bluuebunny/crossref_metadata_2025_split huggingface dataset:
Correlation of protein expression and gene expression in acute leukemia
Authors: Kern Wolfgang , Kohlmann Alexander , Wuchter Christian , Schnittger Susanne , Schoch Claudia , Mergenthaler Susanne , Ratei Richard , Ludwig Wolf‐Dieter , Hiddemann Wolfgang , Haferlach Torsten 
Published: August 2003

AbstractBackgroundFlow cytometry (FC) is a standard method for diagnosing and subclassifying acute myeloid (AML) and acute lymphoblastic (ALL) leukemias and allows the analysis of cell surface and intracellular proteins. In the future, diagnostic procedures may include oligonucleotide microarray analysis (MA) to detect expression patterns of large numbers of specific genes.MethodsFor comparison between methods, we performed FC and MA by using the Affymetrix GeneChip HG‐U133A microarray in parallel and correlated protein expression levels and mRNA abundance of 39 relevant genes in 113 patients with newly diagnosed AML and ALL and four normal bone marrow samples.ResultsIn 1,512 of 2,187 (69.1%) comparisons between methods, congruent results were obtained with regard to positivity or negativity of expression, respectively. Specifically, there was a significant correlation between protein expression and mRNA abundance for genes essential for diagnosing and subclassifying AML and ALL with regard to positivity and expression.ConclusionsThese data suggest that protein expression is highly correlated to mRNA abundance in AML and ALL. Further, expression patterns of specific genes provide important information at diagnosis for patients with AML and ALL that may be used for the discrimination from other leukemias. Cytometry Part B (Clin. Cytometry) 55B:29–36, 2003. © 2003 Wiley‐Liss, Inc.
Sensitivity of hemozoin detection by automated flow cytometry in non‐ and semi‐immune malaria patients
Authors: Grobusch Martin P. , Hänscheid Thomas , Krämer Benedikt , Neukammer Jörg , May Jürgen , Seybold Joachim , Kun Jürgen F. J. , Suttorp Norbert 
Published: August 2003

AbstractBackgroundCell‐Dyn automated blood cell analyzers use laser flow cytometry technology, allowing detection of malaria pigment (hemozoin) in monocytes. We evaluated the value of such an instrument to diagnose malaria in febrile travelers returning to Berlin, Germany, the relation between the instrument's performance and the patients' immune status, and the capacity to increase its sensitivity.MethodsMalaria diagnosis was routinely established by thick‐film microscopy. The patients' immune status was determined by an indirect fluorescent antibody test. Hemozoin detection was performed with a Cell‐Dyn 3000. To assess the capacity for sensitivity increase, the relative frequencies of pigment‐containing monocytes were determined for a subgroup of patients with the Cell‐Dyn 3000 in comparison with a flow cytometer specifically adapted to rare‐event analysis.ResultsOf 403 patients screened microscopically, 107 had malaria. Overall sensitivity with the Cell‐Dyn 3000 reached 48.6% (73.7% in semi‐immune and 28.6% in nonimmune individuals; P < 0.0001). Specificity was 96.2%. The detection limit was at a relative concentration of 2 × 10−4 pigment‐containing monocytes (PCMs). By employing rare‐event flow cytometry, the detection limit decreased to 3.25 × 10−5, thus yielding a considerably increased sensitivity for the subgroup studied.ConclusionsThe correlation between immune status and relative concentration of PCMs explains the failure of the routine instrument for nonimmune patients and its significantly higher sensitivity for semi‐immune individuals. The technique can be significantly improved by rare‐event flow cytometry. Cytometry Part B (Clin. Cytometry) 55B:46–51, 2003. © 2003 Wiley‐Liss, Inc.
Flow cytometry in primary breast carcinomas: Prognostic impact of multiploidy and hypoploidy
Authors: Michels Jean‐Jacques , Duigou Françoise , Marnay Jacques , Denoux Yves , Delozier Thierry , Chasle Jacques 
Published: August 2003

