bluuebunny/crossref_metadata_2025_split huggingface dataset:
Yield of HCV and HIV‐1 NAT after screening of 3.6 million blood donations in central Europe
Authors: Roth W. Kurt , Weber Marijke , Buhr Sylvia , Drosten Christian , Weichert Wolfgang , Sireis Walid , Hedges Doris , Seifried Erhard 
Published: September 2002

BACKGROUND: HCV and HIV‐1 NAT of all blood donations was initiated at our institutions in January 1997 to reduce the residual risk of transfusion‐transmitted virus infections. The yield of NAT after testing more than 3.6 million donations in central Europe is reported. STUDY DESIGN AND METHODS: Automated pipetting instruments were used to pool up to 96 donor samples including those that were antibody reactive. To compensate for dilution of the individual donor samples by pooling, viruses were enriched from the pools by centrifugation at 48,000 ×g. A commercial PCR (Cobas Amplicor, Roche) and an in‐house PCR were applied for HCV and HIV‐1 amplification, respectively. RESULTS: Six HCV and 2 HIV‐1 PCR confirmed‐positive, antibody‐negative donations (yield, 1 in 600,000 and 1 in 1.8 million, respectively) were identified. Thirty‐nine and 11 multiple‐time donors seroconverted for HCV and HIV, respectively, and look‐back procedures were initiated. Archived samples from preseroconversion donations were thawed and retested by single‐sample PCR and remained negative. The recipients of the blood components were traced and tested. All traced recipients were negative for HCV and HIV antibodies. CONCLUSION: The yield of NAT in central European Red Cross blood donors was less than expected from theoretical calculations for American and German multiple‐time donors. Look‐back procedures for HCV and HIV indicated that no donation given before seroconversion of the donor was missed by minipool PCR. Sensitivity of minipool PCR testing after virus enrichment seems to be sufficiently high to close the diagnostic window almost completely.
Change in CD34+ cell concentration during peripheral blood progenitor cell collection: effects on collection efficiency and efficacy
Authors: Ford Clyde D. , Greenwood Jay , Strupp Annie , Lehman Chris M. 
Published: September 2002

BACKGROUND: An understanding of factors affecting CD34+ cell collection efficacy is essential to minimize donor toxicity and cost. STUDY DESIGN AND METHODS: Peripheral blood CD34+ cell (CD34) measurements were determined at various intervals before, during, and after automated cell collection (Cobe Spectra 6.0). The serial mean of multiple, intraprocedural CD34 levels was calculated for each procedure as an estimate of the mean, inlet‐line CD34 level. RESULTS: The CD34+ concentration fell a mean of 30 percent in the first 30 to 70 minutes of collection. The degree of decline was inversely correlated with donor blood volume (BV), but was not due to hemodilution. The mean of the CD34 level before and after collection slightly overestimated the serial mean CD34 level. Cell yields, normalized for the CD34 level before collection, were higher from donors with larger BVs. CONCLUSIONS: The CD34 concentration rapidly decreased to a relative equilibrium level during the collection procedure. The degree of decrease in the CD34 level inversely correlated with the BV of the donor and was consistent with cell pooling in the collection set. The higher equilibrium CD34 levels in donors with larger BVs resulted in increased collection of CD34+ cells, and therefore, large‐volume apheresis should be most efficient in these donors.
Equivalent mobilization and collection of granulocytes for transfusion after administration of glycosylated G–CSF (3 μg/kg) plus dexamethasone versus glycosylated G–CSF (12 μg/kg) alone
Authors: Heuft H.G. , Goudeva L. , Sel S. , Blasczyk R. 
Published: September 2002

