bluuebunny/crossref_metadata_2025_split huggingface dataset:
Spatial pattern analysis of nitrergic neurons in the developing myenteric plexus of the human fetal intestine
Authors: Román V. , Bagyánszki M. , Krecsmarik M. , Horváth A. , Resch B. Á. , Fekete É. 
Published: January 2004

AbstractBackgroundEnteric nervous system precursors derived from the neural crest migrate along defined pathways to colonize the bowel. The individual cells in different environments experience different growth, differentiation, and survival conditions. Hence, the spatial distribution of the neurons is determinant with regard to functional maturation. The question arises as to whether the distribution is random or nonrandom.MethodsNitrergic cells were visualized by means of nicotinamide adenine dinucleotide phosphate diaphorase histochemistry. Stained specimens were photographed, and the borders of the myenteric plexus and the nuclei of the nitrergic neurons were digitalized. Plexus Pattern Analysis software was used to count the nuclei of nitrergic neurons, calculate the proportions of the areas covered by the plexus and the gut wall, and perform randomization analyses.ResultsThe distribution pattern of the nitrergic neurons changed markedly between weeks 14 and 22 of gestation. The nitrergic neurons were randomly distributed at week 14 but were aggregated in the plexus and within the individual ganglia at week 19. The dynamics of these changes exhibited regional differences.ConclusionsThe results suggest that, in addition to the gut wall and the plexus, other intraganglionic constituents may contribute to the aggregation of nitrergic cells and such examinations should be extended to other cell types in the future. © 2004 Wiley‐Liss, Inc.
Measurement of basal, substrate induced and total P‐glycoprotein activity in bronchoalveolar lavage T‐cell subsets
Authors: Donnenberg V. S. , Wilson J. W. , Burckart G. J. , Zeevi A. , Iacono A. , Donnenberg A. D. 
Published: January 2004

AbstractBackgroundP‐glycoprotein (P‐gp) is a member of the ABC transporter superfamily. P‐gp activity can be detected by measuring efflux of fluorescent substrates such as rhodamine 123 (R123). Our objectives were to evaluate P‐gp activity in T cells freshly isolated from bronchoalveolar lavage (BAL) and to develop a strategy to distinguish between basal, in vitro substrate‐induced, and total P‐gp activities.MethodsCells were obtained from blood (n = 44) and BAL (n = 34), stained for expression of CD3, CD4, CD8, and CD14, and incubated with R123 (0.13 μM) ± cyclosporine (5 μM), a specific P‐gp inhibitor. P‐gp activity was detected as median fluorescence intensity (MFI) and percentage of cells falling below a pre‐established cutoff.ResultsBAL T cells displayed significant basal P‐gp activity, which was most apparent when measured as percentage below the cutoff. Induced activity (difference between P‐gp activity measured after load and efflux) was determined equally well when using the MFI or the percentage below cutoff parameter. Total activity was represented by the efflux parameters (MFI or percentage below cutoff) or by the activity‐time area under the curve (AUC) method. The two efflux parameters correlated well but were insensitive to the time‐dependent nature of dye efflux. In the AUC method, two samples with identical R123 brightness or percentage below cutoff after dye efflux can have very different total activities, depending on their basal activity. The AUC method was also most sensitive in distinguishing between P‐gp activity in peripheral blood and resident lung T cells. Application of this methodology to the comparison of P‐gp activity in BAL and peripheral CD8+ T cells best revealed the elevated total P‐gp activity in BAL T cells.ConclusionsWe have systematically evaluated several methodologies for analysis of P‐gp activity and arrived at a novel and robust strategy amenable to standardization and evaluation of the effects of P‐gp modulators in vivo and in vitro. © 2004 Wiley‐Liss, Inc.
Flow cytometer for measurement of the light scattering of viral and other submicroscopic particles
Authors: Steen Harald B. 
Published: January 2004

