01-01

    This paper examines the practice of online discussion in a course specially tailored for in-service teachers who are pursuing their basic degree qualification at a teacher training institute. Analyses of postings to the asynchronous electronic discussion group were made according to the type of postings as proposed by Poole (2000). Four focus…

  19. A PCR amplification strategy for unrestricted generation of chimeric genes.

    PubMed

    Vos, Michel J; Kampinga, Harm H

    2008-09-15

    For analyzing protein function, protein dynamics, or protein-protein interactions, the use of chimeric proteins has become an indispensable tool. The generation of DNA constructs coding for such fused proteins can, however, be a tedious process. Currently used strategies often make use of available endonuclease sites, leading to limitations in the choice of the site of fusion between two genes and problems in maintaining protein secondary structure. We have developed a cloning strategy to get around these disadvantages that is based on a single round of PCR amplification followed by antibiotic-resistant gene complementation. PMID:18555003

  20. Clinical strategy for induction of transplantation tolerance through mixed chimerism.

    PubMed

    Cosimi, A Benedict; Sachs, David H; Sykes, Megan; Kawai, Tatsuo

    2013-01-01

    Following the demonstration that transplant tolerance could be induced in non-human primate recipients treated with non-myeloablative conditioning that resulted in only transient chimerism, we began in 1998 to evaluate this approach first in patients with end stage renal disease (ESRD), secondary to multiple myeloma (MM). A total of 10 patients with ESRD and MM have been treated with this initial protocol. Only 2 recipients developed evidence of reversible renal allograft rejection after stopping immunosuppression. Long-term (up to 14 years) operational renal allograft tolerance has been observed in all 10 patients, even in those with transient hematopoietic chimerism. Control of the MM has been less complete, as recurrent disease developed in five of these patients, three of whom expired. Nevertheless, in view of the essentially 100% 3-5 year mortality typically expected with alternative treatments for this challenging population, it has been suggested that combined kidney and donor bone marrow transplantation (CKBMT) following non-myeloablative conditioning should become the standard therapy for patients with ESRD secondary to MM (27). These encouraging results, as well as the acceptable morbidity observed in cancer patients receiving non-myeloablative human leukocyte antigen (HLA)-mismatched bone marrow transplantation in our center, led us to next evaluate CKBMT in 10 patients of HLA-mismatched transplants. All 10 subjects developed transient chimerism and in seven of these, immunosuppression was successfully discontinued. Four subjects continue to be immunosuppression free for periods of 4.5-11 years, while in three, reinstitution of immunosuppression was advised or accomplished after 5-8 years due to recurrence of original disease or chronic antibody mediated rejection. Donor-specific antibodies were frequently detectable in the earlier recipients. In contrast, no donor-specific antibodies were detected after immunosuppression was discontinued in the last

  1. A PCR amplification strategy for unrestricted generation of chimeric genes.

    PubMed

    Vos, Michel J; Kampinga, Harm H

    2008-09-15

    For analyzing protein function, protein dynamics, or protein-protein interactions, the use of chimeric proteins has become an indispensable tool. The generation of DNA constructs coding for such fused proteins can, however, be a tedious process. Currently used strategies often make use of available endonuclease sites, leading to limitations in the choice of the site of fusion between two genes and problems in maintaining protein secondary structure. We have developed a cloning strategy to get around these disadvantages that is based on a single round of PCR amplification followed by antibiotic-resistant gene complementation.

  2. Chimerism in cattle through microsurgical aggregation of morulae.

    PubMed

    Brem, G; Tenhumberg, H; Kräußlich, H

    1984-11-01

    A cattle chimera was produced by combining four halves of two parent embryos of different breeds (Brown-Swiss x Braunvieh plus Holstein-Friesian x Holstein-Friesian) in one zona pellucida. Parent embryos in the 32-cell morula stage were recovered non-surgically, were bisected, and the combined four halves were transferred non-surgically to recipient heifers. Chimerism of coat colour was used as evidence. Combining of only two half embryos from different parents resulted in five pregnancies carried to term but none of the calves born was a chimera.

  3. Chimeric antigen receptor T-cell therapy for solid tumors

    PubMed Central

    Newick, Kheng; Moon, Edmund; Albelda, Steven M

    2016-01-01

    Chimeric antigen receptor (CAR) T cells are engineered constructs composed of synthetic receptors that direct T cells to surface antigens for subsequent elimination. Many CAR constructs are also manufactured with elements that augment T-cell persistence and activity. To date, CAR T cells have demonstrated tremendous success in eradicating hematological malignancies (e.g., CD19 CARs in leukemias). This success is not yet extrapolated to solid tumors, and the reasons for this are being actively investigated. Here in this mini-review, we discuss some of the key hurdles encountered by CAR T cells in the solid tumor microenvironment. PMID:27162934

  4. Stress generated by customized glass fiber posts and other types by photoelastic analysis.

    PubMed

    Bosso, Kátia; Gonini Júnior, Alcides; Guiraldo, Ricardo Danil; Berger, Sandrine Bittencourt; Lopes, Murilo Baena

    2015-01-01

    Endodontic posts are necessary to provide adequate retention and support when no sufficient remaining structure is available to retain the core. There are different materials and techniques to construct post-and-core, but there is no consensus about which one promotes better stress distribution on the remaining tooth structure. This study aimed to quantify and evaluate the distribution of stress in the root produced by customized glass fiber posts compared to different endodontic posts. Twenty-five simulated roots from photoelastic resin were made and divided into 5 groups: CPC, cast post-and-core; SP, screw post; CF, carbon fiber post; GF, glass fiber post; and CGF, customized glass fiber post. After cementing CPC and SP posts with zinc phosphate cement, and CF, GF and CGF posts with resin cement, resin cores were made for groups 2-5. Specimens were evaluated with vertical or 45° oblique loading. To analyze the fringes, the root was divided into 6 parts: palatal cervical, palatal middle, palatal apical, vestibular cervical, vestibular middle, and vestibular apical. The formed fringes were photographed and quantified. Data were recorded and subjected to two-way ANOVA and Tukey's test (5%). SP (1.95±0.60) showed higher stress (p<0.05) compared to the others (CPC-0.52±0.74; CF-0.50±0.75, GF-0.23±0.48 and CGF-0.45±0.83). All posts showed high stress in apical third (CPC-1.40±0.65; SP-2.30±0.44, CF-1.80±0.45, GF-1.20±0.45, CGF-1.70±1.03) Low stress was found in cervical third (CPC-0.20±0.45; CF-0.00±0.00, GF-0.00±0.00, CGF-0.00±0.00), except by SP (1.90±0.65), which showed statistical difference (p<0.05). Customized post showed high stress concentration at the root and conventional glass fiber posts showed more favorable biomechanical behavior.

  5. A targeted change-detection procedure by combining change vector analysis and post-classification approach

    NASA Astrophysics Data System (ADS)

    Ye, Su; Chen, Dongmei; Yu, Jie

    2016-04-01

    In remote sensing, conventional supervised change-detection methods usually require effective training data for multiple change types. This paper introduces a more flexible and efficient procedure that seeks to identify only the changes that users are interested in, here after referred to as "targeted change detection". Based on a one-class classifier "Support Vector Domain Description (SVDD)", a novel algorithm named "Three-layer SVDD Fusion (TLSF)" is developed specially for targeted change detection. The proposed algorithm combines one-class classification generated from change vector maps, as well as before- and after-change images in order to get a more reliable detecting result. In addition, this paper introduces a detailed workflow for implementing this algorithm. This workflow has been applied to two case studies with different practical monitoring objectives: urban expansion and forest fire assessment. The experiment results of these two case studies show that the overall accuracy of our proposed algorithm is superior (Kappa statistics are 86.3% and 87.8% for Case 1 and 2, respectively), compared to applying SVDD to change vector analysis and post-classification comparison.

  6. Post-Test Analysis of a 10-Year Sodium Heat Pipe Life Test

    NASA Technical Reports Server (NTRS)

    Rosenfeld, John H.; Locci, Ivan E.; Sanzi, James L.; Hull, David R.; Geng, Steven M.

    2011-01-01

    High-temperature heat pipes are being evaluated for use in energy conversion applications such as fuel cells, gas turbine re-combustors, Stirling cycle heat sources; and with the resurgence of space nuclear power both as reactor heat removal elements and as radiator elements. Long operating life and reliable performance are critical requirements for these applications. Accordingly, long-term materials compatibility is being evaluated through the use of high-temperature life test heat pipes. Thermacore, Inc., has carried out a sodium heat pipe 10-year life test to establish long-term operating reliability. Sodium heat pipes have demonstrated favorable materials compatibility and heat transport characteristics at high operating temperatures in air over long time periods. A representative one-tenth segment Stirling Space Power Converter heat pipe with an Inconel 718 envelope and a stainless steel screen wick has operated for over 87,000 hr (10 years) at nearly 700 C. These life test results have demonstrated the potential for high-temperature heat pipes to serve as reliable energy conversion system components for power applications that require long operating lifetime with high reliability. Detailed design specifications, operating history, and post-test analysis of the heat pipe and sodium working fluid are described. Lessons learned and future life test plans are also discussed.

  7. Post-test thermomechanical calculations and preliminary data analysis for the Spent Fuel Test: Climax

    SciTech Connect

    Butkovich, T.R.; Patrick, W.C.

    1985-09-01

    The Spent Fuel Test - Climax (SFT-C) was conducted to evaluate the feasibility of retrievable deep geologic storage of commercially generated, spent nuclear-reactor fuel assemblies. Thermomechanical response of the SFT-C was calculated before the test began using the finite-element structural analysis code ADINA and its companion heat transfer code ADINAT. While we found that the level of agreement between measured and calculated rock displacements was quite good, we needed to revise certain aspects of the heat transfer calculation, material properties, and in situ stresses to incorporate information obtained during and after the heated phase of the test. The post-test calculations reported here were performed using the best available input parameters, thermal and mechanical properties, and power levels that were directly measured or inferred from measurements made during the test. This report documents the results of these calculations and compares those results with selected measurements made during the 3-year heating phase and 6-month cooling phase of the SFT-C.

  8. Post-experimental analysis of a solid oxide fuel cell stack using hybrid seals

    NASA Astrophysics Data System (ADS)

    Thomann, O.; Rautanen, M.; Himanen, O.; Tallgren, J.; Kiviaho, J.

    2015-01-01

    A post-experimental analysis of a SOFC stack is presented. The stack was operated for 1800 h at 700 °C with air and hydrogen and contained hybrid glass-Thermiculite 866 seals. The goal of this work was to investigate the sealing microstructure and possible corrosion during mid-term operation. It was found that hybrid seals could effectively compensate for manufacturing tolerances of cells and other components due to the compliance of the glass layer. Additionally, different interfaces were investigated for corrosion. Corrosion was not observed at two-phase interfaces such as Crofer 22 APU/glass, glass/electrolyte and glass/Thermiculite 866. The three-phase interface between Crofer 22 APU/glass/hydrogen exhibited no corrosion. Some evidence of non-systematic corrosion was found at the Crofer 22 APU/glass/air interface. The possible reasons for the corrosion are discussed. Lastly, dual exposure to humid hydrogen and air of the 0.2 mm Crofer 22 APU interconnect had no detrimental effect on the corrosion compared to air exposure. Overall the hybrid seals used in combination with the thin interconnects were found to be a promising solution due to the low leak rate and limited material interactions.

  9. Scale analysis of pre- and post-midnight ESF bubbles at storm time and quiet time

    NASA Astrophysics Data System (ADS)

    Chen, K. Y.; Su, S. Y.; Yeh, H. C.; Liu, C. H.

    This paper investigates intermediate scale plasma structures observed by ROCSAT-1 in the equatorial F region The empirical mode decomposition EMD method of Hilbert-Huang transform HHT technique is utilized to develop a procedure of scale analysis that allows the mutually correlated components in velocity density and relative density gradient to be identified and extracted Comparing the three data sets good match in wave form is found for velocity and density in scales between kilometers and hundred meters It implies that there are electric fields proportional to density fluctuation -- delta N N in the form similar to what is expected for the Rayleigh Taylor instability In smaller scales velocity and density don t correlate to each other more the good match is then found in velocity and density gradient This is the manifestation of the Boltzmann relation By studying the cases in post-midnight and pre-midnight under storm time and quiet time we find the one-to-one match hold although it is known that ESF bubbles can be driven by different mechanisms under different conditions In other words the spatial structures of electric field in the intermediate scale will always be correlated to the density structures in a manner of delta E sim - delta N N independent of the mechanisms driving the ESF bubbles It is interesting to note that the relation delta V z quad sim delta N N for irregularities in scale of kilometers holds only for ESF occurs within -5 dip latitude while the Boltzmann relation delta

  10. In Silico Analysis of Correlations between Protein Disorder and Post-Translational Modifications in Algae

    PubMed Central

    Kurotani, Atsushi; Sakurai, Tetsuya

    2015-01-01

    Recent proteome analyses have reported that intrinsically disordered regions (IDRs) of proteins play important roles in biological processes. In higher plants whose genomes have been sequenced, the correlation between IDRs and post-translational modifications (PTMs) has been reported. The genomes of various eukaryotic algae as common ancestors of plants have also been sequenced. However, no analysis of the relationship to protein properties such as structure and PTMs in algae has been reported. Here, we describe correlations between IDR content and the number of PTM sites for phosphorylation, glycosylation, and ubiquitination, and between IDR content and regions rich in proline, glutamic acid, serine, and threonine (PEST) and transmembrane helices in the sequences of 20 algae proteomes. Phosphorylation, O-glycosylation, ubiquitination, and PEST preferentially occurred in disordered regions. In contrast, transmembrane helices were favored in ordered regions. N-glycosylation tended to occur in ordered regions in most of the studied algae; however, it correlated positively with disordered protein content in diatoms. Additionally, we observed that disordered protein content and the number of PTM sites were significantly increased in the species-specific protein clusters compared to common protein clusters among the algae. Moreover, there were specific relationships between IDRs and PTMs among the algae from different groups. PMID:26307970

  11. Comparing pre- and post-copulatory mate competition using social network analysis in wild crickets

    PubMed Central

    Fisher, David N.; Rodríguez-Muñoz, Rolando

    2016-01-01

    Sexual selection results from variation in success at multiple stages in the mating process, including competition before and after mating. The relationship between these forms of competition, such as whether they trade-off or reinforce one another, influences the role of sexual selection in evolution. However, the relationship between these 2 forms of competition is rarely quantified in the wild. We used video cameras to observe competition among male field crickets and their matings in the wild. We characterized pre- and post-copulatory competition as 2 networks of competing individuals. Social network analysis then allowed us to determine 1) the effectiveness of precopulatory competition for avoiding postcopulatory competition, 2) the potential for divergent mating strategies, and 3) whether increased postcopulatory competition reduces the apparent reproductive benefits of male promiscuity. We found 1) limited effectiveness of precopulatory competition for avoiding postcopulatory competition; 2) males do not specifically engage in only 1 type of competition; and 3) promiscuous individuals tend to mate with each other, which will tend to reduce variance in reproductive success in the population and highlights the trade-off inherent in mate guarding. Our results provide novel insights into the works of sexual competition in the wild. Furthermore, our study demonstrates the utility of using network analyses to study competitive interactions, even in species lacking obvious social structure. PMID:27174599

  12. High-frequency ultrasound analysis of post-mitotic arrest cell death

    PubMed Central

    Pasternak, Maurice M.; Wirtzfeld, Lauren A.; Kolios, Michael C.; Czarnota, Gregory J.

    2016-01-01

    Non-invasive monitoring of cancer cell death would permit rapid feedback on treatment response. One technique showing such promise is quantitative ultrasound. High-frequency ultrasound spectral radiofrequency analysis was used to study cell death in breast cancer cell samples. Quantitative ultrasound parameters, including attenuation, spectral slope, spectral 0-MHz-intercept, midband fit, and fitted parameters displayed significant changes with paclitaxel-induced cell death, corresponding to observations of morphological changes seen in histology and electron microscopy. In particular, a decrease in spectral slope from 0.24±0.07 dB/MHz to 0.04±0.09 dB/MHz occurred over 24 hours of treatment time and was identified as an ultrasound parameter capable of differentiating post-mitotic arrest cell death from classical apoptosis. The formation of condensed chromatin aggregates of 1 micron or greater in size increased the number of intracellular scatterers, consistent with a hypothesis that nuclear material is a primary source of ultrasound scattering in dying cells. It was demonstrated that the midband fit quantitatively correlated to cell death index, with a Pearson R-squared value of 0.99 at p<0.01. These results suggest that high-frequency ultrasound can not only qualitatively assess the degree of cancer cell death, but may be used to quantify the efficacy of chemotherapeutic treatments. PMID:27226984

  13. In Silico Analysis of Correlations between Protein Disorder and Post-Translational Modifications in Algae.

    PubMed

    Kurotani, Atsushi; Sakurai, Tetsuya

    2015-08-20

    Recent proteome analyses have reported that intrinsically disordered regions (IDRs) of proteins play important roles in biological processes. In higher plants whose genomes have been sequenced, the correlation between IDRs and post-translational modifications (PTMs) has been reported. The genomes of various eukaryotic algae as common ancestors of plants have also been sequenced. However, no analysis of the relationship to protein properties such as structure and PTMs in algae has been reported. Here, we describe correlations between IDR content and the number of PTM sites for phosphorylation, glycosylation, and ubiquitination, and between IDR content and regions rich in proline, glutamic acid, serine, and threonine (PEST) and transmembrane helices in the sequences of 20 algae proteomes. Phosphorylation, O-glycosylation, ubiquitination, and PEST preferentially occurred in disordered regions. In contrast, transmembrane helices were favored in ordered regions. N-glycosylation tended to occur in ordered regions in most of the studied algae; however, it correlated positively with disordered protein content in diatoms. Additionally, we observed that disordered protein content and the number of PTM sites were significantly increased in the species-specific protein clusters compared to common protein clusters among the algae. Moreover, there were specific relationships between IDRs and PTMs among the algae from different groups.

  14. The aetiology of post-traumatic stress following childbirth: a meta-analysis and theoretical framework.

    PubMed

    Ayers, S; Bond, R; Bertullies, S; Wijma, K

    2016-04-01

    There is evidence that 3.17% of women report post-traumatic stress disorder (PTSD) after childbirth. This meta-analysis synthesizes research on vulnerability and risk factors for birth-related PTSD and refines a diathesis-stress model of its aetiology. Systematic searches were carried out on PsycINFO, PubMed, Scopus and Web of Science using PTSD terms crossed with childbirth terms. Studies were included if they reported primary research that examined factors associated with birth-related PTSD measured at least 1 month after birth. In all, 50 studies (n = 21 429) from 15 countries fulfilled inclusion criteria. Pre-birth vulnerability factors most strongly associated with PTSD were depression in pregnancy (r = 0.51), fear of childbirth (r = 0.41), poor health or complications in pregnancy (r = 0.38), and a history of PTSD (r = 0.39) and counselling for pregnancy or birth (r = 0.32). Risk factors in birth most strongly associated with PTSD were negative subjective birth experiences (r = 0.59), having an operative birth (assisted vaginal or caesarean, r = 0.48), lack of support (r = -0.38) and dissociation (r = 0.32). After birth, PTSD was associated with poor coping and stress (r = 0.30), and was highly co-morbid with depression (r = 0.60). Moderator analyses showed that the effect of poor health or complications in pregnancy was more apparent in high-risk samples. The results of this meta-analysis are used to update a diathesis-stress model of the aetiology of postpartum PTSD and can be used to inform screening, prevention and intervention in maternity care.

  15. Post Tyrrhénian deformation analysis in the Sahel coast (Eastern Tunisia): seismotectonic events implication

    NASA Astrophysics Data System (ADS)

    Mejrei, H.; Ghribi, R.; Bouaziz, S.; Balescu, S.

    2012-04-01

    The eastern coast of Tunisia is characterized by Pleistocene coastal deposits considered as a reference of interglacial high sea levels. In this region, the stratigraphy of Tunisian Pleistocene deposits was first established on the basis of geomorphological, lithostratigraphic, biostratigraphic criteria and U/Th data. They have been subdivided into three superimposed formations, from the oldest to the recent "Douira, Rejiche and Chebba" including coastal marine (Strombus bubonius), lagoonal and eolian sediments. These marine formations are organized into parallel bars to the actual shoreline overlaying unconformably the Mio-Pliocene and "Villafranchian" deposits. A luminescence dating method IRSL applied to alkali feldspar grains from the two sandy marines units of the Douira formation demonstrate for the first time the presence of two successive interglacial high sea level events correlative of MIS 7 and MIS 9. These sandy marine units are separated by a major erosional surface and by a continental pedogenised loamy deposit related to a low sea level event which might be assigned to MIS 8. Variations in the height of these marine unit (+13 to +32m) in the Sahel coast reflect a significant tectonic deformations and show precious geomorphological and tectonic markers. An extensive brittle deformations analysis has been carried out in several sites. A detailed analysis of fracturing is based on studies of fault-slip data population and of joint sets. It allows reconstructions of post Tyrrhenian stress regimes which are characterized by N170-016 compression and N095-100 extension. In this paper we present, the combination of IRSL data applied to these raised marine deposits and a reconstruction of tectonic evolution in term of stress pattern evolution since the Tyrrhenian allowed us to assign an accurate the recent tectonic calendar. These reconstituted events will be replaced and will be discussed in the regional setting of sismotectonic activities of the north

  16. Hydraulic analysis of river training cross-vanes as part of post-restoration monitoring

    NASA Astrophysics Data System (ADS)

    Endreny, T. A.; Soulman, M. M.

    2011-07-01

    River restoration design methods are incrementally improved by studying and learning from monitoring data in previous projects. In this paper we report post-restoration monitoring data and simulation analysis for a Natural Channel Design (NCD) restoration project along 1600 m of the Batavia Kill (14 km2 watershed) in the Catskill Mountains, NY. The restoration project was completed in 2002 with goals to reduce bank erosion and determine the efficacy of NCD approaches for restoring headwater streams in the Catskill Mountains, NY. The NCD approach used a reference-reach to determine channel form, empirical relations between the project site and reference site bankfull dimensions to size channel geometry, and hydraulic and sediment computations based on a bankfull (1.3 yr return interval) discharge to test channel capacity and sediment stability. The NCD project included 12 cross-vanes and 48 j-hook vanes as river training structures along 19 meander bends to protect against bank erosion and maintain scour pools for fish habitat. Monitoring data collected from 2002 to 2004 were used to identify aggradation of pools in meander bends and below some structures. Aggradation in pools was attributed to the meandering riffle-pool channel trending toward step-pool morphology and cross-vane arms not concentrating flow in the center of the channel. The aggradation subsequently caused flow splitting and 4 partial point bar avulsions during a spring 2005 flood with a 25-yr return interval. Processing the pre-flood monitoring data with hydraulic analysis software provided clues the reach was unstable and preventative maintenance was needed. River restoration and monitoring teams should be trained in robust hydraulic analytical methods that help them extend project restoration goals and structure stability.

  17. The aetiology of post-traumatic stress following childbirth: a meta-analysis and theoretical framework.

    PubMed

    Ayers, S; Bond, R; Bertullies, S; Wijma, K

    2016-04-01

    There is evidence that 3.17% of women report post-traumatic stress disorder (PTSD) after childbirth. This meta-analysis synthesizes research on vulnerability and risk factors for birth-related PTSD and refines a diathesis-stress model of its aetiology. Systematic searches were carried out on PsycINFO, PubMed, Scopus and Web of Science using PTSD terms crossed with childbirth terms. Studies were included if they reported primary research that examined factors associated with birth-related PTSD measured at least 1 month after birth. In all, 50 studies (n = 21 429) from 15 countries fulfilled inclusion criteria. Pre-birth vulnerability factors most strongly associated with PTSD were depression in pregnancy (r = 0.51), fear of childbirth (r = 0.41), poor health or complications in pregnancy (r = 0.38), and a history of PTSD (r = 0.39) and counselling for pregnancy or birth (r = 0.32). Risk factors in birth most strongly associated with PTSD were negative subjective birth experiences (r = 0.59), having an operative birth (assisted vaginal or caesarean, r = 0.48), lack of support (r = -0.38) and dissociation (r = 0.32). After birth, PTSD was associated with poor coping and stress (r = 0.30), and was highly co-morbid with depression (r = 0.60). Moderator analyses showed that the effect of poor health or complications in pregnancy was more apparent in high-risk samples. The results of this meta-analysis are used to update a diathesis-stress model of the aetiology of postpartum PTSD and can be used to inform screening, prevention and intervention in maternity care. PMID:26878223

  18. Post-Messinian evolution of the Florence Ridge area (Western Cyprus Arc), Part I: Morphostructural analysis

    NASA Astrophysics Data System (ADS)

    Sellier, N. C.; Loncke, L.; Vendeville, B. C.; Mascle, J.; Zitter, T.; Woodside, J.; Loubrieu, B.

