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Sorting
Kinds of Sorting Terms modified by Sorting Selected AbstractsSCHOOL CHOICE AND STUDENT SORTING: EVIDENCE FROM ADACHI WARD IN JAPAN,THE JAPANESE ECONOMIC REVIEW, Issue 4 2009ATSUSHI YOSHIDA We examine whether the school choice programme of public junior high schools in Adachi ward has caused student sorting and has thus increased the differences in scores between the schools. We find that students are sorted in the sense that the students living in the school attendance areas where there is a higher proportion of high-status occupations are more likely to select private schools even after the introduction of the school choice programme, or they select public schools with higher scores. Adachi's average scores relative to the Tokyo average have improved, while the between school differences in scores have not expanded. [source] Sorting nexin-14, a gene expressed in motoneurons trapped by an in vitro preselection methodDEVELOPMENTAL DYNAMICS, Issue 4 2001Patrick Carroll Abstract A gene-trap strategy was set up in embryonic stem (ES) cells with the aim of trapping genes expressed in restricted neuronal lineages. The vector used trap genes irrespective of their activity in undifferentiated totipotent ES cells. Clones were subjected individually to differentiation in a system in which ES cells differentiated into neurons. Two ES clones in which the trapped gene was expressed in ES-derived neurons were studied in detail. The corresponding cDNAs were cloned, sequenced, and analysed by in situ hybridisation on wild-type embryo sections. Both genes are expressed in the nervous system. One gene, YR-23, encodes a large intracellular protein of unknown function. The second clone, YR-14, represents a sorting nexin (SNX14) gene whose expression in vivo coincides with that of LIM-homeodomain Islet-1 in several tissues. Sorting nexins are proteins associated with the endoplasmic reticulum (ER) and may play a role in receptor trafficking. Gene trapping followed by screening based on in vitro preselection of differentiated ES recombinant clones, therefore, has the potential to identify integration events in subsets of genes before generation of mouse mutants. © 2001 Wiley-Liss, Inc. [source] The Vps4 C-terminal helix is a critical determinant for assembly and ATPase activity and has elements conserved in other members of the meiotic clade of AAA ATPasesFEBS JOURNAL, Issue 7 2008Parimala R. Vajjhala Sorting of membrane proteins into intralumenal endosomal vesicles, multivesicular body (MVB) sorting, is critical for receptor down regulation, antigen presentation and enveloped virus budding. Vps4 is an AAA ATPase that functions in MVB sorting. Although AAA ATPases are oligomeric, mechanisms that govern Vps4 oligomerization and activity remain elusive. Vps4 has an N-terminal microtubule interacting and trafficking domain required for endosome recruitment, an AAA domain containing the ATPase catalytic site and a , domain, and a C-terminal , helix positioned close to the catalytic site in the 3D structure. Previous attempts to identify the role of the C-terminal helix have been unsuccessful. Here, we show that the C-terminal helix is important for Vps4 assembly and ATPase activity in vitro and function in vivo, but not endosome recruitment or interactions with Vta1 or ESCRT-III. Unlike the , domain, which is also important for Vps4 assembly, the C-terminal helix is not required in vivo for Vps4 homotypic interaction or dominant-negative effects of Vps4,E233Q, carrying a mutation in the ATP hydrolysis site. Vta1 promotes assembly of hybrid complexes comprising Vps4,E233Q and Vps4 lacking an intact C-terminal helix in vitro. Formation of catalytically active hybrid complexes demonstrates an intersubunit catalytic mechanism for Vps4. One end of the C-terminal helix lies in close proximity to the second region of homology (SRH), which is important for assembly and intersubunit catalysis in AAA ATPases. We propose that Vps4 SRH function requires an intact C-terminal helix. Co-evolution of a distinct Vps4 SRH and C-terminal helix in meiotic clade AAA ATPases supports this possibility. [source] Transmission Electron Microscopy and UV,vis,IR Spectroscopy Analysis of the Diameter Sorting of Carbon Nanotubes by Gradient Density UltracentrifugationADVANCED FUNCTIONAL MATERIALS, Issue 14 2009Romain Fleurier Abstract Diameter separation of single-walled carbon nanotubes is achieved via the density gradient ultracentrifugation process. Statistical analysis of the separated samples is performed using high-resolution transmission electron microscopy (HRTEM). The evolution of the diameter distribution with respect to the gradient density is extracted by analyzing hundreds of HRTEM images, and the results are found to be consistent with those estimated by UV,vis,IR spectroscopy. The efficiency of the separation process can be quantitatively characterized by the standard deviation of the diameter distribution, which is determined from the TEM analyses. This particular study indicated that for electric arc nanotubes dispersed in sodium cholate, diameter sorting is more efficient in the upper part of the gradient. [source] Young children's difficulty with inhibitory control in a social context,JAPANESE PSYCHOLOGICAL RESEARCH, Issue 2 2008YUSUKE MORIGUCHI Abstract:, The authors' prior research has documented that young children's behaviors in the Dimensional Change Card Sorting (DCCS) task can be influenced by their observation of another person performing the task and has suggested that young children committed perseverative errors in a social context. The present study explored whether children who committed perseverative errors in the social context also committed perseverative errors in the standard DCCS task. Three- and 4-year-old children were given the standard DCCS and the observation