AbstractBackgroundLight scattering is an essential parameter in flow cytometry, facilitating functions such as size measurement, discrimination of cell types on the basis of shape and morphology, detection of fluorescence‐negative cells, and gating of fluorescence measurements. Light scattering measurement of viruses is generally not feasible with current flow cytometers due to their small size. The problem is aggravated by the fact that the light scattering of particles in this size range (<200 nm) falls off with roughly the sixth power of their linear dimensions.MethodsA new optical layout using darkfield illumination and detection has been developed. A 532‐nm laser was used for excitation, and scattered light was collected with large aperture optics.ResultsLight scattering histograms of polymer particles with diameters of 70–300 nm were recorded without gating by other parameters. By extrapolation, a detection limit of about 50 nm was obtained. Different species of virus with sizes of approximately 100 nm also were recorded.ConclusionsFlow cytometric light scattering measurement of submicroscopic particles, in a size range that includes many viral species, is now feasible. The results indicate that it may be practically impossible to measure by flow cytometry the light scattering of particles smaller than 40 nm. © 2004 Wiley‐Liss, Inc.
Quantitation of MHC tetramer‐positive cells from whole blood: Evaluation of a single‐platform, six‐parameter flow cytometric method
Authors: Wölfl Matthias , Schalk Stefan , Hellmich Martin , Huster Katharina M. , Busch Dirk H. , Berthold Frank 
Published: January 2004

AbstractBackgroundQuantitation of antigen‐specific T cells provides an insight into the development and dynamics of T‐cell responses in tumor immunology and infectious diseases. Soluble major histocompatibility class I tetramers are widely used to monitor immune responses; however, variations due to handling and analysis are likely to confound comparisons between different experiments and laboratories.MethodsWhole blood from healthy donors was stained with HLA‐A*0201/tetramers specific for an epitope of phosphoprotein 65, the immunodominant antigen in cytomegalovirus infection. With the help of Trucount tubes, a single‐platform, four‐color flow cytometric assay was established to obtain absolute counts of tetramer‐positive cells. Various staining and gating strategies were evaluated.ResultsThe no‐wash method was a quick and straightforward procedure for the quantitation of tetramer‐positive events from whole blood. The level for background staining was low. This information about the intra‐assay–related variation and the physiologic variation will allow validation and interpretation of data in future studies.ConclusionsThe method is highly reliable and can be standardized for multiple experiments. It is therefore suitable for the direct ex vivo analysis of antigen‐specific T cells in a variety of clinical settings such as infectious, autoimmune, or neoplastic diseases and can be implemented as a tool for multicenter studies. © 2004 Wiley‐Liss, Inc.
New method for the analysis of cell cycle–specific apoptosis
Authors: Tao Deding , Wu Jianhong , Feng Yongdong , Qin Jichao , Hu Junbo , Gong Jianping 
Published: January 2004

AbstractBackgroundIn this study, a new method for the analysis of cell cycle specificity of apoptosis was designed by using a modified annexin V and propidium iodide (API) method.MethodsCells of the human promyelocytic HL‐60 line treated with camptothecin (CPT) or ultraviolet light (UV) were labeled with fluorescein isothiocyanate–conjugated annexin V and prefixed with 1% methanol‐free formaldehyde on ice, and their DNA was stained stoichiometrically with propidium iodide in the presence of digitonin. Cellular green and red fluorescences were measured by flow cytometry.ResultsCell cycle specificity of apoptosis obtained by the API method and those analyzed for the presence of DNA strand breaks by using terminal deoxynucleotidyl transferase (TdT) assay were similar: CPT‐ or UV‐induced apoptosis preferentially in S‐ or G1‐phase cells, respectively. When the internucleosomal DNA degradation was prevented by the serine protease inhibitor N‐tosyl‐L‐phenylalanine chloromethyl ketone, apoptotic cells could not be detected by the TdT assay but were identified by the API method.ConclusionsThe API method, similar to the TdT assay, accurately detects the cell cycle phase specificity of apoptosis. Also, the API method appears to detect earlier stages of apoptosis than the TdT assay. © 2004 Wiley‐Liss, Inc.
Comparison of flow cytometry and laser scanning cytometry for the analysis of CD34+ hematopoietic stem cells
Authors: Oswald Joachim , Jørgensen Birgitte , Pompe Tilo , Kobe Fritz , Salchert Katrin , Bornhäuser Martin , Ehninger Gerhard , Werner Carsten 
Published: January 2004

