bluuebunny/crossref_metadata_2025_split huggingface dataset:
Importance of CD117 in the evaluation of acute leukemias by flow cytometry
Authors: Newell James O. , Cessna Melissa H. , Greenwood Jay , Hartung Leah , Bahler David W. 
Published: February 2003

AbstractBackgroundThe issue of which specific antibodies need to be used when evaluating acute leukemias by flow cytometry is controversial.MethodsRecent studies have suggested that antibodies against CD117 or c‐kit are not essential for the assignment of blast lineage by flow cytometry, even though CD117 appears to be a very specific marker for myeloid lineage acute leukemias. We report a case of acute myeloid leukemia M2 subtype with an 8:21 translocation, where the leukemic blasts expressed CD117, CD19, and CD15 but did not show definitive expression of the myeloid markers CD13 or CD33.Results and ConclusionsThis study highlights the importance of CD117 when evaluating acute leukemias by flow cytometry, which was necessary in this case to suggest that the blasts were phenotypically abnormal myeloblasts. In addition, this case presented an unusual acute myeloid leukemia phenotype that will likely be encountered by others and could be difficult to interpret. Cytometry Part B (Clin. Cytometry) 52B:40–43, 2003. © 2003 Wiley‐Liss, Inc.
Normal values of CD4 and CD8 lymphocyte subsets in healthy indian adults and the effects of sex, age, ethnicity, and smoking
Authors: Uppal S. S. , Verma Shashi , Dhot P. S. 
Published: February 2003

AbstractBackgroundInformation on lymphocyte populations (T, B, and natural killer cells) and subpopulations (CD4 and CD8) in India is generally lacking. Measurement of T‐cell subsets is important in India for evaluating disease stage and progression in individuals with the human immunodeficiency virus (HIV). Hence, this study was conducted to provide normal ranges of absolute and percentage values of CD4 and CD8 T‐lymphocyte subsets and the ratio of CD4 to CD8 in normal Indian adults.MethodsFlow cytometric analysis (EPICS‐XL) was used to determine the range of T‐lymphocyte subpopulations in normal Indian blood donors at Command Hospital and the Armed Forces Medical College, Pune, India. The reference population consisted of 94 healthy HIV‐seronegative blood donors. T‐lymphocyte subsets were analyzed with two‐color immunophenotyping of peripheral blood lymphocytes with the use of a lysed whole‐blood technique and enumerated.ResultsFor normal values of various blood components, we found mean values of 2114 cells/μl for total lymphocytes, 865 cells/μl (40.2%) for CD4+ lymphocytes, 552 cells/μl (31.3%) for CD8+ lymphocytes, and 1.7 for the CD4:CD8 ratio. The 95% confidence intervals for the same parameters were 1115–4009 cells/μl, 430–1740 cells/μl (30.75–49.60%), 218–1396 cells/μl (20.06–42.52%), and 0.39–3.02 respectively. Females had significantly higher CD4 counts (P < 0.05), percentage of CD4 lymphocytes (P < 0.01), and CD4:CD8 ratio (P < 0.01). Males had a significantly higher percentage of CD8 lymphocytes (P < 0.01). They also had higher CD8 counts that did not reach significance. Age, ethnicity (Dravidian versus Aryan), smoking, alcohol consumption, and the interval between drawing the blood sample and its analysis were factors that did not produce statistically significant differences in the T‐cell subsets studied.ConclusionsWhen compared with other published series, the CD4 and CD8 values in healthy Indians were no different from those reported in the West. These observations have important clinical implications for the use of T‐lymphocyte subset measurements in India, especially in the management of HIV infection. The normal ranges established by this study can be used as a reference for decisions made in clinical practice. Cytometry Part B (Clin. Cytometry) 52B:32–36, 2003. © 2003 Wiley‐Liss, Inc.
Flow cytometry versus histamine release analysis of in vitro basophil degranulation in allergy to Hymenoptera venom
Authors: Lambert C. , Guilloux L. , Dzviga C. , Gourgaud‐Massias C. , Genin C. 
Published: February 2003