version of the DCCS, and the relationship between them was examined. The results showed that the correlation between these two tasks was significant. Furthermore, 4-year-old children displayed more difficulty in the observation version than in the standard DCCS, whereas 3-year-olds did not. The results are discussed in terms of the development of inhibitory control and social cognition. [source] Quality improvement through consumer sorting and disposalAGRIBUSINESS : AN INTERNATIONAL JOURNAL, Issue 4 2009Peyton Ferrier Sorting allows consumers to capture the value of quality differences. As higher quality goods are removed, the value of the seller's remaining stock falls, lowering the price and profits. Bundling and other marketing mechanisms can discourage sorting and prevent the depreciation of the seller's stock. With comparative statics and simulations, the author shows that sellers can increase expected quality and profits by committing to discard a proportion of their resale stock after sorting occurs. In this manner, sorting acts similarly to agricultural grading. [EconLit Classification: Q1, Q11, Q13, L0, L1, D8, D82]. © 2009 Wiley Periodicals, Inc. [source] Establishment of embryonic stem cells secreting human factor VIII for cell-based treatment of hemophilia AJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 8 2008S. KASUDA Summary.,Background:,Hemophilia A is an X-chromosome-linked recessive bleeding disorder resulting from an F8 gene abnormality. Although various gene therapies have been attempted with the aim of eliminating the need for factor VIII replacement therapy, obstacles to their clinical application remain. Objectives:,We evaluated whether embryonic stem (ES) cells with a tetracycline-inducible system could secrete human FVIII. Methods and results:,We found that embryoid bodies (EBs) developed under conditions promoting liver differentiation efficiently secreted human FVIII after doxycycline induction. Moreover, use of a B-domain variant F8 cDNA (226aa/N6) dramatically enhanced FVIII secretion. Sorting based on green fluorescent protein (GFP),brachyury (Bry) and c-kit revealed that GFP,Bry+/c-kit+ cells during EB differentiation with serum contain an endoderm progenitor population. When GFP,Bry+/c-kit+ cells were cultured under the liver cell-promoting conditions, these cells secreted FVIII more efficiently than other populations tested. Conclusion:,Our findings suggest the potential for future development of an effective ES cell-based approach to treating hemophilia A. [source] Sorting Out "Fair Use" and "Likelihood of Confusion" in Trademark LawAMERICAN BUSINESS LAW JOURNAL, Issue 1 2006Stephanie M. Greene [source] Downstream Fining and Sorting of Gravel Clasts in the Braided Rivers of mid-Canterbury, New ZealandNEW ZEALAND GEOGRAPHER, Issue 2 2004Greg Browne ABSTRACT Gravel clast size dimensions have been determined in the Rakaia, Ashburton, and Rangitata rivers by measuring 100 clasts at representative sample locations along each river. In all rivers, gravel size decreases and sorting improves downstream for mean, D50, and D90 fractions of the bed material. Clast size entering the sea is similar in all rivers (30,40 mm b-axis dimension), despite large variations in transport distance, input size of clasts at their gorges, and discharge. The greatest size reduction occurs in the Rangitata River which has the shortest transport distance and steepest gradient. Rates of downstream clast size reduction are greater than would be assumed from Sternberg's Law, suggesting that additional factors, other than physical abrasion, such as sorting and selective entrainment operate. [source] Sorting out successful failures: Exploratory analyses of factors associated with academic and behavioral outcomes of retained studentsPSYCHOLOGY IN THE SCHOOLS, Issue 4 2001Phillip Ferguson This prospective longitudinal study followed a sample of 106 kindergarten students through 11th grade examining the effects of family characteristics, school readiness, socialization, and student demographics on academic achievement and behavioral adjustment outcomes. These educational outcomes were contrasted among four groups consisting of: 1) early grade retainees; 2) transitionally placed retained students; 3) students recommended for transitional placement, but promoted; and 4) regularly promoted students. While previous studies examining the efficacy of early grade retention focus exclusively on between-group comparisons, this study examines the family and individual characteristics of successful and unsuccessful retained students by including both between-group and within-group effects on academic and behavioral outcomes. The results of this study demonstrate that retained students' initial school readiness, socioeconomic status, mother's level of education, parental value of education, kindergarten personal-social functioning, and chronological age are distinctly associated with subsequent academic or behavioral outcomes. Variables associated with relative educational success following early failure are delineated and research implications are discussed. © 2001 John Wiley & Sons, Inc. [source] Reduction of Oxidative Stress in Bovine Spermatozoa During Flow Cytometric SortingREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2007P Klinc Contents The goal of the study was to investigate the effect of antioxidant supplementation on the quality of frozen-thawed flow cytometrically sorted bull spermatozoa. Twelve ejaculates from two Holstein Friesian bulls were sorted according to the Beltsville Sperm Sexing Technology. Each ejaculate was divided into three parts and processed as (i) unsorted controls, (ii) according to a standard sorting protocol and (iii) in the presence of different antioxidants (S-AO). Cooling and freezing of the samples were performed in the same way for all three groups, except that antioxidants