AbstractBackgroundCharacterization of hematopoietic stem cells (HSCs) by laser scanning cytometry (LSC) was compared with conventional flow cytometry (FCM). The method was evaluated for application in the development of advanced cell culture substrates that were supposed to support the ex vivo expansion of HSC. For this purpose, adherent HSCs were grown in culture on thin polymer films coated with reconstituted collagen I fibrils and subsequently analyzed by LSC.MethodsCD34+ HSCs were isolated from cord blood by immunomagnetic separation and cultivated on polymer films coated with reconstituted collagen I fibrils. Cell surface antigens (CD34, CD29) were stained with antibodies, and nuclei were labeled with a DNA stain (TO‐PRO‐3 iodide) that does not interfere with the fluorochromes of the antibodies. Fluorescence intensity of the adherent cells was measured by means of LSC. Before and after in vitro expansion for time periods of up to 7 days, suspension cells were analyzed with LSC and FCM.ResultsLSC‐based analysis enabled reliable quantification of CD34+ cells with bright antigen expression before cell culture. At this stage, LSC and FCM data for CD34 expression at given HSC samples largely coincided. After in vitro expansion, LSC data deviated from FCM data for cells with dim CD34 antigen expression, whereas the fluorescence intensity of the CD29 antigen remained comparable. The deviation between LSC and FCM data for CD34dim was attributed to the better resolution of weak fluorescence by FCM. Based on the preceding evaluation of the method, LSC analysis could be applied to characterize HSCs cultivated on collagen I–coated polymer films without detachment of the cells from the substrate.ConclusionsLSC‐based analysis allows for the automated evaluation of adherent HSCs. Although resolution of weakly expressed antigens can be achieved more precisely with FCM, the method provides a valuable tool to study interactions of HSCs with bioartificial substrates. © 2004 Wiley‐Liss, Inc.
Flow cytometer electronics
Authors: Snow Christopher “Kit” 
Published: January 2004

AbstractFlow cytometers, like all things electronic, have undergone rapid evolution since the introduction of the first commercial instruments 30 years ago. Cytometers then had so little automation and sophistication that the operator had to be expert in many different scientific disciplines to be able to coax any data at all out of the instrument. But trends in electronic components toward higher speed, smaller size, and lower power consumption have allowed the instruments to become faster, more sensitive, and actually quasi‐intelligent. Automation of complex processes that once required considerable operator expertise, such as running a high‐speed sort or correctly compensating a multicolor experiment, has expanded the number of flow cytometry applications and the number of flow cytometry users; there is no reason to expect that trend to stop. This review discusses the electronic components of a current “generic” flow cytometer from a functional point of view with the hope that a better understanding of what is going on “inside the box” will lead to a better understanding of how operator‐selectable instrument settings can influence the quality of the data produced. © 2004 Wiley‐Liss, Inc.
Simple, semiautomatic assay of cytostatic and cytotoxic effects of antitumor drugs by laser scanning cytometry: Effects of the bis‐intercalator WP631 on growth and cell cycle of T‐24 cells
Authors: Pozarowski Piotr , Huang Xuan , Gong Richard W. , Priebe Waldemar , Darzynkiewicz Zbigniew 
Published: January 2004

AbstractBackgroundCommon assays of drug‐induced cytotoxicity on adherent cells rely on cell trypsinization followed by count of live and dead cells. To estimate the cell cycle effects, cellular DNA content is analyzed by flow cytometry. This procedure is laborious and time consuming. The alternative viability assays, e.g., based on reduction of 3‐(4,5‐dimethylthiazol‐2‐yl) 2,5‐diphenyl tetrazolium bromide, although rapid and convenient, do not provide information about individual cells or cell cycle effects and may be biased by growth imbalance.MethodsThe bladder carcinoma T‐24 cells were seeded onto eight‐chamber microscope slide‐based tissue culture vessels. The novel antitumor drug, the bis‐intercalating anthracycline WP631, was administered at various concentrations to different chamber cultures on the same slide; the control cultures were left untreated. After 24, 48, and 72 h, the cultures were fixed, and cellular DNA was stained with 4,6‐diamidino‐2‐phenyl indole (DAPI). The slides were scanned by laser scanning cytometry (LSC) to obtain the number of attached cells per culture chamber and reveal their cell cycle distribution.ResultsThe cell growth and viability plots in the absence and presence of WP621 were constructed from the frequency of the attached cells per chamber. A 50% reduction in cell number was observed at the 75 nM concentration of WP321. Mitotic and postmitotic cells were identified based on high intensity of maximal pixel of DAPI fluorescence. An increase in proportion of cells in G2 was seen at 75–300 nM of WP631. Relatively few (<12%) apoptotic cells, identified by the presence of DNA strand breaks, remained attached in the WP631‐treated cultures.ConclusionsBecause late apoptotic cells detach during culturing, the cells that remain attached in the multi‐chamber cultures represent predominantly live cells; the deficit in their number compared with the untreated cultures, recorded by LSC during scanning, provides information about the degree of cytostatic and cytotoxic effects of the studied drug. The possibility to demonstrate the cell cycle distribution, including a distinction between G2 and M cells, provides an additional advantage of this assay. Other parameters that may be associated with the cell cycle perturbation or with induction of apoptosis also can be measured in the same cultures by using the multiparameter capabilities of LSC. Each measured cell can be relocated for imaging or measurement after subsequent staining with other probes. © 2004 Wiley‐Liss, Inc.
Comparison of ThinPrep® and conventional preparations in pancreatic fine‐needle aspiration biopsy
Authors: de Luna Regina , Eloubeidi Mohamad A. , Sheffield Matthew V. , Eltoum Isam , Jhala Nirag , Jhala Darshana , Chen Victor K. , Chhieng David C. 
Published: January 2004