    2013-04-01

    The Florence Ridge, part of the western Cyprus arc, is a compressional relief that was eroded during the Messinian salinity crisis while deposition of salt occurred North (Antalya basin) and South (Herodotus abyssal plain). In order to better assess the impact of salt-tectonics in the Florence Ridge region deformations, we conducted a morpho-structural analysis of available multibeam and seismic data (Simed and Prismed II campaigns). It is indeed a crucial issue to distinguish crustal and gravity driven structures in the compressional to strike-slip belts of the eastern Mediterranean. Along the Antalya basin, we mainly observed multi-directional tectonic rafts typical of gravity gliding above salt. On the Florence Ridge itself, the base of salt evolves laterally to a Messinian erosional surface that erodes a series of stacked nappes. This surface is involved in recent faulting. South of the Florence Ridge, a nearly 100 km wide fold belt characterizes the Herodotus abyssal plain. Three different zones parallel to the Florence Ridge appear within this fold belt. Those are respectively from North to South zones A, B and C. Zone A is characterized mainly by small-wavelength folding and faulting. Approaching Zone B, a long extensional graben deforms the seafloor. Zone B stands ~ 100 to 200 m higher than zones A and C. There, salt welding seems common. In the easternmost zone B deep sub-circular bathymetric depressions are associated with extremely thick and fan-shaped depocenters probably emplaced in relation with active sub-salt thrusts. Many evidences suggest post-Messinian uplift in this zone. Zone C shows medium to high wavelength salt-cored folds. Wavelength of those folds increase approaching the distal Nile deep-sea fan. Within zone C, a nearly undeformed domain exists approaching the Eratosthenes seamount. 'En echelon' folds bound this flat domain suggesting lateral salt extrusion at the junction between zone C and the distal Nile deep-sea fan. To conclude

  19. Clinical Factors Associated with Dose of Loop Diuretics After Pediatric Cardiac Surgery: Post Hoc Analysis.

    PubMed

    Haiberger, Roberta; Favia, Isabella; Romagnoli, Stefano; Cogo, Paola; Ricci, Zaccaria

    2016-06-01

    A post hoc analysis of a randomized controlled trial comparing the clinical effects of furosemide and ethacrynic acid was conducted. Infants undergoing cardiac surgery with cardiopulmonary bypass were included in order to explore which clinical factors are associated with diuretic dose in infants with congenital heart disease. Overall, 67 patients with median (interquartile range) age of 48 (13-139) days were enrolled. Median diuretic dose was 0.34 (0.25-0.4) mg/kg/h at the end of postoperative day (POD) 0 and it significantly decreased (p = 0.04) over the following PODs; during this period, the ratio between urine output and diuretic dose increased significantly (p = 0.04). Age (r -0.26, p = 0.02), weight (r -0.28, p = 0.01), cross-clamp time (r 0.27, p = 0.03), administration of ethacrynic acid (OR 0.01, p = 0.03), and, at the end of POD0, creatinine levels (r 0.3, p = 0.009), renal near-infrared spectroscopy saturation (-0.44, p = 0.008), whole-blood neutrophil gelatinase-associated lipocalin levels (r 0.30, p = 0.01), pH (r -0.26, p = 0.02), urinary volume (r -0.2755, p = 0.03), and fluid balance (r 0.2577, p = 0.0266) showed a significant association with diuretic dose. At multivariable logistic regression cross-clamp time (OR 1.007, p = 0.04), use of ethacrynic acid (OR 0.2, p = 0.01) and blood pH at the end of POD0 (OR 0.0001, p = 0.03) was independently associated with diuretic dose. Early resistance to loop diuretics continuous infusion is evident in post-cardiac surgery infants: Higher doses are administered to patients with lower urinary output. Independently associated variables with diuretic dose in our population appeared to be cross-clamping time, the administration of ethacrynic acid, and blood pH. PMID:26961571

  20. Clinical Factors Associated with Dose of Loop Diuretics After Pediatric Cardiac Surgery: Post Hoc Analysis.

    PubMed

    Haiberger, Roberta; Favia, Isabella; Romagnoli, Stefano; Cogo, Paola; Ricci, Zaccaria

    2016-06-01

    A post hoc analysis of a randomized controlled trial comparing the clinical effects of furosemide and ethacrynic acid was conducted. Infants undergoing cardiac surgery with cardiopulmonary bypass were included in order to explore which clinical factors are associated with diuretic dose in infants with congenital heart disease. Overall, 67 patients with median (interquartile range) age of 48 (13-139) days were enrolled. Median diuretic dose was 0.34 (0.25-0.4) mg/kg/h at the end of postoperative day (POD) 0 and it significantly decreased (p = 0.04) over the following PODs; during this period, the ratio between urine output and diuretic dose increased significantly (p = 0.04). Age (r -0.26, p = 0.02), weight (r -0.28, p = 0.01), cross-clamp time (r 0.27, p = 0.03), administration of ethacrynic acid (OR 0.01, p = 0.03), and, at the end of POD0, creatinine levels (r 0.3, p = 0.009), renal near-infrared spectroscopy saturation (-0.44, p = 0.008), whole-blood neutrophil gelatinase-associated lipocalin levels (r 0.30, p = 0.01), pH (r -0.26, p = 0.02), urinary volume (r -0.2755, p = 0.03), and fluid balance (r 0.2577, p = 0.0266) showed a significant association with diuretic dose. At multivariable logistic regression cross-clamp time (OR 1.007, p = 0.04), use of ethacrynic acid (OR 0.2, p = 0.01) and blood pH at the end of POD0 (OR 0.0001, p = 0.03) was independently associated with diuretic dose. Early resistance to loop diuretics continuous infusion is evident in post-cardiac surgery infants: Higher doses are administered to patients with lower urinary output. Independently associated variables with diuretic dose in our population appeared to be cross-clamping time, the administration of ethacrynic acid, and blood pH.

  1. Role of C-terminal domain and transmembrane helices 5 and 6 in function and quaternary structure of major intrinsic proteins: analysis of aquaporin/glycerol facilitator chimeric proteins.

    PubMed

    Duchesne, Laurence; Pellerin, Isabelle; Delamarche, Christian; Deschamps, Stephane; Lagree, Valerie; Froger, Alexandrine; Bonnec, Georgette; Thomas, Daniel; Hubert, Jean-Francois

    2002-06-01

    We previously observed that aquaporins and glycerol facilitators exhibit different oligomeric states when studied by sedimentation on density gradients following nondenaturing detergent solubilization. To determine the domains of major intrinsic protein (MIP) family proteins involved in oligomerization, we constructed protein chimeras corresponding to the aquaporin AQPcic substituted in the loop E (including the proximal part of transmembrane domain (TM) 5) and/or the C-terminal part (including the distal part of TM 6) by the equivalent domain of the glycerol channel aquaglyceroporin (GlpF) (chimeras called AGA, AAG, and AGG). The analogous chimeras of GlpF were also constructed (chimeras GAG, GGA, and GAA). cRNA corresponding to all constructs were injected into Xenopus oocytes. AQPcic, GlpF, AAG, AGG, and GAG were targeted to plasma membranes. Water or glycerol membrane permeability measurements demonstrated that only the AAG chimera exhibited a channel function corresponding to water transport. Analysis of all proteins expressed either in oocytes or in yeast by velocity sedimentation on sucrose gradients following solubilization by 2% n-octyl glucoside indicated that only AQPcic and AAG exist in tetrameric forms. GlpF, GAG, and GAA sediment in a monomeric form, whereas GGA and AGG were found mono/dimeric. These data bring new evidence that, within the MIP family, aquaporins and GlpFs behave differently toward nondenaturing detergents. We demonstrate that the C-terminal part of AQPcic, including the distal half of TM 6, can be substituted by the equivalent domain of GlpF (AAG chimera) without modifying the transport specificity. Our results also suggest that interactions of TM 5 of one monomer with TM 1 of the adjacent monomer are crucial for aquaporin tetramer stability. PMID:11927589

  2. A chimeric Sindbis-based vaccine protects cynomolgus macaques against a lethal aerosol challenge of eastern equine encephalitis virus

    PubMed Central

    Roy, Chad J.; Adams, A. Paige; Wang, Eryu; Leal, Grace; Seymour, Robert L.; Sivasubramani, Satheesh K.; Mega, William; Frolov, Ilya; Didier, Peter J.; Weaver, Scott C.

    2013-01-01

    Eastern equine encephalitis virus (EEEV) is a mosquito-borne alphavirus that causes sporadic, often fatal disease outbreaks in humans and equids, and is also a biological threat agent. Two chimeric vaccine candidates were constructed using a cDNA clone with a Sindbis virus (SINV) backbone and structural protein genes from either a North (SIN/NAEEEV) or South American (SIN/SAEEEV) strain of EEEV. The vaccine candidates were tested in a nonhuman primate (NHP) model of eastern equine encephalitis (EEE). Cynomolgus macaques were either sham-vaccinated, or vaccinated with a single dose of either SIN/NAEEEV or SIN/SAEEEV. After vaccination, animals were challenged by aerosol with a virulent North American strain of EEEV (NA EEEV). The SIN/NAEEEV vaccine provided significant protection, and most vaccinated animals survived EEEV challenge (82%) with little evidence of disease, whereas most SIN/SAEEEV-vaccinated (83%) and control (100%) animals died. Protected animals exhibited minimal changes in temperature and cardiovascular rhythm, whereas unprotected animals showed profound hyperthermia and changes in heart rate post-exposure. Acute inflammation and neuronal necrosis were consistent with EEEV-induced encephalitis in unprotected animals, whereas no encephalitis-related histopathologic changes were observed in the SIN/NAEEEV-vaccinated animals. These results demonstrate that the chimeric SIN/NAEEEV vaccine candidate protects against an aerosol EEEV exposure. PMID:23333212

  3. Generation of transgenic rabbits by the novel technique of chimeric somatic cell cloning.

    PubMed

    Skrzyszowska, M; Smorag, Z; Słomski, R; Katska-Ksiazkiewicz, L; Kalak, R; Michalak, E; Wielgus, K; Lehmann, J; Lipiński, D; Szalata, M; Pławski, A; Samiec, M; Jura, J; Gajda, B; Ryńska, B; Pieńkowski, M

    2006-06-01

    A novel technique of chimeric somatic cell cloning was applied to produce a transgenic rabbit (NT20). Karyoplasts of transgenic adult skin fibroblasts with Tg(Wap-GH1) gene construct as a marker were microsurgically transferred into one, previously enucleated, blastomere of 2-cell non-transgenic embryos, while the second one remained intact. The reconstructed embryos either were cultured in vitro up to the blastocyst stage (Experiment I) or were transferred into recipient-females immediately after the cloning procedure (Experiment II). In Experiment I, 25/102 (24.5%) embryos formed blastocysts from whole embryos and 46/102 (44.12%) embryos developed to the blastocyst stage from single non-operated blastomeres, while the reconstructed blastomeres were damaged and degenerated. Thirteen (12.7%) embryos did not exceed 3- to 4-cell stages and 18 (17.7%) embryos were inhibited at the initial 2-cell stage. Out of 14 blastocysts which were subjected to molecular analysis, the transgene was detected in the cells of 4 blastocysts. In Experiment II, 163/217 (75.0%) embryos were transferred into 9 pseudopregnant recipient-rabbits (an average of 18 embryos per recipient). Four recipient-females (44.4%) became pregnant and delivered a total of 24 (14.7%) pups. Molecular analysis confirmed that two pups (1.2%), one live and one stillborn, showed a positive transgene signal. Live transgenic rabbit NT20 appeared healthy and anatomically as well as physiologically normal. The results of our experiments showed that transgenic adult skin fibroblast cell nuclei, which have been introduced into the cytoplasmic microenvironment of single enucleated blastomeres from 2-cell stage rabbit embryos, are able to direct the development of chimeric embryos not only to the blastocyst stage but also up to term.

  4. Sex Differences in Hookah-Related Images Posted on Tumblr: A Content Analysis.

    PubMed

    Primack, Brian A; Carroll, Mary V; Shensa, Ariel; Davis, Wesley; Levine, Michele D

    2016-01-01

    Hookah tobacco smoking is prevalent, widespread, and associated with large amounts of toxicants. Hookah tobacco smoking may be viewed differently by males and females. For example, females have been drawn to types of tobacco that are flavored, milder, and marketed as more social and exotic. Individuals often use the growing segment of anonymous social networking sites, such as Tumblr, to learn about potentially dangerous or harmful behaviors. We used a systematic process involving stratification by time of day, day of week, and search term to gather a sample of 140 Tumblr posts related to hookah tobacco smoking. After a structured codebook development process, 2 coders independently assessed all posts in their entirety, and all disagreements were easily adjudicated. When data on poster sex and age were available, we found that 77% of posts were posted by females and 35% were posted by individuals younger than 18. The most prominent features displayed in all posts were references to or images of hookahs themselves, sexuality, socializing, alcohol, hookah smoke, and tricks performed with hookah smoke. Compared with females, males more frequently posted images of hookahs and alcohol-related images or references. This information may help guide future research in this area and the development of targeted interventions to curb this behavior. PMID:26890733

  5. Sex Differences in Hookah-Related Images Posted on Tumblr: A Content Analysis.

    PubMed

    Primack, Brian A; Carroll, Mary V; Shensa, Ariel; Davis, Wesley; Levine, Michele D

    2016-01-01

    Hookah tobacco smoking is prevalent, widespread, and associated with large amounts of toxicants. Hookah tobacco smoking may be viewed differently by males and females. For example, females have been drawn to types of tobacco that are flavored, milder, and marketed as more social and exotic. Individuals often use the growing segment of anonymous social networking sites, such as Tumblr, to learn about potentially dangerous or harmful behaviors. We used a systematic process involving stratification by time of day, day of week, and search term to gather a sample of 140 Tumblr posts related to hookah tobacco smoking. After a structured codebook development process, 2 coders independently assessed all posts in their entirety, and all disagreements were easily adjudicated. When data on poster sex and age were available, we found that 77% of posts were posted by females and 35% were posted by individuals younger than 18. The most prominent features displayed in all posts were references to or images of hookahs themselves, sexuality, socializing, alcohol, hookah smoke, and tricks performed with hookah smoke. Compared with females, males more frequently posted images of hookahs and alcohol-related images or references. This information may help guide future research in this area and the development of targeted interventions to curb this behavior.

  6. Mixed chimerism to induce tolerance for solid organ transplantation

    SciTech Connect

    Wren, S.M.; Nalesnik, M.; Hronakes, M.L.; Oh, E.; Ildstad, S.T. )

    1991-04-01

    Chimerism, or the coexistence of tissue elements from more than one genetically different strain or species in an organism, is the only experimental state that results in the induction of donor-specific transplantation tolerance. Transplantation of a mixture of T-cell-depleted syngeneic (host-type) plus T-cell-depleted allogeneic (donor) bone marrow into a normal adult recipient mouse (A + B----A) results in mixed allogeneic chimerism. Recipient mice exhibit donor-specific transplantation tolerance, yet have full immunocompetence to recognize and respond to third-party transplantation antigens. After complete hematolymphopoietic repopulation at 28 days, animals accept a donor-specific skin graft but reject major histocompatibility complex (MHC) locus-disparate third-party grafts. We now report that permanent graft acceptance can also be achieved when the graft is placed at the time of bone marrow transplantation. Histologically, grafts were viable and had only minimal inflammatory changes. This model may have potential future clinical application for the induction of donor-specific transplantation tolerance.

  7. Finite element stress analysis of short-post core and over restorations prepared with different restorative materials.

    PubMed

    Gurbuz, Taskin; Sengul, Fatih; Altun, Ceyhan

    2008-07-01

    The present study was conducted to determine the effect on the distribution of stress with the use of short-post cores and over restorations composed of different materials. The restorative materials used were namely two different composite resin materials (Valux Plus and Tetric Flow), a polyacid-modified resin material (Dyract AP), and a woven polyethylene fiber combination (Ribbond Fiber + Bonding agent + Tetric Flow). Finite element analysis (FEA) was used to develop a model for the maxillary primary anterior teeth. A masticatory force of 100 N was applied at 148 degrees to the incisal edge of the palatal surface of the crown model. Stress distributions and stress values were compared using von Mises criteria. The tooth model was assumed to be isotropic, homogeneous, elastic, and asymmetrical. It was observed that the highest stress usually occurred in the cervical area of the tooth when Tetric Flow was used as the short-post core and over restoration material. The same maximum stress value was also obtained when Ribbond fiber + Tetric Flow material was used for the short-post core. The results of FEA showed that the mechanical properties and elastic modulus of the restorative material influenced the stresses generated in enamel, dentin, and restoration when short-post core restorations were loaded incisally. Resin-based restorative materials with higher elastic moduli were found to be unsuitable as short-post core materials in endodontically treated maxillary primary anterior teeth. PMID:18833762

  8. Evaluation of Temperature and Stress Distribution on 2 Different Post Systems Using 3-Dimensional Finite Element Analysis

    PubMed Central

    Değer, Yalçın; Adigüzel, Özkan; Özer, Senem Yiğit; Kaya, Sadullah; Polat, Zelal Seyfioğlu; Bozyel, Bejna

    2015-01-01

    Background The mouth is exposed to thermal irritation from hot and cold food and drinks. Thermal changes in the oral cavity produce expansions and contractions in tooth structures and restorative materials. The aim of this study was to investigate the effect of temperature and stress distribution on 2 different post systems using the 3-dimensional (3D) finite element method. Material/Methods The 3D finite element model shows a labio-lingual cross-sectional view of the endodontically treated upper right central incisor and supporting periodontal ligament with bone structures. Stainless steel and glass fiber post systems with different physical and thermal properties were modelled in the tooth restored with composite core and ceramic crown. We placed 100 N static vertical occlusal loading onto the center of the incisal surface of the tooth. Thermal loads of 0°C and 65°C were applied on the model for 5 s. Temperature and thermal stresses were determined on the labio-lingual section of the model at 6 different points. Results The distribution of stress, including thermal stress values, was calculated using 3D finite element analysis. The stainless steel post system produced more temperature and thermal stresses on the restorative materials, tooth structures, and posts than did the glass fiber reinforced composite posts. Conclusions Thermal changes generated stresses in the restorative materials, tooth, and supporting structures. PMID:26615495

  9. Finite element stress analysis of short-post core and over restorations prepared with different restorative materials.

    PubMed

    Gurbuz, Taskin; Sengul, Fatih; Altun, Ceyhan

    2008-07-01

    The present study was conducted to determine the effect on the distribution of stress with the use of short-post cores and over restorations composed of different materials. The restorative materials used were namely two different composite resin materials (Valux Plus and Tetric Flow), a polyacid-modified resin material (Dyract AP), and a woven polyethylene fiber combination (Ribbond Fiber + Bonding agent + Tetric Flow). Finite element analysis (FEA) was used to develop a model for the maxillary primary anterior teeth. A masticatory force of 100 N was applied at 148 degrees to the incisal edge of the palatal surface of the crown model. Stress distributions and stress values were compared using von Mises criteria. The tooth model was assumed to be isotropic, homogeneous, elastic, and asymmetrical. It was observed that the highest stress usually occurred in the cervical area of the tooth when Tetric Flow was used as the short-post core and over restoration material. The same maximum stress value was also obtained when Ribbond fiber + Tetric Flow material was used for the short-post core. The results of FEA showed that the mechanical properties and elastic modulus of the restorative material influenced the stresses generated in enamel, dentin, and restoration when short-post core restorations were loaded incisally. Resin-based restorative materials with higher elastic moduli were found to be unsuitable as short-post core materials in endodontically treated maxillary primary anterior teeth.

  10. Post-radiation changes in oral tissues - An analysis of cancer irradiation cases

    PubMed Central

    Pandya, Jay Ashokkumar; Srikant, N.; Boaz, Karen; Manaktala, Nidhi; Kapila, Supriya Nikita; Yinti, Shanmukha Raviteja

    2014-01-01

    Introduction: Radiation, commonly employed as neoadjuvant, primary, and adjuvant therapy for head and neck cancer causes numerous epithelial and stromal changes, prominent among which is fibrosis with its early and late consequences. Very little is known about the true nature of the fibrosed tissue and the type of fibers accumulated. Radiotherapy affects the supporting tumor stroma often resulting in a worsening grade of tumor post-radiation. Aim: To study epithelial, neoplastic, stromal, and glandular changes in oral cavity induced by radiation therapy for oral squamous cell carcinoma (OSCC) using special stains. Materials and Methods: The study included 27 samples of recurrent OSCC following completion of radiotherapy (recurrence within an average span of 11 months), and 26 non-irradiated cases of OSCC. Patients with a history of combined radiotherapy and chemotherapy were not included in the study. The epithelial changes assessed included epithelial atrophy, apoptosis, necrosis, dysplasia, and neoplasia. The connective tissue was evaluated for amount of fibrosis, quality of fibers (using picrosirius red staining), fibrinous exudate, necrosis, pattern of invasion, vessel wall thickening, and salivary gland changes. The aforementioned changes were assessed using light and polarizing microscopy and tabulated. Statistical Analysis: Epithelial and connective tissue parameters were compared between the irradiated and non-irradiated cases using chi square and t-tests. Results: Epithelial and connective tissue parameters were found to be increased in irradiated patients. Pattern of invasion by tumor cells varied from strands and  cords between the two groups studied. The effect of radiation was seen to reflect on the maturity of fibers and the regularity of their distribution. PMID:25136522

  11. Building, Testing, and Post Test Analysis of Durability Heat Pipe No.6

    SciTech Connect

    MOSS, TIMOTHY A.

    2002-03-01

    The Solar Thermal Program at Sandia supports work developing dish/Stirling systems to convert solar energy into electricity. Heat pipe technology is ideal for transferring the energy of concentrated sunlight from the parabolic dish concentrators to the Stirling engine heat tubes. Heat pipes can absorb the solar energy at non-uniform flux distributions and release this energy to the Stirling engine heater tubes at a very uniform flux distribution thus decoupling the design of the engine heater head from the solar absorber. The most important part of a heat pipe is the wick, which transports the sodium over the heated surface area. Bench scale heat pipes were designed and built to more economically, both in time and money, test different wicks and cleaning procedures. This report covers the building, testing, and post-test analysis of the sixth in a series of bench scale heat pipes. Durability heat pipe No.6 was built and tested to determine the effects of a high temperature bakeout, 950 C, on wick corrosion during long-term operation. Previous tests showed high levels of corrosion with low temperature bakeouts (650-700 C). Durability heat pipe No.5 had a high temperature bakeout and reflux cleaning and showed low levels of wick corrosion after long-term operation. After testing durability heat pipe No.6 for 5,003 hours at an operating temperature of 750 C, it showed low levels of wick corrosion. This test shows a high temperature bakeout alone will significantly reduce wick corrosion without the need for costly and time consuming reflux cleaning.

  12. Post-Flight Analysis of Dynamic Data Acquired During the ATV-2 Johannes Kepler Launch

    NASA Astrophysics Data System (ADS)

    Meitzner, R.; Abdoly, K.; Newerla, A.