were added to the TRIS-egg-yolk freezing extender for those semen samples that were already sorted in the presence of antioxidants. The semen quality in frozen-thawed samples was determined by morphology analysis immediately after thawing, motility estimation in a thermo-resistance test after 0, 6, 12 and 24 h incubation at 37°C and Fluorescein isothiocyanate conjugated PNA/propidium iodide (FITC-PNA/PI) staining after 0, 12 and 24 h of incubation at 37°C. There was a significantly higher (p < 0.05) percentage of motile spermatozoa in S-AO samples in comparison to unsorted frozen-thawed control at 0, 6 and 24 h after thawing and compared with normally sorted samples at all times after thawing. The percentage of damaged acrosomes was significantly lower (p < 0.05) in S-AO samples than in the unsorted controls (20.8 ± 6.9% vs 30.3 ± 12.0%). The percentage of morphologically abnormal spermatozoa in this group was significantly lower (p < 0.05) than in the unsorted controls and normally sorted samples (25.8 ± 5.2%, 36.0 ± 12.5% and 35.1 ± 7.4%, respectively). Analysis of frozen-thawed spermatozoa with FITC/PI revealed no significant difference in membrane integrity at 0 and 12 h after sorting, but after 24 h of incubation the S-AO samples had a significantly higher (p < 0.001) percentage of spermatozoa with intact membranes in comparison to unsorted controls and normally sorted semen (40.7 ± 6.3%, 7.8 ± 4.7% and 7.4 ± 4.6%, respectively). The percentage of acrosome-reacted spermatozoa was significantly lower (p < 0.05) in the S-AO samples than in the unsorted controls (14.1 ± 7.5%, 23.4 ± 5.4% and 28.8 ± 6.3% vs 25.9 ± 14.4%, 38.5 ± 16.7% and 79.8 ± 4.1%, for 0, 12 and 24 h after thawing, respectively) and in comparison to normally sorted semen 24 h after thawing (67.3 ± 10.0%). This study demonstrates the highly protective effects of antioxidants on the quality of flow cytometrically sorted frozen-thawed bull spermatozoa. [source] Civic Engagement and Education: An Empirical Test of the Sorting ModelAMERICAN JOURNAL OF POLITICAL SCIENCE, Issue 4 2009David E. Campbell According to the sorting model of education, the impact of education on civic engagement is relative, rather than absolute. Education correlates with greater engagement because it is a marker of social status; the degree of status conferred by your level of education is determined by the average level of education within your environment. This article tests the sorting model by paying strict heed to its assumptions. The analysis confirms the model, but considerably narrows its reach. Sorting applies only to one particular type (electoral activity), only when the educational environment accounts for variation across age and place, and only when one models the interactive relationship between education at the individual and environmental levels. Furthermore, sorting applies more to men than women. The same analytical framework demonstrates that being in a more highly educated environment amplifies the relationship between education and democratic enlightenment (political knowledge and tolerance). [source] Intergenerational Mobility and Marital Sorting,THE ECONOMIC JOURNAL, Issue 513 2006John Ermisch We use data from the German Socio-Economic Panel and the British Household Panel Survey to estimate the extent of intergenerational economic mobility in a framework that highlights the role played by assortative mating. We find that assortative mating plays an important role. On average about 40,50% of the covariance between parents' and own permanent family income can be attributed to the person to whom one is married. This effect is driven by strong spouse correlations in human capital, which are larger in Germany than Britain. [source] Flow Cytometric Sorting of Fresh and Frozen-Thawed Spermatozoa in the Western Lowland Gorilla (Gorilla gorilla gorilla)AMERICAN JOURNAL OF PRIMATOLOGY, Issue 4 2005J.K. O'Brien Abstract We adapted flow cytometry technology for high-purity sorting of X chromosome-bearing spermatozoa in the western lowland gorilla (Gorilla gorilla gorilla). Our objectives were to develop methodologies for liquid storage of semen prior to sorting, sorting of liquid-stored and frozen-thawed spermatozoa, and assessment of sorting accuracy. In study 1, the in vitro sperm characteristics of gorilla ejaculates from one male were unchanged (P>0.05) after 8 hr of liquid storage at 15°C in a non-egg yolk diluent (HEPES-buffered modified Tyrode's medium). In study 2, we examined the efficacy of sorting fresh and frozen-thawed spermatozoa using human spermatozoa as a model for gorilla spermatozoa. Ejaculates from one male were split into fresh and frozen aliquots. X-enriched samples derived from both fresh and frozen-thawed human semen were of high purity, as determined by fluorescence in situ hybridization (FISH; 90.7%±2.3%, overall), and contained a high proportion of morphologically normal spermatozoa (86.0%±1.0%, overall). In study 3, we processed liquid-stored semen from two gorillas for sorting using a modification of methods for human spermatozoa. The sort rate for enrichment of X-bearing spermatozoa was 7.3±2.5 spermatozoa per second. The X-enriched samples were of high purity (single-sperm PCR: 83.7%) and normal morphology (79.0%±3.9%). In study 4 we examined frozen-thawed gorilla semen, and the sort rate (8.3±2.9 X-bearing sperm/sec), purity (89.7%), and normal morphology (81.4%±3.4%) were comparable to those of liquid-stored semen. Depending on the male and the type of sample used (fresh or frozen-thawed), 0.8,2.2% of gorilla spermatozoa in the processed ejaculate were present in the X-enriched sample. These results demonstrate that fresh or frozen-thawed gorilla spermatozoa can be flow cytometrically sorted into samples enriched for X-bearing spermatozoa. Am. J. Primatol. 