BACKGROUND: The aim of this study was to find a regimen for mobilization and collection of granulocytes that combines low‐dose G–CSF administration with satisfactory PMN mobilization and apheresis at a low rate of donor adverse reactions. STUDY DESIGN AND METHODS: In a prospective study, 52 healthy unrelated volunteers received a single subcutaneous injection of glycosylated G–CSF (Lenograstim Chugai‐Pharma, Frankfurt, Germany) at medians of 3.1 (range, 2.4‐3.6) μg per kg plus dexamethasone (8 mg orally; n = 29) or at 11.8 (7.1‐18.5) μg of lenograstim per kg (p ≤ 0.0001) without dexamethasone (n = 23) and underwent standard apheresis using the PMN program of a cell separator (Spectra, COBE [now Gambro] BCT). WBC and PMN mobilization results and apheresis yields were compared and the severity and clinical significance of donor adverse reactions was evaluated. RESULTS: For the low‐dose G–CSF plus dexamethasone versus the high‐dose G–CSF alone group, similar mobilization results were observed for WBCs with 31.3 (19.1‐44.9) × 109 per L versus 27.5 (19.2‐44.0) × 109 per L (p = 0.21, NS) and PMNs with 29.0 (17.6‐42.2) × 109 per L versus 25.2 (16.2‐39.0) × 109 per L (p = 0.08, NS). The PMN apheresis yields were equal with 70 (39‐139) × 109 per unit with low‐dose G–CSF versus 68 (33‐120) × 109 per unit in the high‐dose G–CSF group (p = 0.83, NS). Regarding donor adverse reactions, 7 out of 29 (24%) and 8 out of 23 donors (35%) reported moderate or severe symptoms. The character of these reactions was different; symptoms of greater clinical significance and a higher need for analgesics were observed in the high‐dose G–CSF group. CONCLUSIONS: A Lenograstim dose of 3 μg per kg plus DXM assures effective PMN mobilization and acceptable apheresis components. The combination of glycosylated G–CSF with DXM allows a significant dose reduction in G–CSF for PMN mobilization and collection as compared with higher G–CSF doses alone. In the high‐dose G–CSF mobilization group, adverse reactions were more severe and required more analgesics.
Experience with universal bacterial culturing to detect contamination of apheresis platelet units in a hospital transfusion service
Authors: AuBuchon James P. , Cooper Linda K. , Leach Miriam F. , Zuaro Deborah E. , Schwartzman Joseph D. 
Published: September 2002

BACKGROUND: Bacterial contamination of platelet units poses one of the greatest risks of morbidity and mortality to platelet transfusion recipients. A routine culture of all units (WBC‐reduced apheresis platelet units) was instituted on Day 2 over a 2‐year period to reduce this risk. STUDY DESIGN AND METHODS: A sterile connecting device was used to attach a small transfer pack on the morning of Day 2 after collection, and 10 mL of the unit were transferred to the small bag. After disconnection from the unit, about half of this volume was transferred to an aerobic culture bottle of an automated bacterial detection system. Units were maintained in available inventory until and unless a report was received of growth in the sample. When available, the unit or a retained aliquot was recultured if the initial sample was positive. Units were held up to 2 days beyond their 5‐day outdate and used for transfusion if no other suitable units were available to meet the clinical need or were evaluated with in vitro testing on Day 8. RESULTS: Of 2678 units cultured, 16 (0.6%) were positive on initial culture. Thirteen could be recultured, and all of these samples were negative. Shortly after the 2‐year period of the study, two units (split from the same collection) were documented as growing coagulase‐negative Staphylococci 12 hours after sampling. Units transfused on Day 6 or 7 (n = 40) yielded expected clinical responses, and CCI available on 21 recipients 10 to 60 minutes after transfusion demonstrated acceptable results (mean, 14,400 ± 8800; median, 12,191; 90% > 7500). More than 96 percent of units tested on Day 8 had pH greater than 6.2 and continued to demonstrate swirling. CONCLUSIONS: Routine culturing of apheresis platelet units is feasible, can be accomplished with a low rate of false positivity, and can detect contaminated units. The cost of such a protocol could be mitigated with extension of the storage period, and clinical experience with units held for 6 or 7 days was satisfactory.
Quantification of fetomaternal hemorrhage by fluorescence microscopy is equivalent to flow cytometry
Authors: Ochsenbein‐Imhof Nadin , Ochsenbein A.F. , Seifert B. , Huch Albert , Huch Renate , Zimmermann Roland 
Published: September 2002