AbstractBackgroundThe aims of the present work were to study the prognostic impact of multiploidy and/or hypoploidy in breast cancers and their relation to other classic clinicopathologic prognostic factors (T, grade, receptors, and lymph node status).MethodsFrom 3 January 1990 to 7 January 1999, 1984 previously untreated, invasive breast carcinoma samples were snap frozen for flow‐cytometry.ResultsMultiploid tumors had the same prognosis as the aneuploid ones, and those with one hypoploid peak had a better prognosis than did the other aneuploid tumors. However, the presence of both multiploid and hypoploid peaks was correlated with a poor outcome, even after multivariate analysis. In this series after quality control, 93.4% of the histograms could be evaluated concerning ploidy; of these 81.6% could be assessed concerning S‐phase fraction (SPF) in the entire population and 77.1% in the multiploid population. In the entire population, we performed a multivariate analysis including all relevant prognostic factors remaining after monovariate analysis by using a compound factor (proliferative activity) regrouping SPF and mitotic activity. This analysis showed that lymph node status and proliferative activity correlates with every type of survival, whereas receptor status correlates with all types of survival except recurrence free survival size, correlated with non‐metastasis and overall survival. Grade and age correlated only with overall survival and vascular permeations only with disease‐free survival.ConclusionsSPF is a valuable predictor of survival, can be confidently assessed in multiploid histograms, and thus improves the yield of flow cytometry. When combined with mitotic activity, the prognostic impact of SPF is the same as that of lymph node status. Tumors that are hypoploid and multiploid have a significantly worse prognosis. Cytometry Part B (Clin. Cytometry) 55B:37–45, 2003. © 2003 Wiley‐Liss, Inc.
Comparison of intracellular cytokine production with extracellular cytokine levels using two flow cytometric techniques
Authors: Schuerwegh A. J. , De Clerck L. S. , Bridts C. H. , Stevens W. J. 
Published: August 2003

AbstractBACKGROUNDWe investigated the relation between intracellular cytokine production and extracellular cytokine levels by using two flow cytometric techniques.METHODSA two‐color flow cytometric technique was used to measure interleukin (IL)–1β, IL‐6, tumor necrosis factor α (TNF‐α), IL‐10, and IL‐12 production blocked intracellularly with brefeldin A in lipopolysaccharide (LPS)–stimulated CD14+ monocytes and IL‐2, IL‐4, and IFN‐γ production in phorbol‐12‐mirystate‐13‐acetate (PMA)–stimulated CD3+ T lymphocytes in samples from patients with rheumatoid arthritis. A flow cytometric microsphere‐based immunoassay was performed to detect cytokine secretion in plasma of PMA‐ and LPS‐stimulated whole blood samples.RESULTSThere was a strong linear correlation between extracellular quantitative (pg/ml) and intracellular semiquantitative detection of LPS‐stimulated IL‐1β, IL‐6, IL‐10, and IL‐12 production (r > 0.9). For lymphocytes, extracellularly detected IL‐2 and IFN‐γ correlated well with percentages of cytokine‐producing cells (r > 0.8). The percentages of IL‐4–positive T cells were moderately correlated with the secreted amounts of IL‐4 as detected with the microsphere‐based immunoassay (r = 0.7).CONCLUSIONOverall, there was a good correlation between semiquantitative intracellular detection of cytokines and the secreted amounts of cytokines detected with the microsphere based immunoassay. Cytometry Part B (Clin. Cytometry) 55B:52–58, 2003. © 2003 Wiley‐Liss, Inc.
Comparative immunophenotypic features of EBV‐positive and EBV‐negative atypical lymphocytosis
Authors: Hudnall S. David , Patel Jyoti , Schwab Hanna , Martinez José 
Published: August 2003