66:297,315, 2005. © 2005 Wiley-Liss, Inc. [source] P-96 Fetal endothelial cells in full-term placenta, but not in first trimester placenta, express Fc,RIIb2 mRNAAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 5 2004Mishima Takuya In the human placenta, IgG-transport from maternal to fetal blood starts in the second trimester and continues through the third trimester. There are two cellular barriers for IgG-transport: the first and second barriers are syncytiotrophoblast and fetal endothelial cells (ECs), respectively. Sorting via FcRn is considered the IgG-transporter in the first barrier. The mechanism of IgG-transport in the second barrier is not fully understood. Recently, we reported a novel Fc,RIIb-containing compartment that may serve as an IgG-transporter in the second barrier (Mol Biol Cell 13 (suppl) 548a, 2002). To further investigate the feasibility of the Fc,RIIb-vesicle involvement in IgG-transport in placental ECs, we have studied FcR-expression in the developing human placenta by RT-PCR and direct sequencing analysis. First trimester and full-term placentas were used. We proved that the Fc,RII expressed in villus ECs in full-term placenta was the b2 isoform. Fc,RIIb2 mRNA expression could not be detected in first trimester placenta, while it was expressed in full-term placenta. In contrast, FcRn mRNA expression was detectable in first trimester placenta as well as in full-term placenta. These results seem consistent with our hypothesis that in first trimester placenta IgG is transcytosed via FcRn across the first barrier but IgG cannot be transported in the second barrier that does not express Fc,RIIb2 and that the Fc,RIIb2-positive compartment is critical for IgG-transport in the human placenta. [source] Quality of life and memory performance in patients with temporal lobe epilepsyACTA NEUROLOGICA SCANDINAVICA, Issue 5 2000A. R. Giovagnoli Objective, To explore the contribution of memory performance to quality of life (QOL) in patients with left or right temporal lobe epilepsy (TLE). Subjects and methods, Sixty-five patients with left or right TLE compiled the QOL in Epilepsy-89 Inventory (QOLIE-89), the State-Trait Anxiety Inventory (STAI) and the Hopelessness Scale (BDI) for self-evaluation of QOL and mood. Memory was assessed by tests of verbal and non-verbal memory and the Questionnaire of Memory Efficiency (QME). A neuropsychological battery was also administered to assess general intelligence, attention, visual perception, language, set shifting, word fluency and conceptual-motor tracking. Results, On factor analysis, the neuropsychological battery and mood scales consisted of six factors (Memory, Mental Speed, Mood, Praxis, Sorting and Perception), while the QOLIE-89 consisted of five factors (Psychosocial Satisfaction, Epilepsy-Related Effects, Role, Physical Performance, Cognition). On regression analysis, overall QOLIE-89 score was predicted by the factor Mood and QME score. The QOLIE-89 factor Cognition was predicted by QME score and the Memory, Mental Speed, Perception and Praxis factors of the neuropsychological battery. Conclusion, In TLE patients self-reported memory, as assessed by QME, is an important predictor of QOL, and also correlates with performance on memory tests. This suggests that memory improvement by specific training may help to improve QOL in these patients. [source] Parallel space-filling curve generation through sortingCONCURRENCY AND COMPUTATION: PRACTICE & EXPERIENCE, Issue 10 2007J. Luitjens Abstract In this paper we consider the scalability of parallel space-filling curve generation as implemented through parallel sorting algorithms. Multiple sorting algorithms are studied and results show that space-filling curves can be generated quickly in parallel on thousands of processors. In addition, performance models are presented that are consistent with measured performance and offer insight into performance on still larger numbers of processors. At large numbers of processors, the scalability of adaptive mesh refined codes depends on the individual components of the adaptive solver. One such component is the dynamic load balancer. In adaptive mesh refined codes, the mesh is constantly changing resulting in load imbalance among the processors requiring a load-balancing phase. The load balancing may occur often, requiring the load balancer to perform quickly. One common method for dynamic load balancing is to use space-filling curves. Space-filling curves, in particular the Hilbert curve, generate good partitions quickly in serial. However, at tens and hundreds of thousands of processors serial generation of space-filling curves will hinder scalability. In order to avoid this issue we have developed a method that generates space-filling curves quickly in parallel by reducing the generation to integer sorting. Copyright © 2007 John Wiley & Sons, Ltd. [source] An examination of different fetal specific antibodies and magnetic activated cell sorting for the enrichment of fetal erythroblasts from maternal bloodCONGENITAL ANOMALIES, Issue 3 2002Xiao Xi Zhao ABSTRACT, The aim of the present study was to compare the rates of fetal cells obtained after separation from maternal blood by magnetic activated cell sorting (MACS) using different fetal specific antibodies, and to evaluate the potential role of this method in the prenatal diagnosis of fetal trisomies. Peripheral blood samples were obtained from 42 women carrying chromosomally normal fetuses and from 4 women with aneuploid fetuses (2 cases of 47,XX,+18 and 2 of 47,XY,+21) at 9,20 weeks of gestation. After fetal cells were enriched by MACS with three different monoclonal antibodies (GPA, CD71, CD14), fluorescence in situ hybridization (FISH) with chromosome X, and Y-specific probes was performed to detect the rates of fetal cells in the samples sorted. FISH with chromosome 13-, 18-, and 21-specific probes was carried out to compare proportions of cells with three-signal