    2012-07-01

    An in-flight data acquisition system called TeleMesure Autonome (TMA) has been implemented on ATV-1 Jules Verne (launched in March 2008) and ATV-2 Johannes Kepler (launched in February 2011). The TMA served the main objective to measure dynamic responses on the ATV spacecraft for comparison with coupled load analysis predictions and to verify that the ATV mechanical flight environment has been sufficiently covered by the respective ATV design specifications. The acquired flight data included low frequency sinusoidal, random vibration and shock measurements. Whereas the TMA on ATV-1 Jules Verne failed to properly work after 17 seconds after liftoff the improved TMA on ATV-2 Johannes Kepler performed its tasks successfully for all flight phases. The flight data have been subsequently evaluated by the ATV prime contractor Astrium. As first step of the performed analyses a correction of the acquired data was necessary to remove any artificial content (spikes, mean truncation, offset correction) followed by a visual inspection of the corrected data to ensure data quality. Then standard post processing methods were applied to the data consisting of generating equivalent sinusoidal responses and transfer functions for the low frequency data, power spectral densities for the random data and shock responses for the high frequency shock data. Although it was noted that the quality of the data was limited by the available transmission bandwidth and the amplitude resolution of the data acquisition system the implementation of the TMA on ATV-2 Johannes Kepler has nevertheless turned out successful and valuable data have been acquired for all relevant flight phases.

  13. Radial-Velocity Analysis of the Post-AGB Star, HD101584

    NASA Astrophysics Data System (ADS)

    Díaz, F.; Hearnshaw, J.; Rosenzweig, P.; Guzman, E.; Sivarani, T.; Parthasarathy, M.

    2007-08-01

    This project concerns the analysis of the periodicity of the radial velocity of the peculiar emission-line supergiant star HD 101584 (F0 Ia), and also we propose a physical model to account for the observations. From its peculiarities, HD 101584 is a star that is in the post-AGB phase. This study is considered as a key to clarify the multiple aspects related with the evolution of the circum-stellar layer associated with this star's last phase. The star shows many lines with P Cygni profiles, including H-alpha, Na D lines in the IR Ca triplet, indicating a mass outflow. For HD 101584 we have performed a detailed study of its radial-velocity variations, using both emission and absorption lines over a wide range of wavelength. We have analyzed the variability and found a periodicity for all types of lines of 144 days, which must arise from the star's membership in a binary system. The data span a period of five consecutive years and were obtained using the 1-m telescope of Mt John Observatory, in New Zealand., with the echelle and Hercules high resolution spectrographs and CCD camera. HD101584 is known to be an IRAS source, and our model suggests it is a proto-planetary nebula, probably with a bipolar outflow and surrounded by a dusty disk as part of a binary system. We have found no evidence for HD101584 to contain a B9 star as found by Bakker et al (1996). A low resolution IUE spectrum shows the absence of any strong UV continuum that would be expected for a B star to be in this system.

  14. Comparison of different blood sample processing methods for sensitive detection of low level chimerism by RHD real-time PCR assay.

    PubMed

    Javadi, Ahmad; Verduin, Esther P; Brand, Anneke; Schonewille, Henk

    2013-01-01

    The rhesus D blood group, which is expressed on the red blood cells (RBC) of 85% of the Caucasian population, is one of the most immunogenic RBC antigens, inducing D antibody formation in up to 20-80% of D-negative transfusion recipients and about 10% of pregnancies at risk. Pregnancy-induced D-antibodies can persist for many years, but the mechanisms underlying this persistence are unclear. The LOTUS study, a long-term follow-up study of mothers from severely affected children with hemolytic disease of the fetus and newborn investigates, among other endpoints, whether persistent feto-maternal chimerism is associated with long-term maternal anti-D persistence. We questioned which blood sample processing method should be used to detect low levels of RHD chimerism with the highest sensitivity and specificity using qPCR. After optimization of primer and probe concentrations for singleplex RHD exon 5 and 7 qPCR, sensitivity, specificity and efficiency of RHD and DYS1 qPCR were investigated in artificial chimeric samples. Sensitivity of DYS1 was one log higher (0.0001%) in enriched mononuclear cell fractions as compared with whole blood. Comparable linear sensitivity (0.007%) and mean efficiency (84-99%) for RHD qPCR were observed in all samples regardless whether whole blood or pre- or post-mixing of cellular fractions had been used. We conclude that RHD chimerism using singleplex exon 5 and 7 qPCR is linearly detectable down to 1.0 GE, without an advantage of fraction enrichment.

  15. Chimeric External Control to Quantify Cell Free DNA in Plasma Samples by Real Time PCR

    PubMed Central

    Eini, Maryam; Behzad-Behbahani, Abbas; Takhshid, Mohammad Ali; Ramezani, Amin; Rafiei Dehbidi, Gholam Reza; Okhovat, Mohammad Ali; Farhadi, Ali; Alavi, Parniyan

    2016-01-01

    Background: DNA isolation procedure can significantly influence the quantification of DNA by real time PCR specially when cell free DNA (cfDNA) is the subject. To assess the extraction efficiency, linearity of the extraction yield, presence of co-purified inhibitors and to avoid problems with fragment size relevant to cfDNA, development of appropriate External DNA Control (EDC) is challenging. Using non-human chimeric nucleotide sequences, an EDC was developed for standardization of qPCR for monitoring stability of cfDNA concentration in blood samples over time. Methods: A0 DNA fragment of 167 bp chimeric sequence of parvovirus B19 and pBHA designated as EDC fragment was designed. To determine the impact of different factors during DNA extraction processing on quantification of cfDNA, blood samples were collected from normal subjects and divided into aliquots with and without specific treatment. In time intervals, the plasma samples were isolated. The amplicon of 167 bp EDC fragment in final concentration of 1.1 pg/500 μl was added to each plasma sample and total DNA was extracted by an in house method. Relative and absolute quantification real time PCR was performed to quantify both EDC fragment and cfDNA in extracted samples. Results: Comparison of real time PCR threshold cycle (Ct) for cfDNA fragment in tubes with and without specific treatment indicated a decrease in untreated tubes. In contrast, the threshold cycle was constant for EDC fragment in treated and untreated tubes, indicating the difference in Ct values of the cfDNA is because of specific treatments that were made on them. Conclusions: Spiking of DNA fragment size relevant to cfDNA into the plasma sample can be useful to minimize the bias due to sample preparation and extraction processing. Therefore, it is highly recommended that standard external DNA control be employed for the extraction and quantification of cfDNA for accurate data analysis. PMID:27141267

  16. Repeated Evolution of Chimeric Fusion Genes in the β-Globin Gene Family of Laurasiatherian Mammals

    PubMed Central

    Gaudry, Michael J.; Storz, Jay F.; Butts, Gary Tyler; Campbell, Kevin L.; Hoffmann, Federico G.

    2014-01-01

    The evolutionary fate of chimeric fusion genes may be strongly influenced by their recombinational mode of origin and the nature of functional divergence between the parental genes. In the β-globin gene family of placental mammals, the two postnatally expressed δ- and β-globin genes (HBD and HBB, respectively) have a propensity for recombinational exchange via gene conversion and unequal crossing-over. In the latter case, there are good reasons to expect differences in retention rates for the reciprocal HBB/HBD and HBD/HBB fusion genes due to thalassemia pathologies associated with the HBD/HBB “Lepore” deletion mutant in humans. Here, we report a comparative genomic analysis of the mammalian β-globin gene cluster, which revealed that chimeric HBB/HBD fusion genes originated independently in four separate lineages of laurasiatherian mammals: Eulipotyphlans (shrews, moles, and hedgehogs), carnivores, microchiropteran bats, and cetaceans. In cases where an independently derived “anti-Lepore” duplication mutant has become fixed, the parental HBD and/or HBB genes have typically been inactivated or deleted, so that the newly created HBB/HBD fusion gene is primarily responsible for synthesizing the β-type subunits of adult and fetal hemoglobin (Hb). Contrary to conventional wisdom that the HBD gene is a vestigial relict that is typically inactivated or expressed at negligible levels, we show that HBD-like genes often encode a substantial fraction (20–100%) of β-chain Hbs in laurasiatherian taxa. Our results indicate that the ascendancy or resuscitation of genes with HBD-like coding sequence requires the secondary acquisition of HBB-like promoter sequence via unequal crossing-over or interparalog gene conversion. PMID:24814285

  17. Selective autoantibody production by Yaa+ B cells in autoimmune Yaa(+)- Yaa- bone marrow chimeric mice

    PubMed Central

    1991-01-01

    The accelerated autoimmune syndrome observed in BXSB/MpJ male mice is associated with the presence on the Y chromosome of an as yet unidentified mutant gene, designated Y chromosome-linked autoimmune acceleration (Yaa). To study the mechanisms by which the Yaa gene accelerates and/or induces the production of autoantibodies, we have developed double-congenic bone marrow chimeras containing B cells from autoimmune males carrying the Yaa gene, and from nonautoimmune male or female mice lacking it and differing by the Igh allotype. The analysis of the allotype of total immunoglobulins and anti-DNA antibodies in Yaa+ male-normal female (Yaa-) chimeric mice revealed that the selective activation of B cells from autoimmune Yaa+ male mice was responsible for the hypergammaglobulinemia and autoantibody production. This phenomenon was not due to an anti-HY interaction between female T helper cells and male B cells, because first, Yaa+ B cells were selectively stimulated to produce autoantibodies in Yaa+ male-Yaa- male chimeric mice; and second, normal male and female chimeras failed to develop an autoimmune syndrome. In addition, the fact that both B cell populations in Yaa(+)-Yaa- chimeras similarly responded to a foreign antigen, human IgG, argues against the possibility that the selective activation of Yaa+ B cells may be due to their hyper-responsiveness to T helper signals. We propose that a cognate interaction of T helper cells with Yaa+ B cells, because of possible T cell recognition of a Yaa-related molecule expressed on Yaa+ B cells, may be responsible for the acceleration and/or induction of autoantibodies in BXSB/MpJ mice. PMID:1834759

  18. Repeated evolution of chimeric fusion genes in the β-globin gene family of laurasiatherian mammals.

    PubMed

    Gaudry, Michael J; Storz, Jay F; Butts, Gary Tyler; Campbell, Kevin L; Hoffmann, Federico G

    2014-05-09

    The evolutionary fate of chimeric fusion genes may be strongly influenced by their recombinational mode of origin and the nature of functional divergence between the parental genes. In the β-globin gene family of placental mammals, the two postnatally expressed δ- and β-globin genes (HBD and HBB, respectively) have a propensity for recombinational exchange via gene conversion and unequal crossing-over. In the latter case, there are good reasons to expect differences in retention rates for the reciprocal HBB/HBD and HBD/HBB fusion genes due to thalassemia pathologies associated with the HBD/HBB "Lepore" deletion mutant in humans. Here, we report a comparative genomic analysis of the mammalian β-globin gene cluster, which revealed that chimeric HBB/HBD fusion genes originated independently in four separate lineages of laurasiatherian mammals: Eulipotyphlans (shrews, moles, and hedgehogs), carnivores, microchiropteran bats, and cetaceans. In cases where an independently derived "anti-Lepore" duplication mutant has become fixed, the parental HBD and/or HBB genes have typically been inactivated or deleted, so that the newly created HBB/HBD fusion gene is primarily responsible for synthesizing the β-type subunits of adult and fetal hemoglobin (Hb). Contrary to conventional wisdom that the HBD gene is a vestigial relict that is typically inactivated or expressed at negligible levels, we show that HBD-like genes often encode a substantial fraction (20-100%) of β-chain Hbs in laurasiatherian taxa. Our results indicate that the ascendancy or resuscitation of genes with HBD-like coding sequence requires the secondary acquisition of HBB-like promoter sequence via unequal crossing-over or interparalog gene conversion.

  19. Post-treatment of molasses wastewater by electrocoagulation and process optimization through response surface analysis.

    PubMed

    Tsioptsias, C; Petridis, D; Athanasakis, N; Lemonidis, I; Deligiannis, A; Samaras, P

    2015-12-01

    Molasses wastewater is a high strength effluent of food industry such as distilleries, sugar and yeast production plants etc. It is characterized by a dark brown color and exhibits a high content in substances of recalcitrant nature such as melanoidins. In this study, electrocoagulation (EC) was studied as a post treatment step for biologically treated molasses wastewater with high nitrogen content obtained from a baker's yeast industry. Iron and copper electrodes were used in various forms; the influence and interaction of current density, molasses wastewater dilution, and reaction time, on COD, color, ammonium and nitrate removal rates and operating cost were studied and optimized through Box Behnken's response surface analysis. Reaction time varied from 0.5 to 4 h, current density varied from 5 to 40 mA/cm(2) and dilution from 0 to 90% (v/v expressed as water concentration). pH, conductivity and temperature measurements were also carried out during each experiment. From preliminary experiments, it was concluded that the application of aeration and sample dilution, considerably influenced the kinetics of the process. The obtained results showed that COD removal varied between 10 and 54%, corresponding to an operation cost ranging from 0.2 to 33 euro/kg COD removed. Significant removal rates were obtained for nitrogen as nitrate and ammonium (i.e. 70% ammonium removal). A linear relation of COD and ammonium to the design parameters was observed, while operation cost and nitrate removal responded in a curvilinear function. A low ratio of electrode surface to treated volume was used, associated to a low investment cost; in addition, iron wastes could be utilized as low cost electrodes i.e. iron fillings from lathes, aiming to a low operation cost due to electrodes replacement. In general, electrocoagulation proved to be an effective and low cost process for biologically treated molasses-wastewater treatment for additional removal of COD and nitrogen content and

  20. Post-treatment of molasses wastewater by electrocoagulation and process optimization through response surface analysis.

    PubMed

    Tsioptsias, C; Petridis, D; Athanasakis, N; Lemonidis, I; Deligiannis, A; Samaras, P

    2015-12-01

    Molasses wastewater is a high strength effluent of food industry such as distilleries, sugar and yeast production plants etc. It is characterized by a dark brown color and exhibits a high content in substances of recalcitrant nature such as melanoidins. In this study, electrocoagulation (EC) was studied as a post treatment step for biologically treated molasses wastewater with high nitrogen content obtained from a baker's yeast industry. Iron and copper electrodes were used in various forms; the influence and interaction of current density, molasses wastewater dilution, and reaction time, on COD, color, ammonium and nitrate removal rates and operating cost were studied and optimized through Box Behnken's response surface analysis. Reaction time varied from 0.5 to 4 h, current density varied from 5 to 40 mA/cm(2) and dilution from 0 to 90% (v/v expressed as water concentration). pH, conductivity and temperature measurements were also carried out during each experiment. From preliminary experiments, it was concluded that the application of aeration and sample dilution, considerably influenced the kinetics of the process. The obtained results showed that COD removal varied between 10 and 54%, corresponding to an operation cost ranging from 0.2 to 33 euro/kg COD removed. Significant removal rates were obtained for nitrogen as nitrate and ammonium (i.e. 70% ammonium removal). A linear relation of COD and ammonium to the design parameters was observed, while operation cost and nitrate removal responded in a curvilinear function. A low ratio of electrode surface to treated volume was used, associated to a low investment cost; in addition, iron wastes could be utilized as low cost electrodes i.e. iron fillings from lathes, aiming to a low operation cost due to electrodes replacement. In general, electrocoagulation proved to be an effective and low cost process for biologically treated molasses-wastewater treatment for additional removal of COD and nitrogen content and

  1. Reproducible Analysis of Post-Translational Modifications in Proteomes—Application to Human Mutations

    PubMed Central

    Holehouse, Alex S.; Naegle, Kristen M.

    2015-01-01

    Background Protein post-translational modifications (PTMs) are an important aspect of protein regulation. The number of PTMs discovered within the human proteome, and other proteomes, has been rapidly expanding in recent years. As a consequence of the rate in which new PTMs are identified, analysis done in one year may result in different conclusions when repeated in subsequent years. Among the various functional questions pertaining to PTMs, one important relationship to address is the interplay between modifications and mutations. Specifically, because the linear sequence surrounding a modification site often determines molecular recognition, it is hypothesized that mutations near sites of PTMs may be more likely to result in a detrimental effect on protein function, resulting in the development of disease. Methods and Results We wrote an application programming interface (API) to make analysis of ProteomeScout, a comprehensive database of PTMs and protein information, easy and reproducible. We used this API to analyze the relationship between PTMs and human mutations associated with disease (based on the ‘Clinical Significance’ annotation from dbSNP). Proteins containing pathogenic mutations demonstrated a significant study bias which was controlled for by analyzing only well-studied proteins, based on their having at least one pathogenic mutation. We found that pathogenic mutations are significantly more likely to lie within eight amino acids of a phosphoserine, phosphotyrosine or ubiquitination site when compared to mutations in general, based on a Fisher’s Exact test. Despite the skew of pathogenic mutations occurring on positively charged arginines, we could not account for this relationship based only on residue type. Finally, we hypothesize a potential mechanism for a pathogenic mutation on RAF1, based on its proximity to a phosphorylation site, which represents a subtle regulation difference that may explain why its biochemical effect has failed to

  2. Analysis of Solar Wind Samples Returned by Genesis Using Laser Post Ionization Secondary Neutral Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Veryovkin, I. V.; Calaway, W. F.; Tripa, C. E.; Pellin, M. J.; Burnett, D. S.

    2005-12-01

    A new secondary neutral mass spectrometry (SNMS) instrument implementing laser post ionization (LPI) of ion sputtered and laser desorbed neutral species has been developed and constructed for the specific purpose of quantitative analysis of metallic elements at ultra trace levels in solar wind collector samples returned to Earth by the Genesis Discovery mission. The first LPI SNMS measurements are focusing on determining Al, Ca, Cr, and Mg in these samples. These measurements provide the first concentration and isotopic abundances determinations for several key metallic elements and also elucidate possible fractionation effects between the photosphere and the solar wind compositions. It is now documented that Genesis samples suffered surface contamination both during flight and during the breach of the Sample Return Capsule when it crashed. Since accurate quantitative analysis is compromised by sample contamination, several features have been built into the new LPI SNMS instrument to mitigate this difficulty. A normally-incident, low-energy (<500 eV) ion beam combined with a keV energy ion beam and a desorbing laser beam (both microfocused) enables dual beam analyses. The low-energy ion beam can be used to remove surface contaminant by sputtering with minimum ion beam mixing. This low-energy beam also will be used to perform ion beam milling, while either the microfocused ion or laser beam probes the solar wind elemental compositions as a function of sample depth. Because of the high depth resolution of dual beam analyses, such depth profiles clearly distinguish between surface contaminants and solar wind implanted atoms. In addition, in-situ optical and electron beam imaging for observing and avoiding particulates and scratches on solar wind sample surfaces is incorporated in the new LPI SNMS instrument to further reduce quantification problems. The current status of instrument tests and analyses will be presented. This work is supported by the U. S. Department of

  3. Harnessing Reconsolidation to Weaken Fear and Appetitive Memories: A Meta-Analysis of Post-Retrieval Extinction Effects

    PubMed Central

    Kredlow, M. Alexandra; Unger, Leslie D.; Otto, Michael W.

    2015-01-01

    A new understanding of the mechanisms of memory retrieval and reconsolidation holds the potential for improving exposure-based treatments. Basic research indicates that following fear extinction, safety and fear memories may compete, raising the possibility of return of fear. One possible solution is to modify original fear memories through reconsolidation interference, reducing the likelihood of return of fear. Post-retrieval extinction is a behavioral method of reconsolidation interference that has been explored in the context of conditioned fear and appetitive memory paradigms. This meta-analysis examines the magnitude of post-retrieval extinction effects and potential moderators of these effects. A PubMed and PsycINFO search was conducted through June 2014. Sixty-three comparisons examining post-retrieval extinction for preventing the return of fear or appetitive responses in animals or humans met inclusion criteria. Post-retrieval extinction demonstrated a significant, small-to-moderate effect (g = .40) for further reducing the return of fear in humans and a significant, large effect (g = 0.89) for preventing the return of appetitive responses in animals relative to standard extinction. For fear outcomes in animals, effects were small (g = 0.21) and non-significant, but moderated by the number of animals housed together and the duration of time between post-retrieval extinction/extinction and test. Across paradigms, these findings support the efficacy of this pre-clinical strategy for preventing the return of conditioned fear and appetitive responses. Overall, findings to date support the continued translation of post-retrieval extinction research to human and clinical applications, with particular application to the treatment of anxiety, traumatic stress, and substance use disorders. PMID:26689086

  4. New insights into the electrode mechanism of lithium sulfur batteries via air-free post-test analysis.

    PubMed

    Chen, Lin; Dietz Rago, Nancy L; Bloom, Ira D; Shaw, Leon L

    2016-08-01

    Effects of the volume expansion and shrinkage of Li2S cathodes on electrochemical cycle life are investigated via post-test analysis without exposure to air. The engineered electrodes that confine volume changes within micro-reactors have significantly longer life than the electrodes without the micro-reactor structure, providing the first unambiguous evidence of the importance of confining volume changes for improved battery performance. PMID:27430393

  5. A Post Hoc Analysis of D-Threo-Methylphenidate Hydrochloride (Focalin) Versus D,l-Threo-Methylphenidate Hydrochloride (Ritalin)

    ERIC Educational Resources Information Center

    Weiss, Margaret; Wasdell, Michael; Patin, John

    2004-01-01

    Objective: To evaluate clinical measures of the benefit/risk ratio in a post hoc analysis of a clinical trial of d-threo-methylphenidate hydrochloride (d-MPH) and d,l-threo-methylphenidate hydrochloride (d,l-MPH). Method: Data from a phase III clinical trial was used to compare equimolar doses of d-MPH and d,l-MPH treatment for…

  6. Thermal Analysis of Post-eruption Loops from 80,000 to 1.6 million K

    NASA Technical Reports Server (NTRS)

    Kucera, T.; Landi, E.

    2006-01-01

    We analyze the thermal properties of a set of post eruptive loops which appeared after a prominence eruption on April 30, 2004. The event was observed by TRACE and SOHO/SUMER. The SUMER data was taken from a single slit location with a 90 second cadence and included a number of lines spanning the temperature range 80,000 to 1.6 million K. We perform a differential emission measure analysis of the loops in order to study their thermal evolution.

  7. Characterization of hemopoietic stem cell chimerism in antibody-facilitated bone marrow chimeras

    SciTech Connect

    Francescutti, L.H.; Gambel, P.; Wegmann, T.G.

    1985-07-01

    The authors have previously described a model for bone marrow transplantation that involves preparation of the host with monoclonal antibody against class I or class II antigens instead of irradiation or cytotoxic drugs. This allows engraftment and subsequent repopulation of the host by donor tissue. They have previously reported on chimerism in the peripheral blood of P1----(P1 X P2)F1 animals. In this report, the authors describe the examination of the bone marrow and spleen stem cell chimerism of these antibody-facilitated (AF) chimeras, by determining, with an isozyme assay, the phenotype of methylcellulose colonies grown from stem cells. They have found a correlation between peripheral blood chimerism and the stem cell constitution of both spleen and bone marrow. The peripheral blood chimerism also correlates with the level of chimerism in macrophages derived from peritoneal exudate cells. These findings indicate that assaying the peripheral blood of such chimeras provides an excellent indication of the degree of chimerism at the stem cell level and stands in sharp contrast to the level of chimerism in certain lymphoid compartments.

  8. Feasibility study of preemptive withdrawal of immunosuppression based on chimerism testing in children undergoing myeloablative allogeneic transplantation for hematologic malignancies.