AbstractBackgroundAtypical lymphocytosis is a common peripheral blood abnormality seen not only in Epstein‐Barr virus (EBV)‐associated acute infectious mononucleosis but also in other conditions, including other viral infections, cancer, immune reactions, etc. Despite numerous reports of individual immunophenotypic alterations in EBV‐positive infectious mononucleosis, a detailed comparative analysis of the immunophenotypic changes of peripheral blood lymphocyte subsets in infectious mononucleosis and other forms of atypical lymphocytosis is lacking.MethodsMultiparametric flow immunocytometry with 26 monoclonal antibodies was performed on peripheral blood lymphocytes from 97 cases of atypical lymphocytosis and 37 normal controls. Atypical lymphocytosis was defined as absolute lymphocytosis with >10% atypical lymphocytes. Absolute or relative mean values of various lymphocyte subsets from EBV‐positive cases, EBV‐negative cases, and normal controls were compared with the Student's t‐test.ResultsHighly significant abnormalities detected in atypical lymphocytosis include increases in CD3+/CD8+, CD3−/CD16/56+, CD3+/γδ+, CD8+/CD48−, CD8+/CD57−, CD8+/CD95+, CD4+/CCR5+ CD4+/CD7−, CD4+/CD43−, CD4+/CD48−, and CD4+/CD62L− subsets. In contrast, no change in absolute CD4+ T cell and CD19+ B cell counts is seen. When compared with EBV‐negative cases, EBV‐positive cases are characterized by younger age, and increased numbers of absolute lymphocytes, atypical lymphocytes, CD8+ cells, NK cells, γδ T cells, CD8+/CD45RO+ cells, CD8+/CD57− cells, and CD8+/CD28+ cells.ConclusionsAll forms of atypical lymphocytosis are characterized by a marked increase in activated CD8‐positive T cells, a moderate increase in NK cells, and no increase in CD4‐positive T cells and B cells. Although morphologically indistinguishable, EBV‐associated mononucleosis is characterized by several significant differences in peripheral blood lymphocyte subsets when compared with EBV‐negative atypical lymphocytosis, most notably increased numbers of CD57‐negative CD8 T cells and γδ T cells. Cytometry Part B (Clin. Cytometry) 55B:22–28, 2003. © 2003 Wiley‐Liss, Inc.
Flow rate calibration II: A clinical evaluation study using PanLeucoGating as a single‐platform protocol
Authors: Storie Ian , Sawle Alex , Whitby Liam , Goodfellow Karen , Granger Vivian , Reilly John T. , Barnett David 
Published: August 2003

AbstractBackgroundCD4+ T‐lymphocyte enumeration is vital for monitoring disease progression in individuals positive for the human immunodeficiency virus (HIV), and as a result, there is a need to develop cost‐effective protocols that provide accuracy, precision, and affordability. Recently, PanLeucoGating has been shown to fulfill these requirements; however, although comparable to state‐of‐the‐art single‐platform protocols (SP), there is still a requirement for an accurate total white cell count. To overcome this limitation, we recently developed a flow‐rate based calibration method that enables the PanLeucoGating protocol to be used as a SP approach, and in this study show that this approach can be used for CD4+ T‐lymphocyte enumeration.MethodsA total of 113 HIV samples were analyzed using three protocols: (a) state‐of‐the art SP bead‐based method (MultiSet; predicate protocol), (b) PanLeucoGating protocol used as a dual‐platform (DP) approach, and (c) the newly developed flow rate–based SP approach. We demonstrate that flow rate calibration can be achieved easily and that the method is highly comparable to the state‐of‐the‐art SP method.ResultsA high correlation was observed between the predicate protocol and the SP PanLeucoGating approach over the whole range of CD4 counts tested (r2 = 0.9928; bias 8 cells/μl), including the clinically relevant range (e.g., 0–200 CD4 cells/μl; bias 0 cells/μl). For batched samples, the cost of providing a CD4+ T‐lymphocyte count was reduced to approximately US $1.ConclusionsThe SP PanLeucoGating is a cost‐effective approach to CD4+ T‐lymphocyte enumeration that maintains accuracy and precision. Cytometry Part B (Clin. Cytometry) 55B:8–13, 2003. © 2003 Wiley‐Liss, Inc.
Flow rate calibration I: A novel approach for performing absolute cell counts
Authors: Storie Ian , Sawle Alex , Goodfellow Karen , Whitby Liam , Granger Vivian , Reilly John T. , Barnett David 
Published: August 2003