nuclei in chromosomally normal and abnormal groups. In male infants, X-and Y-positive cells were detected in 80%, 73.3%, and 66.6% of samples after the separation by antibodies CD14, GPA, and CD71, respectively. The percentage of nuclei with three signals was increased in pregnancies with trisomy, ranging between 2% and 5.18%. Pregnancies with normal fetuses showed 0 to 3.7% of nuclei with three signals. The data demonstrate that fetal cell detection varies depending on the antibodies used for cell sorting. This study provides further evidence on the feasibility of screening for fetal chromosomal abnormalities by enriching maternal blood for fetal cells and using FISH. [source] Cellular oxygen sensing, signalling and how to survive translational arrest in hypoxiaACTA PHYSIOLOGICA, Issue 2 2009M. Fähling Abstract Hypoxia is a consequence of inadequate oxygen availability. At the cellular level, lowered oxygen concentration activates signal cascades including numerous receptors, ion channels, second messengers, as well as several protein kinases and phosphatases. This, in turn, activates trans -factors like transcription factors, RNA-binding proteins and miRNAs, mediating an alteration in gene expression control. Each cell type has its unique constellation of oxygen sensors, couplers and effectors that determine the activation and predominance of several independent hypoxia-sensitive pathways. Hence, altered gene expression patterns in hypoxia result from a complex regulatory network with multiple divergences and convergences. Although hundreds of genes are activated by transcriptional control in hypoxia, metabolic rate depression, as a consequence of reduced ATP level, causes inhibition of mRNA translation. In a multi-phase response to hypoxia, global protein synthesis is suppressed, mainly by phosphorylation of eIF2-alpha by PERK and inhibition of mTOR, causing suppression of 5,-cap-dependent mRNA translation. Growing evidence suggests that mRNAs undergo sorting at stress granules, which determines the fate of mRNA as to whether being translated, stored, or degraded. Data indicate that translation is suppressed only at ,free' polysomes, but is active at subsets of membrane-bound ribosomes. The recruitment of specific mRNAs into subcellular compartments seems to be crucial for local mRNA translation in prolonged hypoxia. Furthermore, ribosomes themselves may play a significant role in targeting mRNAs for translation. This review summarizes the multiple facets of the cellular adaptation to hypoxia observed in mammals. [source] Optimization of a flow cytometry-based protocol for detection and phenotypic characterization of multipotent mesenchymal stromal cells from human bone marrowCYTOMETRY, Issue 6 2006Elena A. Jones Abstract Background: To study the biology of rare bone marrow (BM) multipotent mesenchymal stromal cells (MSCs), recognized protocols are needed. Colony-forming unit-fibroblast (CFU-F) assays have historically been used for the enumeration of MSCs. However, the need to isolate and further analyze MSCs requires new strategies based on cell surface markers. The purpose of this work was to verify the phenotype of BM MSCs in vivo and to develop flow cytometry-based methods for their evaluation. Methods: Pre-enrichment with D7-FIB-conjugated microbeads, cell sorting for CD45lowD7-FIB+LNGFR+ cells, and CFU-F assay were used to confirm the phenotype of BM MSCs in vivo. Further phenotypic characterization of MSCs was performed using three-color flow cytometry following pre-enrichment or by direct four-color flow cytometry. The sensitivity of direct flow cytometry/rare event analysis for the accurate enumeration of MSCs was validated using 85 samples from patients with neoplastic BM diseases. Results: In normal BM, a significant correlation was found between the frequencies of CFU-Fs and CD45lowD7-FIB+LNGFR+ cells (n = 19, R = 0.719, P = 0.001). Following cell sorting, ,15% of these cells were clonogenic. The same cells were enriched using LNGFR-based positive selection, CD45/Glycophorin A-based depletion, or plastic adherence. CD45lowD7-FIB+LNGFR+ cells expressed classic makers of cultured MSCs CD73/SH3 and CD105/SH2 and markers of stromal reticular cells CD106/VCAM and alkaline phosphatase. Novel markers were identified including leukemia inhibitory factor receptor and gp130. CD45lowD7-FIB+LNGFR+ cells were increased fourfold in the floating fat fraction of normal BM aspirates. Their frequency was decreased in chronic lymphocytic leukemia (threefold, n = 13, P = 0.049) and chronic myelogenous leukemia (ninefold, n = 11, P = 0.001) compared with that in age-matched controls (n = 26 and n = 31, respectively). Conclusions: This study demonstrates the usefulness of flow cytometry-based methods for the detection, enumeration and further phenotypic analysis of BM MSCs. These findings have broad applications for the future evaluation of BM MSCs in health and disease. © 2006 International Society for Analytical Cytology [source] Near-infrared dyes for six-color immunophenotyping by laser scanning cytometryCYTOMETRY, Issue 3 2002Andreas O.H. Gerstner Abstract Background To adequately analyze the complexity of the immune system and reduce the required sample volume for immunophenotyping in general, more measurable colors for the discrimination of leukocyte subsets are necessary. Immunophenotyping by the laser scanning cytometer (LSC), a slide-based cytometric technology, combines cell detection based on multiple colors with their subsequent visualization without the need for physical cell sorting. In the present study, the filter setting of the LSC was adapted for the measurement of the far-red emitting dye cyanine 7 (Cy7), thereby increasing the number of measurable commercially available fluorochromes. Methods The optical filters of the LSC were replaced,photomultiplier (PMT) 3/allophycocyanin (APC): 740-nm dichroic