    PubMed

    Horn, B; Soni, S; Khan, S; Petrovic, A; Breslin, N; Cowan, M; Pelle-Day, G; Cooperstein, E; Baxter-Lowe, L-A

    2009-03-01

    An increasing percentage of autologous cells (increasing chimerism) in the whole blood (WB) chimerism test following allogeneic transplant is related to a very high risk of relapse. Preemptive immunotherapy may decrease the risk of relapse in some patients. Our prospective multi-institutional study evaluated the feasibility of longitudinal chimerism testing in a central laboratory, compared WB, CD3+ and leukemia-specific lineage chimerism in patients with a variety of hematologic malignancies, and evaluated the feasibility of fast withdrawal of immunosuppression based on WB chimerism results. Centralized chimerism testing was feasible and showed low interassay variability. Increasing mixed chimerism (MC) in WB was not useful as a predictor of relapse in our study. The presence of full donor chimerism in WB, CD3+ and leukemia-specific lineages on all measurements was related to a significantly lower risk of relapse than the presence of MC in either subset (11 vs 71%, respectively; P=0.03). Increasing host chimerism in leukemia-specific lineage heralds relapse, but it was not detected early enough to allow immunotherapy. Further studies correlating lineage-specific chimerism and minimal residual disease are required. The goal of preemptive immunotherapy should be to achieve full donor chimerism in WB in CD3+ and leukemia-specific lineages.

  9. Construction of bioactive chimeric MHC class I tetramer by expression and purification of human-murine chimeric MHC heavy chain and beta(2)m as a fusion protein in Escherichia coli.

    PubMed

    Ren, Ding; Wang, Fang; He, Xiaowen; Jiang, Lei; Li, Dean; Ying, He; Sun, Shuhan

    2006-12-01

    Major histocompatibility (MHC) class I tetramers are used in the quantitative analysis of epitope peptide-specific CD8+ T-cells. An MHC class I tetramer was composed of 4 MHC class I complexes and a fluorescently labeled streptavidin (SA) molecule. Each MHC class I complex consists of an MHC heavy chain, a beta(2)-microglobulin (beta(2)m) molecule and a synthetic epitope peptide. In most previous studies, an MHC class I complex was formed in the refolding buffer with an expressed MHC heavy chain molecule and beta(2)m, respectively. This procedure inevitably resulted in the disadvantages of forming unwanted multimers and self-refolding products, and the purification of each kind of monomer was time-consuming. In the present study, the genes of a human/murine chimeric MHC heavy chain (HLA-A2 alpha1, HLA-A2 alpha2 and MHC-H2D alpha3) and beta(2)m were tandem-cloned into plasmid pET17b and expressed as a fusion protein. The recombinant fusion protein was refolded with each of the three HLA-A2 restricted peptides (HBc18-27 FLPSDFFPSI, HBx52-60 HLSLRGLPV, and HBx92-100 VLHKRTLGL) and thus three chimeric MHC class I complexes were obtained. Biotinylation was performed, and its level of efficiency was observed via a band-shift assay in non-reducing polyacrylamide gel electrophoresis (PAGE). Such chimeric MHC class I tetramers showed a sensitive binding activity in monitoring HLA/A2 restrictive cytotoxic T lymphocytes (CTLs) in immunized HLA/A*0201 transgenic mice. PMID:17046278

  10. Experimental validation of finite element model analysis of a steel frame in simulated post-earthquake fire environments

    NASA Astrophysics Data System (ADS)

    Huang, Ying; Bevans, W. J.; Xiao, Hai; Zhou, Zhi; Chen, Genda

    2012-04-01

    During or after an earthquake event, building system often experiences large strains due to shaking effects as observed during recent earthquakes, causing permanent inelastic deformation. In addition to the inelastic deformation induced by the earthquake effect, the post-earthquake fires associated with short fuse of electrical systems and leakage of gas devices can further strain the already damaged structures during the earthquakes, potentially leading to a progressive collapse of buildings. Under these harsh environments, measurements on the involved building by various sensors could only provide limited structural health information. Finite element model analysis, on the other hand, if validated by predesigned experiments, can provide detail structural behavior information of the entire structures. In this paper, a temperature dependent nonlinear 3-D finite element model (FEM) of a one-story steel frame is set up by ABAQUS based on the cited material property of steel from EN 1993-1.2 and AISC manuals. The FEM is validated by testing the modeled steel frame in simulated post-earthquake environments. Comparisons between the FEM analysis and the experimental results show that the FEM predicts the structural behavior of the steel frame in post-earthquake fire conditions reasonably. With experimental validations, the FEM analysis of critical structures could be continuously predicted for structures in these harsh environments for a better assistant to fire fighters in their rescue efforts and save fire victims.

  11. Radar rainfall estimation in the context of post-event analysis of flash-flood events

    NASA Astrophysics Data System (ADS)

    Delrieu, G.; Bouilloud, L.; Boudevillain, B.; Kirstetter, P.-E.; Borga, M.

    2009-09-01

    This communication is about a methodology for radar rainfall estimation in the context of post-event analysis of flash-flood events developed within the HYDRATE project. For such extreme events, some raingauge observations (operational, amateur) are available at the event time scale, while few raingauge time series are generally available at the hydrologic time steps. Radar data is therefore the only way to access to the rainfall space-time organization, but the quality of the radar data may be highly variable as a function of (1) the relative locations of the event and the radar(s) and (2) the radar operating protocol(s) and maintenance. A positive point: heavy rainfall is associated with convection implying better visibility and lesser bright band contamination compared with more current situations. In parallel with the development of a regionalized and adaptive radar data processing system (TRADHy; Delrieu et al. 2009), a pragmatic approach is proposed here to make best use of the available radar and raingauge data for a given flash-flood event by: (1) Identifying and removing residual ground clutter, (2) Applying the "hydrologic visibility" concept (Pellarin et al. 2002) to correct for range-dependent errors (screening and VPR effects for non-attenuating wavelengths, (3) Estimating an effective Z-R relationship through a radar-raingauge optimization approach to remove the mean field bias (Dinku et al. 2002) A sensitivity study, based on the high-quality volume radar datasets collected during two intense rainfall events of the Bollène 2002 experiment (Delrieu et al. 2009), is first proposed. Then the method is implemented for two other historical events occurred in France (Avène 1997 and Aude 1999) with datasets of lesser quality. References: Delrieu, G., B. Boudevillain, J. Nicol, B. Chapon, P.-E. Kirstetter, H. Andrieu, and D. Faure, 2009: Bollène 2002 experiment: radar rainfall estimation in the Cévennes-Vivarais region, France. Journal of Applied

  12. Statistical analysis of eruptive vent distribution from post-subduction monogenetic fields in Baja California, Mexico

    NASA Astrophysics Data System (ADS)

    Germa, A.; Cañon-Tapia, E.; Connor, L.; Le Corvec, N.

    2012-04-01

    Volcanism in Baja California (BC, Mexico) was active from the end of the subduction of the Farallon plate (12.5Ma) until recently (< 1Ma). Most of this volcanism formed twelve volcanic fields, seven of them being monogenetic, delineating a ~600-km-long array parallel to the Gulf of California. Previous studies on these fields have focused on the compositional diversity of magmatic products. Although geochemistry and ages of few lava flows are constrained, only two studies investigated the spatial distribution of the eruptive vents of San Borja. Within a monogenetic volcanic field, cone alignments and linear arrays are considered to reflect the geometry of feeder dikes formed either parallel to the maximum principal stress (σ1) in the lithosphere or using pre-existing crustal fractures. These intrinsic local structures will be compared with the shape of the field, which could reflect the shape of the source at depth. Using satellite imagery to define the location of eruptive centres on four monogenetic volcanic fields from central Baja California (Jaraguay, San Borja, Santa Clara, and San Ignacio), we completed statistical analyses of their spatial distribution. Using commercially available GIS software, spatial density analysis, and statistical scripts, each volcanic field was analysed for the number and density of vents, clustering, vent spacing and alignment azimuths. Our preliminary results reveal that vent densities are within the range of 0.001 to 0.2 vents / 100 km2. Eruptive vents are generally clustered, with density higher than 0.1 vents/100 km2. A common elongation direction trends N135° to N152° in most clusters and fields. We thus propose a NW-SE direction as the preferred orientation of the maximum principal stress (σ1), direction that needs to be confirmed by azimuths of the vents alignments. Using a combination of different computational methods, this study allows to quantify the influence of tectonic stresses at the deep and shallow level within

  13. [Medicolegal experiences in external post-mortem examinations before cremation--a retrospective analysis of the years 1998-2008].

    PubMed

    Eckstein, Philipp; Schyma, Christian; Madea, Burkhard

    2010-01-01

    The paper presents a retrospective analysis of 16,541 external post-mortem examinations carried out before cremation by the Institute of Forensic Medicine of the University of Bonn between 1998 and 2008 with regard to different variables (age, sex, place of death, cause of death etc.). In more than 50 % of cases, the individuals had died in a hospital followed by their apartment or a nursing home. The first post-mortem examination was mainly performed by hospital doctors followed by emergency doctors and office practitioners. As to the age distribution, the 6th to 8th decade of life was prevalent. In 99.8 %, a natural death was certified in the first external post-mortem and only in 0.2 % the manner of death was determined to be non-natural or unclear. In more than half of the cases, the non-natural deaths were determined by emergency doctors. Deaths wrongly classified in the first external post-mortem were mostly accidents and deaths associated with medical procedures. Although the second external examination before cremation is very useful in detecting previously overlooked signs of homicide, it is only of limited value in homicides with few external traces or for detecting underlying diseases and causes of death. In this respect, the second external examination before cremation can only check the plausibility of the cause of death given by the attending physician. The gold standard for a reliable certification of the manner and cause of death remains the autopsy.

  14. Hypomelanosis of Ito: a manifestation of mosaicism or chimerism.

    PubMed Central

    Donnai, D; Read, A P; McKeown, C; Andrews, T

    1988-01-01

    We describe three patients with the cutaneous manifestations of hypomelanosis of Ito. Two, with unusual abnormalities of their toes, had a mixture of diploid and triploid cells in cultured skin fibroblasts. The published clinical descriptions of hypomelanosis of Ito and diploid-triploid mosaicism are reviewed. Chromosome heteromorphisms, HLA types, and DNA fingerprints were studied in an attempt to elucidate the origin of the disease in our patients. We conclude that hypomelanosis of Ito is a manifestation of a heterogeneous group of disorders, the common factor being the presence of two genetically different cell lines. It can result from chromosomal mosaicism or chimerism, from a postzygotic mutation, or from X inactivation. The risk of recurrence is negligible if the proband is a male; if the proband is female the risk is also low but an X linked mutation must be considered. Images PMID:3236362

  15. Chimeric Antigen Receptors Modified T-Cells for Cancer Therapy

    PubMed Central

    Dai, Hanren; Wang, Yao; Lu, Xuechun

    2016-01-01

    The genetic modification and characterization of T-cells with chimeric antigen receptors (CARs) allow functionally distinct T-cell subsets to recognize specific tumor cells. The incorporation of costimulatory molecules or cytokines can enable engineered T-cells to eliminate tumor cells. CARs are generated by fusing the antigen-binding region of a monoclonal antibody (mAb) or other ligand to membrane-spanning and intracellular-signaling domains. They have recently shown clinical benefit in patients treated with CD19-directed autologous T-cells. Recent successes suggest that the modification of T-cells with CARs could be a powerful approach for developing safe and effective cancer therapeutics. Here, we briefly review early studies, consider strategies to improve the therapeutic potential and safety, and discuss the challenges and future prospects for CAR T-cells in cancer therapy. PMID:26819347

  16. Chimeric Antigen Receptor T Cell Therapy in Hematology.

    PubMed

    Ataca, Pınar; Arslan, Önder

    2015-12-01

    It is well demonstrated that the immune system can control and eliminate cancer cells. Immune-mediated elimination of tumor cells has been discovered and is the basis of both cancer vaccines and cellular therapies including hematopoietic stem cell transplantation. Adoptive T cell transfer has been improved to be more specific and potent and to cause less off-target toxicity. Currently, there are two forms of engineered T cells being tested in clinical trials: T cell receptor (TCR) and chimeric antigen receptor (CAR) modified T cells. On 1 July 2014, the United States Food and Drug Administration granted 'breakthrough therapy' designation to anti-CD19 CAR T cell therapy. Many studies were conducted to evaluate the benefits of this exciting and potent new treatment modality. This review summarizes the history of adoptive immunotherapy, adoptive immunotherapy using CARs, the CAR manufacturing process, preclinical and clinical studies, and the effectiveness and drawbacks of this strategy.

  17. The pharmacology of second-generation chimeric antigen receptors.

    PubMed

    van der Stegen, Sjoukje J C; Hamieh, Mohamad; Sadelain, Michel

    2015-07-01

    Second-generation chimeric antigen receptors (CARs) retarget and reprogramme T cells to augment their antitumour efficacy. The combined activating and co-stimulatory domains incorporated in these CARs critically determine the function, differentiation, metabolism and persistence of engineered T cells. CD19-targeted CARs that incorporate CD28 or 4-1BB signalling domains are the best known to date. Both have shown remarkable complete remission rates in patients with refractory B cell malignancies. Recent data indicate that CD28-based CARs direct a brisk proliferative response and boost effector functions, whereas 4-1BB-based CARs induce a more progressive T cell accumulation that may compensate for less immediate potency. These distinct kinetic features can be exploited to further develop CAR-based T cell therapies for a variety of cancers. A new field of immunopharmacology is emerging.

  18. Chimeric Antigen Receptors Modified T-Cells for Cancer Therapy.

    PubMed

    Dai, Hanren; Wang, Yao; Lu, Xuechun; Han, Weidong

    2016-07-01

    The genetic modification and characterization of T-cells with chimeric antigen receptors (CARs) allow functionally distinct T-cell subsets to recognize specific tumor cells. The incorporation of costimulatory molecules or cytokines can enable engineered T-cells to eliminate tumor cells. CARs are generated by fusing the antigen-binding region of a monoclonal antibody (mAb) or other ligand to membrane-spanning and intracellular-signaling domains. They have recently shown clinical benefit in patients treated with CD19-directed autologous T-cells. Recent successes suggest that the modification of T-cells with CARs could be a powerful approach for developing safe and effective cancer therapeutics. Here, we briefly review early studies, consider strategies to improve the therapeutic potential and safety, and discuss the challenges and future prospects for CAR T-cells in cancer therapy.

  19. Chimeric Antigen Receptor T Cell Therapy in Hematology

    PubMed Central

    Ataca, Pınar; Arslan, Önder

    2015-01-01

    It is well demonstrated that the immune system can control and eliminate cancer cells. Immune-mediated elimination of tumor cells has been discovered and is the basis of both cancer vaccines and cellular therapies including hematopoietic stem cell transplantation. Adoptive T cell transfer has been improved to be more specific and potent and to cause less off-target toxicity. Currently, there are two forms of engineered T cells being tested in clinical trials: T cell receptor (TCR) and chimeric antigen receptor (CAR) modified T cells. On 1 July 2014, the United States Food and Drug Administration granted ‘breakthrough therapy’ designation to anti-CD19 CAR T cell therapy. Many studies were conducted to evaluate the benefits of this exciting and potent new treatment modality. This review summarizes the history of adoptive immunotherapy, adoptive immunotherapy using CARs, the CAR manufacturing process, preclinical and clinical studies, and the effectiveness and drawbacks of this strategy. PMID:26377367

  20. Chimeric elk/mouse prion proteins in transgenic mice.

    PubMed

    Tamgüney, Gültekin; Giles, Kurt; Oehler, Abby; Johnson, Natrina L; DeArmond, Stephen J; Prusiner, Stanley B

    2013-02-01

    Chronic wasting disease (CWD) of deer and elk is a highly communicable neurodegenerative disorder caused by prions. Investigations of CWD are hampered by slow bioassays in transgenic (Tg) mice. Towards the development of Tg mice that will be more susceptible to CWD prions, we created a series of chimeric elk/mouse transgenes that encode the N terminus of elk PrP (ElkPrP) up to residue Y168 and the C terminus of mouse PrP (MoPrP) beyond residue 169 (mouse numbering), designated Elk3M(SNIVVK). Between codons 169 and 219, six residues distinguish ElkPrP from MoPrP: N169S, T173N, V183I, I202V, I214V and R219K. Using chimeric elk/mouse PrP constructs, we generated 12 Tg mouse lines and determined incubation times after intracerebral inoculation with the mouse-passaged RML scrapie or Elk1P CWD prions. Unexpectedly, one Tg mouse line expressing Elk3M(SNIVVK) exhibited incubation times of <70 days when inoculated with RML prions; a second line had incubation times of <90 days. In contrast, mice expressing full-length ElkPrP had incubation periods of >250 days for RML prions. Tg(Elk3M,SNIVVK) mice were less susceptible to CWD prions than Tg(ElkPrP) mice. Changing three C-terminal mouse residues (202, 214 and 219) to those of elk doubled the incubation time for mouse RML prions and rendered the mice resistant to Elk1P CWD prions. Mutating an additional two residues from mouse to elk at codons 169 and 173 increased the incubation times for mouse prions to >300 days, but made the mice susceptible to CWD prions. Our findings highlight the role of C-terminal residues in PrP that control the susceptibility and replication of prions.

  1. Modeling cognition and disease using human glial chimeric mice.

    PubMed

    Goldman, Steven A; Nedergaard, Maiken; Windrem, Martha S

    2015-08-01

    As new methods for producing and isolating human glial progenitor cells (hGPCs) have been developed, the disorders of myelin have become especially compelling targets for cell-based therapy. Yet as animal modeling of glial progenitor cell-based therapies has progressed, it has become clear that transplanted hGPCs not only engraft and expand within murine hosts, but dynamically outcompete the resident progenitors so as to ultimately dominate the host brain. The engrafted human progenitor cells proceed to generate parenchymal astrocytes, and when faced with a hypomyelinated environment, oligodendrocytes as well. As a result, the recipient brains may become inexorably humanized with regards to their resident glial populations, yielding human glial chimeric mouse brains. These brains provide us a fundamentally new tool by which to assess the species-specific attributes of glia in modulating human cognition and information processing. In addition, the cellular humanization of these brains permits their use in studying glial infectious and inflammatory disorders unique to humans, and the effects of those disorders on the glial contributions to cognition. Perhaps most intriguingly, by pairing our ability to construct human glial chimeras with the production of patient-specific hGPCs derived from pluripotential stem cells, we may now establish mice in which a substantial proportion of resident glia are both human and disease-derived. These mice in particular may provide us new opportunities for studying the human-specific contributions of glia to psychopathology, as well as to higher cognition. As such, the assessment of human glial chimeric mice may provide us new insight into the species-specific contributions of glia to human cognitive evolution, as well as to the pathogenesis of human neurological and neuropsychiatric disease.

  2. Chimeric elk/mouse prion proteins in transgenic mice

    PubMed Central

    Tamgüney, Gültekin; Giles, Kurt; Oehler, Abby; Johnson, Natrina L.; DeArmond, Stephen J.

    2013-01-01

    Chronic wasting disease (CWD) of deer and elk is a highly communicable neurodegenerative disorder caused by prions. Investigations of CWD are hampered by slow bioassays in transgenic (Tg) mice. Towards the development of Tg mice that will be more susceptible to CWD prions, we created a series of chimeric elk/mouse transgenes that encode the N terminus of elk PrP (ElkPrP) up to residue Y168 and the C terminus of mouse PrP (MoPrP) beyond residue 169 (mouse numbering), designated Elk3M(SNIVVK). Between codons 169 and 219, six residues distinguish ElkPrP from MoPrP: N169S, T173N, V183I, I202V, I214V and R219K. Using chimeric elk/mouse PrP constructs, we generated 12 Tg mouse lines and determined incubation times after intracerebral inoculation with the mouse-passaged RML scrapie or Elk1P CWD prions. Unexpectedly, one Tg mouse line expressing Elk3M(SNIVVK) exhibited incubation times of <70 days when inoculated with RML prions; a second line had incubation times of <90 days. In contrast, mice expressing full-length ElkPrP had incubation periods of >250 days for RML prions. Tg(Elk3M,SNIVVK) mice were less susceptible to CWD prions than Tg(ElkPrP) mice. Changing three C-terminal mouse residues (202, 214 and 219) to those of elk doubled the incubation time for mouse RML prions and rendered the mice resistant to Elk1P CWD prions. Mutating an additional two residues from mouse to elk at codons 169 and 173 increased the incubation times for mouse prions to >300 days, but made the mice susceptible to CWD prions. Our findings highlight the role of C-terminal residues in PrP that control the susceptibility and replication of prions. PMID:23100369

  3. Principles of Leading Change: An Inductive Analysis from Post-Katrina New Orleans

    ERIC Educational Resources Information Center

    Beabout, Brian R.

    2014-01-01

    Despite over forty years of research on theories of educational change, little is known of the change theories-in-use of school-based administrators, often tasked with implementing externally imposed reform mandates. Capitalizing on the unique case of post-Katrina schooling, this qualitative study examines the ways in which ten principals spoke…

  4. Transformation or Continuation? A Critical Analysis of the Making of the Post-2015 Education Agenda

    ERIC Educational Resources Information Center

    Enns, Charis

    2015-01-01

    As the target date of 2015 draws near for both the Education for All and Millennium Development Goals, consultations are well-underway to begin defining the shape and scope of the post-2015 development agenda. Between September 2012 and March 2013, UN Member States, private sector representatives, multilateral development agencies, epistemic…

  5. Educational Participation Post-16: A Longitudinal Analysis of Intentions and Outcomes

    ERIC Educational Resources Information Center

    Croll, Paul

    2009-01-01

    The issue of levels of participation in post-compulsory education has been emphasised by the current policy initiatives to increase the age to which some form of participation is compulsory. One of the acknowledged weaknesses of research in the field of children's intentions with regard to participation is the lack of longitudinal data. This paper…

  6. Integrated Education and Black Development in Post-apartheid South Africa: A Critical Analysis.

    ERIC Educational Resources Information Center

    Abdi, Ali A.

    2001-01-01

    Focuses on the problems of education and development in post-apartheid South Africa. Argues that there must be a focus on the uneven terrain of educational attainment and long term socio-economic development. Discusses factors hindering educational development. Calls for reconstitution of South Africa's educational programs for all citizens. (CAJ)

  7. Supportive Distortions: An Analysis of Posts on a Pedophile Internet Message Board

    ERIC Educational Resources Information Center

    Malesky, L. Alvin, Jr.; Ennis, Liam

    2004-01-01

    A covert observation of posts on a pro-pedophile Internet message board investigated evidence of distorted cognitions that were supportive of sexually abusive behavior. Implications for the treatment and supervision of members of online communities that support pedophilic interests and behaviors are discussed. The purpose of the present study was…

  8. The Overlap between Emotional Intelligence and Post-Industrial Leadership Capacity: A Construct Validity Analysis

    ERIC Educational Resources Information Center

    Rosch, David M.; Joseph, Dana L.; Newman, Daniel A.

    2011-01-01

    A sample of 276 students enrolled in campus leadership programs completed the Emotional Competence Inventory-University Edition (ECI-U) and the Socially Responsible Leadership Scale (SRLS) as a means to determine the relatedness in college students of emotional intelligence (EI) to the practice of post-industrial leadership skills. Confirmatory…

  9. E3 10C compressor test analysis of high-speed post-stall data

    NASA Technical Reports Server (NTRS)

    Dvorak, S. D.; Hosny, W. M.; Steenken, W. G.

    1986-01-01

    In-stall characteristics from high-speed post-stall transients are determined. The transient, surge-cycle nature of high-speed post-stall operation precludes the possibility of obtaining in-stall characteristics in a steady-state manner, as is possible during low-speed post-stall operation, which is characterized by quasi-steady rotating-stall behavior. Maximum likelihood parameter estimation techniques were used to obtain the quasi-steady high-speed characteristics from transient data. The necessary data was first obtained from a specially instrumented compressor that was tested well beyond its limits of normal operation. The unsteady, post-stall data thus obtained was then digitized and processed through a simplified analytical model to construct the input-output relationship necessary for estimation. In-stall characteristics were determined using this estimation procedure at two different high-speed conditions, 90 and 98.5 percent corrected speed. The estimated characteristics were found to be robust in the presence of measurement noise and unmodelled system dynamics, but the compressor response-time constants, also estimated, were more sensitive to these same disturbances. The experimentally determined low-speed in-stall characteristics and the estimated high-speed in-stall characteristics were then incorporated into a one-dimensional compressor simulation model developed as a parallel effort to the compressor testing and data reduction effort, which yielded predictable results.