BACKGROUND: The quantification of fetal cells in the maternal circulation remains an important goal to determine the amount of anti‐D necessary to prevent active immunization of a D– mother giving birth to a D+ baby. Underestimation of fetomaternal hemorrhage (FMH) results in inefficient anti‐D prophylaxis and maternal immunization; overestimation of FMH results in higher doses of passively transferred anti‐D, higher costs, and the risk of disease transmission. Thus, a reliable method to quantitatively assess FMH is necessary. STUDY DESIGN AND METHODS: Serial dilutions of artificial FMH were quantitatively measured by three different methods: flow cytometry, fluorescence microscopy (each after anti‐D staining), and by the Kleihauer‐Betke test. The accuracy and precision of the three methods were compared by statistical analysis. RESULTS: Fluorescence microscopy and flow cy‐ tometry were comparably accurate and precise in quantifying FMH. In contrast, the accuracy of the Kleihauer‐Betke test was poor, resulting in substan‐ tial overestimation of FMH in the samples with lower fetal cell concentrations. CONCLUSION: Anti‐D flow cytometry and fluorescence microscopy for detection of fetal cells offer equally reliable and precise methods in contrast to the Kleihauer‐Betke test. Fluorescence microscopy may be established as standard to quantify FMH in clinical practice because it is comparable to flow cytometry; in addition, it is time saving and is less expensive.
Combined isolation of CD34+ progenitor cells and reduction of B cells from peripheral blood by use of immunomagnetic methods
Authors: Martín‐Henao G.A. , Picón M. , Rueda F. , Amill B. , Querol S. , Gonzalez‐Barca E. , Ferrá C. , Grañena A. , García J. 
Published: September 2002

BACKGROUND: Malignant cells may contribute to relapse after autologous hematopoietic cell transplantation The effectiveness of a double immunomagnetic purging strategy combining CD34‐positive with B‐negative cell selection to purge peripheral blood progenitor cells (PBPCs) from patients with chronic lymphoproliferative disorders has been analyzed. STUDY DESIGN AND METHODS: Twenty‐two CD34+ cell selections from patients with follicular lymphoma (n = 14), chronic lymphocytic leukemia (n = 6), mantle cell lymphoma (n = 1), and splenic marginal zone lymphoma (n = 1) were performed by use of a magnetic cell selector followed by a negative cell selection step with anti‐CD19 monoclonal antibody bound to immunomagnetic beads. RESULTS: The PBPC components contained median CD34+ cells of 1.24 percent (range, 0.38‐3.92%) and CD19+ cells of 1.83 percent (range, 0.06‐69.7%). After positive selection (n = 22), 49 percent (range, 16‐72%) of CD34+ cells were recovered with a purity of 93 percent (range, 24‐99%). The double‐positive and ‐negative selections (n = 20) yielded 57.5 percent of CD34+ cells (range, 33.4‐79.4%) with a purity of 95 percent (range, 63‐99%). Logarithms of B‐cell reduction in the CD34+‐cell‐enriched B‐cell‐depleted component had a median value of 3.63 (range, 2.74‐4.84 log) and CD19+ and CD5+ cells for chronic lymphocytic leukemia patients with more than 4.56 log (>3.6‐5.6 log). Of 13 PBPC components that had a tumor‐specific clonal signal, 10 became PCR negative after the double‐selection procedure. CONCLUSION: Combined positive and negative magnetic cell selection achieves a high grade of tumor cell reduction with up to 77 percent of the grafts being negative for tumor‐specific clonal signal by PCR analysis. This technique preserves an adequate recovery of progenitor cells able to engraft.
TT virus infection during childhood
Authors: Ohto Hitoshi , Ujiie Niro , Takeuchi Chikako , Sato Akira , Hayashi Akio , Ishiko Hiroaki , Nishizawa Tsutomu , For The Vertical Transmission of Hepatitis Viruses Collabora Hiroaki Okamoto 
Published: September 2002