AbstractBackgroundFlow cytometry (FCM) has been proposed for specific allergy in vitro testing. We investigated its biological significance for allergy to Hymenoptera venoms and compared it with the routinely performed basophil histamine release test (HRT).MethodsBlood samples from 26 allergic and 8 nonallergic donors were incubated with venom at serial concentrations. Basophils were analyzed with anti‐CD45‐PE‐Cyanin 5, Anti‐IgE‐FITC, and Anti‐CD63‐Phycoerythrine. HRT was measured by radioimmunoassay.ResultsFCM was as convenient as HRT for measuring basophil reactivity in at least 87% of allergic and 75% of nonallergic subjects. CD63 outer expression was specifically induced in 91% of releaser subjects (86% on HRT) and in 1 of 10 tests in nonallergic donors, or one of six tests (16% on HRT) in allergic patients tested with an irrelevant allergen. Both methods were concordant in 85.7% of the tests. The three discordant patients had low‐grade reactions and borderline biological responses on FCM (n = 2) or HRT (n = 1).ConclusionsThe dynamic, physiologic significance of CD63, the dose–response curve, and dependency on ethylene‐diaminetetra acetic acid suggested that both tests reflect the same mechanism. Cytometry Part B (Clin. Cytometry) 52B:13–19, 2003. © 2003 Wiley‐Liss, Inc.
B‐cell monoclonal lymphocytosis and B‐cell abnormalities in the setting of familial B‐cell chronic lymphocytic leukemia
Authors: Marti Gerald E. , Carter Patricia , Abbasi Fatima , Washington Glennelle C. , Jain Nisha , Zenger Vincent E. , Ishibe Naoko , Goldin Lynn , Fontaine Laura , Weissman Nancy , Sgambati Maria , Fauget Guy , Bertin Pablo , Vogt Robert F. , Slade Barbara , Noguchi Philip D. , Stetler‐Stevenson M. A. , Caporaso Neil 
Published: February 2003

AbstractBackgroundAmong all hematologic malignancies, B‐cell chronic lymphocytic leukemia (BCLL) has the highest familial clustering (three‐ to sevenfold increase), strongly suggesting a genetic component to its etiology. Familial BCLL can be used as a model to study the early pathogenesis of this disease.MethodsWe examined nine kindreds from the National Cancer Institute's Familial BCLL Registry, consisting of 19 affected members with BCLL and 33 clinically unaffected first‐degree relatives. Flow cytometric immunophenotyping to detect a B‐cell monoclonal lymphocytosis (BCML) was performed. Monoclonality was confirmed by polymerase chain reaction analysis of whole blood DNA. Cell cycle analysis for aneuploidy was conducted.ResultsIn all affected individuals, we observed the classic BCLL CD5/CD19/CD20/CD23 immunophenotypic patterns. Six of the 33 unaffected individuals (18%) had evidence of BCML. Additional individuals (13/33, 39%) showed some other abnormality, whereas 14 individuals (42%) were normal. Based on an estimated prevalence of 0.7% for BCML in the general population, the finding of six subjects (18%) with clonal abnormalities in this relatively modest sample was significantly greater than expected (i.e., 18% vs. 0.7%, P < 5.7 × 10−9).ConclusionsIndividual components of BCML and other B‐cell abnormalities were observed in almost half of the apparently unaffected individuals. Our findings suggested that BCML may be an early detectable abnormality in BCLL. The spectrum of some of these observed abnormalities suggested the involvement of different B‐cell subpopulations or different pathways in clonal evolution. Population‐based, longitudinal studies will be required to determine the incidence of BCML and other B‐cell abnormalities and their relation to disease progression in BCLL and other closely related B‐cell lymphoproliferative disorders. Cytometry Part B (Clin. Cytometry) 52B:1–12, 2003. Published 2003 Wiley‐Liss, Inc.
Separation of proteins in a multicompartment electrolyzer with chambers defined by a bed of gel beads
Authors: Cretich Marina , Pirri Giovanna , Carrea Giacomo , Chiari Marcella 
Published: February 2003

AbstractMulticompartment electrolyzers (MEs) with isoelectric membranes were introduced in 1989 for purifying proteins in an electric field. At the basis of ME technology there are membranes consisting of cross‐linked copolymers of acrylamide and acrylamido monomers bearing protolytic groups. The technology employed for casting the membranes is an extension of the isoelectric focusing in immobilized pH gradient technique for which specific acrylamido monomers, known with the trade name of Immobiline, have been developed. However, the use of continuous membranes presents several disadvantages. Due to the mechanical characteristics of polyacrylamide, the gel must physically adhere onto a rigid support, which prevents it from collapsing. The support must have a highly porous structure in order to be permeable to proteins. The mechanical fragility of the membranes is one of the main problems that hinders the industrial scale application of ME separators. In order to overcome this problem, we propose to substitute the continuous membranes with a bed of gel beads of identical comonomer composition, obtained by an inverse emulsion polymerization process.
Rapid high‐resolution electrophoresis of multimeric von Willebrand Factor using a thermopiloted gel apparatus
Authors: Smejkal Gary B. , Shainoff John R. , Kottke‐Marchant Kandice M. 
Published: February 2003