  10. Adoption and guardianship: A moderated mediation analysis of predictors of post-permanency continuity.

    PubMed

    Testa, Mark F; Snyder, Susan M; Wu, Qi; Rolock, Nancy; Liao, Minli

    2015-03-01

    The original thinking behind permanency planning for foster children was to secure a caregiver's intention to provide a permanent home, not to guarantee it. Little is known, however, about how intentions change after permanence or what effect this change has on post-permanency continuity of care. This study examined the mediating effects of caregivers' thoughts about ending an adoption or guardianship, and how this mechanism may be contingent on primordial and bureaucratic factors that child welfare agencies rely on to ensure family continuity after legal permanence. In 2006, a sample of 346 Illinois caregivers who finalized an adoption or guardianship between 1998 and 2002 completed surveys about child behavior problems, the adequacy of financial assistance, and thoughts about maintaining the permanency relationship. Responses were linked to administrative data that tracked continuity of care through 2012. Simple mediation and moderated mediation hypotheses of the effects of caregiver thoughts on post-permanency continuity were tested. At last observation, 8% of caregivers were no longer living with the child or stopped receiving subsidies on the child's behalf. Thoughts expressed at survey time about ending the permanency relationship mediated the effect of child behavior problems on post-permanency discontinuity rates. This indirect effect was more pronounced among distantly related kin, lone caregivers, and caregivers who felt the subsidy was inadequate to cover their expenses. Our findings suggest that post-permanency services should target a narrow segment of caregivers who express weakened permanency commitments that arise from the challenges of parenting a child with multiple behavioral problems.

  11. Evaluation of an Innovative Post-Arrest Diversion Program: 12-Month Recidivism Analysis

    ERIC Educational Resources Information Center

    Dembo, Richard; Walters, Wansley; Wareham, Jennifer; Burgos, Catherimarty; Schmeidler, James; Hoge, Robert; Underwood, Lee

    2008-01-01

    We report the results of an evaluation study of a diversion program, operated by the Miami-Dade County Juvenile Assessment Center in Florida. The Post-Arrest Diversion (PAD) program represents an innovative approach to treatment and intervention within the juvenile justice system that utilizes standardized psychosocial risks and needs assessment…

  12. Factor Analysis of Aviation Training Measures and Post-Training Performance Evaluations.

    ERIC Educational Resources Information Center

    Booth, Richard F.; Berkshire, James R.

    The purpose of this study was to relate the factor structure of naval air training measures to the performance of Marine pilots in operational squadrons. Five post-training criteria were developed; four were Commanding Officer (C.O.) nominations of junior officers for hypothetical special assignments, and the fifth was a general…

  13. ORNL ADCP POST-PROCESSING GUIDE AND MATLAB ALGORITHMS FOR MHK SITE FLOW AND TURBULENCE ANALYSIS

    SciTech Connect

    Gunawan, Budi; Neary, Vincent S

    2011-09-01

    Standard methods, along with guidance for post-processing the ADCP stationary measurements using MATLAB algorithms that were evaluated and tested by Oak Ridge National Laboratory (ORNL), are presented following an overview of the ADCP operating principles, deployment methods, error sources and recommended protocols for removing and replacing spurious data.

  14. Slovak Preschool Curriculum Reform and Teachers' Emotions: An Analysis of Facebook Posts

    ERIC Educational Resources Information Center

    Kascak, O.; Pupala, B.; Mbugua, T.

    2016-01-01

    The school system in the Slovak Republic, as in other post-communist countries, was first reformed at the beginning of the 1990s. It was not until 2008, when an education law was adopted to set out new parameters for creating a national curriculum at all levels, that fundamental curricular reforms were made. Thus the law also meant that preschool…

  15. Decolonizing Education: A Critical Discourse Analysis of Post-Secondary Humanities Textbooks

    ERIC Educational Resources Information Center

    Harper, Kimberly C.

    2012-01-01

    This dissertation examines nine post-secondary humanities textbooks published between 2001 and 2011 using an approach that includes both qualitative and quantitative methodology to analyze the written and visual content of humanities textbooks. This dissertation engages in current debates that address bias in humanities textbooks and contributes…

  16. Evaluation of chimeric yellow fever 17D/dengue viral replication in ticks.

    PubMed

    Kazimírová, Mária; Mantel, Nathalie; Raynaud, Sandrine; Slovák, Mirko; Ustaniková, Katarína; Lang, Jean; Guy, Bruno; Barban, Veronique; Labuda, Milan

    2012-11-01

    Chimeric yellow fever 17D/DENV-1-4 viruses (CYD-1-4) have been developed as a tetravalent dengue vaccine candidate which is currently being evaluated in efficacy trials in Asia and America. While YF 17D and DENV are mosquito-borne flaviviruses, it has been shown that CYD-1-4 do not replicate after oral infection in mosquitoes and are not transmitted to new hosts. To further document the risk of environmental dissemination of these viruses, we evaluated the replication of CYD-1-4 in ticks, the vector of tick-borne encephalitis virus (TBEV), another member of the flavivirus family. Females of two hard tick species, Ixodes ricinus and Rhipicephalus appendiculatus, were inoculated intracoelomically with CYD-1-4 viruses and parent viruses (DENV-1-4 and YF 17D). Virus persistence and replication was assessed 2, 16, and 44 days post-inoculation by plaque titration and qRT-PCR. CYD-1-4 viruses were detected in I. ricinus ticks at early time points post-inoculation, but with infectious titers at least 100-fold lower than those observed in TBEV-infected ticks. Unlike TBEV, complete viral clearance occurred by day 44 in most ticks except for CYD-2, which had a tendency to decline. In addition, while about 70% of TBEV-infected I. ricinus nymphs acquired infection by co-feeding with infected tick females on non-viremic hosts, no co-feeding transmission of CYD-2 virus was detected. Based on these results, we conclude that the risk of dissemination of the candidate vaccine viruses by tick bite is highly unlikely.

  17. Comparative classification analysis of post-harvest growth detection from terrestrial LiDAR point clouds in precision agriculture

    NASA Astrophysics Data System (ADS)

    Koenig, Kristina; Höfle, Bernhard; Hämmerle, Martin; Jarmer, Thomas; Siegmann, Bastian; Lilienthal, Holger

    2015-06-01

    In precision agriculture, detailed geoinformation on plant and soil properties plays an important role, e.g., in crop protection or the application of fertilizers. This paper presents a comparative classification analysis for post-harvest growth detection using geometric and radiometric point cloud features of terrestrial laser scanning (TLS) data, considering the local neighborhood of each point. Radiometric correction of the TLS data was performed via an empirical range-correction function derived from a field experiment. Thereafter, the corrected amplitude and local elevation features were explored regarding their importance for classification. For the comparison, tree induction, Naive Bayes, and k-Means-derived classifiers were tested for different point densities to distinguish between ground and post-harvest growth. The classification performance was validated against highly detailed RGB reference images and the red edge normalized difference vegetation index (NDVI705), derived from a hyperspectral sensor. Using both geometric and radiometric features, we achieved a precision of 99% with the tree induction. Compared to the reference image classification, the calculated post-harvest growth coverage map reached an accuracy of 80%. RGB and LiDAR-derived coverage showed a polynomial correlation to NDVI705 of degree two with R2 of 0.8 and 0.7, respectively. Larger post-harvest growth patches (>10 × 10 cm) could already be detected by a point density of 2 pts./0.01 m2. The results indicate a high potential of radiometric and geometric LiDAR point cloud features for the identification of post-harvest growth using tree induction classification. The proposed technique can potentially be applied over larger areas using vehicle-mounted scanners.

  18. The Impact of a Science Qualification Emphasising Scientific Literacy on Post-compulsory Science Participation: An analysis using national data

    NASA Astrophysics Data System (ADS)

    Homer, Matt; Ryder, Jim

    2015-06-01

    In 2006 in England an innovative suite of science qualifications for 14-16-year-olds called Twenty-First Century Science (21CS) was introduced. These qualifications have a strong focus on developing scientific literacy in all students whilst simultaneously providing preparation for the study of post-compulsory science for a smaller proportion of students. Claims have been made that such an innovative qualification would impact significantly on post-compulsory science participation-either positively or negatively. Using national data in England to track one cohort of students over 2007-2011, this study compares progression rates to post-compulsory science qualifications in England between 21CS qualifications and more traditional non-21CS qualifications. Methods employed include simple comparisons of proportions progressing from each qualification, and more complex multi-level modelling approaches that take account of both students clustered in schools, and potentially differing demographic and achievement profiles of students in the 2 groups of qualifications. A simple descriptive analysis shows that there is very little difference in overall progression rates between the 2 types of 14-16 science qualification. More fine-grained descriptive analyses show that there are some important differences, based in particular on the interaction between the amount of science studied at ages 14-16, and on the post-16 science qualification chosen (biology, chemistry or physics). Furthermore, sophisticated modelling analyses indicate a consistently negative small to moderate impact on progression from the 21CS qualification. Overall, our findings suggest that the emphasis on scientific literacy within the 21CS qualification suite has not had a major impact on the uptake of post-compulsory science qualifications.

  19. Establishing a Population-Based HLA-Antibody Panel for Flow Cytometric Monitoring of Chimerism in HLA-Haploidentical Stem Cell Transplantation.

    PubMed

    Choe, Wonho; Hwang, Min-A; Jang, Seongsoo; Park, Chan-Jeoung; Chi, Hyun-Sook; Im, Ho Joon

    2016-01-01

    Determining the chimerism in stem cell transplantation (SCT) is important in the monitoring of engraftment. Conventional monitoring methods such as short tandem repeat polymerase chain reaction (STR-PCR) are labor intensive and difficult in showing the dynamics of cell subpopulations. In HLA-haploidentical SCT, flow cytometric analysis using anti-HLA antibody for the mismatched HLA can be useful in observing changes of cell subpopulations and determining chimerism. We designed a specific panel of HLA antibody reagents for the Korean population, and verified its clinical application in flow cytometric monitoring of chimerism after haploidentical stem cell transplantation. A total of 12 anti-HLA-A, -B-antibodies were selected, which could cover 82.5% of HLA-A and 16.5% of HLA-B in Korean population. This HLA panel distinguished donor and recipient cells in 22 of 23 HLA-haploidentical SCT cases. In one case, the patient had HLA-A*02/A*24, B*48/B*61 while the donor had HLA-A*02/A*33, B*44/B*48. The donor type HLA-B*44(+) and CD3(+) T cells, and HLA-B*44(+) and CD56(+) NK cells were seen at day 14 and day 8, respectively. Increased HLA-B*44(+) cells throughout the study period indicated the engraftment of donor stem cells. We were able to design a population specific panel of HLA-antibodies, and verified that flow cytometric analysis using HLA antibody for the detection of chimerism in HLA-haploidentical SCT was a simple and sensitive monitoring technique. This method allowed us to observe the dynamic changes in cell subpopulations after HLA-haploidentical SCT. Flow cytometric analysis can be considered as a strong tool for the monitoring of engraftment in HLA-haploidentical SCT.

  20. Analysis of error introduced during end-user post-processing of airborne laser data (lidar)

    NASA Astrophysics Data System (ADS)

    Smith, Sarah Louise

    The primary aims and objectives of this thesis are to identify the sources and operation of the errors which are introduced during end-user post-processing of airborne laser scanning data. Previous research has concentrated on the errors incorporated during data capture and preliminary supplier processing. The errors which are introduced by the end-users have been largely neglected. As a result, data users cannot currently estimate the errors within, and therefore the quality of, the models they produce. Laser scanning is a remote sensing technique for the capture of height data of the surface of the Earth. It offers competitive capture costs, high accuracy, and is particularly suited to capturing information in complex urban areas. As a result the commercial value of laser scanning data is high. However, in order to realise the potential of this technique, the quality of the datasets derived from the data must be assessed and the errors introduced during modelling understood. For users to make informed decisions regarding the design of their post-processing workflow it is fundamental that they know how and where errors may be introduced. The characteristics of these errors are investigated in this thesis using a range of approaches. End-user post-processing is divided into three techniques in the thesis: data structuring, filtering and segmentation. Each process is investigated hi terms of accuracy and sensitivity, through the comparison of several methods with reference models. New algorithms for filtering and segmenting laser data are presented. The errors created by each process are identified and analysed. The location of errors across the elevation surface are also investigated. It is shown how this information could be used to aid end-users design their post-processing methodology. The methodology for analyzing the errors is presented as a framework which could be used as a standard for ALS models. This thesis shows that the choice of post

  1. Key molecular processes of the diapause to post-diapause quiescence transition in the alfalfa leafcutting bee Megachile rotundata identified by comparative transcriptome analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Insect diapause (dormancy) synchronizes an insect’s life cycle to seasonal changes in the abiotic and biotic resources required for development and reproduction. Transcription analysis of Megachile rotundata diapause termination identified 399 post-diapause upregulated and 144 post-diapause down-reg...

  2. The effect of cochlear implantation and post-operative rehabilitation on acoustic voice analysis in post-lingual hearing impaired adults.

    PubMed

    Hassan, Sabah M; Malki, Khalid H; Mesallam, Tamer A; Farahat, Mohamad; Bukhari, Manal; Murry, Thomas

    2011-10-01

    Post-lingual deaf adults can develop some vocal abnormalities similar to those developed in pre-lingual deaf individuals. The aim of this work was to study the effect of cochlear implantation followed by post-operative rehabilitation on voice acoustics in post-lingual hearing impaired adults with different durations of hearing loss. The study included 35 post-lingual hearing impaired adults who underwent cochlear implantation. Patients were divided into two groups according to the duration of their hearing loss. Each group was further divided into two subgroups according to whether they received auditory rehabilitation or not. Using the Multi-Dimensional Voice Program (MDVP) parameters, comparisons were made between each subgroup of patients and the normal MDVP Saudi database, and between subgroups of patients. Most of the patients in the two groups reported significant improvement in their MDVP results post-implantation. Further, significantly deviant MDVP parameters were reported in the group of patients with longer duration of hearing loss. Patients who received rehabilitation significantly improved more than those who did not. In conclusion, it appears that cochlear implantation improves the auditory control of voice production in post-lingual deaf adults. Also, it is obvious that cochlear implantation at an early stage of hearing loss gives better results on voice control, especially if augmented with auditory rehabilitation. PMID:21331786

  3. Peripheral Red Blood Cell Split Chimerism as a Consequence of Intramedullary Selective Apoptosis of Recipient Red Blood Cells in a Case of Sickle Cell Disease

    PubMed Central

    Marziali, Marco; Isgrò, Antonella; Sodani, Pietro; Gaziev, Javid; Fraboni, Daniela; Paciaroni, Katia; Gallucci, Cristiano; Alfieri, Cecilia; Roveda, Andrea; De Angelis, Gioia; Cardarelli, Luisa; Ribersani, Michela; Andreani, Marco; Lucarelli, Guido

    2014-01-01

    Allogeneic cellular gene therapy through hematopoietic stem cell transplantation is the only radical cure for congenital hemoglobinopathies like thalassemia and sickle cell anemia. Persistent mixed hematopoietic chimerism (PMC) has been described in thalassemia and sickle cell anemia. Here, we describe the clinical course of a 6-year-old girl who had received bone marrow transplant for sickle cell anemia. After the transplant, the patient showed 36% donor hematopoietic stem cells in the bone marrow, whereas in the peripheral blood there was evidence of 80% circulating donor red blood cells (RBC). The analysis of apoptosis at the Bone Marrow level suggests that Fas might contribute to the cell death of host erythroid precursors. The increase in NK cells and the regulatory T cell population observed in this patient suggests that these cells might contribute to the condition of mixed chimerism. PMID:25408852

  4. Chemical analysis of post explosion samples obtained as a result of model field experiments.

    PubMed

    Borusiewicz, Rafal; Zadora, Grzegorz; Zieba-Palus, Janina

    2013-11-15

    Five different explosives were detonated in a series of field experiments. Each experiment (detonation of the charge of each specific explosive) was repeated three times. The experiments were conducted under controlled conditions, exceeding those of research published so far. Detonated charges were uniform in size and, as far as possible, in shape. The explosives used originated from the same batch. Additionally, the same kind of electric detonators were used. Witness plates (sheets of galvanised steel 100 cm × 90 cm × 0.5 mm) were used to collect post-blast residues in a reproducible way. They were placed relatively close to the charge to minimise the influence of the wind. Samples were collected by systematic swabbing of the surface of the plate by acetone moistened cotton swabs. Samples were packed tight, transferred to the laboratory, and extracted with methanol. Extracts were concentrated by solvent evaporation, cleaned by centrifugation, and analysed using HPLC-DAD. Each extract was analysed three times and the mean value of the amount of the given explosive within the extract was calculated. For each of the explosive materials used the results of the repetition of the experiments proved them to be irreproducible. After each detonation of a specific charge different amounts of given explosives were found in post-blast samples. Also, the intuitively expected relationship between the distance from the charge and amount of post-blast residues were not observed. These results are consistent with previously published results of field experiments. The lack of reproducibility may be explained by differences in efficiency of detonation. The efficiency of a detonation may be influenced even by small differences in the shape of the charge as well as by the position and properties of the detonator. The lack of dependency between the amount of the explosive in the post-blast samples and the distance from the charge may be explained by the fact that during detonation

  5. Reproductive history and post-reproductive mortality: A sibling comparison analysis using Swedish register data.

    PubMed

    Barclay, Kieron; Keenan, Katherine; Grundy, Emily; Kolk, Martin; Myrskylä, Mikko

    2016-04-01

    A growing body of evidence suggests that reproductive history influences post-reproductive mortality. A potential explanation for this association is confounding by socioeconomic status in the family of origin, as socioeconomic status is related to both fertility behaviours and to long-term health. We examine the relationship between age at first birth, completed parity, and post-reproductive mortality and address the potential confounding role of family of origin. We use Swedish population register data for men and women born 1932-1960, and examine both all-cause and cause-specific mortality. The contributions of our study are the use of a sibling comparison design that minimizes residual confounding from shared family background characteristics and assessment of cause-specific mortality that can shed light on the mechanisms linking reproductive history to mortality. Our results were entirely consistent with previous research on this topic, with teenage first time parents having higher mortality, and the relationship between parity and mortality following a U-shaped pattern where childless men and women and those with five or more children had the highest mortality. These results indicate that selection into specific fertility behaviours based upon socioeconomic status and experiences within the family of origin does not explain the relationship between reproductive history and post-reproductive mortality. Additional analyses where we adjust for other lifecourse factors such as educational attainment, attained socioeconomic status, and post-reproductive marital history do not change the results. Our results add an important new level of robustness to the findings on reproductive history and mortality by showing that the association is robust to confounding by factors shared by siblings. However it is still uncertain whether reproductive history causally influences health, or whether other confounding factors such as childhood health or risk-taking propensity could

  6. Immunogenicity and antigenicity of a recombinant chimeric protein containing epitopes of poliovirus type 1.

    PubMed

    Pan, X-X; Wang, J; Xia, W-Y; Li, X-F; Yang, L-J; Huang, C; Chen, Y-D

    2016-01-01

    To design a vaccine that simultaneously prevents both rotavirus (RV) and poliovirus (PV), a PV type 1 (PV1) chimeric protein using RV VP6 as a vector (VP6F) was constructed, expressed in Escherichia coli expression system and characterized by SDS-PAGE, Western blot, immunofluorescence assay and neutralization test. The results showed that the chimeric protein reacted with anti-VP6F and anti-PV1 antibodies and elicited production of serum antibodies against the chimeric protein in guinea pigs. Antibodies against the chimeric protein neutralized RV Wa and PV1 infection in vitro. The results provided a relevant possibility of developing novel approaches in the rational design of vaccines effective against both RV and PV. PMID:27640433

  7. Single-born marmosets without hemopoietic chimerism: naturally occurring and induced.

    PubMed

    Gengozian, N; Batson, J S

    1975-01-01

    Marmosets have a high frequency of fraternal twinning, and placental vascular anastomoses between the twin fetuses invariably lead to hemopoietic chimerism. The occasional finding of chimerism in single-born marmosets suggested that in a twin pregnancy one fetus had undergone resorption after contributing hemopoietic stem cells to its twin. In this study non-chimeric single-born marmosets were produced by fallopian tube ligation or surgical relocation of one ovary in breeding females. Further, in an examination of hemopoietic cells from over 50 single-born young from nonoperated females, chimerism occurred less frequently than what one would expect if resorption of a co-twin had occurred after a functional anastomosis had been established. PMID:808628

  8. Chimeric self-sufficient P450cam-RhFRed biocatalysts with broad substrate scope

    PubMed Central

    Robin, Aélig; Köhler, Valentin; Jones, Alison; Ali, Afruja; Kelly, Paul P; O'Reilly, Elaine; Turner, Nicholas J

    2011-01-01

    Summary A high-throughput screening protocol for evaluating chimeric, self-sufficient P450 biocatalysts and their mutants against a panel of substrates was developed, leading to the identification of a number of novel biooxidation activities. PMID:22238522

  9. A chimeric measles virus with a lentiviral envelope replicates exclusively in CD4+/CCR5+ cells

    SciTech Connect

    Mourez, Thomas; Mesel-Lemoine, Mariana; Combredet, Chantal; Najburg, Valerie; Cayet, Nadege; Tangy, Frederic

    2011-10-25

    We generated a replicating chimeric measles virus in which the hemagglutinin and fusion surface glycoproteins were replaced with the gp160 envelope glycoprotein of simian immunodeficiency virus (SIVmac239). Based on a previously cloned live-attenuated Schwarz vaccine strain of measles virus (MV), this chimera was rescued at high titers using reverse genetics in CD4+ target cells. Cytopathic effect consisted in the presence of large cell aggregates evolving to form syncytia, as observed during SIV infection. The morphology of the chimeric virus was identical to that of the parent MV particles. The presence of SIV gp160 as the only envelope protein on chimeric particles surface altered the cell tropism of the new virus from CD46+ to CD4+ cells. Used as an HIV candidate vaccine, this MV/SIVenv chimeric virus would mimic transient HIV-like infection, benefiting both from HIV-like tropism and the capacity of MV to replicate in dendritic cells, macrophages and lymphocytes.

  10. Post-natal growth of the gastrointestinal tract of the Siberian hamster: morphometric analysis.

    PubMed

    Wołczuk, K; Kobak, J

    2014-12-01

    Post-natal growth of the gastrointestinal tract of the Siberian hamster was studied in newborn and 3-, 7-, 14-, 21-, 42- and 90-day-old animals. Morphometric measurements and calculations were carried out: length and internal surface of gastrointestinal tract segments, size (height, width, surface) and density of villi as well as allometric growth rate of the length and internal surface of the segments with respect to the body mass. The fastest growth rate of the gastrointestinal tract segments was noticed during the first 3 days of the post-natal life. Nevertheless, significant regional differences in their growth rate were found. The increase in the length and internal surface of the large intestine was fastest, while the smallest increase was observed in the oesophagus. All segments of the gastrointestinal tract except oesophagus exhibited a positive allometric relationship to the body mass from birth till final weaning, whereas during the post-weaning period, the increase was isometric. Thus, at birth, the gastrointestinal tract segments were relatively smaller compared with those observed in adults, but then, the gastrointestinal tract grew faster than the rest of the body and reached its adult proportions just before the transition to solid food. Most probably, reaching the adult structure of the gastrointestinal tract before the final weaning is an essential condition for the proper growth of an organism after the weaning.

  11. Transcranial direct current stimulation facilitates motor learning post-stroke: a systematic review and meta-analysis.