BACKGROUND: TT virus (TTV) is widespread in the general population, however, the mode of its transmission and the mechanism of maintaining it in the general population are unclear. STUDY DESIGN AND METHODS: To determine the possible mother‐to‐infant route of transmission, 54 infants born to 50 anti‐HCV‐positive mothers were assessed longitudinally. Nucleotide sequences amplified by seminested PCR with primers targeting the N22 variable coding region of genotypes 1 through 6 were compared in mothers and their infants. RESULTS: The prevalence of TTV DNA was 30 percent (15/50; 95% CI, 18‐45) in mothers and 44 percent (24/54; 95% CI, 31‐59) in their infants. TTV DNA was detected during a follow‐up period in 13 (87%; 95% CI, 60–98) of 15 infants born to infected mothers and in 11 (28%; 95% CI, 15‐45) of 39 infants born to DNA‐negative mothers. None of 38 cord blood samples, but one of 14 blood samples, obtained at 1 month of age had detectable TTV DNA. The lowest infection rate at the earliest ages and the subsequent increasing prevalence of infection (22% at 6 months and 33%[43% cumulative rate] at 2 years) is consistent with an age‐dependent acquisition of TTV by nonparenteral routes. In 13–mother‐infant pairs positive for TTV DNA, six showed a high degree of nucleotide sequence similarity (99.1‐100%), whereas the remaining seven pairs differed more than 10 percent from each other (46.8‐89.2%). The viral load of maternal blood was not a plausible risk factor for transmission. Genotype 1, of which pathogenicity failed to be shown by measurement of hepatic enzymes, was more rapidly cleared (88 vs. 8% other genotypes, p < 0.001) among infants. CONCLUSIONS: These observations strongly suggest that the main factor for TTV acquisition in children involves their age‐associated increase in environmental interactions with infectious materials. Genotype 1 might be involved in a weak or a limited pathologic role, which can possibly be diluted by other harmless genotypes.
Controlled study of citrate effects and response to IV calcium administration during allogeneic peripheral blood progenitor cell donation
Authors: Bolan Charles D. , Cecco Stacey A. , Wesley Robert A. , Horne McDonald , Yau Yu Ying , Remaley Alan T. , Childs Richard W. , Barrett A. John , Rehak Nadja N. , Leitman Susan F. 
Published: September 2002

BACKGROUND: Leukapheresis procedures are generally performed at citrate anticoagulation rates extrapolated from shorter plateletpheresis procedures. However, neither the metabolic effects nor the management of associated symptoms have been critically evaluated during leukapheresis in healthy donors. STUDY DESIGN AND METHODS: Symptom assessments (n = 315) and laboratory analyses (n = 49) were performed during 244 procedures performed with and 71 without prophylactic calcium (Ca) chloride or Ca gluconate given at a dose linked to the citrate infusion rate (1.0‐2.2 mg/kg/min). RESULTS: During leukapheresis of 12 to 25 L processed, ionized Ca and ionized magnesium (Mg) decreased as much as 35 and 56 percent, respectively, each exhibiting a tight negative correlation with marked increases in serum citrate levels. Significant increases in urinary Ca and Mg excretion accompanied the renal excretion of a large citrate load. Serum divalent cation levels remained depressed 24 hours after leukapheresis. Symptoms were more frequent in donors who were women, had low initial total Mg levels, and underwent procedures in which larger volumes were processed at higher citrate infusion rates. Ca infusions reduced clinically significant paresthesias by 96 percent and also attenuated decreases in serum potassium. Ca chloride maintained higher Ca levels than Ca gluconate. CONCLUSIONS: Prophylactic Ca infusions safely attenuate the marked metabolic effects of citrate administration and promote faster, more comfortable, leukapheresis procedures.
Transcriptome analysis of
Escherichia coli
using high-density oligonucleotide probe arrays
Authors: Tjaden Brian , Saxena Rini Mukherjee , Stolyar Sergey , Haynor David R. , Kolker Eugene , Rosenow Carsten 
Published: August 2002