AbstractRapid and highly reproducible nonreducing agarose gel electrophoresis (NRAGE) of von Willebrand Factor (vWF) multimers was performed using a thermostated minigel apparatus that monitors and precisely controls internal gel temperature. The substitution of lithium dodecyl sulfate (LiDS) for sodium dodecyl sulfate (SDS) allowed electrophoresis to be performed below the 16°C Krafft point of SDS and facilitated NRAGE of vWF over the entire range of 0–35°C. Internal gel temperature was regulated by a thermocouple probe inserted directly into the gel during electrophoresis which interfaced with a thermopilot that continually measures and adjusts temperature to within ± 0.5°C. At 10°C operative temperature, NRAGE at 1.5% agarose concentration was completed in 20 min at 250 V. Electrophoresis could be performed in only 10 min at 500 V, but at such high voltages, localized temperature fluctuations as much as 6°C resulted in perturbation of banding patterns in those vicinities. In the optimized method, both high molecular weight multimers and proteolytic fragments of vWF were separable suggesting clinical applicability of this system for the diagnosis of von Willebrand Disease and thrombotic thrombocytopenic purpura.
Detection of glycoproteins in polyacrylamide gels and on electroblots using Pro‐Q Emerald 488 dye, a fluorescent periodate Schiff‐base stain
Authors: Hart Courtenay , Schulenberg Birte , Steinberg Thomas H. , Leung Wai‐Yee , Patton Wayne F. 
Published: February 2003

AbstractPro‐Q Emerald 488 glycoprotein stain reacts with periodic acid‐oxidized carbohydrate groups, generating a bright green‐fluorescent signal on glycoproteins. The stain permits detection of less than 5–18 ng of glycoprotein per band, depending upon the nature and the degree of protein glycosylation, making it roughly 8–16‐fold more sensitive than the standard colorimetric periodic acid‐Schiff base method using acidic fuchsin dye (pararosaniline). The green‐fluorescent signal from Pro‐Q Emerald 488 stain may optimally be visualized using charge‐coupled device/xenon arc lamp‐based imaging systems or 470–488 nm laser‐based gel scanners. Though glycoprotein detection may be performed on transfer membranes, direct detection in gels avoids electroblotting and the specificity of staining is better in gels. After detecting glycoproteins with Pro‐Q Emerald 488 dye, total protein profiles may subsequently be detected using SYPRO Ruby protein gel stain. Using computer‐assisted registration techniques, images may then be merged to generate differential display maps.
Fluorescence detection and quantitation of recombinant proteins containing oligohistidine tag sequences directly in sodium dodecyl sulfate‐polyacrylamide gels
Authors: Hart Courtenay , Schulenberg Birte , Diwu Zhenjun , Leung Wai‐Yee , Patton Wayne F. 
Published: February 2003

AbstractTwo fluorophore‐nitrilotriacetic acid conjugates, Pro‐Q Sapphire 365 and Pro‐Q Sapphire 488 oligohistidine gel stains, have been developed for the fluorescence detection of fusion proteins containing oligohistidine tags directly in sodium dodecyl sulfate polyacrylamide gels, without the requirement for electroblotting, reporter enzymes or secondary detection reagents. Pro‐Q Sapphire 365 oligohistidine gel stain exhibits bright‐blue fluorescence (emission maximum = 450 nm) when illuminated with UV‐A or UV‐B light from a standard ultraviolet transilluminator. Pro‐Q Sapphire 488 oligohistidine gel stain exhibits bright‐green fluorescence (emission maximum = 515 nm) when illuminated with visible light from a laser‐based gel scanner equipped with a 470 nm second‐harmonic generation (SHG) or 488 nm argon‐ion laser source. Typically, 25–65 ng of oligohistidine‐tagged fusion protein in whole cell lysates is detectable using either stain. After documenting the fluorescence signal from the Pro‐Q Sapphire dyes, gels may be post‐stained with the red‐fluorescent SYPRO Ruby protein gel stain in order to reveal the total protein pattern.
Microscale analysis of mucin‐type O‐glycans by a coordinated fluorophore‐assisted carbohydrate electrophoresis and mass spectrometry approach
Authors: Robbe Catherine , Capon Calliope , Flahaut Christophe , Michalski Jean‐Claude 
Published: February 2003