AbstractBackgroundReports suggest that flow rate (FR) is constant on bench top flow cytometers. Therefore, if FR is constant, the volume acquired in a fixed time period will also be constant, enabling absolute leucocyte counting using flow rate calibration (FRC).MethodsFR stability was ascertained on a standard FACSCalibur by counting TruCount beads suspended in phosphate buffered saline over 120 s. Studies using two lysing solutions (FACS lysing solution and PharM Lyse) and corresponding sample lysates established a lysing solution calibration factor (CF). Absolute CD4+T‐lymphocyte counts on 10 peripheral blood samples determined using FRC were compared with the predicate method TruCount/MultiTEST, incorporating MultiSET software. Linearity studies were also performed at three different flow rates.ResultsA high degree of linearity over a wide range of counts (50 to >1,600 CD4+T lymphocytes/μl) at all three pressures was observed. Importantly, there was no significant difference from the predicate method when appropriate lysing solution CF was used.ConclusionsUsing a simple calibration procedure and incorporation of an appropriate lysing solution CF, we show that FRC can easily be performed. The technical details that underpin this novel approach for absolute leucocyte enumeration are provided. Cytometry Part B (Clin. Cytometry) 55B:1–7, 2003. © 2003 Wiley‐Liss, Inc.
Patterning of embryonic blood vessels
Authors: LaRue Amanda C. , Mironov Vladimir A. , Argraves W. Scott , Czirók András , Fleming Paul A. , Drake Christopher J. 
Published: August 2003

AbstractMorphometric methods were developed to characterize the geometry of vascular patterns in avian and murine embryos. By using these methods, we found that networks of blood vessels formed during vasculogenesis share similar geometric properties (i.e., mean blood vessel diameters and avascular space diameters) regardless of developmental stage, location, or species in which they form. We also found that endothelial cell density within a unit area of an embryonic vasculature could be used to accurately distinguish between a small diameter, capillary‐like vascular network (low endothelial cell density) and a large diameter, presinusoidal network (high endothelial cell density). Furthermore, we show that endothelial cell size remains constant in small and large diameter vessels, indicating that increased endothelial cell size is not the basis for diversity in vessel diameter. These observations serve as a foundation for future studies seeking to evaluate the effects of agents or genetic mutations on aspects of vasculogenesis. Developmental Dynamics 228:21–29, 2003. © 2003 Wiley‐Liss, Inc.
Timeless in lung morphogenesis
Authors: Xiao Jing , Li Changgong , Zhu Nian‐Ling , Borok Zea , Minoo Parviz 
Published: August 2003

AbstractThe Clock gene, timeless, regulates circadian rhythm in Drosophila, but its vertebrate homolog is critical to embryonic development. Timeless was shown to be involved in murine urethral bud branching morphogenesis. We generated a polyclonal antibody to mouse TIMELESS (mTIM) and studied its distribution and its potential role during lung development, which also requires branching morphogenesis. In the early mouse embryo, TIM was localized to all organs, especially the neural epithelium. In embryonic day (E) 9.5 embryos, TIM was present in both epithelial and mesenchymal cells at the onset of lung morphogenesis. In E15 embryos, TIM decreased in the mesenchyme but remained pronounced in the epithelium of both large and small airways. Later, TIM was localized to a specific subset of epithelial cells with alveolar type 2 phenotype. This finding was verified by immunostaining of isolated alveolar type 2 cells. In the proximal airways, TIM was colocalized with CCSP to nonciliated columnar epithelial cells. Antisense oligonucleotides to mTim specifically inhibited branching morphogenesis of embryonic lungs in explant culture without affecting SpC expression an alveolar type 2 cell marker. In cultured lung cells, expression of TIM is independent of cell cycle and proliferation. These studies indicate that the function of Timeless is highly conserved in organs whose formation requires branching morphogenesis. Developmental Dynamics 228:82–94, 2003. © 2003 Wiley‐Liss, Inc.
Analysis of transcription regulatory regions of embryonic chicken pepsinogen (ECPg) gene
Authors: Watanuki Kumiko , Yasugi Sadao 
Published: August 2003