long pass, and 670-/55-nm bandpass; PMT 4/Cy7: 810-/90-nm bandpass. Peripheral blood leukocytes were stained directly by fluorochrome-labeled antibodies or by indirect staining. The tandem dyes of Cy7 (phycoerythrin [PE]-Cy7, APC-Cy7) and the fluorochromes fluorescein isothiocyanate (FITC), PE, PE-Cy5, and APC were tested alone and in different combinations. Results With the new filter combination and tandem fluorochromes, Cy7 was measurable at 488-nm (argon laser) or 633-nm (helium-neon laser) excitation. Resolution was in the range of FITC for PE-Cy7 but approximately 30% lower for APC-Cy7; spillover into the respective donor fluorochrome channel for both tandem dyes was prominent. A six-color panel for leukocyte subtyping was designed. Conclusions With this adaptation, it is possible to measure the tandem conjugates PE-Cy7 and APC-Cy7. This new setup opens the way for six-color immunophenotyping by LSC. Cytometry 48:115,123, 2002. © 2002 Wiley-Liss, Inc. [source] Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteinsCYTOSKELETON, Issue 3 2001Nathalie F. Worth Abstract Smooth muscle cells (SMC) exhibit a functional plasticity, modulating from the mature phenotype in which the primary function is contraction, to a less differentiated state with increased capacities for motility, protein synthesis, and proliferation. The present study determined, using Western analysis, double-label immunofluorescence and confocal microscopy, whether changes in phenotypic expression of rabbit aortic SMC in culture could be correlated with alterations in expression and distribution of structural proteins. "Contractile" state SMC (days 1 and 3 of primary culture) showed distinct sorting of proteins into subcellular domains, consistent with the theory that the SMC structural machinery is compartmentalised within the cell. Proteins specialised for contraction (,-SM actin, SM-MHC, and calponin) were highly expressed in these cells and concentrated in the upper central region of the cell. Vimentin was confined to the body of the cell, providing support for the contractile apparatus but not co-localising with it. In line with its role in cell attachment and motility, ,-NM actin was localised to the cell periphery and basal cortex. The dense body protein ,-actinin was concentrated at the cell periphery, possibly stabilising both contractile and motile apparatus. Vinculin-containing focal adhesions were well developed, indicating the cells' strong adhesion to substrate. In "synthetic" state SMC (passages 2,3 of culture), there was decreased expression of contractile and adhesion (vinculin) proteins with a concomitant increase in cytoskeletal proteins (,-non-muscle [NM] actin and vimentin). These quantitative changes in structural proteins were associated with dramatic changes in their distribution. The distinct compartmentalisation of structural proteins observed in "contractile" state SMC was no longer obvious, with proteins more evenly distributed throughout the cytoplasm to accommodate altered cell function. Thus, SMC phenotypic modulation involves not only quantitative changes in contractile and cytoskeletal proteins, but also reorganisation of these proteins. Since the cytoskeleton acts as a spatial regulator of intracellular signalling, reorganisation of the cytoskeleton may lead to realignment of signalling molecules, which, in turn, may mediate the changes in function associated with SMC phenotypic modulation. Cell Motil. Cytoskeleton 49:130,145, 2001. © 2001 Wiley-Liss, Inc. [source] Cellular and molecular dissection of pluripotent adult somatic stem cells in planariansDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 1 2010Norito Shibata Freshwater planarians, Plathelminthes, have been an intriguing model animal of regeneration studies for more than 100 years. Their robust regenerative ability is one of asexual reproductive capacity, in which complete animals develop from tiny body fragments within a week. Pluripotent adult somatic stem cells, called neoblasts, assure this regenerative ability. Neoblasts give rise to not only all types of somatic cells, but also germline cells. During the last decade, several experimental techniques for the analysis of planarian neoblasts at the molecular level, such as in situ hybridization, RNAi and fluorescence activated cell sorting, have been established. Moreover, information about genes involved in maintenance and differentiation of neoblasts has been accumulated. One of the molecular features of neoblasts is the expression of many RNA regulators, which are involved in germline development in other animals, such as vasa and piwi family genes. In this review, we introduce physiological and molecular features of the neoblast, and discuss how germline genes regulate planarian neoblasts and what differences exist between neoblasts and germline cells. [source] Single-cell gene profiling of planarian stem cells using fluorescent activated cell sorting and its "index sorting" function for stem cell researchDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 1 2010Tetsutaro Hayashi To achieve an integrated understanding of the stem cell system of planarians at both the cellular and molecular levels, we developed a new method by combining "fluorescent activated cell sorting (FACS) index sorting" analysis and single-cell reverse transcription,polymerase chain reaction (RT,PCR) to detect the gene expression and cell cycle state of stem cells simultaneously. Single cells were collected using FACS, and cDNAs of each cell were used for semi-quantitative RT,PCR. The results were plotted on the FACS sorting profile using the "index sorting" function, which enabled us to analyze the gene expression in combination with cell biological data (such as cell cycle phase) for each cell. Here we investigated the adult stem cells of planarians using this method and obtained findings suggesting that