AbstractThe total glycan moiety was released in a single step from native glycoproteins by a nonreductive β‐elimination procedure. The generated oligosaccharides were further derivatized either with the hydrophobic fluorophore 2‐aminoacridone (AMAC) or the charged 8‐aminonaphthalene‐1,3,6‐trisulfonic acid (ANTS) fluorophore, and the resulting fluorescent derivatives were separated according to their hydrodynamic size or charge with high‐resolution gel electrophoresis. Both N‐ and O‐glycans released by this β‐elimination procedure might be analyzed simultaneously. AMAC derivatization allows a rapid separation of neutral and charged oligosaccharides without prior fractionation. Derivatized oligosaccharide species were then eluted from the gel slices and analyzed by mass spectrometry. This methodology allowed the rapid structural characterization of each glycan in term of monosaccharide composition and sequence. Using this technique we were able to screen several heterogeneous O‐glycan mixtures isolated at the picomolar range from reference glycoproteins or mucins.
Congruence between starch gel and polyacrylamide gel electrophoresis in detecting allozyme variation in pulmonate land slugs
Authors: Geenen Sofie , Jordaens Kurt , Castilho Rita , Backeljau Thierry 
Published: February 2003

AbstractThe predominantly selfing slug species Arion (Carinarion) fasciatus, A. (C.) silvaticus and A. (C.) circumscriptus are native in Europe and have been introduced into North America, where each species consists of a single, homozygous multilocus genotype (strain), as defined by starch gel electrophoresis (SGE) of allozymes. In Europe, the “one strain per species” hypothesis does not hold since polyacrylamide gel electrophoresis (PAGE) of allozymes uncovered 46 strains divided over the three species. However, electrophoretic techniques may differ in their ability to detect allozyme variation. Therefore, several Carinarion populations from both continents were screened by applying the two techniques simultaneously on the same individual slugs and enzyme loci. SGE and PAGE yielded exactly the same results, so that the different degree of variation in North American and European populations cannot be attributed to differences in resolving power between SGE and PAGE. We found four A. (C.) silvaticus strains in North America indicating that in this region the “one strain per species” hypothesis also cannot be maintained. Hence, the discrepancies between previous electrophoretic studies on Carinarion are most likely due to sampling artefacts and possible founder effects.
Sequence‐specific and nonspecific mobilities of single‐stranded oligonucleotides observed by changing the borate buffer concentration
Authors: Biyani Manish , Nishigaki Koichi 
Published: February 2003

AbstractThe suitability of gel electrophoresis to structural analysis of nucleic acids has been examined, using from borate buffer concentrations (in the range from 4.5 to 450 mM. The gel electrophoretic mobility of single‐stranded oligonucleotides was shown to be sequence‐dependent at higher concentrations than 27 mM of borate buffer and nondependent at lower one (less than 9 mM). As a result, each dodecamer had a sequence‐specific critical concentration of Tris‐borate‐EDTA (TBE) buffer at which each seems to change its structural state. At the lower concentration than the critical one, all the dodecamers turned to the state of a finite mobility and migrated in a sharp band. This finding is discussed and rationalized by the assumption that the difference in conformational dynamics of oligonucleotides, due to the difference in their sequence, is mainly responsible for the observed difference in their mobility.
Detection of single‐nucleotide polymorphisms coding for three ovine prion protein variants by primer extension assay and capillary electrophoresis
Authors: Zsolnai Attila , Anton István , Kühn Christa , Fésüs László 
Published: February 2003

AbstractAn alternative method is described for the determination of ovine prion protein allelic variants at codon 136, 154, and 171. The four mutations responsible for amino acid changes are typed simultaneously. The technique utilizes dideoxy chain termination reaction using fluorescently labeled dideoxy nucleotides. The single‐base extended primers are resolved on a capillary electrophoresis instrument. Data obtained by our approach are presented according to genotype distribution in some breeds as a part of the validation procedure.
Application of high‐resolution capillary array electrophoresis with automated fraction collection for GeneCalling™ analysis of the yeast genomic DNA
Authors: Berka Jan , Ruiz‐Martinez Marie C. , Hammond Richard , Minarik Marek , Foret Frantisek , Sosic Zoran , Kleparnik Karel , Karger Barry L. 
Published: February 2003