    PubMed

    Kang, Nyeonju; Summers, Jeffery J; Cauraugh, James H

    2016-04-01

    Transcranial direct current stimulation (tDCS) is an attractive protocol for stroke motor recovery. The current systematic review and meta-analysis investigated the effects of tDCS on motor learning post-stroke. Specifically, we determined long-term learning effects by examining motor improvements from baseline to at least 5 days after tDCS intervention and motor practise. 17 studies reported long-term retention testing (mean retention interval=43.8 days; SD=56.6 days) and qualified for inclusion in our meta-analysis. Assessing primary outcome measures for groups that received tDCS and motor practise versus sham control groups created 21 valid comparisons: (1) 16 clinical assessments and (2) 5 motor skill acquisition tests. A random effects model meta-analysis showed a significant overall effect size=0.59 (p<0.0001; low heterogeneity, T(2)=0.04; I(2)=22.75%; and high classic fail-safe N=240). 4 moderator variable analyses revealed beneficial effects of tDCS on long-term motor learning: (1) stimulation protocols: anodal on the ipsilesional hemisphere, cathodal on the contralesional hemisphere, or bilateral; (2) recovery stage: subacute or chronic stroke; (3) stimulation timing: tDCS before or during motor practise; and (4) task-specific training or conventional rehabilitation protocols. This robust meta-analysis identified novel long-term motor learning effects with tDCS and motor practise post-stroke. PMID:26319437

  12. Transcranial direct current stimulation facilitates motor learning post-stroke: a systematic review and meta-analysis.

    PubMed

    Kang, Nyeonju; Summers, Jeffery J; Cauraugh, James H

    2016-04-01

    Transcranial direct current stimulation (tDCS) is an attractive protocol for stroke motor recovery. The current systematic review and meta-analysis investigated the effects of tDCS on motor learning post-stroke. Specifically, we determined long-term learning effects by examining motor improvements from baseline to at least 5 days after tDCS intervention and motor practise. 17 studies reported long-term retention testing (mean retention interval=43.8 days; SD=56.6 days) and qualified for inclusion in our meta-analysis. Assessing primary outcome measures for groups that received tDCS and motor practise versus sham control groups created 21 valid comparisons: (1) 16 clinical assessments and (2) 5 motor skill acquisition tests. A random effects model meta-analysis showed a significant overall effect size=0.59 (p<0.0001; low heterogeneity, T(2)=0.04; I(2)=22.75%; and high classic fail-safe N=240). 4 moderator variable analyses revealed beneficial effects of tDCS on long-term motor learning: (1) stimulation protocols: anodal on the ipsilesional hemisphere, cathodal on the contralesional hemisphere, or bilateral; (2) recovery stage: subacute or chronic stroke; (3) stimulation timing: tDCS before or during motor practise; and (4) task-specific training or conventional rehabilitation protocols. This robust meta-analysis identified novel long-term motor learning effects with tDCS and motor practise post-stroke.

  13. Postnatal donor lymphocytes enhance prenatally-created chimerism at the risk of graft-versus-host disease.

    PubMed

    Chen, Jeng-Chang; Ou, Liang-Shiou; Yu, Hsiu-Yueh; Kuo, Ming-Ling; Chang, Pei-Yeh; Chang, Hsueh-Ling

    2015-01-01

    The major barrier to clinical application of in utero hematopoietic stem cell transplantation is insufficient chimerism for phenotypic correction of target diseases or induction of graft tolerance. Postnatal donor lymphocyte infusion (DLI) may enhance donor cell levels so as to further facilitate tolerance induction. We created murine mixed chimeras in utero. Chimeras with <10% donor cells were subjected to postnatal DLI to evaluate the effects of DLI on chimerism augmentation and skin tolerance induction. Within one day after DLI, recipients experienced a transient peaking of donor chimerism, which could be as high as 20~40%. However, the transient chimerism peaking didn't benefit donor skin survivals despite immediate skin placement after DLI. In case of fruitful DLI, chimerism augmentation was usually observed after a latent period of 2~4 weeks. Otherwise, chimerism would return to around pre-DLI levels by days 7~14. Peripheral chimerism of >3% could be consistently boosted up to >10%, whereas chimerism of <0.2% hardly showed any significant enhancement. As for chimerism levels of 0.2~3%, chimerism augmentation up to >10% succeeded in 3(15%) of 20 recipients. Notably, chimerism augmentation by postnatal DLI was often associated with unexpected death or graft-versus-host disease (GVHD). In conclusion, transient chimerism augmentation by DLI played no role in facilitating graft tolerance. Substantial augmentation by DLI demanded a threshold chimerism level and posed a serious risk of GVHD to the recipients. It raised the concern about using postnatal DLI to broaden therapeutic horizons of in utero hematopoietic stem cell transplantation.

  14. Deletional and regulatory mechanisms coalesce to drive transplantation tolerance through mixed chimerism.

    PubMed

    Hock, Karin; Mahr, Benedikt; Schwarz, Christoph; Wekerle, Thomas

    2015-09-01

    Establishing donor-specific immunological tolerance could improve long-term outcome by obviating the need for immunosuppressive drug therapy, which is currently required to control alloreactivity after organ transplantation. Mixed chimerism is defined as the engraftment of donor hematopoietic stem cells in the recipient, leading to viable coexistence of both donor and recipient leukocytes. In numerous experimental models, cotransplantation of donor bone marrow (BM) into preconditioned (e.g., through irradiation or cytotoxic drugs) recipients leads to transplantation tolerance through (mixed) chimerism. Mixed chimerism offers immunological advantages for clinical translation; pilot trials have established proof of concept by deliberately inducing tolerance in humans. Widespread clinical application is prevented, however, by the harsh preconditioning currently necessary for permitting BM engraftment. Recently, the immunological mechanisms inducing and maintaining tolerance in experimental mixed chimerism have been defined, revealing a more prominent role for regulation than historically assumed. The evidence from murine models suggests that both deletional and regulatory mechanisms are critical in promoting complete tolerance, encompassing also the minor histocompatibility antigens. Here, we review the current understanding of tolerance through mixed chimerism and provide an outlook on how to realize widespread clinical translation based on mechanistic insights gained from chimerism protocols, including cell therapy with polyclonal regulatory T cells.

  15. Effects of chimerism in sheep-goat concepti that developed from blastomere-aggregation embryos.

    PubMed

    Ruffing, N A; Anderson, G B; Bondurant, R H; Currie, W B; Pashen, R L

    1993-04-01

    Chimeric sheep-goat pregnancies were established in 24 ewes and 29 does by transferring 251 embryos, prepared by the blastomere-aggregation technique, to 52 ewes and 61 does. Fifteen does experienced early pregnancy failure; however, term offspring were delivered by 24 ewes (17 lambs, 3 kids, 6 chimeras) and 14 does (6 lambs, 9 kids, 6 chimeras). (Fetal classifications were based on phenotype, red blood cell isozymes, and lymphocyte antigen expression). RIAs for ovine and caprine placental lactogen detected chimerism in the binucleate cell population of the trophoblast throughout the pregnancies of 2 ewes and 7 does; these pregnancies resulted in the birth of 12 healthy offspring. Histological examinations of intact placentomes from 2 of these recipients revealed a continuous cellular trophoblast apposed to a syncytium as in normal placentas. Chimerism was detected electrophoretically in the membranes of the placentas with binucleate cell chimerism and in 17/28 of the other placentas. Data collected on placental lactogen production, chimerism in the conceptus, and placental morphometry were examined with respect to the stages of the blastomeres aggregated to form the chimeric embryo and with respect to fetal status at delivery. For comparison, analogous data were collected on sheep-goat concepti that developed from embryos prepared by inner cell mass transplantation. PMID:8485255

  16. Does HLA-dependent chimerism underlie the pathogenesis of juvenile dermatomyositis?

    PubMed

    Reed, Ann M; McNallan, Kelly; Wettstein, Peter; Vehe, Richard; Ober, Carole

    2004-04-15

    Juvenile dermatomyositis (JDM) is a multisystem autoimmune disease that at times resembles chronic graft-vs-host disease. This led us to suggest that nonself cells may play a role in the disease process. In this study we examined the relationship between HLA genotype and the presence of maternally derived chimeric cells in JDM patients and healthy controls, and assessed immunologic activity in the chimeric cells. We identified chimeric cells more often in children with JDM (60 of 72) than in their unaffected siblings (11 of 48) or in healthy controls (5 of 29). The presence of chimerism in the JDM patients, their healthy siblings, and unaffected control children was associated with a HLA-DQA1*0501 allele in the mother (p = 0.011). Further, we show that maternally transferred chimeric T cells are responsive to the host's (JDM childs') lymphocytes (33.75 +/- 8.4 IFN-gamma-producing cells from JDM cells vs 5.0 +/- 1.25 from maternal cells), and that this is a memory response. These combined data indicate that chimeric cells play a direct role in the JDM disease process and that the mother's HLA genotype facilitates the transfer and/or persistence of maternal cells in the fetal circulation. PMID:15067086

  17. Effects of pharmacological concentrations of dietary zinc oxide on growth of post-weaning pigs: a meta-analysis.

    PubMed

    Sales, James

    2013-06-01

    Pharmacological dietary zinc (Zn) concentrations of 1,000 to 3,000 mg/kg diet from Zn oxide have been found to increase growth in post-weaning pigs. However, results were inconsistent among studies. A frequentist meta-analysis, in which effects were numerically described with standardized effect sizes (Hedges's g), was conducted in order to identify and quantify the responses in average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G/F) in post-weaning pigs upon dietary Zn supplementation from Zn oxide. The inability of independent continuous variables to explain significant heterogeneity obtained with fixed effect models necessitated the use of random effects models to calculate summary statistics. Dietary Zn supplementation increased (P < 0.05) ADG (mean effect size = 1.086, 95 % confidence intervals = 0.905-1.266, 26 studies, 72 comparisons), ADFI (mean effect size = 0.794, 95 % confidence intervals = 0.616-0.971, 25 studies, 71 comparisons), and G/F (mean effect size = 0.566, 95 % confidence intervals = 0.422-0.710, 24 studies, 70 comparisons). Zinc oxide provided a feasible alternative to in-feed antibiotics to improve growth in post-weaning pigs, and its reduction in diets due to potential environmental pollution will have to be negated by alternative feed additives and management strategies in order to prevent economic losses.

  18. Forced expression of chimeric human fibroblast tropomyosin mutants affects cytokinesis

    PubMed Central

    1995-01-01

    Human fibroblasts generate at least eight tropomyosin (TM) isoforms (hTM1, hTM2, hTM3, hTM4, hTM5, hTM5a, hTM5b, and hTMsm alpha) from four distinct genes, and we have previously demonstrated that bacterially produced chimera hTM5/3 exhibits an unusually high affinity for actin filaments and a loss of the salt dependence typical for TM-actin binding (Novy, R.E., J. R. Sellers, L.-F. Liu, and J.J.-C. Lin, 1993. Cell Motil. & Cytoskeleton. 26: 248-261). To examine the functional consequences of expressing this mutant TM isoform in vivo, we have transfected CHO cells with the full-length cDNA for hTM5/3 and compared them to cells transfected with hTM3 and hTM5. Immunofluorescence microscopy reveals that stably transfected CHO cells incorporate force- expressed hTM3 and hTM5 into stress fibers with no significant effect on general cell morphology, microfilament organization or cytokinesis. In stable lines expressing hTM5/3, however, cell division is slow and sometimes incomplete. The doubling time and the incidence of multinucleate cells in the stable hTM5/3 lines roughly parallel expression levels. A closely related chimeric isoform hTM5/2, which differs only in the internal, alternatively spliced exon also produces defects in cytokinesis, suggesting that normal TM function may involve coordination between the amino and carboxy terminal regions. This coordination may be prevented in the chimeric mutants. As bacterially produced hTM5/3 and hTM5/2 can displace hTM3 and hTM5 from actin filaments in vitro, it is likely that CHO-expressed hTM5/3 and hTM5/2 can displace endogenous TMs to act dominantly in vivo. These results support a role for nonmuscle TM isoforms in the fine tuning of microfilament organization during cytokinesis. Additionally, we find that overexpression of TM does not stabilize endogenous microfilaments, rather, the hTM-expressing cells are actually more sensitive to cytochalasin B. This suggests that regulation of microfilament integrity in vivo

  19. Genomic Convergence Analysis of Schizophrenia: mRNA Sequencing Reveals Altered Synaptic Vesicular Transport in Post-Mortem Cerebellum

    PubMed Central

    Mudge, Joann; Miller, Neil A.; Khrebtukova, Irina; Lindquist, Ingrid E.; May, Gregory D.; Huntley, Jim J.; Luo, Shujun; Zhang, Lu; van Velkinburgh, Jennifer C.; Farmer, Andrew D.; Lewis, Sharon; Beavis, William D.; Schilkey, Faye D.; Virk, Selene M.; Black, C. Forrest; Myers, M. Kathy; Mader, Lar C.; Langley, Ray J.; Utsey, John P.; Kim, Ryan W.; Roberts, Rosalinda C.; Khalsa, Sat Kirpal; Garcia, Meredith; Ambriz-Griffith, Victoria; Harlan, Richard; Czika, Wendy; Martin, Stanton; Wolfinger, Russell D.; Perrone-Bizzozero, Nora I.; Schroth, Gary P.; Kingsmore, Stephen F.

    2008-01-01

    Schizophrenia (SCZ) is a common, disabling mental illness with high heritability but complex, poorly understood genetic etiology. As the first phase of a genomic convergence analysis of SCZ, we generated 16.7 billion nucleotides of short read, shotgun sequences of cDNA from post-mortem cerebellar cortices of 14 patients and six, matched controls. A rigorous analysis pipeline was developed for analysis of digital gene expression studies. Sequences aligned to approximately 33,200 transcripts in each sample, with average coverage of 450 reads per gene. Following adjustments for confounding clinical, sample and experimental sources of variation, 215 genes differed significantly in expression between cases and controls. Golgi apparatus, vesicular transport, membrane association, Zinc binding and regulation of transcription were over-represented among differentially expressed genes. Twenty three genes with altered expression and involvement in presynaptic vesicular transport, Golgi function and GABAergic neurotransmission define a unifying molecular hypothesis for dysfunction in cerebellar cortex in SCZ. PMID:18985160

  20. Post-crisis analysis of an ineffective tsunami alert: the 2010 earthquake in Maule, Chile.

    PubMed

    Soulé, Bastien

    2014-04-01

    Considering its huge magnitude and its location in a densely populated area of Chile, the Maule seism of 27 February 2010 generated a low amount of victims. However, post-seismic tsunamis were particularly devastating on that day; surprisingly, no full alert was launched, not at the national, regional or local level. This earthquake and associated tsunamis are of interest in the context of natural hazards management as well as crisis management planning. Instead of focusing exclusively on the event itself, this article places emphasis on the process, systems and long-term approach that led the tsunami alert mechanism to be ineffectual. Notably, this perspective reveals interrelated forerunner signs of vulnerability.

  1. Hazard Analysis for Post-Fire Debris-Flow Potential in Arizona

    NASA Astrophysics Data System (ADS)

    Youberg, A.; Koestner, K. A.; Schiefer, E.; Neary, D. G.

    2011-12-01

    Several large, devastating wildfires occurred in Arizona during the past 2 years, after a 4-year period without any large wildfires. In June, 2010, the human-caused Schultz Fire near Flagstaff burned 6,100 ha of mostly steep terrain. Subsequent rains from the 4th wettest monsoon on record produced numerous debris flows, significant erosion, and substantial flooding of the downslope residential areas. In May and June of 2011, 3 very large human-caused wildfires (Wallow, Horseshoe 2, and Monument Fires) burned over 320,000 ha, posing serious threats to communities below burned slopes. The Burned Area Emergency Response (BAER) teams, in need of a rapid method to assess the potential for post-fire debris flows, turned to models developed by the USGS for this purpose [Cannon and others, 2010, GSA Bull, 122(1-2), 127-144]. These models, while providing quick results, have not been evaluated for use in Arizona's varied physiographic provinces. Here we use data from the Schultz Fire to compare basin responses with those predicted by the USGS post-fire debris-flow models. Data from the Schultz Fire includes detailed field documentation of debris-flow occurrence and runout distances, 1:12,000 stereo aerial photographs, high-resolution digital elevation models (DEMs) and tipping-bucket rainfall data. These data document debris-flow producing storms, basin response, and the extent of debris-flow runout, and provide estimates of debris-flow volumes. The hydrologic responses from 30 small, steep, upper basins burned by the Schultz Fire were assessed for debris or flood flow occurrences. Nineteen basins produced debris flows during a July 20th storm that had a peak 10-minute intensity of 24 mm. A second storm on August 16th, with a peak 10-minute intensity of 15 mm, produced additional debris flows in several of the same basins. Of the 30 basins assessed, 19 were completely burned; four at high severity and 12 at moderate to high severity. The basin with the smallest burned area

  2. Bioactivity and structural properties of chimeric analogs of the starfish SALMFamide neuropeptides S1 and S2.

    PubMed

    Jones, Christopher E; Otara, Claire B; Younan, Nadine D; Viles, John H; Elphick, Maurice R

    2014-10-01

    The starfish SALMFamide neuropeptides S1 (GFNSALMFamide) and S2 (SGPYSFNSGLTFamide) are the prototypical members of a family of neuropeptides that act as muscle relaxants in echinoderms. Comparison of the bioactivity of S1 and S2 as muscle relaxants has revealed that S2 is ten times more potent than S1. Here we investigated a structural basis for this difference in potency by comparing the bioactivity and solution conformations (using NMR and CD spectroscopy) of S1 and S2 with three chimeric analogs of these peptides. A peptide comprising S1 with the addition of S2's N-terminal tetrapeptide (Long S1 or LS1; SGPYGFNSALMFamide) was not significantly different to S1 in its bioactivity and did not exhibit concentration-dependent structuring seen with S2. An analog of S1 with its penultimate residue substituted from S2 (S1(T); GFNSALTFamide) exhibited S1-like bioactivity and structure. However, an analog of S2 with its penultimate residue substituted from S1 (S2(M); SGPYSFNSGLMFamide) exhibited loss of S2-type bioactivity and structural properties. Collectively, our data indicate that the C-terminal regions of S1 and S2 are the key determinants of their differing bioactivity. However, the N-terminal region of S2 may influence its bioactivity by conferring structural stability in solution. Thus, analysis of chimeric SALMFamides has revealed how neuropeptide bioactivity is determined by a complex interplay of sequence and conformation.

  3. Effective strategies for weight loss in post-partum women: a systematic review and meta-analysis.

    PubMed

    Lim, S; O'Reilly, S; Behrens, H; Skinner, T; Ellis, I; Dunbar, J A

    2015-11-01

    Post-partum weight loss is critical to preventing and managing obesity in women, but the results from lifestyle interventions are variable and the components associated with successful outcomes are not yet clearly identified. This study aimed to identify lifestyle intervention strategies associated with weight loss in post-partum women. MEDLINE, EMBASE, PubMed, CINAHL and four other databases were searched for lifestyle intervention studies (diet or exercise or both) in post-partum women (within 12 months of delivery) published up to July 2014. The primary outcome was weight loss. Subgroup analyses were conducted for self-monitoring, individual or group setting, intervention duration, intervention types, the use of technology as a support, and home- or centre-based interventions. From 12,673 studies, 46 studies were included in systematic review and 32 randomized controlled trials were eligible for meta-analysis (1,892 women, age 24-36 years). Studies with self-monitoring had significantly greater weight lost than those without (-4.61 kg [-7.08, -2.15] vs. -1.34 kg [-1.66, -1.02], P = 0.01 for subgroup differences). Diet and physical activity when combined were significantly more effective on weight loss compared with physical activity alone (-3.24 kg [-4.59, -1.90] vs. -1.63 kg [-2.16, -1.10], P < 0.001 for subgroup differences). Lifestyle interventions that use self-monitoring and take a combined diet-and-exercise approach have significantly greater weight loss in post-partum women.

  4. Stress distribution on dentin-cement-post interface varying root canal and glass fiber post diameters. A three-dimensional finite element analysis based on micro-CT data

    PubMed Central

    LAZARI, Priscilla Cardoso; de OLIVEIRA, Rodrigo Caldeira Nunes; ANCHIETA, Rodolfo Bruniera; de ALMEIDA, Erika Oliveira; FREITAS JUNIOR, Amilcar Chagas; KINA, Sidney; ROCHA, Eduardo Passos

    2013-01-01

    Objective The aim of the present study was to analyze the influence of root canal and glass fiber post diameters on the biomechanical behavior of the dentin/cement/post interface of a root-filled tooth using 3D finite element analysis. Material and Methods Six models were built using micro-CT imaging data and SolidWorks 2007 software, varying the root canal (C) and the glass fiber post (P) diameters: C1P1-C=1 mm and P=1 mm; C2P1-C=2 mm and P=1 mm; C2P2-C=2 mm and P=2 mm; C3P1-C=3 mm and P=1 mm; C3P2-C=3 mm and P=2 mm; and C3P3-C=3 mm and P=3 mm. The numerical analysis was conducted with ANSYS Workbench 10.0. An oblique force (180 N at 45º) was applied to the palatal surface of the central incisor. The periodontal ligament surface was constrained on the three axes (x=y=z=0). Maximum principal stress (σmax) values were evaluated for the root dentin, cement layer, and glass fiber post. Results: The most evident stress was observed in the glass fiber post at C3P1 (323 MPa), and the maximum stress in the cement layer occurred at C1P1 (43.2 MPa). The stress on the root dentin was almost constant in all models with a peak in tension at C2P1 (64.5 MPa). Conclusion The greatest discrepancy between root canal and post diameters is favorable for stress concentration at the post surface. The dentin remaining after the various root canal preparations did not increase the stress levels on the root. PMID:24473716

  5. Whole proteome analysis of post-translational modifications: applications of mass-spectrometry for proteogenomic annotation

    SciTech Connect

    Gupta, Nitin; Tanner, Stephen; Jaitly, Navdeep; Adkins, Joshua N.; Lipton, Mary S.; Edwards, Robert; Romine, Margaret F.; Osterman, Andrei; Bafna, Vineet; Smith, Richard D.; Pevzner, Pavel A.

    2007-09-04

    While bacterial genome annotations have significantly improved in recent years, techniques for bacterial proteome annotation (including post-translational chemical modifications, signal peptides, proteolytic events, etc.) are still in their infancy. At the same time, the number of sequenced bacterial genomes is rising sharply, far outpacing our ability to validate the predicted genes, let alone annotate bacterial proteomes. In this study, we use tandem mass spectrometry (MS/MS) to annotate the proteome of Shewanella oneidensis MR-1, an important microbe for bioremediation. In particular, we provide the first comprehensive map of post-translational modifications in a bacterial genome, including a large number of chemical modifications, signal peptide cleavages and cleavage of N-terminal methionine residues. We also detect multiple genes that were missed or assigned incorrect start positions by gene prediction programs and suggest corrections to improve the gene annotation. This study demonstrates that complementing every genome sequencing project by an MS/MS project would significantly improve both genome and proteome annotations for a reasonable cost.

  6. Transcriptome analysis of differentiating trypanosomes reveals the existence of multiple post-transcriptional regulons

    PubMed Central

    Queiroz, Rafael; Benz, Corinna; Fellenberg, Kurt; Hoheisel, Jörg D; Clayton, Christine

    2009-01-01

    Background Trypanosome gene expression is regulated almost exclusively at the post-transcriptional level, with mRNA degradation playing a decisive role. When trypanosomes are transferred from the blood of a mammal to the midgut of a Tsetse fly, they transform to procyclic forms: gene expression is reprogrammed, changing the cell surface and switching the mode of energy metabolism. Within the blood, trypanosomes can pre-adapt for Tsetse transmission, becoming growth-arrested stumpy forms. We describe here the transitions in gene expression that occur during differentiation of in-vitro cultured bloodstream forms to procyclic forms. Results Some mRNAs showed changes within 30 min of cis-aconitate addition, whereas others responded 12-24 hours later. For the first 12 h after addition of cis-aconitate, cells accumulated at the G1 phase of the cell cycle, and showed decreases in mRNAs required for proliferation, mimicking the changes seen in stumpy forms: many mRNAs needed for ribosomal and flagellar biogenesis showed striking co-regulation. Other mRNAs encoding components of signal transduction pathways and potential regulators were specifically induced only during differentiation. Messenger RNAs encoding proteins required for individual metabolic pathways were often co-regulated. Conclusion Trypanosome genes form post-transcriptional regulons in which mRNAs with functions in particular pathways, or encoding components of protein complexes, show almost identical patterns of regulation. PMID:19857263

  7. Post-Buckling Analysis of Curved Honeycomb Sandwich Panels Containing Interfacial Disbonds

    NASA Technical Reports Server (NTRS)

    Pineda, Evan J.; Bednarcyk, Brett A.; Krivanek, Thomas K.