Abstract
Microarrays traditionally have been used to analyze the expression behavior of large numbers of coding transcripts. Here we present a comprehensive approach for high-throughput transcript discovery in Escherichia coli focused mainly on intergenic regions which, together with analysis of coding transcripts, provides us with a more complete insight into the organism’s transcriptome. Using a whole genome array, we detected expression for 4052 coding transcripts and identified 1102 additional transcripts in the intergenic regions of the E.coli genome. Further classification reveals 317 novel transcripts with unknown function. Our results show that, despite sophisticated approaches to genome annotation, many cellular transcripts remain unidentified. Through the experimental identification of all RNAs expressed under a specific condition, we gain a more thorough understanding of all cellular processes.
Role of transglutaminase 2 in glucose tolerance: knockout mice studies and a putative mutation in a MODY patient
Authors: BERNASSOLA FRANCESCA , FEDERICI MASSIMO , CORAZZARI MARCO , TERRINONI ALESSANDRO , HRIBAL MARTA L. , DE LAURENZI VINCENZO , RANALLI MARCO , MASSA ORNELLA , SESTI GIORGIO , MCLEAN W.H. IRWIN , CITRO GENNARO , BARBETTI FABRIZIO , MELINO GERRY 
Published: August 2002

ABSTRACTTransglutaminase 2 (TGase 2) is a Ca+2‐ dependent enzyme that catalyzes both intracellular and extracellular cross‐linking reactions by transamidation of specific glutamine residues. TGase 2 is known to be involved in the membrane‐mediated events required for glucose‐stimulated insulin release from the pancreatic β cells. Here we show that targeted disruption of TGase 2 impairs glucose‐stimulated insulin secretion. TGase 2‐/‐mice show glucose intolerance after intraperitoneal glucose loading. TGase 2‐/‐mice manifest a tendency to develop hypoglycemia after administration of exogenous insulin as a consequence of enhanced insulin receptor substrate 2 (IRS‐2) phosphorylation. We suggest that the increased peripheral sensitivity to insulin partially compensates for the defective secretion in this animal model. TGase 2‐/‐mouse phenotype resembles that of the maturity‐onset diabetes of young (MODY) patients. In the course of screening for human TGase 2 gene in Italian subjects with the clinical features of MODY, we detected a missense mutation (N333S) in the active site of the enzyme. Collectively, these results identify TGase 2 as a potential candidate gene in type 2 diabetes.—Bernassola, F., Federici, M., Corazzari, M., Terrinoni, A., Hribal, M. L., De Laurenzi, V., Ranalli, M., Massa, O., Sesti, G., Mclean, W. H. I., Citro, G., Barbetti, F., Melino, G. Role of transglutaminase 2 in glucose tolerance: knockout mice studies and a putative mutation in a MODY patient. FASEB J. 16, 1371–1378 (2002)
cDNA array analysis of gene expression profiles in the striata of wild‐type and Cu/Zn superoxide dismutase transgenic mice treated with neurotoxic doses of amphetamine
Authors: KRASNOVA IRINA N. , MCCOY MICHAEL T. , LADENHEIM BRUCE , CADET JEAN LUD 
Published: August 2002