AbstractUse of ThinPrep® preparation for fine‐needle aspiration biopsy (FNA) is gaining popularity. However, there may be a difference in the morphology and the operating characteristics between ThinPrep and conventional methods. The objective of this study was to compare the accuracy of the two methods and to address the pitfalls of ThinPrep preparation in pancreatic FNA. A computer search identified 67 pancreatic FNAs with both conventional smears and ThinPrep preparation during a 19‐mo period. These cases, obtained under endoscopic ultrasound‐guidance, consisted of 47 malignant neoplasms (44 ductal carcinomas, two mucinous neoplasms, and one islet cell tumor) and 20 benign lesions. Direct smears were prepared first and the remaining material was then put into PreservCyt Solution for ThinPrep slides. All slides were reviewed and the cytologic diagnoses were correlated with histologic and clinical follow‐up. Five conventional and 16 ThinPrep specimens were unsatisfactory due to insufficient cellularity. These cases were excluded from the analysis. Among the 62 cases evaluated by conventional preparation, 77% (34) were diagnosed as positive and 14% (seven) atypical/suspicious by conventional smears. For the 51 ThinPrep specimens, 58% (22) were interpreted as positive and 31% (12) atypical/suspicious. The sensitivity, specificity, and accuracy of diagnosing a malignancy were 77%, 100%, and 84% for conventional smears and 58%, 100%, and 67% for ThinPrep preparation, respectively. There were no false positives with either method. However, three benign lesions were interpreted as atypical/suspicious with ThinPrep preparation because of the presence of single atypical cells with distinct nucleoli. One of the two mucinous neoplasms was incorrectly diagnosed with ThinPrep preparation because of lack of mucin. The diagnostic accuracy of pancreatic FNA using ThinPrep is inferior to that of conventional smears. This may be partly due to the use of split sample technique resulting in scant cellularity in ThinPrep preparation and partly due to the differences in morphology between the two preparations. Therefore, the current morphologic criteria may need modification for ThinPrep preparation in pancreatic FNA. Diagn. Cytopathol. 2004;30:71–76. © 2004 Wiley‐Liss, Inc.
Superficial endometriosis of the cervix: A source of abnormal glandular cells on cervicovaginal smears
Authors: Szyfelbein Wanda M. , Baker Patricia M. , Bell Debra A. 
Published: January 2004

AbstractSuperficial endometriosis of the cervix, a benign process which may be associated with atypical glandular cells of undetermined significance (AGUS) on cervicovaginal (CV) smears, is becoming increasingly recognized on colposcopic examination. This study details the clinical, cytologic, and histology features of six cases of superficial cervical endometriosis. All CV smears featured atypical endocervical‐like columnar cells in sheets and strips as well as cells with endometrial characteristics including solid cohesive, crowded, overlapping glandular groups, loss of cellular polarity, and a frequent ragged “feathered” edge appearance with protruding nuclei, occasional rosette formations, and endometrial stroma. Recognition of endometrial stroma in continuity with groups of cells with these features on CV smears may suggest the diagnosis of this benign condition. However, the cytologic features of endometriosis show sufficient overlap with those of precancerous and cancerous glandular lesions that many of these cases will continue to be diagnosed as “atypical glandular cells.” Diagn. Cytopathol. 2004;30:88–91. © 2004 Wiley‐Liss, Inc.
Diagnostic utility of CD10 in differentiating hepatocellular carcinoma from metastatic carcinoma in fine‐needle aspiration biopsy (FNAB) of the liver
Authors: Lin Fan , Abdallah Heba , Meschter Steven 
Published: January 2004