AbstractGenes encoding pepsinogens, zymogens of digestive enzyme pepsins, are expressed specifically in the gland epithelial cells of the vertebrate stomach, and their expression is also developmentally regulated, therefore providing a good model for the analysis of transcriptional regulation of genes. In the development of chicken embryonic stomach, the epithelium invaginates into the mesenchyme and forms glands and gland epithelial cells then begin to express embryonic chicken pepsinogen (ECPg) gene. It has been shown that cGATA5 binds directly GATA binding sites located within 1.1‐kbp upstream of ECPg gene and activates its transcription. To find more precisely the sequences necessary for ECPg gene transcription, we carried out deletion and mutation analysis with 1.1‐kbp upstream region. The results suggest that binding of GATA factor to three GATA binding sites within the upstream region −656 to −419 synergistically regulates ECPg expression in the gland epithelial cells. Developmental Dynamics 228:51–58, 2003. © 2003 Wiley‐Liss, Inc.
Chromatin reorganization accompanying cellular dedifferentiation is associated with modifications of histone H3, redistribution of HP1, and activation of E2F‐target genes
Authors: Williams Leor , Zhao Jing , Morozova Nadya , Li Yan , Avivi Yigal , Grafi Gideon 
Published: August 2003

AbstractThe remarkable regeneration capacity of plant cells is based on their capability to dedifferentiate. We recently reported that cellular dedifferentiation proceeds through two distinct phases, each accompanied by chromatin decondensation: acquisition of competence for fate switch followed by a signal‐dependent reentry into S phase (Zhao et al. [2001] J. Biol. Chem. 276:22772‐22778). The purpose of this study was to (1) characterize changes in chromatin factors associated with chromatin decondensation, and (2) study the relationship between chromatin decondensation and transcriptional activation of pRb/E2F‐regulated genes. We show that plant cells competent for fate switch display a disruption of nucleolar domain appearance associated with condensation of 18S ribosomal DNA, as well as modifications of histone H3 and redistribution of heterochromatin protein 1 (HP1). We further show that the pRb/E2F‐target genes RNR2 and PCNA are condensed and silent in differentiated leaf cells but become decondensed, although not yet activated, as cells acquire competence for fate switch; transcriptional activation becomes evident during progression into S phase, concomitantly with pRb phosphorylation. We propose that chromatin reorganization is central for reversion of the differentiation process leading to resetting of the gene expression program and activation of silent genes. Developmental Dynamics 228:113–120, 2003. © 2003 Wiley‐Liss, Inc.
Vesicle‐associated protein‐A is differentially expressed during intestinal smooth muscle cell differentiation
Authors: Gabetta Vijayalakshmi , Trzyna Wendy , Phiel Christopher , McHugh Kirk M. 
Published: August 2003