    2016-01-01

    A numerical study on the effect of facesheet-core disbonds on the post-buckling response of curved honeycomb sandwich panels is presented herein. This work was conducted as part of the development of a damage tolerance plan for the next-generation Space Launch System heavy lift launch vehicle payload fairing. As such, the study utilized full-scale fairing barrel segments as the structure of interest. The panels were composed of carbon fiber reinforced polymer facesheets and aluminum honeycomb core. The panels were analyzed numerically using the finite element method incorporating geometric nonlinearity. In a predetermined circular region, facesheet and core nodes were detached to simulate a disbond, between the outer mold line facesheet and honeycomb core, induced via low-speed impact. Surface-to-surface contact in the disbonded region was invoked to prevent interpenetration of the facesheet and core elements and obtain realistic stresses in the core. The diameter of this disbonded region was varied and the effect of the size of the disbond on the post-buckling response was observed. Significant changes in the slope of the edge load-deflection response were used to determine the onset of global buckling and corresponding buckling load. Finally, several studies were conducted to determine the sensitivity of the numerical predictions to refinement in the finite element mesh.

  8. Risk assessment of relapse by lineage-specific monitoring of chimerism in children undergoing allogeneic stem cell transplantation for acute lymphoblastic leukemia

    PubMed Central

    Preuner, Sandra; Peters, Christina; Pötschger, Ulrike; Daxberger, Helga; Fritsch, Gerhard; Geyeregger, Rene; Schrauder, André; von Stackelberg, Arend; Schrappe, Martin; Bader, Peter; Ebell, Wolfram; Eckert, Cornelia; Lang, Peter; Sykora, Karl-Walter; Schrum, Johanna; Kremens, Bernhard; Ehlert, Karoline; Albert, Michael H.; Meisel, Roland; Lawitschka, Anita; Mann, Georg; Panzer-Grümayer, Renate; Güngör, Tayfun; Holter, Wolfgang; Strahm, Brigitte; Gruhn, Bernd; Schulz, Ansgar; Woessmann, Wilhelm; Lion, Thomas

    2016-01-01

    Allogeneic hematopoietic stem cell transplantation is required as rescue therapy in about 20% of pediatric patients with acute lymphoblastic leukemia. However, the relapse rates are considerable, and relapse confers a poor outcome. Early assessment of the risk of relapse is therefore of paramount importance for the development of appropriate measures. We used the EuroChimerism approach to investigate the potential impact of lineage-specific chimerism testing for relapse-risk analysis in 162 pediatric patients with acute lymphoblastic leukemia after allogeneic stem cell transplantation in a multicenter study based on standardized transplantation protocols. Within a median observation time of 4.5 years, relapses have occurred in 41/162 patients at a median of 0.6 years after transplantation (range, 0.13–5.7 years). Prospective screening at defined consecutive time points revealed that reappearance of recipient-derived cells within the CD34+ and CD8+ cell subsets display the most significant association with the occurrence of relapses with hazard ratios of 5.2 (P=0.003) and 2.8 (P=0.008), respectively. The appearance of recipient cells after a period of pure donor chimerism in the CD34+ and CD8+ leukocyte subsets revealed dynamics indicative of a significantly elevated risk of relapse or imminent disease recurrence. Assessment of chimerism within these lineages can therefore provide complementary information for further diagnostic and, potentially, therapeutic purposes aiming at the prevention of overt relapse. This study was registered at clinical.trials.gov with the number NC01423747. PMID:26869631

  9. Chimeric Bovine Respiratory Syncytial Virus with Attachment and Fusion Glycoproteins Replaced by Bovine Parainfluenza Virus Type 3 Hemagglutinin-Neuraminidase and Fusion Proteins

    PubMed Central

    Stope, Matthias B.; Karger, Axel; Schmidt, Ulrike; Buchholz, Ursula J.

    2001-01-01

    Chimeric bovine respiratory syncytial viruses (BRSV) expressing glycoproteins of bovine parainfluenza virus type 3 (BPIV-3) instead of BRSV glycoproteins were generated from cDNA. In the BRSV antigenome cDNA, the open reading frames of the major BRSV glycoproteins, attachment protein G and fusion protein F, were replaced individually or together by those of the BPIV-3 hemagglutinin-neuraminidase (HN) and/or fusion (F) glycoproteins. Recombinant virus could not be recovered from cDNA when the BRSV F open reading frame was replaced by the BPIV-3 F open reading frame. However, cDNA recovery of the chimeric virus rBRSV-HNF, with both glycoproteins replaced simultaneously, and of the chimeric virus rBRSV-HN, with the BRSV G protein replaced by BPIV-3 HN, was successful. The replication rates of both chimeras were similar to that of standard rBRSV. Moreover, rBRSV-HNF was neutralized by antibodies specific for BPIV-3, but not by antibodies specific to BRSV, demonstrating that the BRSV glycoproteins can be functionally replaced by BPIV-3 glycoproteins. In contrast, rBRSV-HN was neutralized by BRSV-specific antisera, but not by BPIV-3 specific sera, showing that infection of rBRSV-HN is mediated by BRSV F. Hemadsorption of cells infected with rBRSV-HNF and rBRSV-HN proved that BPIV-3 HN protein expressed by rBRSV is functional. Colocalization of the BPIV-3 glycoproteins with BRSV M protein was demonstrated by confocal laser scan microscopy. Moreover, protein analysis revealed that the BPIV-3 glycoproteins were present in chimeric virions. Taken together, these data indicate that the heterologous glycoproteins were not only expressed but were incorporated into the envelope of recombinant BRSV. Thus, the envelope glycoproteins derived from a member of the Respirovirus genus can together functionally replace their homologs in a Pneumovirus background. PMID:11533200

  10. Expression and purification of chimeric peptide comprising EGFR B-cell epitope and measles virus fusion protein T-cell epitope in Escherichia coli.

    PubMed

    Wu, Meizhi; Zhao, Lin; Zhu, Lei; Chen, Zhange; Li, Huangjin

    2013-03-01

    Chimeric peptide MVF-EGFR(237-267), comprising a B-cell epitope from the dimerization interface of human epidermal growth factor receptor (EGFR) and a promiscuous T-cell epitope from measles virus fusion protein (MVF), is a promising candidate antigen peptide for therapeutic vaccine. To establish a high-efficiency preparation process of this small peptide, the coding sequence was cloned into pET-21b and pET-32a respectively, to be expressed alone or in the form of fusion protein with thioredoxin (Trx) and His(6)-tag in Escherichia coli BL21 (DE3). The chimeric peptide failed to be expressed alone, but over-expressed in the fusion form, which presented as soluble protein and took up more than 30% of total proteins of host cells. The fusion protein was seriously degraded during the cell disruption, in which endogenous metalloproteinase played a key role. Degradation of target peptide was inhibited by combined application of EDTA in the cell disruption buffer and a step of Source 30Q anion exchange chromatography (AEC) before metal-chelating chromatography (MCAC) for purifying His(6)-tagged fusion protein. The chimeric peptide was recovered from the purified fusion protein by enterokinase digestion at a yield of 3.0 mg/L bacteria culture with a purity of more than 95%. Immunogenicity analysis showed that the recombinant chimeric peptide was able to arouse more than 1×10(4) titers of specific antibody in BALB/c mice. Present work laid a solid foundation for the development of therapeutic peptide vaccine targeting EGFR dimerization and provided a convenient and low-cost preparation method for small peptides.

  11. Chimeric cellulase matrix for investigating intramolecular synergism between non-hydrolytic disruptive functions of carbohydrate-binding modules and catalytic hydrolysis.

    PubMed

    Wang, Yuguo; Tang, Rentao; Tao, Jin; Wang, Xiaonan; Zheng, Baisong; Feng, Yan

    2012-08-24

    The conversion of renewable cellulosic biomass is of considerable interest for the production of biofuels and materials. The bottleneck in the efficient conversion is the compactness and resistance of crystalline cellulose. Carbohydrate-binding modules (CBMs), which disrupt crystalline cellulose via non-hydrolytic mechanisms, are expected to overcome this bottleneck. However, the lack of convenient methods for quantitative analysis of the disruptive functions of CBMs have hindered systematic studies and molecular modifications. Here we established a practical and systematic platform for quantifying and comparing the non-hydrolytic disruptive activities of CBMs via the synergism of CBMs and a catalytic module within designed chimeric cellulase molecules. Bioinformatics and computational biology were also used to provide a deeper understanding. A convenient vector was constructed to serve as a cellulase matrix into which heterologous CBM sequences can be easily inserted. The resulting chimeric cellulases were suitable for studying disruptive functions, and their activities quantitatively reflected the disruptive functions of CBMs on crystalline cellulose. In addition, this cellulase matrix can be used to construct novel chimeric cellulases with high hydrolytic activities toward crystalline cellulose. PMID:22778256

  12. Chimeric TALE recombinases with programmable DNA sequence specificity.

    PubMed

    Mercer, Andrew C; Gaj, Thomas; Fuller, Roberta P; Barbas, Carlos F

    2012-11-01

    Site-specific recombinases are powerful tools for genome engineering. Hyperactivated variants of the resolvase/invertase family of serine recombinases function without accessory factors, and thus can be re-targeted to sequences of interest by replacing native DNA-binding domains (DBDs) with engineered zinc-finger proteins (ZFPs). However, imperfect modularity with particular domains, lack of high-affinity binding to all DNA triplets, and difficulty in construction has hindered the widespread adoption of ZFPs in unspecialized laboratories. The discovery of a novel type of DBD in transcription activator-like effector (TALE) proteins from Xanthomonas provides an alternative to ZFPs. Here we describe chimeric TALE recombinases (TALERs): engineered fusions between a hyperactivated catalytic domain from the DNA invertase Gin and an optimized TALE architecture. We use a library of incrementally truncated TALE variants to identify TALER fusions that modify DNA with efficiency and specificity comparable to zinc-finger recombinases in bacterial cells. We also show that TALERs recombine DNA in mammalian cells. The TALER architecture described herein provides a platform for insertion of customized TALE domains, thus significantly expanding the targeting capacity of engineered recombinases and their potential applications in biotechnology and medicine.

  13. Chimerical pyrene-based [7]helicenes as twisted polycondensed aromatics.

    PubMed

    Buchta, Michal; Rybáček, Jiří; Jančařík, Andrej; Kudale, Amit A; Buděšínský, Miloš; Chocholoušová, Jana Vacek; Vacek, Jaroslav; Bednárová, Lucie; Císařová, Ivana; Bodwell, Graham J; Starý, Ivo; Stará, Irena G

    2015-06-01

    Chimerical pyrene-based dibenzo[7]helicene rac-1 and 2H-pyran[7]helicene (M,R,R)-(-)-2, in which two pyrene subunits are fused to the [7]helicene/[7]heterohelicene scaffold, were synthesised by means of Ni(0) - or Co(I) -mediated [2+2+2] cycloisomerisation of dipyrenyl-acetylene-derived triynes. Pyrene-based dibenzo[7]helicene 1 was obtained in enantioenriched form by enantioselective cycloisomerisation under Ni(0) /QUINAP catalysis (57 % ee) or in enantiopure form by racemate resolution by liquid chromatography on a chiral column. 1,3-Allylic-type strain-controlled diastereoselective cycloisomerisation was employed in the synthesis of enantiopure (M,R,R)-(-)-2. Physicochemical properties of 1 and 2 encompassing the helicity assignment, stability to racemisation, X-ray crystal structure, UV/Vis, experimental/calculated electronic circular dichroism and fluorescence spectra were studied. Accordingly, comparison of the X-ray crystal structure of (M,R,R)-(-)-2 with calculated structures (DFT: B3LYP/cc-pVDZ, B97D/cc-pVDZ) indicated that its helical backbone is slightly over-flattened owing to intramolecular dispersion forces between tert-butylated pyrene subunits. Both 1 and 2 are fluorescent (with quantum yields in dichloromethane of ΦF =0.10 and 0.17, respectively) and are suggested to form intramolecular excimer states upon excitation, which are remarkably stabilised and exhibit large Stokes shifts (296 and 203 nm, respectively).

  14. Radial symmetry in a chimeric glutamate receptor pore

    NASA Astrophysics Data System (ADS)

    Wilding, Timothy J.; Lopez, Melany N.; Huettner, James E.

    2014-02-01

    Ionotropic glutamate receptors comprise two conformationally different A/C and B/D subunit pairs. Closed channels exhibit fourfold radial symmetry in the transmembrane domain (TMD) but transition to twofold dimer-of-dimers symmetry for extracellular ligand binding and N-terminal domains. Here, to evaluate symmetry in open pores we analysed interaction between the Q/R editing site near the pore loop apex and the transmembrane M3 helix of kainate receptor subunit GluK2. Chimeric subunits that combined the GluK2 TMD with extracellular segments from NMDA receptors, which are obligate heteromers, yielded channels made up of A/C and B/D subunit pairs with distinct substitutions along M3 and/or Q/R site editing status, in an otherwise identical homotetrameric TMD. Our results indicate that Q/R site interaction with M3 occurs within individual subunits and is essentially the same for both A/C and B/D subunit conformations, suggesting that fourfold pore symmetry persists in the open state.

  15. Chimeric animal models in human stem cell biology.

    PubMed

    Glover, Joel C; Boulland, Jean-Luc; Halasi, Gabor; Kasumacic, Nedim

    2009-01-01

    The clinical use of stem cells for regenerative medicine is critically dependent on preclinical studies in animal models. In this review we examine some of the key issues and challenges in the use of animal models to study human stem cell biology-experimental standardization, body size, immunological barriers, cell survival factors, fusion of host and donor cells, and in vivo imaging and tracking. We focus particular attention on the various imaging modalities that can be used to track cells in living animals, comparing their strengths and weaknesses and describing technical developments that are likely to lead to new opportunities for the dynamic assessment of stem cell behavior in vivo. We then provide an overview of some of the most commonly used animal models, their advantages and disadvantages, and examples of their use for xenotypic transplantation of human stem cells, with separate reviews of models involving rodents, ungulates, nonhuman primates, and the chicken embryo. As the use of human somatic, embryonic, and induced pluripotent stem cells increases, so too will the range of applications for these animal models. It is likely that increasingly sophisticated uses of human/animal chimeric models will be developed through advances in genetic manipulation, cell delivery, and in vivo imaging.

  16. The promise and potential pitfalls of chimeric antigen receptors.

    PubMed

    Sadelain, Michel; Brentjens, Renier; Rivière, Isabelle

    2009-04-01

    One important purpose of T cell engineering is to generate tumor-targeted T cells through the genetic transfer of antigen-specific receptors, which consist of either physiological, MHC-restricted T cell receptors (TCRs) or non MHC-restricted chimeric antigen receptors (CARs). CARs combine antigen-specificity and T cell activating properties in a single fusion molecule. First generation CARs, which included as their signaling domain the cytoplasmic region of the CD3zeta or Fc receptor gamma chain, effectively redirected T cell cytotoxicity but failed to enable T cell proliferation and survival upon repeated antigen exposure. Receptors encompassing both CD28 and CD3zeta are the prototypes for second generation CARs, which are now rapidly expanding to a diverse array of receptors with different functional properties. First generation CARs have been tested in phase I clinical studies in patients with ovarian cancer, renal cancer, lymphoma, and neuroblastoma, where they have induced modest responses. Second generation CARs, which are just now entering the clinical arena in the B cell malignancies and other cancers, will provide a more significant test for this approach. If the immunogenicity of CARs can be averted, the versatility of their design and HLA-independent antigen recognition will make CARs tools of choice for T cell engineering for the development of targeted cancer immunotherapies.

  17. Utilizing Chimeric Antigen Receptors to Direct Natural Killer Cell Activity

    PubMed Central

    Hermanson, David L.; Kaufman, Dan S.

    2015-01-01

    Natural killer (NK) cells represent an attractive lymphocyte population for cancer immunotherapy due to their ability to lyse tumor targets without prior sensitization and without need for human leukocyte antigens-matching. Chimeric antigen receptors (CARs) are able to enhance lymphocyte targeting and activation toward diverse malignancies. CARs consist of an external recognition domain (typically a small chain variable fragment) directed at a specific tumor antigen that is linked with one or more intracellular signaling domains that mediate lymphocyte activation. Most CAR studies have focused on their expression in T cells. However, use of CARs in NK cells is starting to gain traction because they provide a method to redirect these cells more specifically to target refractory cancers. CAR-mediated anti-tumor activity has been demonstrated using NK cell lines, as well as NK cells isolated from peripheral blood, and NK cells produced from human pluripotent stem cells. This review will outline the CAR constructs that have been reported in NK cells with a focus on comparing the use of different signaling domains in combination with other co-activating domains. PMID:25972867

  18. Construction of murine coronavirus mutants containing interspecies chimeric nucleocapsid proteins.

    PubMed Central

    Peng, D; Koetzner, C A; McMahon, T; Zhu, Y; Masters, P S

    1995-01-01

    Targeted RNA recombination was used to construct mouse hepatitis virus (MHV) mutants containing chimeric nucleocapsid (N) protein genes in which segments of the bovine coronavirus N gene were substituted in place of their corresponding MHV sequences. This defined portions of the two N proteins that, despite evolutionary divergence, have remained functionally equivalent. These regions included most of the centrally located RNA-binding domain and two putative spacers that link the three domains of the N protein. By contrast, the amino terminus of N, the acidic carboxy-terminal domain, and a serine- and arginine-rich segment of the central domain could not be transferred from bovine coronavirus to MHV, presumably because these parts of the molecule participate in protein-protein interactions that are specific for each virus (or, possibly, each host). Our results demonstrate that targeted recombination can be used to make extensive substitutions in the coronavirus genome and can generate recombinants that could not otherwise be made between two viruses separated by a species barrier. The implications of these findings for N protein structure and function as well as for coronavirus RNA recombination are discussed. PMID:7636993

  19. Protective and immunological behavior of chimeric yellow fever dengue vaccine.

    PubMed

    Halstead, Scott B; Russell, Philip K

    2016-03-29

    Clinical observations from the third year of the Sanofi Pasteur chimeric yellow fever dengue tetravalent vaccine (CYD) trials document both protection and vaccination-enhanced dengue disease among vaccine recipients. Children who were 5 years-old or younger when vaccinated experienced a DENV disease resulting in hospitalization at 5 times the rate of controls. On closer inspection, hospitalized cases among vaccinated seropositives, those at highest risk to hospitalized disease accompanying a dengue virus (DENV) infection, were greatly reduced by vaccination. But, seronegative individuals of all ages after being vaccinated were only modestly protected from mild to moderate disease throughout the entire observation period despite developing neutralizing antibodies at high rates. Applying a simple epidemiological model to the data, vaccinated seronegative individuals of all ages were at increased risk of developing hospitalized disease during a subsequent wild type DENV infection. The etiology of disease in placebo and vaccinated children resulting in hospitalization during a DENV infection, while clinically similar are of different origin. The implications of the observed mixture of DENV protection and enhanced disease in CYD vaccinees are discussed. PMID:26873054

  20. Long-term assessment of particulate matter using CHIMERE model

    NASA Astrophysics Data System (ADS)

    Monteiro, A.; Miranda, A. I.; Borrego, C.; Vautard, R.; Ferreira, J.; Perez, A. T.

    Particulate matter (PM) and aerosols have became a critical pollutant and object of several research applications, due to their increasing levels, especially in urban areas, causing air pollution problems and thus effects on human health. The main purpose of this study is to perform a first long-term air quality assessment for Portugal, regarding aerosols and PM pollution. The CHIMERE chemistry-transport model, forced by the MM5 meteorological fields, was applied over Portugal for 2001 year, with 10 km horizontal resolution, using an emission inventory obtained from a spatial top-down disaggregation of the 2001 national inventory database. The evaluation model exercise shows a model trend to overestimate particulate pollution episodes (peaks) at urban sites, especially in winter season. This could be due to an underprediction of the winter model vertical mixing and also to an overestimation of PM emissions. Simulated inorganic components (ammonium and sulfate) and secondary organic aerosols (SOA) were compared to measurements taken at Aveiro (northwest coast of Portugal). An underestimation of the three components was verified. However, the model is able to predict their seasonal variation. Nevertheless, as a first approach, and despite the complex topography and coastal location of Portugal affected by sea salt natural aerosols emissions, the results obtained show that the model reproduces the PM levels, temporal evolution, and spatial patterns. The concentration maps reveal that the areas with high PM values are covered by the air quality monitoring network.

  1. Designing chimeric antigen receptors to effectively and safely target tumors.

    PubMed

    Jensen, Michael C; Riddell, Stanley R

    2015-04-01

    The adoptive transfer of T cells engineered to express artificial chimeric antigen receptors CARs) that target a tumor cell surface molecule has emerged as an exciting new approach for cancer immunotherapy. Clinical trials in patients with advanced B cell malignancies treated with CD19-specific CAR-modified T cells (CAR-T) have shown impressive antitumor efficacy, leading to optimism that this approach will be useful for treating common solid tumors. Because CAR-T cells recognize tumor cells independent of their expression of human leukocyte antigen (HLA) molecules, tumors that escape conventional T cells by downregulating HLA and/or mutating components of the antigen processing machinery can be eliminated. The ability to introduce or delete additional genes in T cells has the potential to provide therapeutic cell products with novel attributes that overcome impediments to immune mediated tumor elimination in immunosuppressive tumor microenvironments. This review will discuss recent concepts in the development of effective and safe synthetic CARs for adoptive T cell therapy (ACT).

  2. Protective and immunological behavior of chimeric yellow fever dengue vaccine.

    PubMed

    Halstead, Scott B; Russell, Philip K

    2016-03-29

    Clinical observations from the third year of the Sanofi Pasteur chimeric yellow fever dengue tetravalent vaccine (CYD) trials document both protection and vaccination-enhanced dengue disease among vaccine recipients. Children who were 5 years-old or younger when vaccinated experienced a DENV disease resulting in hospitalization at 5 times the rate of controls. On closer inspection, hospitalized cases among vaccinated seropositives, those at highest risk to hospitalized disease accompanying a dengue virus (DENV) infection, were greatly reduced by vaccination. But, seronegative individuals of all ages after being vaccinated were only modestly protected from mild to moderate disease throughout the entire observation period despite developing neutralizing antibodies at high rates. Applying a simple epidemiological model to the data, vaccinated seronegative individuals of all ages were at increased risk of developing hospitalized disease during a subsequent wild type DENV infection. The etiology of disease in placebo and vaccinated children resulting in hospitalization during a DENV infection, while clinically similar are of different origin. The implications of the observed mixture of DENV protection and enhanced disease in CYD vaccinees are discussed.

  3. Impact of hematopoietic chimerism at day +14 on engraftment after unrelated donor umbilical cord blood transplantation for hematologic malignancies

    PubMed Central

    Moscardó, Federico; Sanz, Jaime; Senent, Leonor; Cantero, Susana; de la Rubia, Javier; Montesinos, Pau; Planelles, Dolores; Lorenzo, Ignacio; Cervera, Jose; Palau, Javier; Sanz, Miguel A.; Sanz, Guillermo F.