AbstractThe differential diagnosis between hepatocellular carcinoma (HCC) and metastatic carcinoma, especially in moderate–poorly differentiated (MPD) HCC and poorly differentiated carcinoma, can be challenging in fine‐needle aspiration biopsy (FNAB) of the liver. Recent studies demonstrate that canalicular staining for CD10 appears to be a highly specific marker for hepatocytic differentiation. The objective of this study was to test the utility of CD10 in differentiating HCC from metastatic carcinoma in FNAB of the liver. Formalin‐fixed, paraffin‐embedded cell blocks of 55 cases (22 HCC, 23 metastases, and 10 benign hepatic lesions) of FNAB of the liver were immunostained using monoclonal antibody against CD10, with microwave oven antigen retrieval, followed by a standard ABC method. Nineteen (86%) of 22 HCC cases were positive for CD10 with a canalicular staining pattern. Among them, 9 (82%) of 11 well‐differentiated (WD) HCC and 10 (91%) of 11 MPD HCC were positive for CD10. Three (13%) of 23 metastatic carcinomas were positive for CD10, demonstrating a contrasting cytoplasmic and membranous staining pattern. The three positive cases were metastatic renal cell carcinoma (RCC), choriocarcinoma, and adenocarcinoma of the lung. All 10 cases of benign hepatic lesions showed positivity for CD10 with a canalicular and focal membranous staining pattern. In conclusion, CD10 appears to be a useful marker in discriminating between HCC and metastatic carcinoma when applied to FNAB of the liver. CD10 does not provide discrimination between WD HCC and benign hepatocytes. Diagn. Cytopathol. 2004;30:92–97. © 2004 Wiley‐Liss, Inc.
Synthesis, Structure, and Redox Properties of a New Aqua Ruthenium Complex Containing the Tridentate [9]aneS3 and the Didentate 1,10‐Phenanthroline Ligands
Authors: Sala Xavier , Poater Albert , Romero Isabel , Rodríguez Montserrat , Llobet Antoni , Solans Xavier , Parella Teodor , Santos Teresa M. 
Published: January 2004

AbstractThe synthesis of a new Ru−H2O complex [Ru(phen)(H2O)([9]aneS3)](ClO4)2, 2, (phen = 1,10‐phenanthroline, [9]aneS3 = 1,4,7‐trithiacyclononane) is described. The complex is characterized by UV/Vis and 1‐D and 2‐D NMR spectroscopy in solution. The complex has been further characterized in the solid‐state by X‐ray diffraction analysis which displays a distorted octahedral coordination geometry around the Ru metal center. The cyclic voltammetric experiments of the complex show a pH‐dependent chemically irreversible wave at Ep,a = 0.82 V, at pH = 7.0, indicating the low stability of the higher oxidation states. Nevertheless, before the complex decomposes, it is capable of electrocatalytically oxidizing benzyl alcohol (BzOH) to benzaldehyde with a second‐order rate constant of 19.8 M−1s−1. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Yttrium β‐Diketonate Glyme MOCVD Precursors: Effects of the Polyether Length on Stabilities, Mass Transport Properties and Coordination Spheres
Authors: Malandrino Graziella , Lo Nigro Raffaella , Fragalà Ignazio L. , Benelli Cristiano 
Published: January 2004

AbstractThe glyme adducts of trishexafluoroacetylacetonato yttrium, [Y(hfa)3·monoglyme], [Y(hfa)3·diglyme], [Y(hfa)3·(H2O)2·triglyme] and [Y(hfa)2·tetraglyme]+[Y(hfa)4]− {CH3(OCH2CH2)nOCH3, n = 1 monoglyme, 2 diglyme, 3 triglyme and 4 tetraglyme}, were synthesized and characterized by elemental analysis, mass spectrometry, and IR, 1H and 13C NMR spectroscopy. Additionally, [Y(hfa)3·monoglyme] and [Y(hfa)3·diglyme] were characterized by single‐crystal X‐ray diffraction studies. The mass‐transport properties of these adducts were investigated by thermogravimetric analysis, differential scanning calorimetry, evaporation‐rate measurements and chemical vapour‐deposition experiments. There is evidence of high volatility and very good thermal stabilities with a residue lower than 2−4 %. Vaporization‐rate experiments proved that all adducts are well‐suited for metal‐organic chemical vapour‐deposition experiments. The [Y(hfa)3·monoglyme] complex was successfully applied in the low‐pressure MOCVD process of YBaCuO HTc superconductor using a multimetal molten single source. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)