ABSTRACTAmphetamine (AMPH) is a drug of abuse that causes the degeneration of striatal dopamine terminals in mammals. Superoxide radicals seem to participate in AMPH‐induced damage because its toxicity is attenuated in Cu/Zn superoxide dismutase transgenic (SOD‐tg) mice. To provide a detailed analysis of molecular changes associated with AMPH toxicity, we used cDNA arrays consisting of 1176 genes to detect differential changes in gene expression in the striata of wild‐type and SOD‐tg mice treated with neurotoxic doses of the drug. We found 42 genes that showed >1.8‐fold changes in at least two consecutive time points during the course of the study and were differentially affected by AMPH in the two genotypes. Specifically, more transcription factors and genes involved in responses to injury/inflammation were affected in wild‐type mice after AMPH administration. Some of these stimulant‐induced superoxide‐dependent alterations in gene expression might affect neuronal functions and promote neuronal damage. Other changes might help to provide some degree of protection against AMPH toxicity. These results support the view that the use of global array analysis of gene expression will help to identify novel molecular mediators of AMPH‐induced neurodegeneration.—Krasnova, I. N., McCoy, M. T., Ladenheim, B., Cadet, J. L. cDNA array analysis of gene expression profiles in the striata of wild‐type and Cu/Zn superoxide dismutase (SOD) transgenic mice treated with neurotoxic doses of amphetamine. FASEB J. 16, 1379–1388 (2002)
Development of hepatocytes from embryonic stem cells after transfection with the HNF‐3β gene
Authors: Ishizaka Shigeaki , Shiroi Akira , Kanda Seiji , Yoshikawa Masahide , Tsujinoue Hirohisa , Kuriyama Shigeki , Hasuma Tadayoshi , Nakatani Kazuki , Takahashi Kenichi 
Published: August 2002

ABSTRACTWe have attempted to generate embryonic stem (ES) cell‐derived hepatocytes expressing liver‐specific functional properties by use of ES cell technology. It was found that ES cells are allowed to differentiate into hepatocytes possessing high metabolic activities when hepatocyte nuclear factor (HNF)‐3β‐transfected ES cells are cultured in α‐MEM medium supplemented with 10% fetal bovine serum (FBS) and fibroblast growth factor (FGF)‐2 in the three‐dimensional cell culture system at 5% CO2. The differentiated cells induced albumin, triacylglycerol, urea, and glycogen synthesis as well as further expression of metabolic proteins and serum factors as markers of hepatocytic differentiation for at least 4 months. The cells differentiated from HNF‐3β‐transfected ES cells also had hepatocyte‐like ultrastructural characteristics, including several endoplasmic reticula, mitochondrion, and glycogen. Our findings indicate that generation of hepatocytes maintaining high metabolic functions developed from mouse ES cells will facilitate the study of the basic mechanism for hepatogenesis and will certainly provide new opportunities for tissue transplantation.
Persistence of microscopic human cancers in mice: alterations in the angiogenic balance accompanies loss of tumor dormancy
Authors: UDAGAWA TATURO , FERNANDEZ ANTONIO , ACHILLES EIKE‐GERT , FOLKMAN JUDAH , D'AMATO ROBERT J. 
Published: August 2002

ABSTRACTSome human tumor lines do not form visible tumors when inoculated into immunosuppressed mice. The fate of these human tumor lines was followed by transfecting them with green fluorescence protein before inoculating them into mice. Although the tumor lines failed to grow progressively, they formed small dormant microscopic foci maintained at constant mass by balanced proliferation and apoptosis. Transfecting the cells with either VEGF165 or activated c‐Ha‐ras induced loss of dormancy, which correlated with a shift in the angiogenic balance toward increased vascularity with reduced tumor cell apoptosis. These results support a model in which loss of dormancy is controlled in part by a switch to an angiogenic phenotype. These tumor lines may serve as models for investigating the cellular mechanisms controlling dormancy and identifying those factors that promote the loss of balanced proliferation and apoptosis. Finally, these models may prove useful in the design and testing of therapies directed toward eradicating dormant tumors and preventing tumor recurrence.—Udagawa, T., Fernandez, A., Achilles, E.‐G., Folkman, J., D'Amato, R. J. Persistence of microscopic human cancers in mice: alterations in the angiogenic balance accompanies loss of tumor dormancy. FASEB J. 16, 1361–1370 (2002)