AbstractGastrointestinal (GI) smooth muscle diseases represent a major health concern affecting in excess of 2 million people each year. Little is currently known regarding the molecular mechanisms controlling either normal or pathogenic GI smooth muscle development. In an effort to identify the specific gene products responsible for modulating GI smooth muscle cell (SMC) differentiation, we performed differential display on distinct intestinal SMC (ISMC) phenotypes. This analysis identified over 40 unique transcripts that appeared to be differentially expressed in distinct SMC phenotypes. One such transcript that appeared to be preferentially expressed in immature smooth muscle myocytes was identified as vesicle‐associated membrane protein, associated protein A (VAP‐A). Northern blot analysis confirmed that VAP‐A was expressed threefold higher in immature smooth muscle myocytes when compared with both smooth muscle myoblasts and mature smooth muscle myocytes. VAP‐A mRNA was differentially expressed during normal rat development and showed peak levels of expression in the intestine during late embryogenesis and early neonatal development. These observations provide the first evidence that VAP‐A–mediated membrane trafficking may play an important role in modulating ISMC differentiation. Developmental Dynamics 228:11–20, 2003. © 2003 Wiley‐Liss, Inc.
HOXB5 expression is spatially and temporarily regulated in human embryonic gut during neural crest cell colonization and differentiation of enteric neuroblasts
Authors: Fu Ming , Chi Hang Lui Vincent , Har Sham Mai , Nga Yin Cheung Annie , Kwong Hang Tam Paul 
Published: August 2003

AbstractHOX genes from paralogous groups 4 and 5 are particularly relevant to the gut neuromusculature development because these genes are expressed at the splanchnic mesoderm surrounding the gut diverticulum, and at the level of the neural tube from where the vagal neural crest cells (NCCs) originate. In this study, we examined the migration and differentiation of NCCs, and investigated the expression patterns of HOXB5 in human embryonic guts. Human embryos of gestational week‐4 to ‐8.5 were studied. Vagal NCCs enter the esophagus, migrate, and colonize the entire gut in a rostrocaudal manner between week‐4 and week‐7. The migrating NCCs in gut express HOXB5. Two separate and discontinuous mesenchymal expression domains of HOXB5 were detected in the gut: the distal domain preceding the migratory NCCs; and the proximal domain overlapping with the NCCs. The two expression domains shift caudally in parallel with the rostrocaudal migration of NCCs between week‐4 and week‐5. Neuron and glia differentiation of NCCs are concomitant with HOXB5 down‐regulation in NCCs and the mesenchyme. By week‐7, myenteric plexuses have formed; HOXB5 expression is switched on in the plexuses. We found that (1) the migratory route of NCCs in human embryonic gut was similar to that in mice and chicks; and (2) the expression pattern of HOXB5 correlated with the migration and differentiation of NCCs, suggesting a regulatory role of HOXB5 in the development of NCCs. Developmental Dynamics 228:1–10, 2003. © 2003 Wiley‐Liss, Inc.
Pak1 and its T212 phosphorylated form accumulate in neurones and epithelial cells of the developing rodent
Authors: Zhong Julia L. , Banerjee Monisha D. , Nikolic Margareta 
Published: August 2003

AbstractThe serine/threonine kinase Pak1 is a target of the RhoGTPases Rac and Cdc42 and an important regulator of cell morphology and migration. Recent work from several laboratories has indicated that Pak1 controls microtubule dynamics as well as the organisation of F‐actin microfilaments. Pak1 is phosphorylated on T212 by the p35/Cdk5 or cyclin B1/Cdc2 kinase in postmitotic neurones and mitotic cells, respectively. To understand its function during development, we have carried out a detailed temporal and spatial analysis of Pak1 expression and phosphorylation on T212. In the embryonic forebrain, Pak1 and Pak1T212(PO4) were seen to accumulate in the corpus callosum, intermediate zone, lateral olfactory tracts, and anterior commissures. Epithelial cells of the mouse embryo lung, kidney, intestine, and skin also exhibited high levels of Pak1 and Pak1T212(PO4), suggesting a previously unsuspected role in epithelial differentiation. Pak1T212(PO4) was undetectable in all adult tissues. Together, these data indicate a specific, developmentally regulated role of the Pak1 kinase. Developmental Dynamics 228:121–127, 2003. © 2003 Wiley‐Liss, Inc.