the stem cells might undergo commitment during S to G2/M phase. This method could be a powerful and straightforward tool for examining the stem cell biology of not only planarians but also other organisms, including vertebrates. [source] Expression of Gpr177, a Wnt trafficking regulator, in mouse embryogenesisDEVELOPMENTAL DYNAMICS, Issue 7 2010Hsiao-Man Ivy Yu Abstract Wls/Evi/Srt encoding a multipass transmembrane protein has been identified as a regulator for proper sorting and secretion of Wnt in flies. We have previously demonstrated that Gpr177 is the mouse ortholog required for axis determination. Gpr177 is a transcriptional target of Wnt that is activated to assist its subcellular distribution in a feedback regulatory loop. We, therefore, proposed that reciprocal regulation of Wnt and Gpr177 is essential for the Wnt-dependent developmental and pathogenic processes. Here, we examine the expression pattern of Gpr177 in mouse development. Gpr177 is expressed in a variety of tissues and cell types during organogenesis. Furthermore, Gpr177 is a glycoprotein primarily accumulating in the Golgi apparatus in signal-producing cells. The glycosylation of Gpr177 is necessary for proper transportation in the secretory pathway. Our findings suggest that the Gpr177-mediated regulation of Wnt is crucial for organogenesis in health and disease. Developmental Dynamics 239:2102,2109, 2010. © 2010 Wiley-Liss, Inc. [source] Tissue surface tensions guide in vitro self-assembly of rodent pancreatic islet cellsDEVELOPMENTAL DYNAMICS, Issue 8 2007Dongxuan Jia Abstract The organization of endocrine cells in pancreatic islets is established through a series of morphogenetic events involving cell sorting, migration, and re-aggregation processes for which intercellular adhesion is thought to play a central role. In animals, these morphogenetic events result in an islet topology in which insulin-secreting cells form the core, while glucagon, somatostatin, and pancreatic polypeptide-secreting cells segregate to the periphery. Isolated pancreatic islet cells self-assemble in vitro into pseudoislets with the same cell type organization as native islets. It is widely held that differential adhesion between cells of the pancreatic islets generates this specific topology. However, this differential adhesion has never been rigorously quantified. In this manuscript, we use tissue surface tensiometry to measure the cohesivity of spherical aggregates from three immortalized mouse pancreatic islet cell lines. We show that, as predicted by the differential adhesion hypothesis, aggregates of the internally segregating INS-1 and MIN6 beta-cell lines are substantially more cohesive than those of the externally segregating ,-TC line. Furthermore, we show that forced overexpression of P-cadherin by ,-TC cells significantly perturbs the sorting process. Collectively, the data indicate that differential adhesion can drive the in vitro organization of immortalized rodent pancreatic islet cells. Developmental Dynamics 236:2039,2049, 2007. © 2007 Wiley-Liss, Inc. [source] Molecular characterization of conditionally immortalized cell lines derived from mouse early embryonic inner earDEVELOPMENTAL DYNAMICS, Issue 4 2004John A. Germiller Abstract Inner ear sensory hair cells (HCs), supporting cells (SCs), and sensory neurons (SNs) are hypothesized to develop from common progenitors in the early embryonic otocyst. Because little is known about the molecular signals that control this lineage specification, we derived a model system of early otic development: conditionally immortalized otocyst (IMO) cell lines from the embryonic day 9.5 Immortomouse. This age is the earliest stage at which the otocyst can easily be separated from surrounding mesenchymal, nervous system, and epithelial cells. At 9.5 days post coitum, there are still pluripotent cells in the otocyst, allowing for the eventual identification of both SN and HC precursors,and possibly an elusive inner ear stem cell. Cell lines derived from primitive precursor cells can also be used as blank canvases for transfections of genes that can affect lineage decisions as the cells differentiate. It is important, therefore, to characterize the "baseline state" of these cell lines in as much detail as possible. We characterized seven representative "precursor-like" IMO cell populations and the uncloned IMO cells, before cell sorting, at the molecular level by polymerase chain reaction (PCR) and immunocytochemistry (IHC), and one line (IMO-2B1) in detail by real-time quantitative PCR and IHC. Many of the phenotypic markers characteristic of differentiated HCs or SCs were detected in IMO-2B1 proliferating cells, as well as during differentiation for up to 30 days in culture. These IMO cell lines represent a unique model system for studying early stages of inner ear development and determining the consequences of affecting key molecular events in their differentiation. Developmental Dynamics 231:815,827, 2004. © 2004 Wiley-Liss, Inc. [source] FACS-array gene expression analysis during early development of mouse telencephalic interneuronsDEVELOPMENTAL NEUROBIOLOGY, Issue 4 2008Eric D. Marsh Abstract Cortical interneuron dysfunction has been implicated in multiple human disorders including forms of epilepsy, mental retardation, and autism. Although significant advances have been made, understanding the biologic basis of these disorders will require a level of anatomic, molecular, and genetic detail of interneuron development that currently does not exist. To further delineate the pathways modulating interneuron development we performed fluorescent activated cell sorting (FACs) on genetically engineered mouse embryos that selectively express green fluorescent protein (GFP) in developing interneurons followed by whole genome microarray expression profiling on the isolated cells. Bioinformatics analysis revealed