AbstractCapillary array instrument was applied to transcript profiling of the yeast genomic DNA using GeneCalling™ chemistry. The instrument integrated a 12‐capillary array for DNA separation with a replaceable sieving matrix, laser‐induced fluorescence detection and an automated microfraction collector. The DNA fractions, exiting the separation capillaries, were continuously deposited in a 1536‐well collection plate made of agarose gel. DNA fragments recovered from selected fractions were cloned and then sequenced. Over 80% of theoretically predicted fragments could be recovered in the collected fractions, cloned and sequenced with an average redundancy of threefold. Excellent correlation of the experimentally obtained sequences with the theoretically predicted gene fragments demonstrated the suitability of capillary array electrophoresis for micropreparative recovery of DNA fragments. This approach, useful especially for rapid DNA expression profiling of unknown genes for nonsequenced organisms, demonstrates the practical capability of the prototype multicapillary fraction collector.
Simplex optimization of electrokinetic injection of DNA in capillary electrophoresis using dilute polymer solution
Authors: Catai Jonatan Ricardo , Carrilho Emanuel 
Published: February 2003

AbstractIn DNA analysis by capillary electrophoresis with polymer solutions there are many variables that can be optimized. However, electric field strength, polymer solution concentration and temperature of analysis are the most relevant ones. These are the variables most responsible for the fragment resolution and analysis time. Optimization of such parameters can be obtained simultaneously using chemometric techniques, reaching the optimum working conditions with few experiments. In this work, we have studied the influence of the sample composition and electrokinetic injection conditions in the reproducibility and the quality of the DNA separation results. A simplex optimization has been carried out and the optimum condition was reached with nine experiments.
Detection of Escherichia coli O157:H7 bacteria by a combination of immunofluorescent staining and capillary electrophoresis
Authors: Kourkine Igor V. , Ristic‐Petrovic Mirjana , Davis Edward , Ruffolo Carmel G. , Kapsalis Andreas , Barron Annelise E. 
Published: February 2003

AbstractAs the number of incidents of bacterial infections continues to rise around the globe, simpler, faster, and more sensitive diagnostic techniques are required to improve the safety of the food supply and to screen for potential bacterial infections in humans. We present here direct and indirect approaches for the detection of bacteria, which are based upon a combination of immunofluorescent staining and capillary electrophoresis. In the direct approach, Escherichia coli O157:H7 bacteria stained with fluorescein‐tagged specific antibodies are detected by CE, while in the indirect approach fluorescein‐tagged specific antibodies to E. coli are first captured by E. coli O157:H7 bacteria and then released and detected by CE. We have identified suitable bacteria staining and CE protocols, which involved a 10 mM Tris‐borate‐EDTA (TBE) buffer, 0.25 μg antibody/1 million bacteria, and capillaries dynamically coated with poly‐N‐hydroxyethylacrylamide (polyDuramide). We have also successfully detected the presence of E. coli O157:H7 in contaminated meat. The total time required for analysis was 6–8 h, which is less than that realized in most commercial assays presently available.
Simultaneous separation of fifteen approved protease and reverse transcriptase inhibitors for human immunodeficiency virus therapy by capillary electrophoresis
Authors: Tuan Nguyen Duc , Gutleben Wolfgang , Scherer Karin , Stoiber Heribert , Falkensammer Barbara , Dierich Manfred P. , Zemann Andreas 
Published: February 2003

AbstractIn the present investigation, a novel approach towards a complete separation of all 15 protease and reverse transcriptase inhibitors which are currently approved for use in highly active antiretroviral therapy in a single analytical run is presented. The developed method employs an acidic background electrolyte with sodium polyanethol sulfonate (SPAS) as polyanionic electroosmotic flow (EOF) modifier to establish a strong cathodic EOF, sodium dodecyl sulfate (SDS) as pseudostationary selector, and acetonitrile and ethanol as organic modifiers. Separation of the analytes is based on two different mechanisms. The more basic analytes are protonated at the prevailing pH conditions and thus migrate in front of the cathodic EOF, whereas the less basic and neutral analytes interact with the SDS and are retained after the EOF. By optimizing electrolyte pH, the amount of solvents and SDS concentrations in the background electrolyte it is possible to completely separate all compounds of interest.