    2009-01-01

    Background Cord blood transplant is a feasible treatment alternative for adult patients with hematologic malignancies lacking a suitable HLA-matched donor. However, the kinetics of myeloid recovery is slow, and primary graft failure cannot be detected easily early after transplantation. We investigated the impact of hematopoietic chimerism status from unselected marrow cells 14 days after transplantation on predicting engraftment after a cord blood transplant. Design and Methods Seventy-one adult patients with hematologic malignancies undergoing single-unit unrelated donor cord blood transplantation after a myeloablative conditioning regimen were included in the study. All patients received conditioning regimens based on busulfan, thiotepa and antithymocyte globulin. Chimerism status was assessed analyzing short tandem repeat polymorphisms. Results The cumulative incidence of myeloid engraftment at 1 month was significantly lower in patients with mixed chimerism than in those with complete donor chimerism (55% vs. 94%; p<0.0001). For patients achieving myeloid recovery, the median time of engraftment was 16 days when donor chimerism at day + 14 was higher than 90%, compared with 24 days when donor chimerism was below this level (p<0.001). A donor chimerism level of 65% was found to be the best cut-off point for predicting primary graft failure, with a sensitivity of 97% and a specificity of 80%. The incidence of primary graft failure was 67% for patients with less than 65% donor chimerism at day +14 as compared to only 2% for those with more than 65% donor chimerism (p<0.001). Patients with mixed chimerism also had a lower cumulative incidence of platelet engraftment than those with complete chimerism (62% vs. 89%; p=0.01). Conclusions Donor-recipient chimerism status at day +14 predicts engraftment after a single-unit cord blood transplant in adults. PMID:19483157

  4. Effect of different adhesion strategies on fiber post cementation: Push-out test and scanning electron microscopy analysis

    PubMed Central

    Saraiva, Letícia Oliveria; Aguiar, Thaiane Rodrigues; Costa, Leonardo; Correr-Sobrinho, Lourenço; Muniz, Leonardo; Mathias, Paula

    2013-01-01

    Aim: The aim of this study was to investigate the effect of phosphoric acid etching and the dentin pre-treatment with sodium hypochlorite (NaOCl) on the push-out bond strength between fiber post and root canal dentin. Materials and Methods: Root canals of 48 human incisors were selected, post spaces were prepared and assigned to four groups: G1-37% phosphoric acid (15 s); G2-5.25% NaOCl (2 min) +37% phosphoric acid (15 s); G3-37% phosphoric acid (60 s); and G4-5.25% NaOCl (2 min) +37% phosphoric acid (60 s). Fiber post cementation was performed with two-step etch-and-rinse adhesive system/dual-cured resin cement according to the manufacturer's recommendation. After 24 h, each root was sectioned transversally into three slices (cervical, middle and apical) and the bond strength of each section was determined using a push-out bond strength test. Morphology analysis of the bonded interface was evaluated using a scanning electron microscopy. Push-out strength data (MPa) were analyzed by Analysis of Variance and Tukey-Kramer (α = 0.05). Results: Considering the NaOCl pre-treatment, no statistically significant differences were observed among groups; however, when the phosphoric acid was applied during 60 s in the apical portion without NaOCl pre-treatment, the bond strength was statistically significant increased. Conclusion: The NaOCl pre-treatment did not improve the bond strength of adhesive luting cement to root canal dentin. The findings suggest that the use of 37% phosphoric acid for 60 s may have a beneficial effect on bond strength in the apical root third. PMID:24403786

  5. Combined Use of Post-Ion Mobility/Collision-Induced Dissociation and Chemometrics for b Fragment Ion Analysis

    NASA Astrophysics Data System (ADS)

    Zekavat, Behrooz; Miladi, Mahsan; Becker, Christopher; Munisamy, Sharon M.; Solouki, Touradj

    2013-09-01

    Although structural isomers may yield indistinguishable ion mobility (IM) arrival times and similar fragment ions in tandem mass spectrometry (MS), it is demonstrated that post-IM/collision-induced dissociation MS (post-IM/CID MS) combined with chemometrics can enable independent study of the IM-overlapped isomers. The new approach allowed us to investigate the propensity of selected b type fragment ions from AlaAlaAlaHisAlaAlaAla-NH2 (AAA(His)AAA) heptapeptide to form different isomers. Principle component analysis (PCA) of the unresolved post-IM/CID profiles indicated the presence of two different isomer types for b4 +, b5 +, and b6 + and a single isomer type for b7 + fragments of AAA(His)AAA. We employed a simple-to-use interactive self-modeling mixture analysis (SIMPLISMA) to calculate the total IM profiles and CID mass spectra of b fragment isomers. The deconvoluted CID mass spectra showed discernible fragmentation patterns for the two isomers of b4 +, b5 +, and b6 + fragments. Under our experimental conditions, calculated percentages of the "cyclic" isomers (at the 95 % confidence level for n = 3) for b4 +, b5 +, and b6 + were 61 (± 5) %, 36 (± 5) %, and 48 (± 2) %, respectively. Results from the SIMPLISMA deconvolution of b5 + species resembled the CID MS patterns of fully resolved IM profiles for the two b5 + isomers. The "cyclic" isomers for each of the two-component b fragment ions were less susceptible to ion fragmentation than their "linear" counterparts.

  6. Application of response surface methodology (RSM) to the optimization of a post-column luminol chemiluminescence analysis of silyl peroxides.

    PubMed

    Baj, Stefan; Słupska, Roksana; Krawczyk, Tomasz

    2013-01-15

    The possibility of the utilization of chemiluminescence post-column luminol oxidation (CL) in a HPLC system for silyl peroxides analysis has been investigated. The conditions of HPLC separation for 12 silyl peroxides, representing bissilyl and alkyl-silyl peroxides, as well as their potential impurities, were established. Optimal chemiluminescent post-column reaction conditions were found using central composite design (CCD) and response surface methodology (RSM). The interaction effects of four of the most important operating variables - the concentrations of luminol, hemin, sodium hydroxide and the post-column solution flow rate - on the light intensity were evaluated. The optimized conditions for analysis were the same for bissilyl and alkyl-silyl peroxides for the base concentration (0.03 M), the luminol concentration (0.4 g L(-1)) and the hemin concentration (0.3 g L(-1)). The only differences occurred in a reagent flow rate (for bissilyl peroxide -0.3 mL min(-1) and for alkyl-silyl peroxides -0.9 mL min(-1)). Under optimal conditions, the detection limits were in the 0.07-0.16 nM range for bissilyl, and 0.53-1.01 for alkyl-silyl peroxides. The calibration curves were linear in the 0.25-3 nM range for bissilyl and the 2.5-25 range for alkyl-silyl peroxides. Intra-day and inter-day precision was lower than 5.5% for each tested concentration level. A mechanism of luminol oxidation by silyl peroxides involving a hydrolysis step with the formation of hydrogen peroxide or hydroperoxide was proposed.

  7. Antiviral agent based on the non-structural protein targeting the maturation process of HIV-1: expression and susceptibility of chimeric Vpr as a substrate for cleavage by HIV-1 protease.

    PubMed

    Serio, D; Singh, S P; Cartas, M A; Weber, I T; Harrison, R W; Louis, J M; Srinivasan, A

    2000-06-01

    The processing of precursor proteins (Gag and Gag-pol) by the viral protease is absolutely required in order to generate infectious particles. This prompted us to consider novel strategies that target viral maturation. Towards this end, we have engineered an HIV-1 virion associated protein, Vpr, to contain protease cleavage signal sequences from Gag and Gag-pol precursor proteins. We previously reported that virus particles derived from HIV-1 proviral DNA, encoding chimeric Vpr, showed a lack of infectivity, depending on the fusion partner. As an extension of that work, the potential of chimeric Vpr as a substrate for HIV-1 protease was tested utilizing an epitope-based assay. Chimeric Vpr molecules were modified such that the Flag epitope is removed following cleavage, thus allowing us to determine the efficiency of protease cleavage. Following incubation with the protease, the resultant products were analyzed by radioimmunoprecipitation using antibodies directed against the Flag epitope. Densitometric analysis of the autoradiograms showed processing to be both rapid and specific. Further, the analysis of virus particles containing chimeric Vpr by immunoblot showed reactivities to antibodies against the Flag epitope similar to the data observed in vitro. These results suggest that the pseudosubstrate approach may provide another avenue for developing antiviral agents.

  8. Non-Occupational Post-Exposure Prophylaxis for HIV: 10-Year Retrospective Analysis in Seattle, Washington

    PubMed Central

    McDougal, Sarah J.; Alexander, Jeremiah; Dhanireddy, Shireesha; Harrington, Robert D.; Stekler, Joanne D.

    2014-01-01

    Despite treatment guidelines in place since 2005, non-occupational post-exposure HIV prophylaxis (nPEP) remains an underutilized prevention strategy. We conducted a retrospective chart review of patients presenting to a publicly-funded HIV clinic in Seattle, Washington for nPEP between 2000 and 2010 (N = 360). nPEP prescriptions were provided for 324 (90%) patients; 83% of prescription decisions were appropriate according to Centers for Disease Control and Prevention guidelines, but only 31% (N = 111/360) of patients were considered “high risk.” In order to use limited resources most efficiently, public health agencies should target messaging for this high-cost intervention to individuals with high-risk HIV exposures. PMID:25140868

  9. Phase-retrieval analysis of pre- and post-repair Hubble Space Telescope images.

    PubMed

    Krist, J E; Burrows, C J

    1995-08-01

    Phase-retrieval measurements of point-spread functions from the pre- and post-repair Hubble Space Telescope are presented. The primary goal was to determine the aberrations present in the second wide-field and planetary camera (WFPC2) to align and validate its corrective optics. With both parametric model-fitting techniques and iterative (Gerchberg-Saxton) methods, accurate measurements have been obtained of the WFPC2 and Hubble Space Telescope optics, including improved maps of the zonal errors in the mirrors. Additional phase-retrieval results were obtained for the aberrated, prerepair cameras and the corrected faint-object camera. The information has been used to improve models produced by point-spread-function simulation programs. On the basis of the measurements a conic constant for the primary mirror of κ = -1.0144 has been derived.

  10. Linear MHD stability analysis of post-disruption plasmas in ITER

    NASA Astrophysics Data System (ADS)

    Aleynikova, K.; Huijsmans, G. T. A.; Aleynikov, P.

    2016-05-01

    Most of the plasma current can be replaced by a runaway electron (RE) current during plasma disruptions in ITER. In this case the post-disruption plasma current profile is likely to be more peaked than the pre-disruption profile. The MHD activity of such plasma will affect the runaway electron generation and confinement and the dynamics of the plasma position evolution (Vertical Displacement Event), limiting the timeframe for runaway electrons and disruption mitigation. In the present paper, we evaluate the influence of the possible RE seed current parameters on the onset of the MHD instabilities. By varying the RE seed current profile, we search for subsequent plasma evolutions with the highest and the lowest MHD activity. This information can be applied to a development of desirable ITER disruption scenario.

  11. Space shuttle post-entry and landing analysis. Volume 1: Candidate system evaluations

    NASA Technical Reports Server (NTRS)

    Crawford, B. S.; Duiven, E. M.

    1973-01-01

    The general purpose of this study is to aid in the evaluation and design of multi-sensor navigation schemes proposed for the orbiter. The scope of the effort is limited to the post-entry, energy management, and approach and landing mission phases. One candidate system based on conventional navigation aids is illustrated including two DME (Distance Measuring Equipment) stations and ILS (Instrument Landing System) glide slope and localizer antennas. Some key elements of the system not shown are the onboard IMUs (Inertial Measurement Units), altimeters, and a computer. The latter is programmed to mix together (filter) the IMU data and the externally-derived data. A completely automatic, all-weather landing capability is required. Since no air-breathing engines will be carried on orbital flights, there will be no chance to go around and try again following a missed approach.

  12. Post-flight Analysis of Mars Science Laboratory Entry Aerothermal Environment and Thermal Protection System Response

    NASA Technical Reports Server (NTRS)

    White, Todd Richard; Mahazari, Milad; Bose, Deepak; Santos, Jose Antonio

    2013-01-01

    The Mars Science Laboratory successfully landed on the Martian surface on August 5th, 2012. The rover was protected from the extreme heating environments of atmospheric entry by an ablative heatshield. This Phenolic Impregnated Carbon Ablator heatshield was instrumented with a suite of embedded thermocouples, isotherm sensors, and pressure transducers. The sensors monitored the in-depth ablator response, as well as the surface pressure at discrete locations throughout the hypersonic deceleration. This paper presents a comparison of the flight data with post-entry estimates. An assessment of the aerothermal environments, as well as the in-depth response of the heatshield material is made, and conclusions regarding the overall performance of the ablator at the suite locations are presented.

  13. Contraband tobacco on post-secondary campuses in Ontario, Canada: analysis of discarded cigarette butts

    PubMed Central

    2013-01-01

    Background No studies to date have assessed young adults’ use of First Nations/Native tobacco, a common form of contraband tobacco in Canada. This study examined the proportion of First Nations/Native cigarette butts discarded on post-secondary campuses in the province of Ontario, and potential differences between colleges and universities and across geographical regions. Methods In 2009, discarded cigarette butts were collected from high-traffic smoking locations at 12 universities and 13 colleges purposively selected to represent a variety of institutions from all 7 health service regions across Ontario. Cigarette butts were identified as First Nations/Native tobacco if they were: known First Nations/Native brands; had names not matching domestic and international legally-manufactured cigarettes; had no visible branding or logos. Results Of 36,355 butts collected, 14% (95% CI = 9.75–19.04) were First Nations/Native. Use of this tobacco was apparent on all campuses, accounting for as little as 2% to as much as 39% of cigarette consumption at a particular school. Proportions of First Nations/Native butts were not significantly higher on colleges (M = 17%) than universities (M = 12%), but were significantly higher in the North region. Conclusions The presence of cheap First Nations/Native (contraband) tobacco on post-secondary campuses suggests the need for regulation and public education strategies aimed to reduce its use. Strategies should account for regional variations, and convey messages that resonate with young adults. Care must be taken to present fair messages about First Nations/Native tobacco, and avoid positioning regulated tobacco as a healthier option than contraband. PMID:23577796

  14. The cost of post-abortion care in developing countries: a comparative analysis of four studies.

    PubMed

    Vlassoff, Michael; Singh, Susheela; Onda, Tsuyoshi

    2016-10-01

    Over the last five years, comprehensive national surveys of the cost of post-abortion care (PAC) to national health systems have been undertaken in Ethiopia, Uganda, Rwanda and Colombia using a specially developed costing methodology-the Post-abortion Care Costing Methodology (PACCM). The objective of this study is to expand the research findings of these four studies, making use of their extensive datasets. These studies offer the most complete and consistent estimates of the cost of PAC to date, and comparing their findings not only provides generalizable implications for health policies and programs, but also allows an assessment of the PACCM methodology. We find that the labor cost component varies widely: in Ethiopia and Colombia doctors spend about 30-60% more time with PAC patients than do nurses; in Uganda and Rwanda an opposite pattern is found. Labor costs range from I$42.80 in Uganda to I$301.30 in Colombia. The cost of drugs and supplies does not vary greatly, ranging from I$79 in Colombia to I$115 in Rwanda. Capital and overhead costs are substantial amounting to 52-68% of total PAC costs. Total costs per PAC case vary from I$334 in Rwanda to I$972 in Colombia. The financial burden of PAC is considerable: the expense of treating each PAC case is equivalent to around 35% of annual per capita income in Uganda, 29% in Rwanda and 11% in Colombia. Providing modern methods of contraception to women with an unmet need would cost just a fraction of the average expenditure on PAC: one year of modern contraceptive services and supplies cost only 3-12% of the average cost of treating a PAC patient. PMID:27045001

  15. The cost of post-abortion care in developing countries: a comparative analysis of four studies

    PubMed Central

    Vlassoff, Michael; Singh, Susheela; Onda, Tsuyoshi

    2016-01-01

    Over the last five years, comprehensive national surveys of the cost of post-abortion care (PAC) to national health systems have been undertaken in Ethiopia, Uganda, Rwanda and Colombia using a specially developed costing methodology—the Post-abortion Care Costing Methodology (PACCM). The objective of this study is to expand the research findings of these four studies, making use of their extensive datasets. These studies offer the most complete and consistent estimates of the cost of PAC to date, and comparing their findings not only provides generalizable implications for health policies and programs, but also allows an assessment of the PACCM methodology. We find that the labor cost component varies widely: in Ethiopia and Colombia doctors spend about 30–60% more time with PAC patients than do nurses; in Uganda and Rwanda an opposite pattern is found. Labor costs range from I$42.80 in Uganda to I$301.30 in Colombia. The cost of drugs and supplies does not vary greatly, ranging from I$79 in Colombia to I$115 in Rwanda. Capital and overhead costs are substantial amounting to 52–68% of total PAC costs. Total costs per PAC case vary from I$334 in Rwanda to I$972 in Colombia. The financial burden of PAC is considerable: the expense of treating each PAC case is equivalent to around 35% of annual per capita income in Uganda, 29% in Rwanda and 11% in Colombia. Providing modern methods of contraception to women with an unmet need would cost just a fraction of the average expenditure on PAC: one year of modern contraceptive services and supplies cost only 3–12% of the average cost of treating a PAC patient. PMID:27045001

  16. Structure-Function Analysis of CCL28 in the Development of Post-viral Asthma*

    PubMed Central

    Thomas, Monica A.; Buelow, Becky J.; Nevins, Amanda M.; Jones, Stephanie E.; Peterson, Francis C.; Gundry, Rebekah L.; Grayson, Mitchell H.; Volkman, Brian F.

    2015-01-01

    CCL28 is a human chemokine constitutively expressed by epithelial cells in diverse mucosal tissues and is known to attract a variety of immune cell types including T-cell subsets and eosinophils. Elevated levels of CCL28 have been found in the airways of individuals with asthma, and previous studies have indicated that CCL28 plays a vital role in the acute development of post-viral asthma. Our study builds on this, demonstrating that CCL28 is also important in the chronic post-viral asthma phenotype. In the absence of a viral infection, we also demonstrate that CCL28 is both necessary and sufficient for induction of asthma pathology. Additionally, we present the first effort aimed at elucidating the structural features of CCL28. Chemokines are defined by a conserved tertiary structure composed of a three-stranded β-sheet and a C-terminal α-helix constrained by two disulfide bonds. In addition to the four disulfide bond-forming cysteine residues that define the traditional chemokine fold, CCL28 possesses two additional cysteine residues that form a third disulfide bond. If all disulfide bonds are disrupted, recombinant human CCL28 is no longer able to drive mouse CD4+ T-cell chemotaxis or in vivo airway hyper-reactivity, indicating that the conserved chemokine fold is necessary for its biologic activity. Due to the intimate relationship between CCL28 and asthma pathology, it is clear that CCL28 presents a novel target for the development of alternative asthma therapeutics. PMID:25556652

  17. Functional analysis of Scr during embryonic and post-embryonic development in the cockroach, Periplaneta americana.

    PubMed

    Hrycaj, Steven; Chesebro, John; Popadić, Aleksandar

    2010-05-01

    The cockroach, Periplaneta americana represents a basal insect lineage that undergoes the ancestral hemimetabolous mode of development. Here, we examine the embryonic and post-embryonic functions of the hox gene Scr in Periplaneta as a way of better understanding the roles of this gene in the evolution of insect body plans. During embryogenesis, Scr function is strictly limited to the head with no role in the prothorax. This indicates that the ancestral embryonic function of Scr was likely restricted to the head, and that the posterior expansion of expression in the T1 legs may have preceded any apparent gain of function during evolution. In addition, Scr plays a pivotal role in the formation of the dorsal ridge, a structure that separates the head and thorax in all insects. This is evidenced by the presence of a supernumerary segment that occurs between the labial and T1 segments of RNAiScr first nymphs and is attributed to an alteration in engrailed (en) expression. The fact that similar Scr phenotypes are observed in Tribolium but not in Drosophila or Oncopeltus reveals the presence of lineage-specific variation in the genetic architecture that controls the formation of the dorsal ridge. In direct contrast to the embryonic roles, Scr has no function in the head region during post-embryogenesis in Periplaneta, and instead, strictly acts to provide identity to the T1 segment. Furthermore, the strongest Periplaneta RNAiScr phenotypes develop ectopic wing-like tissue that originates from the posterior region of the prothoracic segment. This finding provides a novel insight into the current debate on the morphological origin of insect wings.

  18. Top-Down Analysis of Highly Post-Translationally Modified Peptides by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Guerrero, Andres; Lerno, Larry; Barile, Daniela; Lebrilla, Carlito B.

    2015-03-01

    Bovine κ-caseinoglycomacropeptide (GMP) is a highly modified peptide from κ-casein produced during the cheese making process. The chemical nature of GMP makes analysis by traditional proteomic approaches difficult, as the peptide bears a strong net negative charge and a variety of post-translational modifications. In this work, we describe the use of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR MS) for the top-down analysis of GMP. The method allows the simultaneous detection of different GMP forms that result from the combination of amino acid genetic variations and post-translational modifications, specifically phosphorylation and O-glycosylation. The different GMP forms were identified by high resolution mass spectrometry in both negative and positive mode and confirmation was achieved by tandem MS. The results showed the predominance of two genetic variants of GMP that occur as either mono- or bi-phosphorylated species. Additionally, these four forms can be modified with up to two O-glycans generally sialylated. The results demonstrate the presence of glycosylated, bi-phosphorylated forms of GMP never described before.

  19. Effect of precut sphincterotomy on post-endoscopic retrograde cholangiopancreatography pancreatitis: A systematic review and meta-analysis

    PubMed Central

    Choudhary, Abhishek; Winn, Jessica; Siddique, Sameer; Arif, Murtaza; Arif, Zainab; Hammoud, Ghassan M; Puli, Srinivas R; Ibdah, Jamal A; Bechtold, Matthew L

    2014-01-01

    AIM: To conduct a systemic review and meta-analysis to investigate the role of early precut technique. Multiple randomized controlled trails (RCTs) have reported conflicting results of the early precut sphincterotomy. METHODS: MEDLINE/PubMed, EMBASE, Cochrane Central Register of Controlled Trials and Database of Systematic Reviews, and recent abstracts from major conference proceedings were searched (June 2013). Randomized and non-randomized studies comparing early precut technique with prolonged standard methods were included. Pooled estimates of post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP), cannulation and adverse events were analyzed by using odds ratio (OR). Random and fixed effects models were used as appropriate. Publication bias was assessed by funnel plots. Heterogeneity among studies was assessed by calculating I2 measure of inconsistency. RESULTS: Seven randomized and seven non-randomized trials met inclusion criteria. Meta-analysis of RCTs showed a decrease trend for PEP with early precut sphincterotomy but was not statistically significant (OR = 0.58; 95%CI: 0.32-1.05; P = 0.07). No heterogeneity was noted among the studies with I2 of 0%. CONCLUSION: Early precut technique for common bile duct cannulation decreases the trend of post-ERCP pancreatitis. PMID:24744601

  20. Altered post-mortem metabolism identified in very fast chilled lamb M. longissimus thoracis et lumborum using metabolomic analysis.

    PubMed

    Warner, Robyn D; Jacob, Robin H; Rosenvold, Katja; Rochfort, Simone; Trenerry, Craige; Plozza, Tim; McDonagh, Matthew B

    2015-10-01

    The aim of this experiment was to use metabolomic techniques to investigate the energy metabolism in lamb M. longissimus thoracis et lumborum subjected to very fast chilling (VFC) post-mortem. The tissue was prepared by 2 different operators and subjected to very fast chilling (less than 0°C within 1.5h of slaughter) or typical chilling regimes (Control; 0°C within 22h of slaughter). Non-targeted metabolomic analysis ((1)H NMR) and targeted analysis ((31)P NMR, HPLC-PDA and HPLC-MS/MS) were used to examine the change in muscle metabolites post-mortem. One VFC treatment, which resulted in a colder core temperature and more tender meat, had higher levels of glycolytic intermediate metabolites pre-rigor as well as more of the end-products of adenosine and nicotine nucleotide metabolism pre-rigor, relative to conventionally chilled treatments. In conclusion, VFC to less than 0°C within 1.5h of slaughter causes considerable changes in metabolism and rigor onset, which are associated with tender meat. PMID:26163181