expression of both predicted and unexpected genes in developing cortical interneurons. Two unanticipated pathways discovered to be up regulated prior to interneurons differentiating in the cortex were ion channels/neurotransmitters and synaptic/vesicular related genes. A significant association of neurological disease related genes to the population of developing interneurons was found. These results have defined new and potentially important data on gene expression changes during the development of cortical interneurons. In addition, these data can be mined to uncover numerous novel genes involved in the generation of interneurons and may suggest genes/pathways potentially involved in a number of human neurological disorders. © 2008 Wiley Periodicals, Inc. Develop Neurobiol, 2008. [source] The biology and functional morphology of Arca noae (Bivalvia: Arcidae) from the Adriatic Sea, Croatia, with a discussion on the evolution of the bivalve mantle marginACTA ZOOLOGICA, Issue 1 2008Brian Morton Abstract In the Croatian Adriatic, Arca noae occurs from the low intertidal to a depth of 60 m; it can live for > 15 years and is either solitary or forms byssally attached clumps with Modiolus barbatus. The shell is anteriorly foreshortened and posteriorly elongate. The major inhalant flow is from the posterior although a remnant anterior stream is retained. There are no anterior but huge posterior byssal retractor muscles and both anterior and posterior pedal retractors. The ctenidia are of Type B(1a) and the ctenidial,labial palp junction is Category 3. The ctenidia collect, filter and undertake the primary sorting of potential food in the inhalant water. The labial palps are small with simple re-sorting tracks on the ridges of their inner surfaces. The ciliary currents of the mantle cavity appear largely concerned with the rejection of particulate material. The mantle margin comprises an outer and an (either) inner or middle fold. The outer fold is divided into outer and inner components that secrete the shell and are photo-sensory, respectively. The latter bears a large number of pallial eyes, especially posteriorly. The inner/middle mantle fold of A. noae, possibly representative of simpler, more primitive conditions, may have differentiated into distinct folds in other recent representatives of the Bivalvia. [source] Sedimentological, modal analysis and geochemical studies of desert and coastal dunes, Altar Desert, NW MexicoEARTH SURFACE PROCESSES AND LANDFORMS, Issue 4 2007J. J. Kasper-Zubillaga Abstract Sedimentological, compositional and geochemical determinations were carried out on 54 desert and coastal dune sand samples to study the provenance of desert and coastal dunes of the Altar Desert, Sonora, Mexico. Grain size distributions of the desert dune sands are influenced by the Colorado River Delta sediment supply and wind selectiveness. The desert dune sands are derived mainly from the quartz-rich Colorado River Delta sediments and sedimentary lithics. The dune height does not exert a control over the grain size distributions of the desert dune sands. The quartz enrichment of the desert dune sands may be due to wind sorting, which concentrates more quartz grains, and to the aeolian activity, which has depleted the feldspar grains through subaerial collisions. The desert dune sands suffer from little chemical weathering and they are chemically homogeneous, with chemical alteration indices similar to those found in other deserts of the world. The desert sands have been more influenced by sedimentary and granitic sources. This is supported by the fact that Ba and Sr concentration values of the desert sands are within the range of the Ba and Sr concentration values of the Colorado River quartz-rich sediments. The Sr values are also linked to the presence of Ca-bearing minerals. The Zr values are linked to the sedimentary sources and heavy mineral content in the desert dunes. The Golfo de Santa Clara and Puerto Peñasco coastal dune sands are influenced by long shore drift, tidal and aeolian processes. Coarse grains are found on the flanks whereas fine grains are on the crest of the dunes. High tidal regimens, long shore drift and supply from Colorado Delta River sediments produce quartz-rich sands on the beach that are subsequently transported into the coastal dunes. Outcrops of Quaternary sedimentary rocks and granitic sources increase the sedimentary and plutonic lithic content of the coastal dune sands. The chemical index of alteration (CIA) values for the desert and coastal dune sands indicate that both dune types are chemically homogeneous. The trace element values for the coastal dune sands are similar to those found for the desert dune sands. However, an increase in Sr content in the coastal dune sands may be due to more CaCO3 of biogenic origin as compared to the desert dune sands. Correlations between the studied parameters show that the dune sands are controlled by sedimentary sources (e.g. Colorado River Delta sediments), since heavy minerals are present in low percentages in the dune sands, probably due to little heavy mineral content from the source sediment; grain sizes in the dune sands are coarser than those in which heavy minerals are found and/or the wind speed might not exert a potential entrainment effect on the heavy mineral fractions to be transported into the dune. A cluster analysis shows that the El Pinacate group is significantly different from the rest of the dune sands in terms of the grain-size parameters due to longer transport of the sands and the long distance from the source sediment, whereas the Puerto Peñasco coastal dune sands are different from the rest of the groups in terms of their geochemistry, probably caused by their high CaCO3 content and slight decrease in the CIA value. Copyright © 2006 John Wiley & Sons, Ltd. [source] | ||||||||||||||||||||||||||||||||||||||||||||||